PRINCIPLES OF

SEED SCIENCE
AND TECHNOLOGY
4th Edition
PRINCIPLES OF SEED SCIENCE
AND TECHNOLOGY
4th Edition

by

Lawrence o. Copeland
Department of Crap & Soi! Sciences
Michigan State University

Miller B. McDonald
Department ofHorticulture & Crap Science
lJhioState University

SPRINGER SCIENCE+BUSINESS MEDIA, LLC

Library of Congress Cataloging-in-Publication Data

Copeland, L. O. (Lawrence 0.),1936-

Principles of seed science and technology / by Lawrence O. Copeland, Miller B.
McDonald. - 4th ed.
p. cm.
lncludes bibliographical references.
ISBN 978-1-4613-5644-8 ISBN 978-1-4615-1619-4 (eBook)
DOI 10.1007/978-1-4615-1619-4
1. Seeds. 2. Seed technology. 1. McDonald, M. B. II. Title.

SB1l7 .C73 2001

631.5'21-dc21
2001029245

Copyright © 2001 by Springer Science+Business Media New York

OriginalIy published by Kluwer Academic Publishers in 2001
Softcover reprint ofthe hardcover 4th edition 2001
AlI rights reserved. No part of this publication may be reproduced, stored in a
retrieval system or transmitted in any form or by any means, mechanical,
photocopying, recording, or otherwise, without the prior written permission
ofthe publisher, Springer Science+Business Media, LLC.
Printed on acid-free paper.
Contents
Preface
vii
Introduction ix

Flowering Processes in Plants 1

Seed Formation and Development 17

The Chemistry of Seeds 39

Seed Ecology 58

Seed Germination 72

Seed Viability and Viability Testing 124

Seed Dormancy 140

Seed Vigor and Vigor Testing 165

Seed Storage and Deterioration 192

Seed Production 231

Seed Conditioning and Handling 252

Seed Drying 268

Seed Enhancements 277

VI

Seed Certification 297

Seed Testing 316

Seed Pathology and Pathological Testing 354

Seed Marketing 380

Seed Legislation and Law Enforcement 390

 Preface
This Fourth Edition of Principles of Seed Science and Technology, like the fIrst three
editions, is written for the advanced undergraduate student or lay person who desires an
introduction to the science and technology of seeds. The fIrst nine chapters present the seed as
a biological system and cover its origin, development, composition, function (and sometimes
nonfunction), performance and ultimate deterioration. The last nine chapters present the
fundamentals of how seeds are produced, conditioned, evaluated and distributed in our modern
agricultural society. Two new chapters have been added in this fourth edition, one on seed
ecology and the second on seed drying. Finally, revisions have been made throughout to reflect
changes that have occurred in the seed industry since publication of the Third Edition.
Because of the fundamental importance of seeds to both agriculture and to all of society,
we have taken great care to present the science and technology of seeds with the respect and
feeling this study deserves. We hope that this feeling will be communicated to our readers.
Furthermore, we have attempted to present information in a straight-forward, easy-ta-read
manner that will be easily understood by students and lay persons alike. Special care has been
taken to address both current state-of-the-art as well as future trends in seed technology.
We believe this Fourth Edition represents a new level in presenting information that
appeals to advanced undergraduate students as well as to those desiring more fundamental
information on seed form and function. At the same time, it continues to have the strengths of
the fIrst three editions, in its readability as well as its comprehensive coverage of the broader
area of seed science and technology.
 Introduction
One for the buzzard,
One for the crow,
One to rot,
and One to grow!

The farmer hopes for better seed germination rates than the gardener in this old poem by
Fay Yauger. But the fact is that most plants compensate beautifully by producing seeds in great
abundance to assure survival of their species despite the formidable odds. And each seed that
does survive is capable of producing a plant, more seeds, and still more plants and seeds to
come. Deep within the seed are its own development forces, nutritive elements, and time and
place mechanisms that signal the next growth stage. The seed itself is designed to disperse and
scatter, using other forces ofnature--wind, water, insects, birds, and animals.
Seed husbandry formed the basis for early agriculture and eventual civilization. As people
learned to plant, harvest, and preserve the seeds of certain grasses for winter, they abandoned
the nomadic life to build permanent settlements. All the major civilizations throughout history
have been founded on the culture of cereal grains, because these staples have high food value
and are easily stored. The Mesopotamians planted wheat along the banks of the Tigris and
Euphrates. The Chinese grew rice along the banks of the Hwang Ho and Yangtze. And the
Mayans cultivated corn along the dry flat plains of the Yucatan.

Seeds for Survival and Subsistence

Seeds have been, and still are, the mainstay of the world's diet. The Poaceae, or large-
seeded grasses, collectively known as cereals, contribute more food seeds than any other plant
family. Cereal grains comprise by far the greatest share of all cultivated seeds. They provide
people with their most important source of carbohydrates, as well as some protein and other
vital substances. As in ancient times, rice, wheat, and corn are the three major grains. Oat,
barley, sorghum, millet, and rye are other important food and feed grains.
The second most critical food family, the Fabaceae, provides crops such as peanut,
soybean, lentil, bean, pea, and chickpea. Legume seeds generally contain more protein than
cereal seeds, and the protein has a better balance of amino acids for human nutrition than that
in cereals.
In addition to being used directly as foods, seeds play other roles in human diets. Many
seeds are used whole or ground as spices. The popular beverages coffee, cola, and cocoa are
derived from seeds. Beers and ales are brewed from barley, and whiskey and gin are distilled
from fermented mashes of cereal grains. Edible oils are obtained from seeds of corn, soybean,
canola/rapeseed, cotton, peanut, coconut, palm, sunflower, and safflower. And seeds are used
in the manufacture of some drugs and medicines.
x Introduction

The Mayans cultivated corn along the dry, flat plains ofthe Yucatan. Improved varieties ofcorn are
now the major cereal grain ofthe United States, with production exceeding 6 billion bushels annually.

In addition to being the most important source of food for human beings, seeds serve many
commercial functions as well. Cotton, a major fiber, is spun from the cellulose hairs that
surround the seeds of cotton plants. Seeds are also used in the manufacture of soaps, paints,
varnishes, linoleum, jewelry, buttons, and many other products.
The beginnings of the seed lie deep within the burgeoning flower, where tiny structures
begin to grow and develop to form the integument, the seed coat. This outer covering provides
protection for the mature seed; it may even contribute nutritive support for the embryonic seed.
This package also furnishes the ideal environment for the development of the embryo and the
endosperm of the young seed.

From Orchids to Coconuts

Seed size is every bit as varied as the end products of the seed. An orchid species boasts
the smallest known seed-a dust-like particle hardly visible to the naked eye. Seeds oftobacco
and Kingston velvet bent grass are so minute that one-half million of them weigh only an ounce.
Perennial plants (usually woody) claim the largest seeds. including acorns, walnuts. and
coconuts.
Introduction Xl

Seed shape runs the gamut from round, oval, triangular, elliptic, and elongated to irregular.
Predominant colors are black and brown, but hues of red, yellow, purple, green, and white are
also found. Some seeds are multicolored. Seed surfaces may be smooth, rough, or textured, with
silky hairs, cottony masses, hooks, bristles, or wing-like structures. The intricacies of seed
surfaces are often functional, serving as the mechanisms that ensure seed dispersal and survival.

'Blowin' in the Wind

Not all seeds are distributed in brightly colored packets from Burpee Seed Company. The
wind is probably the most effective agent in seed dissemination. It blows and scatters seeds in
various ways-the cottony masses attached to cottonwood seeds, the hairy tufts ofthe dandelion
and milkweed, the wing-like appendages of the ash and maple seed, and the dust-like seed of the
orchid. When the Russian thistle, the legendary tumbleweed, breaks off at the soil surface, the
wind catches at the plant, blowing it across the western plains; it disperses seeds as it tumbles.
Since seeds of almost all species will float on water, many that land in streams will sail
from their home site. Farmers and gardeners extend the territory of useful plant seeds. Animals

The legendary tumbleweed breaks off at the ground surface and tumbles across the fields with the
wind, dispersing seeds as it goes. Some C!fthe weeds have lodged against a barbed wire fence in this
field in southwestern Nebraska.
xii Introduction
and insects may also help seeds along their travels. Squirrels and packrats are notorious seed
gatherers, while ants seem to be the most active seed collectors in the insect world. Innumerable
plant species have fruits that are eaten by birds and animals. If the seeds pass through the
digestive tracts, they may be transported great distances before being dropped to the ground to
give rise to new plants.
Some tenacious seeds manage to adhere to people's clothing or to animal's furry coats.
Those who have tried to disengage the seeds of the beggar-tick from their clothes, cheatgrass
from their socks, or cockleburs from the coat of their Irish setter know just how effective this
dispersal method can be. Other seeds come equipped with spines or thorns that enable them to
attach themselves to animals' feet and spread that way. The fruit of the mistletoe scatters its seed
in a remarkable fashion. It is equipped with a propulsion mechanism that ejects the seed into the
air. The seed is covered with a sticky substance that enables it to adhere to branches of trees and
to the feet of birds. Birds carry the seed from one tree to another, where it germinates and begins
to grow.
Just as seeds feature external structures that aid in dissemination, so they are equipped to
germinate and resume their growth at a time and place that ensures the survival of their kind.
Some seeds possess the ability to germinate and grow the instant they are dispersed. Others
display almost uncanny mechanisms that prevent their germination until the time and place are
right for continued growth. Some, especially weed seeds, may lie dormant in the soil for many
years before they germinate.

Birds relish the purplish-black/roils o/the mulberry tree, and they drop the seeds in/encerows and
hedges, giving rise to new trees that grow as weeds.
Introduction Xlll

When seed pods oj the milkweed dry and split open in the Jail, the white hairs attached to the brown
seeds spread out into rounded tufts and are wafted away in the breeze, carrying some ojthe seeds a
long distance.

In many ways, the seed is a microcosm of life itself. The seed is a neatly Mapped package
containing a living organism capable of exhibiting almost all of the processes found in the
mature plant. By studying the seed or the germinating seedling, we have gained much of our
knowledge about growth regulators, respiration, cell division, morphogenesis, photosynthesis,
and other processes.
But most of the seed research has been done injust the past century. While humankind no
longer prays to the goddesses of grain (Demeter the Greek, and Ceres the Roman), we have a
long way to go to unravel all the mysteries of the seed.
XlV Introduction

The winged seeds ofthe Ural maple may blow many yards in a strong wind, but most reach the ground
near the tree.
 1
Flowering
Processes
In Plants

Plant growth originates within the buds in regions known as meristems. In the meristems,
cell division and elongation occur, and these processes produce tissues that soon develop into
specific plant parts. Vegetative meristems give rise to parts such as stems, leaves, and roots,
while reproductive meristems give rise to floral organs that ultimately produce fruits and seeds.
Within every meristem are minute primordia that resemble knobby outgrowths or ribbed
inverted cones. Although hardly distinguishable to the naked eye, the configurations of the
primordia become visible when the bud scales are removed and examined under magnification.
As growth proceeds, the configurations enlarge and differentiate into recognizable plant organs.

FLORAL INDUCTION

The ability to support reproductive processes requires tremendous energy. Often, many
crops do not begin to form flowers, and eventually seeds, until after substantial vegetative
growth has occurred. In some cases, as with most annuals, this is at the end of the life cycle. In
other cases, the plant may not become reproductive until after several growing seasons as with
many fruit trees. During this phase, in which the plant is unable to form flowers because it does
not possess sufficient vegetative structure, it is said to be in ajuvenile state. However, at some
point, enough vegetative growth occurs and plants reach sexual maturity and are able to flower.
After that stage, certain external (or internal) stimuli can trigger floral induction, a physiological
change that permits the development of reproductive primordia. This change may precede actual
flowering by several days, weeks, or even months.

Temperature Stimuli

For floral induction to occur, many plants require exposure to low temperatures. This
process has been called vernalization. In its narrowest sense, vernalization means the promotion

L. O. Copeland et al., Principles of Seed Science and Technology

© Kluwer Academic Publishers 2001
2 Flowering Processes in Plants
of flowering in some winter cereals by cold treatment ofthe moistened or germinating seeds. In
a broader sense, vernalization means the induction of flowering in any winter annual, biennial,
or even perennial species through exposure to low temperatures. For example, rye (Secale
cereale), a winter annual, and perennial ryegrass (Lolium perenne) both must undergo
prolonged exposure to low temperatures before they can produce flowers. Sugar beets and
carrots are examples of biennial species that grow vegetatively the fIrst year, after which they
are vernalized by exposure to winter temperatures. The optimum temperature for vernalization
is between loe and 7°e (Figure 1.1). These temperatures must be experienced by the vegetative
meristems for periods of between 10 and 100 days before a reproductive meristem is initiated
when the crop is returned to warm temperatures.
In chrysanthemum and tomato, floral induction is accomplished by repeated exposure to
low night temperatures, separated by periods of higher temperature. This phenomenon occurs
in many plants and has been called thermoperiodism.

Day-Length Stimuli

In many species, floral induction occurs in response to day length, or photoperiod. Thus,
plant species have been categorized according to their day-length requirements as short-day,
long-day, intermediate-day, or day-neutral; however, it is really the length ofthe night, or dark
period, that is the critical factor that influences flowering. Table 1.1 provides examples of crops
which require photoperiod and vernalization to induce flowering.
The photoperiod requirements for flowering may be qualitative or quantitative. Some short-
day plants such as the Biloxi variety of soybean and cocklebur are unable to flower except
under short-day treatments; in other short-day species, such as sunflower, flowering is hastened
by the appropriate short-day conditions, although it can eventually occur without them.

1.0

5
.~
0.8
.~
";i
c:
\"
CII 0.6
>
CII
>
.~

1'0 0.4
"ii
ex:
0.2

-10 -5 o 5 10 15 20

Temperature during vernalization (OC)

Figure 1.1. Vernalization response offlowering in winter cereals (based on data for "Petkus" rye
(from Salisbury 1963).
Flowering processes in plants 3
Table 1.1. Photoperiodic and Vernalization Responses of Some Agricultural Species.
Short-Day Plants Day-Neutral Plants Long-Day Plants
Obligate soybean soybean oat
photoperiodic rice cotton annual ryegrass
response dry bean potato canary grass
maize rice red clover
coffee sunflower timothy grass
tobacco spinach
radish
Facultative soybean cabbage
photoperiodic cotton spring barley
response sugarcane spring wheat
rice spring rye
potato potato
sunflower sunflower
red clover
Positive onion onion winter oat
vernalization carrot winter barley
requirement broadbean perennial ryegrass
winterwheat
sugarbeet

Since the original discovery of photoperiod control of flowering by Garner and Allard in
1920 and the discovery of temperature or thermal induction by a Russian scientist, Lysenko
(1932), there has been a widespread search for the existence of a universal flowering hormone,
jlorigen, in plants. However, it now appears that flowering is controlled not by one, but by
several different hormone-like substances.
Phytochrome. With plant responses other than flowering-for example, seed
germination, bud dormancy, stem elongation, and petiole development-research has shown
almost identical responses to light in different plant parts, suggesting that plant reactions are
controlled by the same light-receptive substance. In 1959, this substance was fmally isolated,
identified, and named phytochrome.
Two photoreversible forms of phytochrome exist in plants. P R phytochrome is receptive
to red light [600-680 nanometers (nrn)] and inhibits flowering while P FR phytochrome is
receptive to far-red light (700-760 nrn) and induces flowering. The conversion from P FR
phytochrome to P R phytochrome takes place in the dark, but at a much slower rate than that
induced by far-red light. This is the basis for the "day-length," or photoperiodic light response,
as well as the response to light quality (color, or wavelength) in the control of flowering. By
successive exposures to red and far-red light, flowering of light-sensitive plants can be
repeatedly induced or inhibited.

Chemical Stimuli

Certain natural and synthetic chemical substances can cause floral induction. Some are
auxinlike compounds-for example, indoleacetic acid, naphthaleneacetic acid, or the common
herbicide, 2,4-dichlorophenoxyacetic acid (2,4-D). At certain concentrations, gibberellic acid
4 Flowering Processes in Plants
may also cause floral induction. It promotes flowering of long-day plants held under short-day
conditions; however, it inhibits flowering of short-day plants under the same conditions. It has
been demonstrated that the gibberellin content increased markedly during floral induction of
Hyoscyamus niger; this is consistent with the effects of gibberellic acid in promoting floral
induction.
Other substances known to cause flowering or to increase flower production include
cytokinins, ethylene, acetylene, ethylene chlorohydrin, and 2,3,5-triiodobenzoic acid. In
contrast, maleic hydrazide inhibits flowering.
With our growing knowledge about plant flowering responses and increasing capability for
producing synthetic hormones, it is often convenient and commercially feasible to manipulate
flowering and fruit development in the commercial production of certain crops.

Nutritional Status

In floral induction, the nutritional status of a plant is also important, since construction of
the flowering parts is dependent on food availability and translocation. The carbon-nitrogen ratio
is particularly influential; in species, such as holly, that bear male and female flowers on
separate plants, a high nitrogen-to-carbon ratio favors pistillate rather than staminate flowers.
In tomatoes, carbohydrate deficiencies cause microspore degeneration, leading to pollen sterility;
however, a nitrogen deficiency has no such effect.

FLORAL INITIATION

Following floral induction, which may be triggered by external stimuli,floral initiation is

the morphological expression of the induced state and usually occurs more or less deeply within
the meristems of a plant. In monocotyledonous species, or flowering plants in which a single
embryonic seed leaf appears at germination, floral initiation begins in specialized meristems
called dermatogens, which also give rise to the epidermis. In dicotyledonous species, or
flowering plants in which a pair of embryonic seed leaves appear at germination, floral initiation
occurs in the lateral, terminal, or axillary buds.
Early in their development, reproductive meristems are similar to vegetative meristems,
appearing as knobby or ribbed configurations on an inverted cone or pedestal. As development
procedes, these configurations develop into recognizable flower parts. The structure,
development, and closure of the carpels to form the ovary can be traced in Figures 1.2 and 1.3.

FLORAL MORPHOLOGY

The typical flower of an angiosperm, or plant whose seeds are enclosed in an ovary, is
composed of petals, sepals, stamens, and a pistil. The petals, often the most conspicuous,
collectively are called the corolla. Sepals, usually (but not always) less conspicuous, are known
collectively as the calyx. The stamens are the male pollen-bearing organs, and each consists of
an anther and filament. The pistil, sometimes called the gynoecium, is the female part of the
flower and consists of the stigma, which receives the pollen, the style, and the ovary. The ovary
may be composed of one or more carpels, which may be considered as highly modified leaflike
structures. When only one carpel forms the ovary, it is termed simple and usually contains only
one locule, or cavity. A compound ovary is made up of two or more united carpels and may
Flowering processes in plants 5

ovary wall
(pericarp)

A B
Figure 1.2. Arrangement offruit into locules: (A) afruit arrangement with three locules, (B) other
arrangements.

A B c
Figure 1.3. (A) a simple carpel with one locule, or cavity, (8) a compound carpel with one locule, (C)
a compound carpel with two locules.
6 Flowering Processes in Plants
contain one or more locules, depending on their arrangement (Figure 1.3). The outermost wall
ofthe ovary is called the pericarp.
The manner in which the seeds are attached to the placenta within the ovary locules is
called placentation. Placentation occurs in one of three basic forms (Figure 1.4). Parietal
placentation occurs when the seeds are attached to the ovary wall, usually to both sides ofthe
seam where the carpels fuse to form the ovary. Axile placentation occurs in flowers with ovaries
divided by partitions, called septa, in which the placental attachment arises along the central
axis of the ovary. When no septa are present in the ovary and the seeds are attached along the
central axis, the placentation is termed free central; modifications of this occur in the case of
basal or apical placentation.
Flowers having pistils, stamens, petals, and sepals are termed complete. Incomplete
flowers lack any of these four parts. Flowers containing both stamens (male) and pistils (female)
are termed perfect; unisexual flowers, which are either pistillate or staminate, are called
imperfect. Species such as corn, that have both male and female flowers on the same plant, are
known as monoecious; those that have unisexual flowers on different plants such as holly are
dioecious.

MEGASPOROGENESIS

The seeds of angiosperms originate from meristematic tissue of the ovary wall called ovule
primordia. In species with simple ovaries, these primordia are usually located near the suture
of the ovary wall where the carpel is fused. In species with more than one carpel, or with
polycarpellate ovaries, the seeds form at the fusion of the carpels or along the septa, or central
carpel axes, depending on the type of placentation (Figure 1.4). In some fruits (e.g., tomato),
a well-developed placenta arises from which many ovule primordia develop.
Within the nucellus, or specialized tissue ofthe carpel, one cell, known as the archesporial
cell, develops special characteristics that distinguish it from adjacent cells. As this cell increases
in size, its nucleus becomes larger and its cytoplasm grows more dense in preparation for cell
division. The first division results in a megaspore mother cell and a parietal cell. Usually the
parietal cell remains undivided and soon deteriorates; however, in some species, it undergoes
further division and contributes to seed formation.
The megaspore mother cell is diploid (2N), having the same number of chromosomes as the
parent plant. However, it soon undergoes a two-step cell division known as meiosis (Figure 1.5).

Parietal Axile Free Central (basal)

Figure 1.4. Types o/placentation.

Flowering processes in plants 7
This process gives rise to four megaspores, each having one-half the chromosome complement
of the mother plant; these are thus haploid (l N) cells. Normally, only one megaspore is
functional, while the other three degenerate.

MEGAGAMETOGENESIS

The development ofthe female gametophyte, or embryo sac, from the functional megaspore
is known as megagametogenesis, which is a process of successive nuclear divisions within an
enlarging cell that becomes the embryo sac. Three successive free nuclear divisions (mitosis)
occur (Figure 1.6), culminating in eight haploid (IN) nuclei. Soon these nuclei arrange
themselves within the enlarging embryo sac and cell walls form, resulting in three antipodal cells
at one end, two polar nuclei (without cell walls) near the center, and the egg apparatus
(composed of the egg between two synergid cells) at the other end. After the two polar nuclei
fuse to form a diploid (2N) nucleus, the resulting seven-celled structure is known as the mature
female gametophyte (embryo sac), or megagametophyte, which is ready to receive the mature
male gametophyte.
This describes the normal embryo sac development as it occurs in most species. Variations
to this pattern occur in certain species, especially in the polar nuclei and antipodal development.
With few exceptions, the egg apparatus development is as described.

stigma ovule archesporial

/ primordia cell (2N)
-style

Oval}' wall
(pericarp)

ovary

B
ovule
A primordia

MEIOSIS I MEIOSIS II (:J

8.· 8·.·~· ---",~:.;....-tetrad of
CD megaspores (I N)
c o
Figure 1.5. Megasporogenesis: (A) location of ovule development, (B) cutaway section ofthe lower
region of the ovary wall (pericarp), showing origin ofthe archesporial cell; note that it is larger than
surrounding cells, having a larger nucleus and denser cytoplasm, (C) cell division during
megasporogenesis, (D) cutaway section of lower part of the ovary, showing location of the four
megaspores, three of which normally degenerate.
8 Flowering Processes in Plants

functional
megaspore

..
filA polar--+..a.....:....,...
nuclei
.' '.:
"':: ,: .. ,
.' '
.. '.'. ~ .
VsifJJ
2 3 initial
A arrangement B

Figure 1.6. Megagametogenesis: (A) three normal mitotic nuclear divisions leading to one large cell
enclosing eight nuclei. Later, cell walls enclose the nuclei and the entire structure becomes the female
gametophyte, or embryo sac. (B) mature female gametophyte.

The egg cell comprises most of the egg apparatus. It is a complete cell containing a haploid
(IN) nucleus with surrounding cytoplasm enclosed in a thin wall, or fellicle. The egg cell is
positioned near the small opening (micropyle) of the ovule formed by the surrounding
integuments. A small vacuole may be present near the point of attachment away from the
micropyle.

THE DEVELOPING OVULE

Ovule development (Figure 1.7) occurs within the ovary, which provides a location for the
nurture and development of the female gametophyte, its sexual fusion with the male
gametophyte, and embryo development, survival, and eventual regrowth. Ovule growth begins
as a small outgrowth within the nucellus. As megasporogenesis and megagametogenesis
continue, the region of the nucellus that is to become the ovule enlarges and differentiates into
definite morphological characteristics. Secondary outgrowths, or collars (integuments), soon
appear around the periphery of the nucellar outgrowths and envelop it. These usually consist of
the inner and outer integuments and ultimately become the testa (seed coat) of the mature ovule.
The developing ovule is commonly attached to the placenta by thefuniculus. The scar on
the ovule where the funiculus detaches at maturity is known as the hilum. The point where the
integuments meet at the nucellar apex is the micropyle, and the region of integumentary origin
and attachment, usually opposite the micropyle, is the chalaza. Between the chalaza and the
hilum of many species is an area known as the raphe. The raphe may be visible on the seed coat
of some species.

The Nucellus

The nucellus provides tissue for the origin and nurture of the female gametophyte, from
the archesporia I cell to the mature megagametophyte. It originates from ovary tissue and pro-
vides the site of archesporial cell origin. Subsequently, part of it becomes trapped within the
Flowering processes in plants 9

integument
ovarian locule
ovarian tocule
I
outer integument

nucellus with a fUniculus

funiculus
tetrad of megaspores
inner integument' megagametophyte,
A B or embryo sac

petal (corolla)
anther } stamen
ftlament
ovule stigma}
style pistil
ovary
petal (corolla)
~"-:--+- egg

'/
placenta D
C
Figure 1. 7. Ovule development and its location in the flower: (A) longitudinal section through the
ovary showing the developing ovule, (B) a later stage, (C) a still later stage showing the mature female
gametophyte, (D) a generalized diagram of a complete flower showing the location of the ovule.

integuments as the ovule continues to develop. Normally, no further growth occurs, and the
nuceUus is at least partially consumed, since it supplies nutritive support to the developing
embryo sac. However, in some species it undergoes considerable development and contributes
substantially to the storage tissue as the perisperm. Examples of species with well-developed
perisperm are sugar beets (Beta vulgaris) and leafY spurge (Euphorbia esula).

Integuments

The nature and thickness of the integuments vary considerably among species, depending
on their role in contributing to embryo sac and ovule development. In Apiaceae, the inner
integument is completely absorbed and only two or three cellular layers of the outer integument
persist. In Asteraceae, most cells of both integuments are absorbed, leaving only a thin layer of
crushed integumentary tissue on the inner side of the pericarp. Practically no integumentary
10 Flowering Processes in Plants

tissue remains in the fully developed corn caryopsis, and in Symplocarpus, both integuments and
endosperm are completely consumed by the developing embryo, leaving it naked inside the
pericarp.
Integumentary Outgrowths. Two types of integumentary outgrowths may occur in certain
species, giving rise to special structures not found in most sOOds. A third integument, or aril,
may either arise from the base of the nucellus or split off from the outer integument. Elymus,
for example, has a well-developed aril. Another type of integumentary outgrowth, a caruncle,
arises as a proliferation of the outer integument in the region of the micropyle. Seeds of
Euphorbia esula have a well-developed but fragile caruncle that extends back over the seed and
appears to have no function. Still another type of appendage arises from the seed coat over the
area of the raphe in some species (e.g., Stylophorum and Trillium) and is known as the
strophiole.
Integumentary Tapetum. In some species, the cells of the inner integument serve as
nutritional support for the developing embryo sac and later harden and act as a protective layer
for the ovule. In Lobelia, the cells of the inner integument take on a pronounced radial
elongation and become binucleate before becoming hardened as the integumentary tapetum.

Micropyle

The micropyle is an integumentary pore or opening in the ovule through which the pollen
tube grows to fertilize the egg cell of the female gametophyte. The micropyle may assume one
of several configurations, depending on the closure of the inner and outer integuments (Figure
1.8).

Epistase

Epistase is the development of well-defmed nucellar or integumentary tissue in the

micropylar region of the seed of certain species. In Castalia (water lily) and Costus (spiral flag)
species, epidermal cells of the nucellus proliferate and form a plug beneath the micropyle, which
remains after the rest of the nucellar tissue is gone. Cells adjacent to the micropyle may show
a marked radial elongation. Another type of epistase, an operculum, develops when cells of the
integument proliferate and form a tightly compacted micropylar plug, as in species of Lemna
and Acorus.

Figure 1.8. Types of micropyle arrangements showing different closure of the inner and outer
integuments.
Flowering processes in plants 11
Mature ovules are classified into five different types based on their arrangement within the
ovary (F igure 1.9). The difference in arrangement begins at the time of archesporial development
and becomes well defmed by the time of embryo sac maturity. Ovule types have been
determined for most well-known plant species and serve as a means of plant classification.
The effect of the ovule arrangement is often visible externally. For example, the relative
position ofthe hilum (funicular detachment scar), chalaza, and micropyle of many legumes
can be easily seen.

MICROSPOROGENESIS AND MICROGAMETOGENESIS

The period of flower development when the stigma is ready to receive the pollen is known
as anthesis. Pollen is usually produced in four sacs, or microsporangia (Figure 1.10), of the
anther, although occasionally fewer sporangia may occur. Within the sporangia, certain cells
become the microspore mother cells and undergo a two-step reduction division (meiosis), or
microsporogenesis, to yield four microspores, each of which is haploid (1 N). Each ofthe four
microspores is normally functional and undergoes two divisions, known as microgametogenesis,
giving rise to a microgametophyte, or mature pollen grain.

FRUIT DEVELOPMENT

To understand seeds and seed formation, one must have a basic knowledge of fruit
development and morphology. The botanical definition of fruit is much broader than that
conveyed by popular usage of the term. Actually, a fruit is a mature or ripened ovary that
usually contains one or more ovules that develop into true seeds. Legume pods, peppers, and
cereal grains are fruits, as are apples, oranges, and peaches.
The pericarp, or ovary wall of angiosperm fruits, is composed of three different layers
which are more or less distinct in various species: the exocarp, or outer layer; the mesocarp, or
middle layer; and the endocarp or inner layer. The relative development of each in various
species contributes to the overall fruit structure and morphology.

A B c D E

Figure 1.9. Types of ovules as seen in vertical longitUdinal section: (A) atropous (or
orthotropous)-nucellar apex points away from the funiculus as in Polygonaceae, (B) anatropous- ovule
completely inverted so that nucellar apex is turned toward the funiculus as in Sympetalae, (C)
campylotropous-ovule is slightly curved, with the nucellar apex andfunicular end pointed slightly
downward as in Fabaceae, (D) hemianatropous-ovule is straight with axis lying perpendicular to the
funiculus, as in Ranunculaceae, (£) amphitropous-ovule has a pronounced curve, with the nucellar
apex near the funiculus, as in Botomaceae (From P. Maheshwari 1950).
12 Flowering Processes in Plants
micros pore mother

~1--~.eIl

CROSS SECTION, microsporangium

YOUNG ANTHER
mlCrospores

tube cell
pollen
grain GERMINA TION OF POLLEN
Figure 1.10. The anther and the pollen grain. Each microspore mother cell within a microsporangium
divides to form a tetrad of microspores that soon separate. The nucleus of each microspore then
divides, and a tube cell and generative cell are formed within the wall of the microspore, which
subsequently develops into a pol/en grain. Following pollination, the pollen grain germinates,
producing a pollen tube, and the generative cell gives rise to two male gametes (From Wilson and
Loomis 1952).

FRUIT TYPES

Pseudocarpic fruit consists of one or more ripened ovaries attached or fused to modified
bracts or other non floral structures. Examples: burdock, sandbur.
Multiple fruit is composed of the ovaries of more than one flower. Each unit of these fruits
may be berries, drupes, or nutlets. Examples: fig, mulberry, pineapple.
Aggregatefruit is composed of several ovaries ofa single flower. Each unit of these fruits
may be a berry, drupe, or nutlet. Examples: strawberry, raspberry, blackberry.
Simple fruit is derived from a single pistal.
A. Fleshy fruits have a fleshy or leathery pericarp.
1. Berry has a fleshy pericarp. Examples: grape, tomato, gooseberry, huckleberry.
2. Pepo has a hard rind but no internal separations, or septa. Examples: watermelon,
cantaloupe, squash, cucumber.
3. Pome has a floral cup that forms a thick outer fleshy layer and a papery inner pericarp
(endocarp) forming a multiseeded core. Examples: apple, pear, quince.
Flowering processes in plants 13
4. Drupe is also called stone fruit, and has a stony endocarp, a thick, leathery, or fleshy
mesocarp, and a thin exocarp. The pit is usually one~seeded, but occasionally several
one~seeded pits are present. Examples: cherry, coconut, walnut, peach, plum, olive.
5. Hesperidia are berrylike fruits with papery internal separations, or septa, and a leathery,
separable rind. Examples: orange, lemon, lime, grapefruit.
B. Dry fruit has a thin pericarp that is dry at maturity.
1. Dehiscent fruits split open at maturity and releases mature seed.
a. Legume has a simple (single) pistil that splits open at maturity along two sutures.
Examples: bean, pea, soybean, black locust.
b. Follicle has a simple (single) pistil that splits open at maturity along one suture.
Examples: milkweed, larkspur, spirea.
c. Capsule has a compound pistil that splits open at maturity in one of four ways:
Loculicidal-splitting open through the midrib of the carpel into the locules.
Examples: iris, tulip.
Circumscissle-splitting open at the middle so that the top comes off like a lid (also
called pyris). Examples: plantain, portulaca.
Septicidal-splitting along the septa. Examples: yucca, azalea.
Poricidal-splitting open at pores near the top, releasing mature seeds. Example:
poppy.
d. Silique and Silicle are characteristic of the mustard family, with two valves which
at maturity split away from a persistent central partition. A fruit that is several times
longer than wide is termed silique, while a silicle is broad and short.
2. lndehiscent fruits do not open at maturity to release the seeds.
a. Achene is a small one-seeded fruit in which the seed is attached to the pericarp at
only one point and may be rather loose inside the pericarp. Examples: dandelion,
buttercup, sunflower, dock.
b. Utricle is similar to an achene except that it has an inflated papery pericarp.
Example: Russian thistle.
c. Caryopsis is similar to an achene except that the entire seed coat is tightly fused with
the pericarp. Example: grasses.
d. Samara is similar to an achene except that the pericarp develops a thin, flat, winglike
appendage. This is a characteristic of some woody species. Examples: ash, elm, tree
of heaven. Double samaras occur in the fruit of maple.
e. Nut is a dry on~seeded fruit from a compound pistil that has a very hard and tough
pericarp that is usually wholly or partially enclosed in an involucre. Examples:
acorn, hazel, filbert, chestnut.
f. Nutlet is a small, dry fruit composed of on~half a carpel, enclosing single seed. It
is developed by folding and splitting of the carpels into a compound pistil.
Examples: members ofLamiaceae (mint family) and Boraginaceae (forget-me-not
family).
g. Schizocarp has two fused carpels separating at maturity to form one-seeded
mericarps. Example: members of Apiaceae (carrot family).
14 Flowering Processes in Plants
FLORAL TAXONOMY

The arrangement of the floral axis determines the type of inflorescence (structure of a
flower), and is a stable species characteristic. The main stalk of the inflorescence is the
peduncle. Lateral stalks supporting the individual flowers are called pedicels. Inflorescences
may be determinate or indeterminate. Determinate inflorescences are those in which the axis
terminates as a flower. Indeterminate inflorescences terminate in a vegetative bud, which
continues to grow and produce flowers throughout the growing season, resulting in flowers of
different maturity within the same inflorescences (see Figure 1.11).

Determinate Flowers

Solitary flower-The simplest expression of a determinate inflorescence. Example: com

cockle.
Simple cymt>-The simplest branched determinate inflorescence where the lateral flowers
develop later than the terminal flower. Example: chickweed.
Compound cymt>-A determinate inflorescence where there is secondary branching and
each lateral unit becomes a simple cyme. Example: bouncing bet.
Scorpioid cymt>-A determinate inflorescence in which the lateral buds on one side are
suppressed during growth, resulting in a curved or coiled arrangement. Example: Heliotropium
curassavicum.
Glomeruit>-A very compact compound cyme. Example: saxifrage.

Indeterminate Flowers

Racemt>-The basic type of inflorescence in which pedicels arise laterally on a long central
peduncle. Examples: pennycress and field or garden bean.
Paniclt>-An inflorescence in which the lateral branches arising from the peduncle produce
flower-bearing branches instead of single flowers. Example: oats.
Spikt>-An inflorescence in which the flowers arising along the peduncle are essentially
sessile, or stalkless, and are attached to the peduncle. Example: wheat.
Catkin-A modified type of spike with a single unisexual flower arising from the peduncle.
Example: red alder
Spadix-A special kind of spike covered by a spathe. Example skunk cabbage.
Corymb-An inflorescence in which the lower pedicels arising from the peduncle are
successively longer than the upper ones, giving a round or flat-topped appearance. Example:
bitter cherry.
Umbel-An inflorescence similar to a corymb except that the lateral branches arising from
the peduncle originate from the same location. Example (simple umbel): pennywart. A
compound umbel is similar except that each pedicel is branched, bearing multibranched
individual flowers. Example: wild carrot.
Head-An inflorescence where the peduncle and the pedicels are tightly clustered,
surrounded by a group offlowerlike bracts called involucre. Example: sunflower.
Flowering processes in plants 15

Detenninate Inflorescences

Simple Cyme Compound Cyme

Scorpioid Cyme

Indeterminate Inflorescences

Panicle Catkin
Raceme Spike

Corymb Simple Umbel Compound Umbel Head Spadix

Figure 1.11. Types of determinate and indeterminate iriflorescences (From Dennis 1967).
16 Flowering Processes in Plants
Questions

1. What is the difference between floral induction and floral initiation?

2. Do you think that phytochrome and florigen are the same?
3. What is the difference between a complete flower and a perfect flower?
4. Can a dioecious plant have complete flowers?
5. What is the relationship between a peduncle and a pedicel?
6. What is the difference between a caryopsis and an achene?
7. What is the difference between a schizocarp and a mericarp?

General References

Boke, N. H. 1947. Development of the adult shoot apex and floral initiation in Vinca rosea.
American Journal ofBotany 34:433-439.
_ _ . 1948. Development of the perianth in Vinca rosea L. American Journal of Botany.
35:413-423.
_ _ . 1949. Development of the stamens and carpels in Vinca rosea L. American Journal of
Botany 36:535-547.
Bonnett, O. T. 1935. The development of the barley spike. Journal ofAgricultural Research
51:451-457.
_ _. 1936. The development ofthe wheat spike. Journal ofAgricultural Research 53:445-
451.
_ _ . 1937. The development of the oat spike. Journal ofAgricultural Research 54 :92 7-931.
Dennis, L. 1. 1967. Manual of Introductory Taxonomy and Field Botany. Corvallis, OR.:
Oregon State University Bookstores.
Gamer, W. W. and H. A. Allard. 1920. Effect of the relative length of day and night and other
factors of the environment on growth and reproduction in plants. Journal of Agricultural
Research 18:553-606.
Lysenko, T. D., 1932. Fundamental results of research on vernalization of agricultural plants.
Bull. Jarovizacci, No.4, 1-57. Quoted by Maximow, 1934.
Maheshwari, P. 1950. An Introduction to the Embryology of the Angiosperms. New York:
McGraw-Hill Book Company.
Maximow, N. A. 1934. The theoretical significance of vernalization. Imperial Bulletin of
Genenetics. Aberystwyth (Wales) Herbage Publication Series Bulletin 16.
Salisbury, F. B. 1961. Photoperiodism and the flowering process. Annual Review of Plant
Physiology 2:293-326.
_ _. 1963. The Flowering Process. New York: Pergamon Press.
Searle, N. E. 1965. Physiology of flowering. Annual Review of Plant Physiology 16:97-118.
Siegelman, W. and W. L. Butler. 1965. Properties of phytochrome. Annual Review of Plant
Physiology 16:383-392.
Stratford, G. A. 1965. Plant hormones II: Florigen and gibberellins. Essentials of Plant
Physiology. London: Heineman Educational Books, Ltd.
 2
Seed Formation
and Development

SEED FORMATION

Seed formation begins with the fusion of a male and female gamete, a process known as
fertilization. Fertilization, or syngamy, can occur when both male and female gametophytes are
fully mature. This usually occurs in a dual fusion process known as double fertilization (Figure
2.1). When the pollen grain lands on the stigma, it germinates by sending out a pollen tube,
which grows down the style, through the micropyle and into the embryo sac, with the tube
nucleus closely following the tube apex downward. The tube nucleus soon degenerates, but the
two pollen sperm cells enter the embryo sac, one fusing with the diploid (2N) polar nucleus to
form a triploid (3N) endosperm nucleus and the other fusing with the egg cell to form a diploid
(2N) zygote, or fertilized egg.
The process of fertilization is very important because it not only results in the formation
of a seed but also dictates the level of genetic diversity present in the zygote. Fertilization in
angiosperms typically occurs either by self- or cross-fertilization.

Self-Fertilization

Self-fertilization occurs when pollen from the anthers of a flower is transferred to the stigma
of the same flower, resulting in fertilization. In most cases, this occurs in flowers that do not
open until after pollination and fertilization are complete.

Cross-Fertilization

Cross-fertilization occurs when pollen from one flower is transferred to the stigma of
another flower to cause fertilization. The flowers can be on the same or different plants. In most
agricultural crops, cross-fertilization occurs by two principal methods: wind (anemophily) and
insects (entomophily). Unlike self-fertilization, where progeny are genetically similar, cross-
fertilization results in progeny that are more dissimilar. This evolutionary approach produces
a population of individuals that are more adaptable to a wide array of environmental conditions.

L. O. Copeland et al., Principles of Seed Science and Technology

© Kluwer Academic Publishers 2001
18 Seed Formation and Development

tube
nucleus

A /-L. . '
sperm'cens
, . , (gametes)
pollen
tube

Figure 2.1. Doublefertilization: (A) pol/en tube with its two sperm cells and tube nucleus approaching
the micropyle, (B) sperm cells approaching egg and polar nuclei, (C) double fertilization has occured.

ASEXUAL REPRODUCTION IN PLANTS

In addition to sexual reproduction, many plants are able to reproduce asexually. There are
two types of asexual reproduction in plants: vegetative propagation and apomixis. Vegetative
propagation may be carried out by stolons, rhizomes, tubers, tillers, bulbs, bulbils, or corms.
Apomixis is the production of seeds and vegetative propagules by asexual methods. The main
features of apomixis are: (I) it substitutes asexual reproduction for sexual reproduction, (2) it
occurs in parts of the plant normally concerned with the sexual process (flowering parts), and
(3) it occurs without fusion of egg and sperm cells.
There are two types of apomixis: vegetative proliferation and agamospermy. Vegetative
proliferation (also termed vivipary) is the conversion of the spikelet, above the glumes, into a
leafY shoot. Agamospermy is apomixis through seed production in which substitutions occur for
meiosis (reduction division) and fertilization or both. Agamospermy may occur through
adventitious embryony or gametophytic apomixis.
Seed Formation and Development 19
Adventitious embryony is the development of a diploid (2N) embryo from nucellar or
integumentary tissue (sporophyte tissue). In gametophytic apomixis, unreduced embryo sacs
(gametophytes) are formed without meiosis through the processes of apospory or dip/ospory.
Reduced embryo sacs may also be developed through meiosis.
Tn apospory, diploid embryo sacs are formed in the nucellus or inner integuments by
mitotic divisions. In diplospory, the diploid embryo sac is formed by the megaspore mother cell.
Embryos are formed from the diploid egg cell without fertilization through the processes of
parthenogenesis and pseudogamy.
In diploid pseudogamy, endosperm development requires the stimulus of pollination, and
the egg cell develops without fertilization; in diploid parthenogenesis, pollination is not required
for embryo development. Haploid (IN) embryos are formed from haploid eggs through haploid
parthenogenesis.
These relationships are diagrammed in Figures 2.2 and 2.3. Figure 2.2 shows the general
relationships between the various types of asexual reproduction and Figure 2.3 contrasts sexual
seed production with agamospermy, showing substitutions for meiosis and fertilization.

SEED DEVELOPMENT

In most species, seed formation follows the normal pattern already described. It begins
within the minute embryo sac which, with certain exceptions, is about the same in shape, size,
and arrangement. In spite of initial similarities, the seed develops according to the genetic
specifications for each species, which are coded in the DNA of each cell.
The embryo sac may be ellipsoidal, elongated, or variously bent in shape. The longitudinal
axis extends from the chalaza to the micropyle and through the antipodal cells, the endosperm
nucleus, and the egg apparatus. Morphologically, the micropyle is at the upper end of the
embryo sac.
The embryo sac is a biochemical and biophysical system of considerable complexity. As
a growing, differentiating structure, it requires a constant nutritive supply, which is provided
through the chalaza, establishing a polar gradient from the antipodal to the micropylar end.
Nutrition is also obtained from the nucellus and integumentary layers directly through the wall
ofthe embryo sac.

EMBRYOGENY

After sexual fusion, or syngamy, a brief period of reorganization occurs, during which the
large vacuole adjacent to the zygote gradually disappears, with the zygote cytoplasm becoming
more homogeneous and the nucleus larger. The duration of this period varies among species, but
it is usually about four to six hours before the zygote begins to divide (Table 2.1). Lines of
polarity in preparation for future division and growth already exist in the embryo sac, having
been established in the unfertilized egg. The still undivided zygote typically elongates along the
horizontal axis and small vacuoles become evenly distributed throughout the cytoplasm.
 tv
[ASEXUAL REPRODUCTION o

VEGETATIVE 1 l
PROPAGATION ~

VEGETATNE
PROLIFERATION
(VIVIPARy)

--
Apospory I
Diplospory
------ i ----- __ ___ - MeiosIs
I
" -'
", tI
,-' /-' -
DIPLOID HAPLOID
EMBRYO SAC EMBRYO SAC
-' , i
Diploid .., 'Diploid Haploid ~
parthenogenesis pseudogamy parthenogenesis It
" -' " I ~
"" -'
RHIZOMES, ETC. BULBILS. ETC.
~
§.

SEEDS NOT FORMED SEEDS FORMED

~
~
;€
SPOROPHYTIC DEVELOPMENT GAMETOPHYTIC DEVELOPMENT a-
]
Figure 2.2. Mechanisms of asexual reproduction (Courtesy of D. F. Grabe). ~
Seed Formation and Development 21

Steps in Seed Formation

Sexual Asexual (agamospermy)

( FLOWER I
,ovL ,
MEGASPORE MOTHER CELL
I
MEGASPORE
MOTHER
CELL
/: I
I I
I \ I
(Meiosis)
\ I
I \ I
(Diplospory) (Apospory)
HAPLOID MEGASPORE \ I
\ I
\ I
, t
I HAPLOID EMBRYO SAC I
I
DIPLOID EMBRYO SAC I
I 1\
I / \
(Fertilization) I \
I / \
J I \
I I \
( ZYdOTE I /
(Parthenogenesis) (pseudogamy)
\
I \
/ \
~ ~
EMBRYO (SEED) (EMBRYO) (EMBRYOI

!
[ MATURE PLANT ~ MATURE PLANT ~
I FLOtR I
Figure 2.3. Steps in seed formation through sexual vs. asexual methods (Courtesy of D. F. Grabe).
22 Seed Formation and Development
Table 2.1. Examples of Time Sequence of Embryo Development of Two Species.

A. Hordeum distichon palmella

Time elapsed Growth of Development Development
after pollination pollen tube of embryo of endosperm
5 min. Pollen germinated
10 min. Male gametes inside
pollen tube
45 min. Entry of pollen tube One male gamete in Other male gamete in
into embryo sac contact with egg contact with polar
nuclei
5 hr. Male nucleus forming Male nucleus and polar
a sector of the egg nuclei in process of
nucleus fusion
6 hr. Male sector of zygote First division of primary
nucleus becoming endosperm nucleus
more and more
diffuse
10 hr. Second division of
primary endosperm
nucleus
13 hr. Prophase of first Four endosperm nucleL
division of zygote
15 hr. First division of zygote Eight endosperm
nearing completion nuclei
B. Taraxacum kok-saghys
15 min. Entry of pollen tube
into embryo sac
45 min. Discharge of pollen
tube and approach
of male gametes
toward egg and
secondary nucleus
1 hr., 15 min. Syngamy Triple fusion
3 hr., 50 min. First di vision of primary
endosperm nucleus
5 hr. First division of zygote
6 hr., 15 min. Two-celled proembryo Two-nucleate
endosperm
8 hr., 15 min. Four-celled embryo Four-nucleate
endosperm
24 hr., 45 min. Several-celled Multicellular
proembryo endosperm
Data from Maheshwari (1950).

Types of Embryo Development

The first few cell divisions of the zygote form the proembryo. Plant species may be
classified according to the pattern of cell division, which results in different proembryo types.
The first division almost always occurs at right angles to the longitudinal axis, resulting in a
terminal cell next to the micropyle and a basal cell at the distal end. Depending on the pattern
of subsequent divisions, proembryos are classified as crucifer, asterad, solanad, caryophyllad,
chenopodiad, or pipered.
Seed Formation and Development 23
1. The first division of zygote is transverse.
A. Terminal cell of proembryo divides by a longitudinal wall.
a. Crucifer-basal cell plays only a minor role (or none) in embryo development.
b. Asterad-both the basal and terminal cells contribute to embryo development.
B. Terminal cell of the proembryo divides by a transverse wall.
a. Solanad-basal cell plays only a minor part (or none) in the development of
the embryo.
b. Caryophyllad-basal cell undergoes no further division, and the suspensor,
if present, is always derived from the terminal cell.
c. Chenopodiad-basal cell and terminal cell both contribute to embryo
development.
2. The first wall of the zygote is longitudinal-or nearly so-Pipered

Although the mature embryos of monocotyledons and dicotyledons appear considerably

different, their patterns of embryogeny are similar. The proembryo is divided into the suspensor
and embryo proper. The suspensor forms into a chain of cells, pushing the embryo proper up
into the center of the ovule in contact with the available food supply. The expression constancy
ofdestination suggests that each part of the mature embryo inevitably arises from special parts
of the proembryo; however, proembryos may vary greatly in size and shape.
Figure 2.4 shows the embryo growth of typical dicotyledonous and monocotyledonous
specIes.

Laws of Embryouy

Four laws of embryony (embryogeny) have been established to describe embryo

development:

1. Law ofParsimony: No more cells are produced than are absolutely necessary.
2. Law of Origin: In any species the sequence of cell formation is established in such a
way and with such regularity that the origin of any cell can be specified in terms
of, or related to, the earlier units of the sequence.
3. Law of Numbers: The number of cells produced by different cell generations varies
with the species and depends on the rapidity of the segmentation in the cells of the
same generation.
4. Law of Destination: In the course of normal embryonic development, the cells are
formed by divisions in clearly determined directions, and most appear to occupy
positions in accordance with the role they must play.

ENDOSPERM DEVELOPMENT

The endosperm serves as the principal nutritive support for the embryo of many species
(especially monocotyledons) during both seed development and germination. In angiosperms,
the endosperm normally originates from triple fusion of a sperm cell nucleus from the pollen
tube with the diploid polar nucleus (following fusion of the two polar nuclei) of the embryo sac;
therefore, its nuclear complement is triploid (3N). In gymnosperms, the endosperm is normally
haploid (IN), since it develops from one cel1 of the female gametophyte.
24 Seed Formation and Development

6\ ·

l\
two-cell stage

~~Jffi\"
s~penw~ffi

shoot apex

developing
cotyledon
endosperm

mature
embryo
suspensor
cotyledons
epicotyl
hypocotyl
radicle ~----.
A. Dicot scutellar node
disintegrading / ' root apex
suspensor coleorhiza

Figure 2.4. Typical embryo development: (A) dicotyledonous species, (8) monocotyledonous
species.

In seeds of many species, especially dicotyledons, the endosperm develops only a few cells,
while in others it may be highly modified and hardly recognizable. In Orchidaceae, it is
completely suppressed. Triple fusion occurs in Orchidaceae, but the products soon degenerate
after one or two cell divisions. In most dicotyledonous species, the endosperm is formed but is
almost completely consumed during seed development so that the mature seed is composed
almost entirely of embryo. Considerable speculation exists about the status of the endosperm.
It has been called an anomalous embryo, since the ovum and the two polar nuclei are genetically
identical. Regardless oftheir genetic similarity, the fusion ofthe two polar nuclei and subsequent
fusion with one sperm cell yield the endosperm, while the fusion of the egg with the other sperm
cell nucleus yields the zygote.
One ofthe principal endosperm functions is to provide nutrition for the developing embryo;
therefore, its composition is compatible with the embryo's needs. But the endosperm must also
draw its nutritive support from the embryo sac and surrounding tissues. The net effect is to
surround the embryo with a rich nutritive tissue from which it can draw for development and
growth. This creates competition for nutrients, both within and outside the embryo sac.
Seed Formation and Development 25

Types of Endosperm Development

Division of the primary endosperm nucleus yields micropylar and chalazaI chambers, one
or both of which may contribute to the mature endosperm. When only one develops, the other
is crushed and soon degenerates. Endosperm development may be one ofthree types, depending
on the sequence of nuclear division and cell wall formation (Figure 2.5).
Cellular Endosperm. In this type of endosperm, each nuclear division is accompanied by
cell wall formation.
Nuclear Endosperm. This endosperm type is characterized by nuclear divisions
unaccompanied by cell wall formation. The nuclei may remain free or may later be separated
by cell walls that form in one of three ways: (I) one to three layers of cell wall may form around
the periphery, with free nuclei inside, (2) a cell wall may form in the micropylar area, with the
rest remaining in a free-cell state, or (3) the entire endosperm may be filled with walled cells.
All three endosperm conditions may exist in the same family.
Helobial Endosperm. The helobial endosperm is intermediate between the nuclear and
cellular types. Free nuclear divisions occur, but cell wall formation accompanies nuclear
division in some parts of the endosperm as well.

Endosperm Haustoria

A remarkable feature of the developing endosperm is its capacity to take nutrients from
surrounding tissue for its own development. Nutrient-gathering outgrowths, called haustoria
(Figure 2.6), may develop at either the micropylar and chalazal ends and reach into the nucellar,
integumentary, or even ovary tissue. Haustoria often branch into several prominent lobes, or
diverticulae. When local food supply is exhausted, the haustoria lobes terminate and become
crushed by further endosperm and embryo growth .

..:..-_- endosperm nucleus

>--~.L:T~- zygo te

~~ proembryo

B c o
Figure 2.5. Diagram of types of endosperm development: (A) ovule after fertilization, showing
primary endosperm nucleus and zygote, (B) cellular endosperm, (C) nuclear endosperm, (D) helobial
endosperm.
26 Seed Formation and Development

B c
Figure 2.6. Haustoria arrangements in the developing endosperm of three different species: (A)
Centranthera hispida, (B) Nemophila, (C) Impatiens royle (From Maheshwari 1950).

The Mature Endosperm

Monocotyledonous endosperms usual1y reach their maximum morphological development

at physiological maturity and remain to comprise a major part of the seed. In dicotyledonous
species, the endosperm may either not develop or may be used up by the developing embryo and
comprise none or only a small part of the mature seed.
Seeds with little or no endosperm are exalbuminous, while those with a well-developed
endosperm (or perisperm) are known as albuminous. Some species have a well-developed
chalazosperm, in which both the nucellus and endosperm disappear during development and
chalazal tissue proliferates and forms storage tissue.
The outermost layers ofthe endosperm are known as the aleurone layer. During endosperm
development, aleurone cells become thickened and filled with protein granules. These layers
function both as storage tissue and for secretion of hydrolytic enzymes, which upon activation
during germination help break down storage tissues.

OVERALL SEED DEVELOPMENT

Seed development can be illustrated by the changes that occur in barley, which are typical
of most grasses and cereal grains. Endosperm development of barley is of the cellular type, in
which the first few divisions ofthe primary endosperm nucleus give free nuclei. Cell walls form
Seed Formation and Development 27
about two days after fertilization, beginning with changes at the periphery of the endosperm
which later become the aleurone. During early endosperm growth, the proembryo also begins
to grow and differentiate; however, its contribution to the overall seed morphology is
overshadowed by that of the endosperm.
Cell organelles-plastids, mitochondria, ribosomes, and golgi complexes-become
recognizable immediately after initial cell formation, followed by the endoplasmic reticulum.
After about three weeks, starch and protein granules completely dominate endosperm
composition.
Morphological Development. Morphological development of the seed occurs
concurrently with cytological, chemical, and weight changes noted below. Such morphological
changes can also be illustrated by those occurring in barley. These have been described by
Briggs (1978) and are shown in Figures 2.7-2.10, beginning with development of the
reproductive meristem and culminating in the development of the mature caryopsis.
Changes in Weight. After sexual fusion, the developing seed begins to increase in
weight as a result of nutrient and water intake associated with rapidly accelerating cell division
and elongation. Typically in monocots, the developing endosperm accounts for most of the
weight increase, with the testa-pericarp weighing somewhat less, and the embryo's weight almost
negligible. The developing barley seed undergoes a sharp increase in dry weight until about 35-
40 days after fertilization. Immediately after fertilization, most of the dry weight is in the seed
coat; however, after about eight days, its weight is surpassed by the endosperm, which later
becomes the major seed component.
Chemical Changes. Immediately after fertilization, seed development begins and the seed
becomes the primary recipient (sink) for the assimilates within the plant. There are three general
stages during seed formation. The fITst stage is when 80% of the seed growth occurs. It is
characterized by numerous cell divisions and elongation and dramatic increases in seed weight
as nutrition is supplied through the funiculus by the parent plant. The second stage occurs when
the funiculus degenerates and the seed is separated from the parent plant. At this point, the seed
has achieved its maximum dry weight and seed quality, a stage known as physiological
maturity. The third stage is when the seed undergoes further desiccation after physiological
maturity. This stage is influenced by a variety of weather conditions such as rainfall, high
temperatures, and by exposure to field pathogens that increase and decrease seed moisture
content and cause reductions in seed quality. Eventually, seeds reach harvest maturity, which
is the moisture content (usually 15-20%) at which mechanical harvesting ofthe seed is possible.
Figure 2.11 illustrates these three growth stages.
In monocotyledonous seeds, the major carbohydrate in the endosperm and the entire seed
is starch. The carbohydrate content increases rapidly as the endosperm develops, somewhat at
the expense of the testa-pericarp tissue, where it decreases slightly. Sucrose and reducing sugar
levels, initially high in the young endosperm, decrease rapidly as the starch content rises.
However, both sucrose and reducing sugars increase in the testa-pericarp during early seed
development and then decrease rather sharply as the seed matures.
Immediately after fertilization, the endosperm nitrogen of the barley seed is about 50%
protein in form. As development procedes, the protein nitrogen increases rapidly for about 20
days, after which it remains constant. Amide form of nitrogen increases slightly, so its relative
28 Seed Formation and Development
proportion in the endosperm remains constant. The testa-pericarp nitrogen content follows a
similar trend, although at a slower rate, since the total growth rate of these tissues is slower.
Negligible change in the DNA and RNA of the testa-pericarp occurs during seed
development, since they are nucleotides of the nucleus and cytoplasm, and any marked increase
in their occurrence is a reflection of cell division. In contrast, DNA and RNA increase rapidly
with increased cell division during early embryo and endosperm growth, but level off with
increased cell expansion.

(01 (bl leI

~
Single
ridge 'A Single
ridges

Idl leI

Glume
initial
Glume Floret
initials initial ~~...s.1
lemma--"l:....:o-.-

Collar

AW,
I'll
Ihl
ary
Glul1l4t Ov~r Stamen
• Paleo {
'. och~1a /
~?".;fI
".<. "' ... Lateral floret
t
Rochilla Lateral floret row

Figure 2. 7. Stages in the differentiation of the ear of a two-rowed barley: (A) single ridges present;
these willform leafprimordia. The apex is lenthening; (B) the apex elongatedfurther, but still with
single ridges; (C) double ridgesforming. Initials ofthe lateraljlorets are detectable; (D) the initials
of the sterile glumes associated with the median floret are visible; (E) a further stage; the collar is
visible; (F) a more advanced ear, viewedfrom the side; (G) a developing medianjloret, viewedfrom
the rachis side; (H) a more advanced triad ofspikeletsjrom a two-rowed barley (From Briggs 1978).
Seed Formation and Development 29
Nitrogen is also found in the developing seed in the form of amino acids and protein-bound
phosphorus. The endosperm amino acid content increases rapidly during the first two or three
weeks of seed development. This period corresponds to the time when the endosperm is high in
RNA content, which directs amino acid synthesis.

ENVIRONMENTAL EFFECTS

The environment in which the seed forms affects its development. This is often illustrated
by changes in seed size and weight which are influenced by such factors as soil fertility,
moisture, temperature, light, and position on the plant.

Outer side ~ lobe of ovary,

with hairs

~~I
Base
Inner

(€jlbl
integument
Placenta
IC) Chalazal
tissue Embryo-
sac Micropyle

Stigmatic Lemma
hairs
Ovary

Style

Rachilla

Figure 2.8. Successive stages in the development of the barley ovary: (A-D) successive stages in
median, vertical section. The arrow indicates the approximate position ofthe embryo sac mother cell
as the tissue rotates; (E) a view of the ovary apex (at 9(], to sections A-D), showing the developing
stigmatic hairs on the underdeveloped styles; (F) a longitudinal section of half of the ear, indicating
the relative position of the various parts of the spikelet (From Briggs 1978).
30 Seed Formation and Development

Soil Fertility

Generally, plants that have been fertilized with the three major elements (N, P, K) produce
larger seeds than those which have not been fertilized. The increase in seed size is due to a
greater seed development rate during the seed filling period as a consequence of increased
nutrient availability. This is true for soybean (Boswell and Anderson 1976) and tomato (Varis
and George 1985) seeds. Of all nutrients which affect seed development, nitrogen clearly has
the greatest effect. It increases seed size in perennial ryegrass (Ene and Bean 1975), soybean
(Ham et al. 1975), and com (Eck 1984), although it has also been shown to decrease seed size
in wheat (Frederick and Marshall 1985) and tomato (George et al. 1980). These differences in
response to nitrogen might be attributed to the time at which the nitrogen was applied. Earlier
applications of nitrogen have been shown to produce greater seed weight in wheat (Larger and
Liew 1973) and rice (Humphreys et al. 1987).
Production factors also can influence seed development. Increased competition for limited
nutrients by weeds or from crops as a consequence of narrow row spacings and increased
numbers of seeds/row result in decreased seed size. The intensity of the competition can vary
within (Westermann and Crothers 1977) and among species (Elmore and lackobs 1984).

Moisture

Prolonged droughts and reduced soil moisture content decrease seed size (e.g., soybean and
flax), particularly when these effects occur during flowering and seed fill (Meckel et al. 1984).
If drought occurs only before flowering, its primary effect is on a reduction in seed number,
while seed size is unchanged. This has been observed in soybean (Wright et al. 1984), corn (Eck
1986), clover (Andrews et al. 1977), and wheat (Sionit et al. 1980). The lack of soil moisture
may reduce photosynthesis, which shortens the seed filling period, thereby reducing seed size.

Strand of
pollen-conducting
tissue
Lateral
procambial
strand
Tip of outer
integument
ditR....-t-r--Antipodal cells
Outer} integuments
/NIH--+--Inner
Polar nuclei
Egg cell and two
synergids

Figure 2.9 The route of the pollen tube from the stigmatic hair to the egg cell (From Briggs 1978).
Seed Formation and Development 31

Upper lobe of
of ovary with
epithelial hairs

Outer epidermis Apical cone of tile

Inner epidermis outer integument

",---+-.a.... Ovary wall

Photosynthetic
cells
Outer
integument
Inner
integument
...u4---r-Nucellus -t--rl---f----,,*,
Embryo-sac -\--t""'.----\+

o (b)
Major
provascular strand

Figure 2.10. The mature ovary, before anthesis: (AJ a style with the apical stigmatic hairs,
each composed offour columns ofcells around a central lumen, and the simple basal hairs:
(B) the ovary in transverse section (C) the ovary in longitudinal section (From Briggs 1978).

Temperature

High temperatures during seed development results in development of smaller seeds, while
low temperatures favor larger seeds in orchardgrass and perennial ryegrass (Shimzu et al.
1979), bean (Siddique and Goodwin 1980), wheat (Wardlaw et al. 1989), lupin (Downes and
Gladstone 1984), flax (Green 1986), and sorghum (Kiniry and Musser 1988). Annual
variations in the environment also influence seed size in soybean (Egli et al. 1987), sugar beet
(Wood et al. 1980), and birdsfoot trefoil (McGraw et al. 1986).

Light

In general, reduced light to the parent plant results in smaller seeds. This effect has been
found in carrot (Gray et al. 1986), pea (Gubbels 1981), soybean (Egli et al. 1987), com (Kiniry
and Richie 1985), and clover (Collins et al. 1978). Partial shading results in decreased seed
32 Seed Formation and Development

~~ to) Ib)
fr
@0 W
Ic) Ie)

Figure 2.11. The appearance of the developing seed at various stages after fertilization. The
diagrams span the first 10 days' development. Beyond this time, growth in width and depth continues,
but first the palea then the lemma adhere to the pericarp (ovary wall). (A) The ovary in surface view
and median longitudinal section (L-S) one day after fertilization; (B) median L-S of ovary two days
after fertilization; the embryo sac is lengthening; (C) four days afierfertilization; surface view, median
L-S and dorsal and ventral views of the separated embryo sac. Note the tiny size ofthe embryo; (D)
six days after fertilization-four views; (E) the day-old seed in plane and side view, and in median L-S
and the embryo (L-S, greatly enlarged). Note the decreasing size ofthe ovary tip (From Briggs 1978).
Seed Formation and Development 33

200 Beeson 1976 ~M H,M 100

I
I 90
180 I

160 80
:0
0. 140 70 J
..s
"'0 120 602:?
00-

;:)
Q)
Q) ~
~ 100 50 '0
E
1: cu
co
.~ 80 40 o
C
>. cu
o 60 3O~
cu
a..
Seed weight
40 20

20 10

15 25 35 45 55 65 75 85 95
Time after flowering (days)

Figure 2.12. Changes in seed dry weight and moisture content (fresh weight basis) during seed
development and maturation. PM = physiological maturity; HM = harvest maturity (From TeKrony
et al. 1980).

weight (Jenner 1979; Peet and Kramer 1980). Short days have also been shown to reduce seed
size in pea (Reid 1979) and perennial ryegrass (Bean 1980). These fmdings are probably
attributed to the lack oflight which causes decreased photosynthesis and results in smaller seeds.

Position on the Plant

The position of the seed within the inflorescence can affect seed development rate. For
example, distal seeds in a wheat spike have slower growth rates and shorter seed filling periods
than proximal seeds (Simmons and Crookston 1979). Com seeds at the tip of the ear are usually
smaller than those at the base, which has been attributed to inadequate photosynthate supply
(Hanft et al. 1986). Sorghum seeds at the base of the panicle tend to be smaller and have a
slower growth rate than those elsewhere (Muchow 1990). Physical removal of other
reproductive sinks such as flowers or seeds also increases the size of the remaining seeds on/in
the inflorescence as in soybean (Egli et a1. 1987), wheat (Radley 1978), carrot (Gray et a1.
1986), com (Kiniry et al. 1990), and clover (Rincker et al. 1977).
34 Seed Formation and Development
The inflorescence structure can also affect seed size. In the Asteraceae, the composite
flowering head produces both ray and disk flowers, with ray flowers producing larger seeds.
Interestingly, the environment alters the ratio of ray to disk flowers and thus modifies the seed
size of the crop (Venable and Levin 1985; McGinley 1989). In some cases, these two seed types
have differing germination requirements (Forsyth and Brown 1982). Smaller seeds are also
produced from smaller fruit or fruit that matures later in the growing season. Such trends have
been reported in cotton (Leffler et at. 1977), sugar beet (Malik and Shakara 1977), rape (Clarke
1979), carrot (Verkaar and Schenkeveld 1984), and geranium (Roach 1986).
The physiological mechanism( s) governing seed development remain( s) largely unknown.
Some studies have attempted to relate seed development with hormonal levels. Endogenous
abscisic acid (ABA) levels, for example, have been correlated with seed weight. Large-seeded
soybean genotypes have 50% more ABA than small-seeded genotypes (Schussler et at. 1984).
Injecting ABA into wheat seeds increased their absorption of photosynthate (Dewdney and
McWha 1979). ABA increases the import of sucrose into barley seeds (Teitz and Dingkuhn
1981), and adding ABA to bean seeds increased their seed size (Clifford et al. 1987). While
these studies suggest that ABA may have a direct role in regulating seed development, further
work is still required to better understand this complex phenomenon.

Questions

1. In diploid plants, is the nucellar tissue of diploid or haploid origin?

2. What is the archesporia I cell?
3. Explain why the megagametophyte is sometimes called a macrospore.
4. What is the origin of the perisperm in some seeds? Name some seeds that have a well
developed perisperm.
5. What are some other names for the seed integuments?
6 Name the structure or tissue from which the integuments originate?
7. Can you cite two types of epistase?
8. What is anthesis? How is this related to human health?
9. Name at least one apomictic species?
10. What is the function of the endosperm in seeds?
11. Why do some seeds have no endosperm tissue at maturity?
12. What is the function ofthe albumin layer ofthe endosperm?
13. Why does the DNA content of the embryo increase faster during early seed development,
and level off later?
14. Would you expect the DNA content of pericarp and seed coat tissues to increase as rapidly
as endosperm and embryo? Why, or why not?
15. How long does the zygote remain at rest before it starts preparing for cell division?

General References

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Seed Formation and Development 35
Bhatnager, S. B., and B. M. Hohri. 1972. Development of angiosperm seeds. In: Seed Biology,
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Boswell, F. C., and D. E. Anderson. 1976. Long-term residual fertility and current N-P-K
application effects on soybeans. Agronomy Journal 68:315-318.
Bradbury, D., 1. M. CuH, and M. M. MacMasters. 1956a. Structure of the mature wheat kernel.
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__ . 1956b. Structure of the mature wheat kernel. II. Microscopic structure ofthe pericarp,
seed coat, and other coverings of the endosperm and germ of hard red winter wheat. Cereal
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__ .1956b. Structure of the mature wheat kernel. IV. Microscopic structure of the germ of
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_ _ .1986. Effects of water deficits on yield, yield components, and water use efficiency of
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36 Seed Formation and Development
Ene, B. N., and E. W. Bean. 1975. Variations in seed quality between certified seed lots of
perennial ryegrass and their relationships to nitrogen supply and moisture status during seed
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Seed Formation and Development 37
Leffler, H. R., C. D. Elmore, and 1. D. Hesketh. 1977. Seasonal and fertility-related changes in
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38 Seed Formation and Development
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Singh, H., and B. M. Johri. 1972. Development of gymnosperm seeds. In: Seed Biology, Vol.
1. ed. T. T. Kozlowski, pp. 21-75. New York: Academic Press.
Singh, R. P. 1954. Structure and development of seeds in Euphorbiaceae: Ricinus communis
L. Phytomorphology 4: 118-123.
Sionit, N., H. Hellmers, and B. R. Strain. 1980. Growth and yield of wheat under CO2
enrichment and water stress. Crop Science 20:687-690.
Sripleng, A, and F. H. Smith. 1960. Anatomy ofthe seed of Convolvulus arvensis. American
Journal of Botany 47:386-392.
Tekrony, D. M., D. B. Egli and J. Balles. 1980. The effect of the field production environment
on soya bean seed quality. In; Seed Production (ed. P. D. Hebblethwaite, Butterworths, London.
Teitz, A., and M. Dingkuhn. 1981. Regulation of assimilate transport in barley by the abscisic
acid content of young caryopses. Z. Pjlanzenphysiol. 104:475-479.
Thompson, R. C. 1933. A morphological study of flower and seed development in cabbage.
Journal ofAgricultural Research 47:215-232.
Varis, S., and R. A. T. George. 1985. The influence of mineral nutrition of fruit yield, seed yield
and quality in tomato. Journal of Horticultural Science 60:373-376.
Varner,1. E. 1965. Seed development and germination. In: Plant Biochemistry, ed. J. Bonner
and 1. E. Varner. pp. 763-792. New York: Academic Press.
Venable, D. L., and D. A. Levin. 1985. Ecology ofachene dimorphism in Heterotheca latifolia
1. Achene structure, germination and dispersal. Journal of Ecology 73: 133-145.
Verkaar, H. 1., and A J. Schenkeveld. 1984. On the ecology of short-lived fortes in chalk
grasslands: Semelparity and seed output of some species in relation to various levels of nutrient
supply. New Phytology 98:673-682.
Wardlaw, I. F., I. A Dawson, and P. Munibi. 1989. The tolerance of wheat to high
temperatures during reproductive growth. II. Grain development. Australian Journal of
Agricultural Research 40: 15-24.
Westermann, D. T., and S. E. Crothers. 1977. Plant population effects on the seed yield compo-
nents of beans. Crop Science 17:493-496.
Winter, D. M. 1960. The development of the seed ofAbutilon theophrasti, I. Ovule and embryo.
American Journal of Botany 47: 8-14.
Wood, D. W., R. K. Scott, and P. C. Longden. 1980. The effects of mother-plant temperature
on seed quality in Beta vulgaris L. (sugar beet). In: Seed Production, ed. P. D. HebbJethwaite,
pp. 257-270, London: Butterworth.
Wright, D. L., F. M. Shokes, and R. K. Sprenkel. 1984. Planting method and plant population
influence on soybeans. Agronomy Journal 76:921-924.
 3
The Chemistry
of Seeds
A knowledge of the chemical composition of seeds is essential for several reasons: (1) seeds
are a basic source of food for both people and animals, (2) they are an important source of
medicine and drugs, (3) they contain various antimetabolites that adversely affect human and
animal nutrition, (4) they are an important source of raw materials useful for various industrial
purposes, and (5) they contain reserve food supplies and growth substances that influence seed
germination and seedling vigor, seed storage, and longevity.
Most of our knowledge ofthe chemical composition of seeds concerns cultivated species,
since these comprise a large share of the food supply and also provide many industrial raw
materials. Information about the seeds of wild species is relatively scarce; however, the search
for new sources of food and raw materials is gradually yielding more information about seeds
of wild plants. Moreover, since seeds are a challenging subject for scientific study, much
information about both domestic and wild species is being accumulated simply because of our
thirst for knowledge.
Aside from the normal chemical constituents found in an plant tissue, seeds contain extra
amounts of chemical substances stored as food reserves to accommodate germination. These
reserve foods are stored primarily as carbohydrates, fats (or oils), and proteins. In addition,
seeds contain other chemical substances, some of which play minor storage roles, but most of
which serve as growth substances and metabolism controls. Compared to other plant parts, the
mineral content of most seeds is remarkably low and tends to be centered in the hulls and
structural tissue. Seeds with a relatively high mineral composition include bean, cotton,
sunflower, soybean, and cereal grains with the hulls intact.

THE INFLUENCE OF GENETIC FACTORS

The chemical composition of seeds is basically determined by genetic factors and varies
among different species and seed parts (Tables 3.1 and 3.2). However, it is influenced by
environmental and cultural practices.

L. O. Copeland et al., Principles of Seed Science and Technology

© Kluwer Academic Publishers 2001
40 Chemistry ofSeeds
Table 3.1. Average Chemical Composition of Seeds.

Plant % Protein % Fat (Lipid)

Acorn (red oak) 3.2 10.7
Barley (Pacific coast states) 8.7 1.9
Bean, Mung 23.6 0.2
Bean, Navy 22.9 1.4
Bean, Pinto 22.5 1.2
Beechnuts 15.0 30.6
Buckwheat 10.3 2.3
Chickpeas 20.3 4.3
Cottonseed kernel (without hull) 38.4 33.3
Flaxseed 24.0 35.9
Kafir grain 11.0 2.9
Mustard, Wild 23.0 38.8
Oats 12.0 4.6
Peas 23.4 1.2
Peanut (without hulls) 30.4 47.7
Rape 20.4 43.6
Rice (rough grain) 7.9 1.8
Rye 12.6 1.7
Soybean 37.9 18.0
Sunflower 16.8 25.9
Vetch 29.6 0.8
Wheat 13.2 1.9
From Morrison (1961).

Table 3.2. Chemical Composition of Different Parts of Seeds.

Chemical Entire Seed Endosperm Embryo Pericarp-Testa

Starch 74.0 87.B 9.0 7.0
Sugars 1.8 .8 10.4 .5
Oil (Lipid) 3.9 .8 31.1 1.2
Protein B.2 7.2 18.9 3.8
Ash 1.5 .5 11.3 1.0
*lncludes only selected chemical components.
From Earle et al. (1956).

Because of genetic factors, the chemical composition of seeds varies widely among species
and even among varieties. Through crossing and selection, plant breeders have been able to
manipulate the chemical composition of many domesticated crops and increase their value as
food, fiber, or raw material. Modern varieties of rapeseed, soybean, high lysine corn, sorghum,
and wheat have been bred and developed for higher content of oils, protein, or carbohydrates,
and represent a significant improvement over earlier varieties. The effectiveness of breeding and
selection techniques in increasing the protein and oil content of crop seeds is shown by work
with soybean (Shannon et al. 1972) and corn (Dudley and Lambert 1968). More recently, it is
demonstrated by the high oil and protein levels of specialty varieties of crops such as corn,
soybean, and canola that are becoming commercially available.
Chemistry of Seeds 41

ENVIRONMENTAL INFLUENCES

Many environmental factors influence the chemical composition of seeds, and because of
the interrelationships of these factors, it is sometimes difficult to determine the cause of
variability. A two-year study of eight com hybrids in three Michigan locations revealed a range
in protein content from 7.44 to 12.88% within hybrids (Norden et a1. 1952). Similar
environmental influence on the protein content of chickpeas has been reported in the former
Soviet Union (Ivanov 1933). Wheat protein content varies depending on geographic location
of the crop (Baenziger et a1. 1985). Environmental conditions from year to year similarly
influence the chemistry of peanut (Ketring et a1. 1978) and pea (Gubbels 1981) seeds. Among
the environmental factors that modify seed chemistry are water availability, temperature, soil
fertility, and cultural practices.

Water

Availability of water influences chemical composition of seed. For example, we have long
recognized that the protein nitrogen content and quality of seed are lower in years of high rainfall
than in drier years and in irrigated land compared to dry land. A study by Greaves and Carter
(1923) showed that high rates of irrigation decreased the nitrogen content of seeds in wheat,
barley, and oats grown in Utah (Table 3.3). This decrease in nitrogen content occurred despite
observed increases in phosphorus, potassium, calcium, and magnesium, which are not readily
soluble in water. Similar studies for both wheat and sorghum have shown that the nitrogen
content of the mature seed decreases linearly with the amount of water supplied during seed
development (Stone and Tucker 1968; Stone et a1. 1964; Mathers et a1. 1960). These studies
serve as clear examples ofthe ways in which high-moisture environments, whether created by
rainfall or irrigation, can influence the mineral composition of seeds. Yet, we know relatively
little about why these changes occur. It is not clear, for example, whether the primary effect of
excess moisture is on mineral absorption by the roots, or on the rate of seed fill with
carbohydrates and the concomitant dilution of the basic cell constituents. Further studies are
needed to answer these questions. In contrast, plants exposed to low soil moisture or drought
conditions tend to increase in seed protein content. This has been found in wheat (Karathanasis

Table 3.3. Effect ofIrrigation on the

Mineral Content of Wheat, Barley, and Oat Seed.
------------------------------------------------
Increase ( + ) or decrease ( -) over controls, %
Element Wheat Barley Oats
Nitrogen - 21 -19 -40
Phosphorus + 55 +30 +35
Potassium + 35 +14 +31
Calcium +155 +41 +22
Magnesium + 32 + 9 +65
From Greaves and Carter (1923).
42 Chemistry of Seeds

et al. 1980), perennial ryegrass (Ene and Bean 1975), com (Francois et al. 1986), soybean
(Pikaard and Cherry 1984), and bean (Robinson 1983) seeds.

Temperature

Although few studies have been conducted to demonstrate the influence oftemperature on
seed structure and composition, those studies that have been done show an association. Howell
and Carter (1958) found that the oil content of soybean seeds depended on the temperature
during pod development (Figure 3.1). Seeds that matured at 21°C contained 19.5% oil, whereas
those that matured at 30°C contained 22.3% oil. Soybeans also develop a higher oil content
when they are planted early in the season and, therefore, mature under a warmer temperature
than when they are planted later and mature during cooler weather. Canvin (1965) examined the
influence of temperature on fatty acid content of developing rapeseeds and showed that as the
temperature increased, oleic acid levels increased and erucic acid levels decreased. This study
has special significance since erucic acid has a bitter taste and is an undesirable component of
rapeseeds that are to be used as feed.
In sunflowers, low temperature during seed development favors the production of the
preferred linoleic acid, and high temperatures enhance the quantity of oleic acid in the oil (Harris
et al. 1980). As a result, sunflower seeds grown for high oil content are planted late so that

23

22

.....l 21
<3
E-<
Z
U.l
~ 20
rJ.l
Q..,

19

18

70 77 85
DA Y TEMPERATURE

Figure 3.1. Oil content ofsoybean seed produced with day temperatures as shown during periods of
podjilling; night temperature was 65° F in all cases (From Howell and Carter /958).
Chemistry of Seeds 43

flowering occurs later in the season (Owen 1983; Jones 1984). Unger (1986) showed that the
best quality sunflower oil was produced during the cooler conditions of late summer. Similar
responses have been reported for flaxseed (Green 1986). Beyond oil content, temperature of seed
development also influences protein and carbohydrate quality. For example, high temperatures
increase seed protein content in wheat (Campbell et al. 1981), and cool temperatures lower
protein content in soybean (Redford et al. 1977) seeds. Other studies have shown that high night
temperatures accelerate seed development and maturation in rice, producing "chalky" kernels.
At low night temperatures, the seeds develop the desired "milky white" kernels.

Soil Fertility

Perhaps the easiest environmental parameter to control in an evaluation of factors that

affect seed chemical composition is the mineral nutrition that the mother plant receives. In most
instances, seeds that are mineral deficient tend to perform poorly compared to normal seeds
unless they are planted in a soil that is nutritionally adequate and provides the missing essential
element or elements. Many studies have been conducted to evaluate the influence of nitrogen,
phosphorus, and potassium on seed quality. Some of these are cited below.
Com plants grown under either high nitrogen fertilization or low plant populations produce
seed with a higher protein content than those produced under low nitrogen availability or high
plant populations (Wolfson and Shearer 1981). Similar results with rice indicate that when
lower seeding rates are used, each rice plant has greater access to available nitrogen, which is
then absorbed and supplied to the seeds (DeDatta et al. 1972). Added soil nitrogen also increases
protein content in wheat (Glenn et al. 1985), rice (Allen and Terman 1978), and cotton (Elmore
et al. 1979) seeds. The application of foliar urea to wheat plants increased seed protein content
as well (Altman et al. 1983). Scott (1969) found that excess nitrogen application could have an
indirect and detrimental influence on sugar beet seed quality. When excess nitrogen was applied,
the ripening ofthe crop was delayed and the seeds from plots fertilized with nitrogen were less
mature than seeds from plots that received no nitrogen. In this instance, the reduction in
germination was a nitrogen-induced maturity effect.
Other studies indicate the importance of phosphorus for production of high quality seeds.
For example, Harrington (1960) demonstrated that phosphorus nutrition of parent plants failed
to influence seed performance of the progeny. Austin (1966) showed that freshly harvested seed
from phosphorus-deficient watercress plants had lower (and slower) germination than seeds
from nondeficient plants. Other studies have shown that added phosphorus to the parent plant
increases seed phosphorus content in pea (Peck et a!. 1980), soybean (Cassman et al. 1981),
and wheat (Porter and Paulsen 1983). Low-phosphorus seed produces smaller plants than
nondeficient seed.
Few studies have been conducted on the influence of potassium on seed quality and
composition. Harrington (1960) reported that potassium-deficient plants ofCapsicumjrutescens
gave a higher proportion of abnormal seeds with dark-colored embryos and seed coats. Both
normal and abnormal seeds from such plants had a lower germination than control seeds, and
their viability declined more rapidly in storage.
Other essential elements added to the parent plant also will be found in increased
concentrations in the seed. This is true for calcium in peanut (Coffelt and Hallock 1986);
manganese, zinc, and boron in soybean (Parker et a1. 1981; Raboy and Dickinson 1984;
44 Chemistry ofSeeds

Touchton and Boswell 1975); copper in wheat (Loneragan et al. 1980); and cadmium and
selenium in lettuce and wheat (Cary 1981).

Cultural Practices

Still other environmental factors associated with plant morphology and production
practices are known to modifY seed chemical content. For example, chemical composition of the
seed can be influenced by its position on the plant. The relative proportions of seven fatty acids
have been shown to vary dependent on the position of the seed in the pod of rape (Diepenbrock
and Geisler 1979). First and second seeds in a wheat spikelet have higher nitrogen concentration
than third and fourth seeds (Simmons and Moss 1978). The time during the season when the
seeds develop also influences their chemistry such as the oil content in rape seed (Auld et al.
1984). Soybean seeds which mature later in the growing season have higher protein levels
(Gbikpi and Crookston 1981). Competition within the field also modifies seed chemical content
since there is more competition among plants for the limited nutrients. As competition increases;
protein content declines while oil content increases in sunflower (Robinson et al.1980; Majid and
Schneiter 1987), rice (Nandisha and Mahadevappa 1984), and wheat and barley (Read and
Warder 1982). These results suggest that dense plantings for oil crops may be an advantage as
long as no reduction in yield is obtained. Interestingly, there appears to be an inverse
relationship in oil-containing seeds between oil and protein content; as the protein content
increases, oil content declines. This is true in soybean (Poole et al. 1983), sunflower (Mashers
and Stewart 1982), and in meadowfoam (Crane et at. 1981).

CARBOHYDRATE STORAGE IN SEEDS

Carbohydrates are the major storage substance in seeds of most cultivated plants. Cereals
and grasses are especially rich in carbohydrates and low in fats and proteins. Peas and beans
are moderately high in carbohydrates, with lesser amounts of proteins and a low fat content.
Seeds from many trees, such as chestnut, buckeye, and some oak species have a relatively high
carbohydrate content and are quite low in fats and proteins.
Starch and hemicellulose are the major forms of carbohydrate storage in seeds. Other
carbohydrates that occur in nonstorage forms are pectins and mucilages.

Starch

Seeds are composed largely of metabolically inactive food reserves that are stored until
needed during germination. Starch is stored in two related forms, amylose and amylopectin,
which are two polymers ofD-glucose, one linear and the other branched (Figure 3.2). Starch is
the principal, most widespread storage carbohydrate of seeds.
Amylose is composed of 200 to 1000 glucose units linked by a-l,4 glucosidic bonds, with
a molecular weight ranging from 10,000 to 100,000. The molecule has a helical structure with
six glucose rings to one revolution. Amylose stains blue when exposed to iodine and is 100%
digestible by a-amylase.
Amylopectin, in contrast, is a much larger molecule consisting of 20-25 glucose units per
branch, linked by both a-I,4 and a-I,6 glucosidic bonds with a molecular weight ranging from
Chemistry ofSeeds 45

50,000 to 1,000,000. Amylopectin is only about 50% digestible by the enzyme a-amylase and
stains purplish-red when exposed to iodine.
Both amylose and amylopectin are hydrolyzed by the enzymes a- and ~-amylase during
normal metabolism and germination. Alpha-amylase possesses a molecular weight of 60,000
and requires the cation Ca++ for activation and stabilization against proteolytic destruction and
thermal denaturation of the enzyme. This enzyme attacks both amylose and amylopectin
randomly, cleaving both the a-I,4 and a-I,6 linkages. Although the action pattern varies, there
is a general amylolysis (amylase catalyzed breakdown) of the starch molecule to large amounts
of dextrin molecules to maltose and finally glucose. The enzyme is formed de novo in the
aleurone of barley and the scutellum of com. In com, a-amylase accounts for 90% of the
amylolytic activity and ~-amylase for the remaining 10%.

0---

H OH H OH H OH
CH 2 0H Amylose
0
H H

H OH
°l.
CH 2

0---

H OH H OH H OH

Amylopectin

Figure 3.2. Chemical structure q( amylose and amylopectin showing linear (a-I,4 and 1,6 links)
configuration.
46 Chemistry ofSeeds
Beta-amylase hydrolyzes both amylose and amylopectin from the nonreducing end by
breaking the 0.-1,4 linkage and forming maltose. In the case of amylose, almost complete
hydrolysis occurs. However, since B-amylase is unable to bypass the branched chain point or
0.-1,6 linkage of amylopectin, this molecule is only partially hydrolyzed, yielding the residual
{J-limit dextrin. This means that only the outer portions ofthe amylopectin molecule are attacked
by B-amylase. This enzyme is already formed and is present in situ in seeds. In wheat seeds, B-
amylase is present in an active latent form, apparently chemically bound to the wheat glutenin
by disulfide linkages. During germination, the secretion of a substance capable of releasing the
glutenin-bound enzyme may occur, accounting for the observed B-amylase activity.
In seeds, most starch is laid down in discrete subcellular bodies called starch grains that
range from 2 to 100J.! in size within the endosperm. Many starch grains appear to form around
a central point, the hilum, around which shells of the polysaccharide are deposited. These shells
probably reflect a diurnal periodicity in the synthesis and deposition of starch, since they are
absent from starch grains of seeds developing in continuous light under experimental conditions.
The shape of the starch grain appears to depend on the amylose content-the less angular,
rounded grains having relatively high amylose levels.
Most starch grains are composed of about 50-75% amylopectin and 20-25% amylose.
However, some seeds such as rice may be high in amylose (up to 37%) and are classified as
starchy. The difference in appearance between amylopectin and amylose starch forms is
illustrated in waxy corn (almost 100% amylopectin) versus normal corn (50-75% amylopectin)
(Kerr 1950).

HemiceUulose

Other than starch, the major form of carbohydrate storage in seeds is hemicellulose.
Hemicellulose is a widely used but poorly defmed class of polysaccharides usually found in the
cell walls of plants, although in certain seeds they are found as reserve food materials. This
defmition includes xylans, mannans, and galactans, plus a number of similar but less common
polysaccharides. They are usually found in the thickened tertiary layers of cell walls of the
endosperm or cotyledons instead of in the interior region of the endosperm. They are composed
principally of mannans with small amounts of sugar (glucose, galactose, arabinose) as side
chains on the main linear polymers of mannose residues. Hemicelluloses are particularly
characteristic of seeds of many of the palm species, such as date palm and South American and
Polynesian ivory nut palms. Seeds of the date palm have a small cylindrical embryo embedded
in a sizable horny endosperm of hemicellulose. Hemicellulose has also been reported in the
endosperm or cotyledon of several other species (Mitchell 1930).

Other Seed Carbohydrates

Mucilages. Apart from starch and hemicellulose, the amount of carbohydrates found in
seeds is similar to that found in other parts ofthe plant. An exception might be the mucilages,
which infrequently may be found in rather large amounts. A well known example is the seed of
buckhorn plantain, which is covered with a thick coating of mucilage that becomes sticky when
wet and tends to cling to material that it touches, providing the seed with a dispersal
mechanism. However, seed conditioners also use the sticky coat characteristic when cleaning
Chemistry afSeeds 47
buckhorn seed from certain crop seed (Chapter 11). Mucilages from buckhorn plaintain seed
may be separated commercially for industrial use.
Mucilages are complex carbohydrates consisting principally of polyuronides and
galacturonides that chemically resemble the pectic compounds and hemicelluloses. Physically,
they are similar to the gums found in the bark and stems of many plants.
Pectic Compounds. These compounds occur in seeds and in other plant parts mainly as
components of the cell wall and the middle lamella. The three principal pectic compounds are
pectic acid, pectin, and protopectin. Pectic acid is a long, straight-chain substance composed of
about 100 galacturonic acid molecules. Pectin differs from pectic acid by esterification of many
of the carboxyl groups and has a greater number of galacturonic residues per chain. Pectins
form viscous colloidal sols in water that set into firm gels under the proper conditions; they are
used as setting agents in jams and jellies. Protopectins differ from pectins in their larger
molecular chain. They occur in the primary cell wall and the middle lamella where they bind the
cell walls together. When protopectins are converted into pectins, they are instrumental in the
softening of ripening fruits.

LIPID STORAGE IN SEEDS

The value of seed-borne lipids (Table 3.4) for food and industrial uses has contributed
greatly to our knowledge of the composition of oily seeds. Seed oils have tremendous versatility

Table 3.4. Percentage of Fats and Oils in the Dry Matter of Different Plant Species.

Percentage Percentage
Species fat or oil Species fat or oil
Coconut 65 Spurge 35-45
Brazil nut 70 Rapeseed 33-43
Castor bean 60 Sesame seed 50-55
Sunflower seed 45-50 Colza seed 43-53
Flaxseed 30-35 Madia sativa seed 32
Cottonseed 15-20 Kafir seed 2.5
Peanut 40-50 Feterita seed 2.4
Hemp 30-35 Milo seed 2.3
Walnut 50-65 Corn seed 2.1
Cacao bean 40-50 Wheat seed 1.8
Poppy seed 40-50 Field pea 1.5
Pumpkin seed 41 Bean 2.8
Almond 40-50 Rye seed 1.9
Soybean 15-20 Rice seed 2.5
Cantaloupe seed 30 Buckwheat seed 1.1
From Miller (1931).
48 Chemistry of Seeds

Rl COOH CH 2 0H CH 2 OCOR 1
I I
R2 COOH + HOCH • R2 COOCH +
I I
R3 COOH CH 2 0H CH 2 OCOR 3

3 fatty acids glycerol lipid molecule water

( triglyceride)

Figure 3.3. How three fatty acids combine with glycerol to form a lipid

for industrial uses. In contrast to animal fats, their highly unsaturated chemical nature has
caused increased interest in them for health purposes (e.g., canoIa).
Except for certain fruits, the occurrence of high lipid concentrations differentiates seeds
from other plant organs. High lipid content is usually associated with decreased protein content
(for example, in soybeans, rapeseed, cotton). However, in some species, such as certain oaks,
it is associated with high carbohydrate levels.
Lipids are plant or animal substances that are insoluble in water, but soluble in ether,
chloroform, benzene, or other organic solvents. They are esters of either fatty acids or glycerol
or their various hydrolytic products. They are known as glycerides, or, more specifically,
triglycerides, because each glycerol molecule is combined with three fatty acid molecules
(Figure 3.3). Aside from those described by Bloor (1928), the characteristics oflipids are: (1)
total fatty acid complement, (2) their glyceride structure, and (3) certain other substances that
may be associated with the glycerides as impurities or as part of the lipid molecules.
The term lipid is often used interchangeably with/at and oil, although it is actually the
generic term for both. Oils are distinguished from fats only in that they remain in liquid form
at ordinary room temperatures, whereas fats remain in solid form. Oils are often designated as
fatty oils to differentiate them from essential oils, which are chemically unrelated.

Fatty Acids

The fatty acids are so named because they are a constituent of natural fats, and in the free
state they resemble fats in physical properties. Free fatty acids are seldom found in plant parts
other than in germinating or deteriorating seeds as a result of fat hydrolysis. Fatty acids are
saturated or unsaturated, depending on the type of carbon linkage in the molecule. The
unsaturated fatty acids contain one or more double-bond links, which means the compound can
take up hydrogen atoms to form a saturated compound. The unsaturated fatty acids are most
common in seeds with oleic acids (one double bond) and linoleic acids (two double bonds),
accounting for 60% of the weight of all the lipids present in most oilseed crops (Table 3.5).
Saturated fatty acids are also present and contain an even number (n) of carbon atoms (n usually
being between 4 and 24). Palmitic acid (n = 14) is the most common saturated fatty acid in
oilseeds.
Chemistry o/Seeds 49
Table 3.5. Fatty Acid Content (%) of Seeds and Seed Products of Several Plant Species.

Fat or oil Lauric Myristic Palmitic Stearic Oleic Linoleic Linolenic

Coconut 45 20 5 3 6
Palm kernel 55 12 6 4 10
Olive 14.6 75.4 10
Peanut 8.5 6 51.6 26
Cottonseed 23.4 31.6 45
Rapeseed/canola 2 4 60 17 10
Maize 6.0 2 44.0 48
Linseed 3 77 17.0
Soybean 11.0 2 20.0 64 3.0
Cantaloupe
seed oil 0.3 10.2 4.5 27.2 56.6
Sunflower oil 3.5 2.9 33.4 57.5
Hubbard squash
seed oil 13.0 6.0 37.0 44.0
Modified from Miller (1931).

Glycerol and Other Alcohols

Glycerol and other alcohols are combined with fatty acids to form different kinds of lipids.
Of these, trihydroxy alcohol and glycerol or glycerine are most often involved and form esters
(glycerides) with many different fatty acids.

Classification of Lipids

Lipids may be classified as (a) simple, (b) compound, or (c) derived. The simple lipids
include esters of fatty acids and glycerol or various other alcohols. Among these simple lipids
are fats and fatty oils. Compound lipids are esters of the fatty acids containing additional
chemical groups. Phospholipids are compound lipids in which one ofthe three fatty acid units
is replaced with phosphoric acid combined with choline. Derived lipids are derived from simple
and compound lipids by hydrolysis and are soluble in fat solvents. They include various fatty
acids and large molecular alcohols that are soluble in the fat solvents and may combine with the
fatty acids. Among these is cholesteroL
The great majority of seed lipids are simple lipids, which include fats, fatty oils, and waxes.
Waxes are simple lipids that are fatty acid esters of some alcohol other than glycerol. The term
is used only in a chemical sense, since they occur both as liquids and as solids. Waxes are less
soluble in ordinary fat solvents than fats. They can be saponified (see below), but only with
difficulty. They are usually found as coverings for the leaves, fruits, and seeds of many plants.

Hydrolysis of Lipids

When oilseeds with a high lipid content germinate, there is a rapid disappearance of lipid
with a concomitant rise in sucrose content. Concurrently, lipase activity rises sharply and
participates in the stepwise hydrolysis of triglycerides to diglycerides, monoglycerides, and
50 Chemistry ofSeeds
finally free glycerol and free fatty acids. Fatty acids are further oxidized by both u- and, p-
oxidation during the course of germination. These processes are further discussed in Chapter
5. Most lipids are stored in oil storage bodies called spherosomes that range in size from 0.2 to
6.0J! in diameter. Many of the enzymes essential for fatty acid biosynthesis or for triglyceride
hydrolysis are present in the spherosomes.

PROTEIN STORAGE IN SEEDS

Proteins are nitrogen-containing molecules ofhuge size and exceedingly complex structure,
the greater part of which yield amino acids upon hydrolysis of the peptide bond (Figure 3.4).
Proteins are so important to both plant and animal life that all physiological reactions of living
cells revolve around their physical and chemical properties and those of related compounds.
Aside from water, they are the principal component of all protoplasm of both plant and animal
cells.
Proteins comprise a valuable food storage component in seeds of most plant species.
Soybeans are one of the few species known in which proteins comprise more of the reserve food
supply than do fats or carbohydrates. Most of the plants known to have high-protein seed are
legumes having nitrogen-fixing capacity. However, many high protein species are nonlegumes.
Seed storage proteins are less complex than protoplasmic proteins and less likely to be tied up
with lipids and other prosthetic groups, although they are structurally similar.
The great majority of seed proteins are metabolically inactive and serve merely as food
reserves for use by the growing embryo during germination. Metabolically active proteins
constitute a small amount of the total, but they are extremely important to the developing and
germinating seed. As enzymes, they catalyze all metabolic processes in digestion, translocation,
and utilization of stored food reserves; no growth can occur without them. The nucleoproteins
are another extremely important form of active protein and are molecules of enormous size (the
molecular weight may number in the millions). They are formed by protein-nucleic acid linkage
and are a huge conglomerate molecule composed of a protein, a pentose sugar, a cyclic nitrogen
compound (purine or pyrimidine), and phosphoric acid. Ifthe pentose sugar is deoxyribose, the
resulting nucleoprotein is called deoxyribonucleic acid (DNA); ifthe pentose sugar is D-ribose,
the nucleoprotein is called ribonucleic acid (RNA). Both these forms have been emphasized for
their function in protein synthesis and their crucial role in the structure and function of
chromosomes, genes, and life itself.

H R OH H OH H OH
I I I R I -H 2 0 I R ~, R I
N-C-C + N-C-C .. N-C-C-N-C-c
I H • IHI IHIIHI
H 0 H 0 H ' o H, 0

Figure 3.4. Formation ofa peptide bond Two amino acids combine with loss of water, resulting in
a dipeptide. The peptide link in the dipeptide is indicated within the tinted area at right.
Chemistry of Seeds 51
Proteins are stored in the seed in units known as protein bodies that are 1-2011 in diameter
bounded by a lipoprotein unit membrane. These are somewhat like starch grains in size and
shape and are usually a mixture of different proteins. They may be visualized as the kind of
units that are deposited in the albuminous aleurone layers of cereal seeds and that, during seed
germination, play an important role both as food reserves and as hydrolytic enzymes to aid in
starch breakdown.
An insight into seed proteins has been provided by the work of Osborne (1924) with wheat
seeds. He purified four proteins from wheat and classified them on the basis of their solubility,
two of which were metabolically active (globulin and albumin), and two that were nonactive
(glutelin and prolamine). Osborne considered these to be the only proteins occurring in seeds.
Crocker and Barton (1957) described them as follows:
Seed albumins are soluble in water at neutral or slightly acid pH and are coagulated by
heat. Examples are leucosins of cereal grains, legumelin of various pulse seeds, and ricin of rice.
These are mainly enzymes.
Seed globulins are soluble in saline solutions, but not in water and are generally more
difficult to coagulate with heat than are animal globulins. Their solubility is modified by
combined acids and concentration of saline solutions. Globulins are found predominately in dicot
seeds such as legumes. Examples are legumin, vignin, glycinin, vicilin, and arachin.
Seed glutelins are soluble in aqueous or saline solutions or ethyl alcohol, but can be
extracted with strong acid or alkaline solutions. Glutelins are found in most cereal seeds.
Examples are glutenin of wheat and oryzenin of rice.
Seed prolamines are soluble in 70-90% ethyl alcohol, but not in water; however, the salts
with acids and bases are soluble in water. They are found only in cereal seeds; gliadin of wheat
and rye and zein in maize are examples. Upon hydrolysis they yield proline, glutamic acid, and
ammOnIa.
[n general, glutelins and prolamines form the major components of seed protein that is
metabolically inactive and associated with structural architecture. Seed proteins generally have
a high nitrogen and proline content and are low in lysine, tryptophan, and methionine.

OTHER CHEMICAL COMPOUNDS FOUND IN SEEDS

Tannins

Although one usually thinks of tannins as occurring in other plant parts, especially the
bark, they also occur in seeds, particularly in the seed coat structures. They are found in the
testa of cocoa and bean seed (Bonner and Varner 1965).
Tannins have been used since the dawn of civilization for removing hair from animal skins
during the tanning process. They are naturally occurring compounds of high molecular weight
(500-3000), containing enough phenolic, hydroxyl, or other suitable groups to enable them to
form effective cross-links between proteins and other macromolecules. This characteristic gives
them a unique ability to tie up proteins and inhibit their enzymatic activity and is the basis for
their use in the tanning process.
52 Chemistry of Seeds
Alkaloids

The death of Socrates provides a clue about the nature of alkaloids; Socrates died from
drinking a cup of hemlock containing the alkaloid poison coniine. Better-known alkaloids that
occur in plants or their seeds are morphine from poppy fruits, strychnine from seeds of
Strychnos nux vomica, atropine from the deadly nightshade, and colchicine from meadow
saffi'on. Other extremely common alkaloids are caffeine from coffee and tea, nicotine from
tobacco leaves, and theobromine from cacao.
Alkaloids are complex cyclic compounds containing nitrogen. Most are white solids;
however, nicotine is a liquid at room temperatures.

Glucosides

While most glucosides are found in the vegetative organs of plants, some do appear in
seeds. Examples of glucosides in seeds and vegetative plant parts are salicin in the bark and
leaves of wiHows; amygdalin in seeds of almonds, peaches, and plums; sinigrin in black
mustard; aesculin in horse chestnut seed; and quercitron in the bark of oak (Bonner and Varner
1965).
Glucosides are formed by a reaction between a sugar (usually glucose) and one or more
nonsugar compounds. In their pure state, they are mostly crystalline, colorless, bitter, and
soluble in either water or alcohol. Some ofthe glucosides, such as saponin (which comes from
tung seed), are highly poisonous to both man and animals.

Phytin

Phytin, the insoluble mixed potassium, magnesium, and calcium salt of myoinositol
hexaphosphoric acid, is the major form of phosphorus storage in seeds. In cereal seeds, phytin
is generally associated with the protein bodies in the aleurone layer and is more or less absent
from the protein bodies of the cotyledons. During the germination of seeds, phosphatases that
hydrolyze phytin increase severalfold. The phytase activity is highest in the scutellum and
aleurone layers. Since a large proportion of the seed's phosphate, magnesium, and potassium are
present in phytin, much of the seed's subsequent metabolism is dependent on the hydrolysis of
phytin and the concomitant release of magnesium and potassium ions. In lettuce seeds, 50% of
the total phosphorus is tied up in phytin.

Hormones

The term hormone is used to designate certain organic compounds that, when present in
minute quantities, exert important regulatory effects on the metabolism of either plants or
animals. They are of enormous importance in both plants and animals. A well-known animal
hormone is adrenal in, secreted by the adrenal glands, which exerts a marked influence on both
the heart and vascular system. Many important plant hormones also occur in seeds. They are
variously designated as phytohormones, growth hormones, growth substances, and growth
regulators.
Gibberellins. The presence of gibberellins in higher plants was demonstrated by Radley
(1956), who was able to induce tall growth in dwarf peas by giving them an extract from normal
Chemistry of Seeds 53
tall plants. It is now agreed that gibberellins are a normal constituent of green plants, as well as
seeds. They appear to have a major physiological role in both seed development and seed
germination. They also exert an influence on floral induction and initiation.
The best-known gibberellin is gibberellic acid (GA3)' although at least 50 others have been
identified. Gibberellins are produced commercially by fermentation of fungal cultures
(Gibberella spp.).
Cytokinins. Another group of compounds that occurs in seeds and exerts hormonal
influence is the cytokinins (kinins). They were first discovered in coconut milk (Van Overbeek
et a1. 1941). Fifteen years later, kinetin was purified and the chemical structure identified. The
first naturally occurring cytokinin to be extracted from seeds was zeatin.
Cytokinins appear to be necessary for cell growth and differentiation; perhaps this is the
basis for their influence in promoting seed germination. They have also been shown to inhibit
senescence in leaves (Richmond and Lang 1957) and to regulate the flow of chemicals through
the plant system (Mothes et a1. 1959).
Inhibitors. There is growing conviction among plant physiologists that dor-
mancy-whether in seeds, buds, tubers, or other plant parts-is regulated by a balance, or
interaction, between endogenous inhibitors and growth promoters such as gibberellins and
auxins. Dormin (Cornforth et al. 1965; Thomas et al. 1965) and abscisin II (Ohkuma 1965)
were preliminary names given to abscisic acid (ABA) which is reportedly instrumental in
inhibiting the formation of the leaf abscission layer and, particularly, the onset of winter
dormancy in deciduous plants. Another inhibitor that influences seed dormancy is coumarin.
Ethylene gas can both inhibit and promote seed germination, and is considered a hormone
(Crocker et a1. 1935). as is the growth regulator, maleic hydrazide (Meyer et a1. 1960). Chapter
7 contains a comprehensive discussion of germination inhibitors.

Vitamins

From the standpoint of human physiology, vitamins qualify as growth regulators and
cannot be sharply distinguished from hormones, since they are necessary for the diet but are
required in small, often minute, quantities. Although the role of vitamins in animal life and their
almost universal occurrence in green plants have long been recognized, their role in plant growth
is not so well known. Chemically, vitamins represent a very heterogeneous group.
In contrast to animals, who are dependent on green plants for their vitamin requirements,
green plants are self-sufficient for their vitamin needs (although certain parts of a plant may
depend on other parts for their vitamin supply). All known vitamins or their immediate
precursors are synthesized in higher plants. Whether all are necessary in a plant's metabolism
is not certain. However, specific roles for certain vitamins have been determined. Thiamine
appears to be necessary for the developing embryo and endosperm of seeds in some species. It
is also needed for normal root development. The basis for both needs seems to be the role of
thiamine in continued cell division, which is rapidly occurring within the seed. In the case of
both roots and the developing seed, thiamine is produced in the vegetative plant parts or by the
cotyledon and translocated into the areas where needed. Biotin and ascorbic acid are apparently
involved in respiration processes in seeds. The role of biotin is not known, but ascorbic acid
appears to be responsible for regulating the oxidation-reduction potential during germination.
54 Chemistry of Seeds
Questions

1. What types of chemical compounds are found in seeds?

2. List the factors that influence the chemical composition of seeds.
3. Do you think seeds are more important as food or as raw materials for industrial
products?
4. Make a list of seeds that are useful in the production of (a) drugs and (b) industrial
products.
5. How does the nutritional value of seeds differ from the food value of other sources of
protein, carbohydrates, and oils?
6. Why is the presence of phytates in seeds important to the role of seeds as human and
animal food?
7. Describe three environmental factors that can contribute to alterations in seed chemical
composition.
8. Detail the differences between amylose and amylopectin and emphasize how the
enzymes, u- and, ~-amylase, contribute to their hydrolysis.
9. What are the two most common unsaturated fatty acids found in most oil seeds? Name
the four principal classifications of seed proteins and list the differences between each
group.

General References

Allen, S. E., and G. L. Terman. 1978. Yield and protein content of rice as affected by rate,
source, method, and time of applied N. Agronomy Journal 70:238-242.
Altman, D. W., W. L. McCuistion, and W. E. Kronstad. 1983. Grain protein percentage, kernel
hardness, and grain yield of winter wheat with foliar applied urea. Agronomy Journal 75 :87-91.
Auld, D. L., B. L. Bettis, and M. J. Dial. 1984. Planting date and cultivar effect on winter rape
production. Agronomy Journal 76:197-200.
Austin, R. B. 1966. The growth of watercress Rorippa nasturtium-aquaticum L. (Hayck) from
seed as affected by the phosphorus nutrition of the mother plant. Plant and Soil 24: 113-120.
Baenziger, P. S., R. L. Clements, M. S. Mcintosh, W. T. Yamazaki, T. M. Sterling, D. 1.
Sammons, and J. W. Johnson. 1985. Effect of cultivar, environment, and their interaction and
stability analyses on milling and baking qualities of soft red winter wheat. Crop Science 25 :5-8.
Bloor, W. R. 1928. Biochemistry of fats. Chemical Review 2:243-300.
Bonner, J., and J. E. Varner, eds. 1965. Plant Biochemistry. New York: Academic Press.
Campbell, C. A., H. R. Davidson, and G. E. Winkleman. 1981. Effect of nitrogen, temperature,
growth stage and duration of moisture stress on yield components and protein content of
Manitou spring wheat. Canadian Journal of Plant Science 61 :549-563.
Canvin, D. T. 1965. The effect of temperature on the oil content and fatty acid composition of
the oils from several oil seed crops. Canadian Journal of Botany 43 :63-69.
Cary, E. E. 1981. Effect of selenium and cadmium additions to soil on their concentration in
lettuce and wheat. Agronomy Journal 73 :703-706.
Cassman, K. G., A. S. Whitney, and R. L. Fox. 1981. Phosphorus requirements of soybean and
cowpea as affected by mode ofN fertilization. Agronomy Journal 73: 17-22.
Coffelt, T. A., and D. L. Hallock. 1986. Soil fertility responses of Virginia-type peanut
cuItivars. Agronomy Journal 78: 131-13 7.
Chemistry afSeeds 55
Cornforth, 1. W., B. V. Milborrow, G. Ryback, and P. F. Wareing. 1965. Identity of sycamore
"dormin" with abscisin II. Nature 205:1269-1270.
Crane, J. M., W. Calhoun, and T. A. Ayres. 1981. Seed and oil characteristics of fertilized
meadowfoam. Agronomy Journal 73:255-256.
Crocker, W.,andL. V. Barton. 1957. Physiology ofSeeds. Waltham, Mass.: ChronicaBotanica
Company.
Crocker, W., A. E. Hitchcock, and P. W. Zimmerman. 1935. Similarities in the effects of
ethylene and the plant auxins. Contributions from Boyce Thompson Institute 7:231-248.
DeDatta, S. K., W. N. Obcemea, and R. K. Jana. 1972. Protein content of rice grain as affected
by nitrogen fertilizer and some triazines and substituted ureas. Agronomy Journal 64:785-788.
Diepenbrock, W., and G. Geisler. 1979. Compositional changes in developing pods and seeds
of oilseed rape (Brassica napus L.) as affected by pod position on the plant. Canadian Journal
of Plant Sciences 59:819-830.
Dudley, 1. W., and R. J. Lambert. 1968. Genetic variability after 65 generations of selection in
Illinois high oil, low oil, high protein, and low protein strains of Zea mays L. Crop Science
9:179-181.
Earle, F. R., 1. 1. Curtice, and J. E. Hubbard. 1956. Composition of the component parts of the
corn kernel. Cereal Chemistry 23 :507.
Elmore, C. D., W.1. Spurgeon, and W. O. Thorn. 1979. Nitrogen fertilization increases Nand
alters amino acid concentration of cottonseed. Agronomy Journal 71 :713-716.
Ene, B. N., and E. W. Bean. 1975. Variations in seed quality between certified seed lots of
perennial rye grass and their relationships to nitrogen supply and moisture status during seed
development. Journal ofBritish Grassland Society 30: 195-199.
Francois, L. E., E. V. Maas, T. 1. Donovan, and V. L. Youngs. 1986. Effect of salinity on grain
yield and quality, vegetative growth, and germination of semi-dwarf and durum wheat.
Agronomy Journal 78:1053-1058.
Gbikpi, P. 1., and R. K. Crookston. 1981. Effect of flowering date on accumulation of dry
matter and protein in soybean seeds. Crop Science 21 :652-655.
Glenn, D. M., A. Carey, F. E. Bolton, and M. Vavra. 1985. Effect ofN fertilizer on protein
content of grain, straw and chaff tissues in soft white winter wheat. Agronomy Journal 77 :229-
232.
Greaves, J. E., and E. G. Carter. 1923. The influence of irrigation water on the composition of
grains and the relationship to nutrition. Journal of Biological Chemistry 58:531-541.
Green, A. G. 1986. Effect of temperature during seed maturation on the oil composition of low-
linolenic genotypes oftlax. Crop Science 26:961-965.
Gubbels, G. H. 1981. Quality, yield and seed weight of green field peas under conditions of
applied shade. Canadian Journal of Plant Science 61 :213-2] 7.
Harrington, J. F. 1960. Germination of seeds from carrot, lettuce, and pepper plants grown
under severe nutrient deficiencies. Hilgardia 20:219-255.
Harris, H. C., I. R. McWilliam, and V. 1. Bofinger. 1980. Prediction of oil quality of sunflower
from temperature probabilities in eastern Australia. Australian Journal of Agricultural
Research 31:477-488.
Howell, R. W., and 1. L. Carter. 1958. Physiological factors affecting composition of soybeans.
II. Responses of oil and other constituents of soybeans under controlled conditions. Agronomy
Journal 50:664-667.
56 Chemistry ofSeeds
Ivanov, N. N. 1933. Cause of the chemical variation of chickpea seeds. Bulletin of Applied
Botanical Genetics, Plant Breeding Senes 3(1):3-1 1. (Chemical Abstracts 27:5370, 1933).
Jones, O. R. 1984. Yield, water use efficiency, and oil concentration and quality of dryland
sunflower grown in the southern high plains. Agronomy JounnaI76:229-235.
Karathanasis, A. D., V. A. Johnson, G. A. Peterson, D. H. Sander, and R. A. Olsen. 1980.
Relation of soil properties and other environmental factors to grain yield and quality of winter
wheat grown at international sites. Agronomy Journal 72:329-336.
Kerr, R. W., ed. 1950. Chemistry and Industry of Starch. 2d ed. New York: Academic Press.
Ketring, D. L., C. E. Simpson, and O. D. Smith.1978. Physiology of oil seeds. VII. Growing
season and location effects on seedling vigor and ethylene production by seeds of three peanut
cultivars. Crop Science 18:409-413.
Loneragan, J. F., K. Snowball, and A. D. Robson. 1980. Factors influencing variability in
manganese content of seeds, with emphasis on barley (Hordeum vulgare) and white lupine
(Lupinus alb us). Australian Journal ofAgricultural Research 41:29-37.
Malid, H. R., and A. A. Schneiter. 1987. Yield and quality of semidwarf and standard-height
sunflower hybrids grown at five plant populations. Agronomy Journal 79:681-684.
Mathers, A. C., and B. A. Stewart. 1982. Sunflower nutrient uptake, growth, and yield as
affected by nitrogen or manure, and plant population. Agronomy Journal 74:911-915.
Mathers, A. C., F. G. Viets, Jr., M. E. Jensen, and W. H. Sletten. 1960. Relationship of nitrogen
and grain sorghum yield under three moisture regimes. Agronomy Journal 52:443-446.
Meyer, B. S., D. B. Anderson, and R. H. Bohning. 1960. Introduction to Plant Physiology.
New York: D. Van Nostrand Company.
Miller, E. C. 1931. Plant Physiology. New York: McGraw-Hill.
Mitchell, E. M. 1930. A microchemical study of hemic elluloses of endosperms and cotyledons.
American Journal ofBotany 17: 117-138.
Morrison, F. B. 1961. Feeds and Feeding. Ithaca, N.Y.: Morrison Publishing Company.
Mothes, K., L. E. Engelbrecht, and O. Kulajewa.1959. Uber die Wirkung des Kinetins auf
Stickstoffverteilung und Eiweiss-synthese in isolierten Blattern. Flora: Oder Allgemeine
Botanische Zeitung 147:445-464.
Nandisha, B. S., and M. Mahadevappa. 1984. Influence of mother-plant nutrition and spacing
on planting value of rice seeds (Oryza sativa L.). Seed Research 12:25-32.
Norden, A. J., E. C. Rossman, and E. J. Benne. 1952. Some factors that affect protein content
of com. Michigan Agricultural Experiment Station Bulletin 34 :210-225.
Ohkuma, K. 1965. Synthesis of some analogs of abscisin II. Agricultural and Biological
Chemistry 29:962-964.
Osborne, T. B. 1924. Monographs on Biochemistry: The Vegetable Proteins. London:
Longmans, Green, and Company.
Owen, D. F. 1983. Differential response of sunflower hybrids to planting date. Agronomy
Journal 75:259-262.
Parker, M. B., F. C. Boswell, K. Ohki. L. M. Shumand, and D. O. Wilson. 1981. Manganese
effects on yield and nutrient concentration in leaves and seed of soybean cultivars. Agronomy
Journal 73:643-646.
Peck, N. H., D. L. Grunes, R. M. Welch, and G. E. MacDonald. 1980. Nutritional quality of
vegetable crops as affected by phosphorus and zinc fertilizers. Agronomy Journal 72:528-534.
Pikaard, C. S., and J. H. Cherry. 1984. Maintenance ot normal or supranormal protein
accumulation in developing ovules of Glycine max (L.) Merr. during PEG-induced water
stress.Plant PhySiology 75:176-180.
Chemistry ojSeeds 57
Poole, W. D., G. W. Randall, and G. E. Ham. 1983. Foliar fertilization of soybeans. 1. Effect
of fertilizer sources, rates and frequency of application. Agronomy Journal 75: 195-200.
Porter, M. A., and G. M. Paulsen. 1983. Grain protein response to phosphorus nutrition of
wheat. Agronomy Journal 75:303-305.
Raboy, V., andD. B. Dickinson. 1984. Effect of phosphorus and zinc nutrition on soybean seed
phytic acid and zinc. Plant Physiology 75:1094-1098.
Radford, R. L., C. Chavengsaksongkram, and T. Hymowitz. 1977. Utilization of nitrogen to
sulphur ratio for evaluating sulphur-coating amino acid concentration in seed of Glycine max
and G. soya. Crop Science 17:273-277.
Radley, M. 1956. Occurrence of substances similar to gibberellic acid in higher plants. Nature
178:1070-1071.
Read, D. W. L., and F. G. Warder. 1982. Wheat and barley responses to rates of seeding and
fertilizer in southwestern Saskatchewan. Agronomy Journal 74:33-36.
Richmond, A. E., and A. Lang. 1957. Effect of kinetin on protein content and survival of
detachedXanthium leaves. Science 125:650-651.
Robinson, R. G. 1983. Yield and composition offield bean and adzuki bean in response to
irrigation, compost, and nitrogen. Agronomy Journal 75 :31-35.
Robinson, R. G., 1. H. Ford, W. E. Lueschen, D. L. Rabas, L. 1. Smith, D. D. Warnes, and J.
V. Wiersma. 1980. Response of sunflower to plant population. Agronomy Journal 72 :869-871.
Scott, R. K. 1969. The effect of sowing and harvesting dates, plant population and fertilizers
on seed yield and quality of direct-drilled sugar beet seed crops. Journal ofAgricultural Science
70:373-385.
Shannon, J. G., J. R. Wilcox, and A. H. Probst. 1972. Estimated gains from selection for protein
and yield in the F4 generation of six soybean populations. Crop Science 12:824-826.
Simmons, S. R., and D. N. Moss. 1978. Nitrogen and dry matter accumulation by kernels
formed at specific florets in spikelets of spring wheat. Crop Science 18:139-143.
Stone,1. F., and B. B. Tucker. 1968. Nitrogen content of grain as influenced by water supply
to the plant. Agronomy Journal 61 :76-78.
Stone, 1. F., R. H. Griffin, II, and B. J. Ott. 1964. Irrigation studies of grain sorghum in the
Oklahoma Panhandle, 1958 to 1962. Oklahoma Agricultural Experiment Station Bulletin B-
619.
Thomas, T. H., P. F. Wareing, and P. M. Robinson. 1965. Action of the sycamore "dormin" as
a gibberellin antagonist. Nature 205:1270-1272.
Touchton, J. T., and F. C. Boswell. 1975. Effects of boron application on soybean yield,
chemical composition and related characteristics. Agronomy Journal 67:417-420.
Unger, P. W. 1986. Growth and development of irrigated sunflower in the Texas high plains.
Agronomy Journal 78:508-515.
Van Overbeek, l, M. E. Conklin, and A. F. Blakeslee. 1941. Factors in coconut milk essential
for growth and development of very young Datura embryos. Science 94:350-351.
Wolfson, J. L., and G. Shearer. 1981. Amino acid composition of grain protein of maize grown
with and without pesticides and standard commercial fertilizers. AgronomyJournal 73 :611-613.
4
Seed
Ecology
Seed ecology is the study of the ecological strategies plants utilize to ensure their
reproduction by seed. These strategies include timing of seed production; allocation of nutrients
to few or many seeds, big seeds or small ones; mechanisms of seed dispersal; understanding the
seed bank; the environmental conditions that impose dormancy and induce germination; and
factors that influence successful seedling establishment. Detailed considerations of seed ecology
can be found in Baskin and Baskin (1998), Fenner (1985, 1992), and Leck, et al. (1989). Seed
ecology is complex because it involves the interaction of many environmental factors with a
changing biological entity, and differing plants employ varying ecological strategies that make
this topic compelling and rewarding. A knowledge of seed ecology enhances (1) effective
planning for the control of weeds, (2) successful propagation of native economically important
trees, shrubs, vines, forbs and grasses, and (3) reclamation of damaged ecosystems (Baskin and
Baskin 1998).

Reproductive Strategies

Each plant species has a characteristic germination season to optimize successful

reproduction and survival of the species. Thompson and Grime (1979) summarized these by
classirying plants according to their germination strategies (Figure 4.1). These include:

Type 1. Autumn-germinating species whose transient seed banks are present throughout
the summer only. These are typically large-seeded species that germinate over a wide
range of temperature and light conditions.
Type 2. Spring-germinating species whose transient seed banks are present during the
winter only. Often these have a chilling requirement, which imposes winter dormancy. It
is like Type 1 in being relatively large-seeded and not requiring light for germination.
Type 3. Species in which most of the seeds germinate soon after dispersal (usually in late
summer), but in which a small proportion becomes incorporated into a persistent seed
bank. These species tend to have small seeds that germinate only over a restricted range
of temperatures and are often light requiring.

L. O. Copeland et al., Principles of Seed Science and Technology

© Kluwer Academic Publishers 2001
Seed Ecology 59

Typel~ a

Typey
I 1

Type~?t> It
c
12#

I I I I I I I

TypelV ~_~
Wffi I I I I I I I I I ) ) ) I )
~~
) I I ) ! I I
AM J J A SJ NO J F M A M J J A SO NO J F M A

Figure 4.1. Four types ofseed banksfoundfor herbaceous species in contrasting habitats in England
Unshaded areas, seeds alive but not capable ofgerminating at 20115°C; shaded areas, seeds capable
of germinating at 20115°C. Type L transient seeds germinate in autumn; Type II, transient seeds
germinate in spring; Type III, persistent seeds germinate primarily in autumn and the seed reserve is
small; and type IV, persistent seeds germinate in autumn and the seed reserve is large (from Thompson
and Grime 1979).

Type 4. Species in which only a few of the seeds germinate soon after dispersal. Most of
the seeds enter the persistent seed bank that is large in relation to the annual
production. These species differ only in degree from those of Type 3, but represent the
extreme case of species in which the seed bank strategy is most strongly developed. These
are typically annual and perennial herbs and shrubs.

Successful plants must first establish a significant vegetative structure to capture sunlight
and utilize a vast root system to facilitate mineral uptake that supports continued photosynthesis
and vegetative growth. This vegetative structure is generally more significant in perennial than
biennial and vegetative crops. However, at some point, resources typically sequestered for
vegetative growth are allocated to reproduction to ensure survival of the species. Flowering
plants accomplish this reproduction either sexually by seed production or asexually by rhizomes,
buds, etc. (Chapters 1 and 2). Seeds, however, provide several evolutionary advantages.
Because they are a product of sexual recombination, every seed is genetically unique, producing
a population of individuals that respond best to changing or differing environments. This is
contrasted with asexual reproduction in which the offspring are clones that have the same
genetic composition as the parent plant. While asexual reproduction is successful under stable,
unchanging environmental conditions, such a reproductive strategy is less flexible to new and
changing environments.
60 Seed Ecology

There are two basic sexual reproduction strategies found in plants: monocarpic and
polycarpic. Monocarpic plants are those in which seeds are produced only once in the life of
the plant, followed by its death. These plants can be annual, biennial or perennial. Annuals
produce seeds as quickly as possible. Biennials spend their first growing season amassing large
reserves of energy stored in vegetative tissues subsequently used for reproduction the second
growing season. Perennials, such as bamboos, delay reproduction for several years and then
allocate a "big bang" of seed production followed by plant death (Gadgil and Bossert 1970).
Polycarpic plants are those in which seeds are produced repeatedly for an indefinite period.
This can occur annually, as with dandelion, or irregularly at intervals of several years, as with
many forest trees. Each of these reproductive strategies is beneficial for particular
environments. For example, in an unstable environment, annuals are not favored because they
produce little vegetative structure to support subsequent seed production. In contrast, perennials
can delay seed production under stress conditions so that increased resources are allocated to
vegetative tissues that can be utilized later under more favorable conditions for seed formation,
thereby increasing fecundity. Biennials represent a compromise between these two reproductive
strategies for fast production of a few seeds and delayed production of many seeds.
Allocation of Plant Resources. The level of allocation of plant resources to seed
production is an important concept in seed ecology. Plants have phenotypic plasticity, which
means they can respond to an environment by determining the relative allocation of plant
resources to vegetative growth or sexual reproduction. For example, early colonizers in a site
are usually smaller than those that succeed them, and they usually have a higher fraction of their
biomass allocated to seed production (Fenner 1985). Increasing plant density often favors
sexual reproduction, a useful reproductive tactic for colonizing new sites after local
consolidation has been accomplished.
Seed Size and Number. Another reproductive strategy is whether a plant produces fewer,
but bigger or many, but smaller seeds (Harper, et a!. 1970). Seed weight tends to be related to
the size of the parent plant. On a global basis, Levin and Kerster (1974) showed that trees,
shrubs and herbaceous plants have mean seed weights of328, 69 and 7 mg, respectively. Thus,
bigger plants usually produce heavier seeds.
These fmdings are ecologically consistent for plants in a successional hierarchy. Generally,
small seeds are advantageous for plants requiring dispersal of many seeds and large seeds are
advantageous for plants requiring successful seedling establishment. For example, plants of
transient, widely scattered open sites are favored by producing small seeds that permit wide
dispersal to other sites, thereby minimizing competition from surrounding plants. Plants
growing in stable environments with closed vegetation, such as trees, require less emphasis on
dispersal and more on large seed production to ensure successful seedling establishment in a
highly competitive, established environment.

Seed Dispersal

Seed rain is the total contribution of seeds by the parent plant to the soil seed bank. Seed
rain is the equivalent of seed production. The spatial distribution of dispersed seeds around their
source is called a seed shadow (Janzen 1969). Often, the source is a maternal plant, but the
term can also include the distribution of seeds around multiple parents.
Seed Ecology 61
The ecological consequences of seed dispersal are important (Howe and Smallwood 1982).
Seed dispersal patterns contribute to the genetic structure of plant populations and to the
potential for genetic drift and responses to natural selection. Long-distance seed dispersal by
wind, animals, and even ocean currents has determined the composition of the flora of many
isolated islands. The initial colonization ofMt. St. Helen's following its volcanic eruption in
Washington state was primarily by wind-dispersed species (Dale 1989).
Why are Seeds Dispersed? Seed dispersal is advantageous to plants for several reasons.
First, it enables the germinating seedling to avoid established competitive conditions around the
mature parent plant. Second, dispersal enhances the avoidance of natural enemies such as
pathogens, predators, parasites and herbivores that accumulate around an established site.
Third, sibling competition for the same environmental resources is minimized. Fourth, some
plants have special seed dormancy traits that must be satisfied before germination can occur.
Seed dispersal enhances the probability that at least some of these seeds will encounter the
appropriate conditions to break seed dormancy. Thus, if the dispersal of seeds includes enough
safe seed sites, species with widespread seed shadows should be more evolutionarily successful
than those with restricted seed shadows.
How are Seeds Dispersed? The concepts of seed dispersal are discussed by Murray
(1986) and Fahn and Werker (1972). Animals (zoochory; chory from chorein - to wander),
wind (anemochory), water (hydrochory), and the plant itself (autochory) assist seed dispersal.
Van der Pilj (1972) divided zoochory into three groups: (1) endozoochorous plants where
the seeds are ingested by animals and pass through the digestive tract without damage, (2)
epizoochorous plants where the seeds adhere to the fleece, coat or feathers of animals and then
loosen and fall to the ground and (3) synzoochorous plants where the seeds are collected and
cached for feeding.
In endozoochory, various animals such as fish, reptiles, birds and rodents eat seeds. These
seeds typically possess attractive features to animals such as color, odor, and abundance of
storage materials and large size. Many fruits, for example, dramatically change color from
green to red, blue or black as they mature, which contrasts them against the green foliage. This
morphological change assures easy visibility to the foraging animal, signifies that the fruit is ripe
and simultaneously ensures that the seeds are mature and capable of reproduction. Other fruits
such as Citrus emit appealing aromas that attract animals. The type of storage material
(carbohydrates, lipids, proteins, etc.) and the size of the fruit are also important nutritional
attributes that animals consider when foraging for food.
In epizoochory, seeds develop at least two animal-adhering mechanisms that assist their
dispersal: (1) hooklike spines and (2) sticky substances. Hooklike spines are usually more
common in the fruit than the seed (Figure 4.2), but they can also be found on seeds (Figure 4.3).
The hairs and bristles of some seeds that assist in wind dispersal also cling to animals. Sticky
substances such as mucilage extruded from moistened seed coats allow the seed to adhere to a
passing animal in the morning when dews are present, and then be deposited elsewhere when the
mucilage dries later in the day. Mucilage production is common in seeds from the Brassicaceae,
Lamiaceae, Asteraceae and Plantaginaceae. Genera that produce abundant mucilage include
Salvia and Plantago.
In synzoochory, seeds are collected and stored by animals for nutritional reasons. In some
cases, the nutritional part of the seed is a fleshy appendage called an elaisome that can be easily
detached from the remainder of the seed, which is hard and inedible. Examples of this include
Acacia and Viola seeds where ants (myrmechory) harvest the seeds, carry them to their nests
62 Seed Ecology

Position of
abscission layer

10mm

B c
Upper
seed
" / 1,
: "I
....
Hook

Air
callities

Lower
seed

Figure 4.2. The fruit ofXanthium occidentale: (A) the bur; (B) transverse section ofthe bur showing
unequal seeds in chambers with air cavities; (C) hooked spine (from Murray 1986).
Seed Ecology 63

Figure 4.3. A cypsela o/Eidens pi/osa (from Carlquist 1966).

where the elaisome or attractive appendages are separated from the seed, and the undesirable
seeds discarded (Figure 4.4). The value of this approach is not in the distance the seeds are
transported, which may be only a few meters, but in the reduction of the risk to predation and
in the enhanced quality of the germination micro-site in which the seed is deposited (Beattie
1985).
In anemochory, wind is the most effective of all seed dispersal agents. Appropriately,
seeds have developed specialized structures to assist their dispersal by air. These include small
seed size, balloons, plumage and wings. Seed size is important in wind dispersal because very
small seeds can be transported in air just like dust. These seeds often contain undeveloped
embryos and small storage reserves and seldom exceed 3 to 4 J..Lg. Examples include members
of Gypsophila, Pyro/a, Sedum, Campanu/a, Digitalis and Orobanche genera. In other seeds,
such as orchids, a balloon-like sack surrounds the small seed, which is only one to a few layers
thick. This causes the specific weight of the seed to decrease and the surface area to increase
facilitating movement of the seed through the air. Plumed seeds have trichomes or hairs
covering their surface. Examples include Asclepias, Salix and Populus. The cypsela of the
Asteraceae also produces a feathery pappus (Figure 4.5). Winged seeds can also possess one-
sided wings such as maple or ash seeds or wings that surround the seed such as beech and elm
seeds and are flattened in one plane or curved. The shape of the wing determines how the seed
moves in the wind. The wings are usually large, but small in weight and relatively rigid.
In hydrochory, seeds are dispersed in water because they possess a specialized air-filled
tissue that allows the seed to be buoyant in water. This tissue can contain (1) spongy, inter-
cellular spaces filled with air, (2) air-filled cells in differing parts of the fruit such as the
lignified air-filled cells of the mesocarp in coconut and (3) an aril that encloses the seed as an
air-filled sack that functions as a swim bladder.
In autochory, plants have developed mechanisms to disperse their own seeds ballistically.
This can occur by the explosive opening of the fruits or the springing of a trip lever in a fruit
that ejects the seed (Figure 4.6).
64 Seed Ecology

Figure 4.4. Ants picking up seeds of Viola nuttallii. The seed bears an attractive and edible
appendage. Ants carry the entire seed back to the nest, eat the appendage and discard the seed.
Dispersal ofseed by ants is very common in some floras, but the advantage of ant dispersal may vary
greatly among species or regions (e.g., escape from predators or other destructive agents or deposition
in an especially favorable site for germination and growth) (from Beattie 1985).

Figure 4.5. Cypsela of a member ofthe Asteraceae showing the feathery pappus that assists in wind
dispersal (from Fahn 1967).
Seed Ecology 65

Figure 4.6. Depression of the calyx tube and subsequent expulsion of the seed in Salvia lyrata (from
Brodie 1955).

Seed Banks

A reserve of viable, ungerminated seeds in a soil is called a seed bank (Roberts 1981).
Seed banks are classified as transient (none of the seed produced in a given year remain viable)
and persistent (seeds are 1 or more years old). In transient seed banks, seeds lose dormancy in
a short period or encounter conditions favorable for germination. Non-dormant seeds retain
their viability in the transient seed bank for only short periods. As a result, most seed ecologists
focus their attention on the persistent seed bank because it contributes germinating seedlings
over a longer duration and has more effect on the plant population of a particular location.
The input and output of a typical seed bank is illustrated in Figure 4.7. The input is the
total contribution of the seed rain in the soil. These seeds (1) can be lost to predation, decay
and senescence leading to seed death, (2) if appropriate environmental stimuli exist to break
dormancy, they enter the transient (active) seed bank and germinate, or (3) remain in the
persistent (dormant) seed bank with the potential to contribute seedlings in the future. It is these
seeds and the environmental conditions necessary to cause them to germinate that are the subject
of interest to seed ecologists.

Dormancy

Ecologically, dormancy is considered to be a mechanism that delays germination until

conditions are suitable for seedling establishment. For example, in desert climates, water is
unpredictable and seeds must respond to an appropriate environmental cue that signals that
sufficient water is available to enable successful germination and seedling establishment.
Thus,seed dormancy is an important survival trait in many desert species (Gutterman 1993).
66 Seed Ecology
In contrast, many tropical rainforest trees produce seeds (referred to as recalcitrant seeds, see
Chapter 7) that are short-lived and possess little or no dormancy. This lack of dormancy
suggests that their tropical rainforest habitat is predictable and that dormancy would confer no
selective advantage. The mechanisms that impose seed dormancy are discussed further in
Chapter 7.

Germination

Seeds in the soil seed bank are ready to germinate provided they encounter the appropriate
environmental cue(s) (Figure 4.7). Such cues enable seeds to maximize the likelihood of
survival in patchy and unpredictable environments. Appropriate dormancy-breaking conditions
must be present followed by satisfactory post-germination environments for successful seedling
establishment.
One of the most common environmental cues to ensure successful seedling establishment
is gaps in vegetation. These are openings that minimize competition from other plants and
maximize availability ofnutrients and environmental resources. Gaps produce specific cues that
signal a seed that the environment is conducive for successful seedling establishment. Examples
include a covering of vegetation that acts as an effective temperature buffer by insulating the
soil against large diurnal fluctuations (Thompson, et al. 1977). When gaps are present, the sun
strikes the soil in the day causing it to warm. In the evening, more rapid loss of soil temperature
occurs creating large fluctuations in soil temperature. These alternating temperatures provide
a signal to seeds that successful germination and seedling establishment are possible. Another
gap detection cue is light quality. Direct sunlight has a greater red/far-red ratio compared to
light passing through a leafed canopy. Thus, light-sensitive seeds detect the difference in light
quality, prompting increased seed germination in gaps (Vazquez-Yanes and Smith 1982).

Seed rain

Seedlings

Predation Dormant
seed
bank

Decay and
senescence

Death
Figure 4.7. Flow chart for the dynamics of a seed population in a seed bank (from Harper 1977).
Seed Ecology 67
In addition to gaps, chemicals present in the soil stimulate germination. Of these, the
nitrate ion has the greatest effect (Karssen and Hilhorst 1992). Since the concentration of nitrate
in the field fluctuates seasonally due to the changing activity of soil microorganisms, an ability
to respond to this ion could act as another cue to promote germination of seeds during the most
favorable growing season. Since the rate of mineralization of nitrogen depends on temperature,
a higher production of nitrate and ammonium can be expected during the summer. In general,
concentrations as low as 0.1 to 0.5 mol tt N03 stimulate germination. These concentrations are
consistent with levels found naturally in the soil (Young and Aldag 1982).
Other factors that influence soil nitrate levels include soil disturbances that can release
considerable bound nitrate into the soil solution. Established plants may incorporate available
nitrate, lower the nitrate content of the soil around their root systems and cause seeds in the
immediate environment to remain dormant (Pons 1989). It has also been noted that many
nitrate-dependent seeds are light requiring (Karssen and Hilhorst 1992). While the soil nitrogen
content can influence germination, increasing the endogenous nitrate content of the seed may
also alleviate this requirement. For example, a positive relationship between endogenous nitrate
content and germination in water was found for Chenopodium album (Saini, et al. 1985) and
Sisymbrium ojficinale (Hilhorst 1990b). Seed nitrate levels below 0.1 J..Llllole g,t did not
stimulate germination. How nitrate breaks seed dormancy is unknown. However, it has been
postulated that nitrate binds directly to the phytochrome receptor protein such as PFR allowing
it to become physiologically active (Hilhorst 1990a,b; Karssen and Hilhorst, 1992).
The germination of seeds in the soil seed bank can also be regulated by allelopathy.
Allelopathy refers to the detrimental effects of higher plants of one species on the germination,
growth or development of plants of another species (Putnam 1985; Rice, 1984). Extensive
evidence exists that allelopathy contributes to interactive relationships among plants, such as
those that determine species composition in community structure, species placement in
succession, etc. Inhibitors of seed germination have been detected in extracts from decaying
sorghum and sunflower plants (Karssen and Hilhorst 1992). Another example of the role of
allelopathy in species placement in plant succession was provided by Numata (1982) who
identified cis-dehydroxy-matricriaster (cis-DME) as the allelopathic compound in Solidago
altissima which succeeds Ambrosia artimisifolia in old-field succession. This compound
survived in the soil for several months without decomposition and as little as 2.5% of the dry
weight of Solidago plant material successfully inhibited germination of Ambrosia seeds.
Allelochemicals are also known to leak from seeds in addition to vegetative tissues.
Picman and Picman (1984) showed that Parthenium hysterophorus contained two major water
soluble sesquiterpene lactones primarily in the seed coat that represented 8% of the seed dry
weight and were autotoxic to the plant's own seeds. However, because they are water soluble,
they may act as a rain gauge by governing the quantity of inhibitor leached from the vicinity of
the seed.

Seedling Establishment

A seedling is considered fully established when it is independent of its storage reserves.

The early stages of seedling growth represent the highest mortality risk for developing seedlings.
Mortality rates primarily due to desiccation of greater than 80% in the first year have been
reported for nine species of herbaceous perennials in chalk grasslands (Silvertown and Dickie
1981). Competition from neighboring plants also represents a major hazard faced by colonizing
68 Seed Ecology
seedlings with the greatest mortality occurring during favorable growing periods. For example,
chickweed seedling mortality was greatest in the spring after plants had resumed growth (Mack
1976). When seeds are buried, biotic factors such as predation, disease and competition also
create conditions that enhance seedling mortality.
Factors that enhance seedling survival include seed size and rapidity of seedling growth.
One of the most successful adaptations for ensuring successful seedling establishment is the
development of a large seed. Large seeds generally produce larger seedlings that provide a
competitive advantage over small seeds. This has been shown in Lupinus texensis (Schall
1980), Mirabilis hirsuta (Weis 1982) and Raphanus raphanistrum (Stanton 1984). Four
biennial species of different seed sizes were planted into fields that were 1, 5 and 15 years old.
The small-seeded species established only in the one-year-old field while the large-seeded species
survived in all fields (Gross and Werner 1982). Similar fmdings were reported for monocarpic
perennial plants (Gross 1984). Seeds from plants adapted to closed vegetation such as trees
produce larger seeds than those from open sites (Fenner 1985). Such seedlings from climax
forest trees, for example, will spend years in conditions of poor sunlight, so ample energy
reserves are necessary to permit seedling survival and rapid seedling growth. Large seeds may
also provide an advantage in arid environments because they enable more rapid root elongation.
Since the sand in which the seed germinates dries out quickly, the seedling must have rapid root
elongation ahead of the descending drying front. Larger seeds can produce shoots that grow
more quickly and emerge from greater depths (Buckley 1982).
Seedling morphology can also determine the competitiveness of a plant for light. For
example, Fenner (1983) showed that large seeds from the Asteraceae family tended to have
higher shoot/root ratios, suggesting that the capture of light was more important than the uptake
of soil nutrients. Species from closed grassland vegetation tended to show a high degree of
morphological plasticity to extend hypocotyls, cotyledons, internodes, petioles or laminae of
early leaves (Grime and Jeffrey 1965). Early attainment of plant height can result in the
seedling escaping the shade at ground level. The value of occupying space by early germination
and establishment has been shown by Cook (1980). Early Viola seedlings produced larger
seedlings compared to those produced 15 days later in a wood, and the probability of late
seedling death was also increased. Thus, there is a strong selection pressure for rapid
germination response in established fields or closed canopies to ensure successful seedling
estab lishment.
The type of germination may also confer a selective ecological advantage. For example,
in epigeal germination, the hypocotyl elongates and exposes the cotyledons to light, allowing
them to function in photosynthesis. In hypogeal germination, the storage reserves remain below
ground and function only in providing nutrition to the developing seedling.

Questions

1. Define seed ecology and describe the various ecological strategies plants use in ensuring
their survival.
2. Plants can be classified according to the timing of their seasonal germination. Identify at
least four germination strategies and describe how each is different.
3. Contrast the advantages and disadvantages of asexual and sexual reproduction from an
evolutionary perspective.
Seed Ecology 69
4. Defme monocarpic and polycarpic plants and describe the advantages and disadvantages
each possesses in their reproductive strategies.
5. From an ecological perspective, contrast the advantages/disadvantages of small vs. large
seed production.
6. Differentiate seed rain, seed shadow, and seed bank.
7. IdentifY four reasons why seed dispersal is important for evolutionary success.
8. Describe three ways that animals disperse seeds.
9. What are transient and persistent seed banks? Which is of most interest to seed ecologists
and why?
10. IdentifY at least two environmental stimuli provided by gaps in vegetation that provide
cues that conditions are suitable for successful seedling establishment.
11. Detail the environmental mechanisms that modifY soil nitrate content and influence
seasonal seed germination.
12. What is allelopathy? How does it influence seed ecology?
13. IdentifY the environmental factors responsible for creating the greatest mortality risk
during seedling establishment.

Literature Cited

Baskin, C. C., and J. M. Baskin. 1998. Seeds: Ecology, Biogeography, and Evolution of
Dormancy and Germination. Academic Press, NY.
Beattie, A. 1. 1985. The Evolutionary Ecology of Ant-Plant Mutualisms. Cambridge
University Press, Cambridge.
Brodie, H. J. 1955. Springboard dispersal operated by rain. Canadian Journal of Botany
33:156-164.
Buckley, R. C. 1982. Seed size and seedling establishment in tropical and dunecrest plants.
Biotropica 14:314-315.
Carlquist, S. 1966. The biota of long-distance dispersal. II. Loss of dispersability in Pacific
Compositae. Evolution 20:30-54.
Conn, J. S. and M. L. Farris. 1987. Seed viability and dormancy of 17 weed species after 21
months in Alaska. Weed Science 35:524-529.
Cook, R. E. 1980. Germination and size-dependent mortality in Viola blanda. Oecologia
47: 115-117.
Dale, V. H. 1989. Wind dispersed seeds and plant recovery on Mt. 8t. Helens debris
avalanche. Canadian Journal of Botany 67: 1434-1441.
Fahn, A. 1967. Plant Anatomy. Pergamon, Oxford.
Fahn, A. and E. Werker. 1972. Anatomical mechanisms of seed dispersal. In: Seed Biology
(ed. T. T. Kozlowski). Pp. 151-221. Academic Press, NY.
Fenner, M. 1983. Relationships between seed weight, ash content, and seedling growth in
twenty-four species of Compositae. New Phytology 95 :697 -706.
Fenner, M. 1985. Seed Ecology. Chapman & Hall, NY.
Fenner, M. 1992. Seeds: The Ecology of Regeneration in Plant Communities. CAB
International, Wallingford.
Gadgil, P. M. and W. H. Bossert. 1970. The life historical consequences of natural selection.
American Naturalist 104: 1-24.
70 Seed Ecology
Grime, 1. P. and D. W. Jeffrey. 1965. Seedling establishment in vertical gradients of sunlight.
Journal of Ecology 53 :621-642.
Gross, K. L. 1984. Effects of seed size and growth form on seedling establishment of six
monocarpic perennial plants. Journal ofEcology 72:369-387.
Gross, K. L. and P. A. Werner. 1982. Colonizing abilities of 'biennial' plant species in relation
to ground cover: implications for their distributions in a successional sere. Ecology 63 :921-
931.
Gutterman, Y. 1993. Seed Germination in Desert Plants. Springer Verlag, NY.
Harper,1. L. 1977. Population Biology of Plants. Academic Press, NY.
Harper, 1. L., P. Lovell and K. Moore. 1970. The shapes and sizes of seeds. Annual Review
of Ecological. Systematics 1:327-356.
Hilhorst, H. W. M. 1990a. Dose-response analysis of factors involved in germination and
dormancy of seeds of Sisymbrium officinale. I. Phytochrome. Plant Physiology 94:1090-
1095.
Hilhorst, H. W. M. 1990b. Dose-response analysis of factors involved in germination and
dormancy of seeds of Sisymbrium officinale. II. Nitrate. Plant Physiology 94: 1096-11 02.
Howe, H. F. and J. Smallwood. 1982. Ecology of seed dispersal. Annual Review ofEcological
Systematics 13:201-228.
Jansen, D. H. 1969. Seed-eaters vs. seed size, number, toxicity and dispersal. Evolution 23:1-
27.
Karssen, C. M. and H. W. M. Hilhorst. 1992. Effeet of chemical environment on seed
germination. In: Seeds: The Ecology ofRegeneration in Plant Communities (ed. M. Fenner).
Pp. 327-348. CAB International, Wallingford.
Leek, M. A., V. T. Parker and R. L. Simpson. 1989. Ecology of Soil Seed Banks. Academic
Press, NY.
Levin, D. A. andH. W. Kerster. 1974. Gene flow in seed plants. Evolutionary Biology 7:139-
220.
Mack, R. H. 1976. Survivorship of Cerastium atrovirens at Aberffiaw, Anglesey. Journal
ofEcology. 64:309-312.
Murray, D. R. 1986. Seed Dispersal. Academic Press, NY.
Numata, M. 1982. Weed-ecological approaches to allelopathy. In: Biology and Ecology of
Weeds (eds. W. Holzner and N. Numata), pp. 169-173. Junk, The Hague.
Picman, 1. and A. K. Picman. 1984. Autotoxicity in Parthenium hysterophorus and its
possible role in control of germination. Biochemical Systematics and Ecology 12:287-292.
Pons, T. L. 1989. Breaking of seed dormancy as a gap detection mechanism. Annals of
Botany 63:139-143.
Putnam, A. R. 1985. Weed allelopathy. In: Weed Physiology, Vol. 1, Reproduction and
Ecophysiology (ed. S. O. Duke). Pp. 131-155. CRC Press, Boca Raton, FL.
Rice, E. L. 1984. Allelopathy, 2nd edn. Academic Press, NY.
Roberts, H. A. 1981. Seed banks in soil. Advances in Applied Biology 6:1-55.
Saini, H. S., P. K. Bassi and M. S. Spencer. 1985. Seed germination of Chenopodium album
L.: Further evidence for the dependence of the effects of growth regulators on nitrate
availability. Plant, Cell and Environment 8:707-711.
Schall, B. A. 1980. Reproductive capacity and seed size in Lupinus texensis. American
Journal of Botany 67:703-709.
Seed Ecology 71
Silvertown, J. W. and 1. B. Dickie. 1981. Seedling survivorship in natural populations of nine
perennial chalk grassland plants. New Phytology 88:555-558.
Stanton, M. L. 1984. Seed variation in wild radish: effect of seed size on components of
seedling and adult fitness. Ecology 65: 11 05-1112.
Thompson, K., J. P. Grime and G. Mason. 1977. Seed germination in response to diurnal
fluctuations to temperature. Nature 267: 147-149.
Thompson, K. and 1. P. Grime. 1979. Seasonal variation in the seed banks of herbaceous
species in ten contrasting habitats. Journal of Ecology 67:893-921.
Van der Pilj, L. 1972. Principles of Seed Dispersal in Higher Plants. Springer-Verlag, NY.
Vazques-Yanes, C. and H. Smith. 1982. Phytochrome control of seed germiantion in the
tropical rain forest pioneer trees Cecropia obtusifolia and Piper auritum and its ecological
significance. New Phytology 92:477-485.
Weis,1. M. 1982. The effects of propagule size on germination and seedHng growth in
Mirabilis hirsuta. Canadian Journal o/Botany 60:1868-1874.
Wulff, R. D. 1986. Seed size variation in Desmodium paniculatum. I. Factors affecting seed
size. Journal of &ology 74:87-97.
Young, J. L. and R. W. Aldag. 1982. Organic forms of nitrogen soils. In: Nitrogen in
Agricultural Soils (ed. F. 1. Stevenson), pp.43-66. ASA, CSSA, SSSA, Madison, WI.
5
Seed
Germination

In the germination process, the seed's role is that of a reproductive unit; it is the thread of
life that assures the survival of all plant species. Furthermore, because of its role in stand
establishment, seed germination remains a key to modern agriculture. Thus, especially in a world
acutely aware of the delicate balance between food production and world population, a
fundamental understanding of germination is essential for maximum crop production.

DEFINITION OF GERMINATION

Various definitions of seed germination have been proposed, and it is important to

understand their distinctions. To the seed physiologist, germination is defined as the emergence
of the radicle through the seed coat. To the seed analyst, germination is "the emergence and
development from the seed embryo of those essential structures which, for the kind of seed in
question, are indicative of the ability to produce a normal plant under favorable conditions"
(AOSA 2000). Others consider germination to be the resumption of active growth by the embryo
resulting in the rupture of the seed coat and the emergence of a young plant. This definition
presumes that the seed has been in a state of quiescence (see Chapter 7), or rest, after its
formation and development. During this period of rest, the seed is in a relatively inactive state
and has a low metabolic rate. It can remain in this state until environmental conditions trigger
the resumption of growth. Regardless of which defmition is preferred, it should be emphasized
that one cannot actually see the process of germination unfold. Therefore, all defmitions include
some measure of seedling development, even though this occurs subsequent to the germination
event.
Some seeds are capable of germination only a few days after fertilization and long before
their normal harvesting time; others are dormant and require an extended rest period or
additional development before germination can occur. Depending on the species, this period may
last for only a few days or for as long as several years. Regardless of the length of time between
maturity and resumption of growth, seed germination may be characterized by several general
processes that are discussed in this chapter.

L. O. Copeland et al., Principles of Seed Science and Technology

© Kluwer Academic Publishers 2001
Seed Germination 73

MORPHOLOGY OF GERMINATION
Based on the fate of the cotyledons or storage organs, two kinds of seed germination occur,
and neither appears to be related to seed structure. These two types are illustrated by the
germination of bean and pea seeds. Although these seeds are similar in structure and are in the
same taxonomic family, their germination patterns are quite different.

Epigeal Germination
Epigeal germination (F igure 5.1) is characteristic of bean and pine seeds and is considered
evolutionarily more primitive than hypogeal germination (described below). During germination,
the cotyledons are raised above the ground where they continue to provide nutritive support to
the growing points. During root establishment, the hypocotyJ begins to elongate in an arch that
breaks through the soil, pulling the cotyledon and enclosed plumule through the ground and
projecting them into the air. Afterward, the cotyledons open, plumule growth continues and the
cotyledons wither and fall to the ground.

HypogeaJ Germination
Hypogeal germination (F igure 5.1) is characteristic of pea seeds, all grasses such as com,
and many other species. During germination, the cotyledons or comparable storage organs
remain beneath the soil while the plumule pushes upward and emerges above the ground. In
hypogeal germination, the epicotyl is the rapidly elongating structure. Regardless oftheir above-
ground or below-ground locations, the cotyledons or comparable storage organs continue to
provide nutritive support to the growing points throughout germination.
The co/eoptile, a temporary sheath enclosing the plumule, is associated with hypogeal
germination of many species (e.g., grasses). It provides protection and rigidity to the emerging
plumule as it pushes through the soil and is exposed to light. Then it stops growing and
disintegrates as the plumule breaks through and continues to grow.

REQUIREMENTS FOR GERMINATION

Seed Maturity

Seeds of most species are capable of germinating long before physiological maturity
(Holmes 1953; Harrington 1959; Bowers 1958; Giri 1967; Williams 1972; Hill and Watkin
1975, Pegler 1976). For example, smooth bromegrass seeds are able to germinate only a few
days after fertilization (Grabe 1956). In other cases, maximum seed germination can only be
obtained if the seed is allowed to dry down slowly as it matures. This is true, for example,in
soybeans in which the ability to synthesize germination requiring enzymes such as malate
synthase and isocitrate lyase developed during the slow maturation process (Adams et al. 1983).
Table 5.1 shows the relationship between seed maturity and germination capability of sow thistle
and Canada thistle.
74 Seed Germination

A. Epigeal Germination of Bean Seed

6
7

8
3 9

4
5
9 ~7 ; J

10

B. Hypogeal Germination of Com Seed

13
15
9
16

1. Seed coat 8. Radicle 15. Endosperm

2. Micropyle 9. Cotyledon (scutellum) 16. Coleoptile
3. Hilum 10. Primary root 17. Coleorhiza
4. Raphe 11. Secondary root 18. Seminal root
5. Chalaza 12. Leaf 19. Adventitious roots
6. Plumule 13. Pericarp 20. Epicotyl
7. Hypocotyl 14. Point of caryopsis
attachment
Figure 5.1. Two patterns ofseed germination and germination structures present in a dicot (bean) and
a monaco! (corn).
Seed Germination 75
Table 5.1. Germination of Perennial Sow Thistle and Canada
Thistle Seeds Harvested at Different Stages of Maturity
(From Kinch and Termunde 1957).

Days After Percent of Germination

Flower Opening Perennial Sow Thistle Canada Thistle
2 o a
3 o o
4 4 o
5 o
6 34 19
7 66 37
8 70 76
9 83 88
10 90
11 80

Environmental Factors

Water. Water is a basic requirement for germination. It is essential for enzyme activation,
breakdown, translocation, and use of reserve storage material. In their resting state, seeds are
characteristically low in moisture and relatively inactive metabolically. That is, they are in a
state of quiescence. Thus, quiescent seeds are able to maintain a minimum level of metabolic
activity that assures their long-term survival in the soil and during storage.
Moisture availability is described in various ways. Field capacity moisture is about
optimum for germination in soil; however, germination varies among species (Table 5.2) and
may occur at soil moistures near the permanent wilting point. In other situations, soils are either
very high in salt concentrations or are located in semiarid regions making water less available
for seed germination. Even under such conditions, however, a number of crops, such as basin
wild rye and tall wheatgrass, among others, are still able to germinate when many other species
cannot (Young and Evans 1981; Ries and Hoffman 1983; Roundy 1985; Dudeck and Peacock
1985).

Table 5.2. The Germination Response of Seeds of Different Species Subjected to 14 Days of
Moisture Stress (From Eslick and Vogel 1959).

Species Exhibiting Species Exhibiting

Increased Germination Species Not Affected Decreased Germination
western wheat grass crested wheat grass big bluegrass
standard crested wheat grass tall wheat grass orchard grass
Ruby Valley milk vetch Russian wild rye mountain bromegrass
broadleaf bird's-foot trefoil timothy switchgrass
tall oat grass Kentucky bluegrass
red fescue alsike clover
smooth bromegrass white sweet clover
ladino clover sickle milk vetch
red clover
76 Seed Germination
In other situations, the initial stages of germination may even proceed with moisture
available from a high-humidity environment, although such conditions are not adequate for
complete germination. This is often demonstrated by a phenomenon known as precocious
germination or sprouting when seeds actually germinate in the head or pod following rains or
high-humidity conditions (Figure 5.2). The physiological factors that regulate precocious seed
germination are being intensively investigated because this phenomenon causes annual soft white
winter wheat crop yield losses that result in millions of dollars in lost crop revenue in the Pacific
Northwest, as well as the soft white wheat areas of Michigan, New York, and Ontario, Canada.
High moisture levels may inhibit germination. For example, when moisture content was
increased from 20% to 40%, dwarf bean germination was reported to decrease significantly
(Ensor 1967). Excess moisture or flooding is also known to retard the germination of sugar beet
seed (Snyder 1975), com (Fausey and McDonald 1985), and several other species (Heydecker
et a1. 1969).

80

/
/'
.--_............\ ,
If
e_e-·-·--'---e
\

/ \: ..-.......!tX
'-.l\,
I A-
c: 40 I .-- ",. \
.~-.,..~
'II
§
(J
60
o
~ 20
/ \, ,~e
$proufinq .,
.......~ n ..,:"
.. -
0 . .
A r
L ! Non visible I I
I I
I
I I
l.-t Visible I I ~
U I I ~

B a-Non visible n:1 ~ ~

Visible
n
: : M
f·l ~ :" t.
t-
--------------------~Q~
o 16 32 48 64
Days after onthesis

Figure 5.2. Degree ofsprouting in two lines ofwheat (A and B), grown in the field, in 1977. Curves
show changes in water content during the development and maturation ofthe seed Vertical columns
show the percentage ofseeds sprouting on the ear. Sprouting is visible when the co/eoptile emerges
through the pericarp, and is invisible when it does not emerge. Water content: A-line.. ..... : B-line
II (from Mitchell et al. 1980).
Seed Germination 77
Air (Oxygen and Carbon Dioxide). Air is composed of about 20% oxygen, 0.03 % carbon
dioxide, and about 80% nitrogen gas. If one provides different proportions of these three gases
under experimental conditions, it soon becomes clear that oxygen is required for germination of
most species. Carbon dioxide (C02) concentrations higher than 0.03 % retard germination, while
nitrogen gas has no influence (Table 5.3).
Respiration increases sharply during seed germination. Since respiration is essentially an
oxidative process, an adequate supply of oxygen must be available. If the oxygen concentration
is reduced substantantially below that of air, germination of most seeds is retarded. For example,
Van Toai et al. (1988) demonstrated that com seed germination declined with decreasing oxygen
concentrations below that of ambient air. AI-Ani et al. (1985) indicated that maximum seed
germination for most crops including wheat, sorghum, com, soybean, and sunflower occurred
at oxygen concentrations close to those of ambient air. The reasons for decreased germination
under anaerobic conditions remain unknown. It has been postulated that anoxic conditions lead
to the production of increased ethanol in cells which is toxic to normal metabolism (Thomson
and Greenway 1991). Other studies which have examined the ethanol toxicity hypothesis have
been unable to substantiate this physiological process in seeds (Van Toai et al. 1985; Martin et
a1. 1991). However, there are some notable exceptions to this postulate. Rice (Takahashi 1985),
barnyard grass (Kennedy et al. 1987), and other aquatic plants germinate under water where
oxygen is present only in limited concentrations. Rice seeds can germinate even in the complete
absence of oxygen although the seedlings are weak and abnormal. Low oxygen levels have been
shown to stimulate coleoptile growth while inhibiting root growth in this crop (Bertani et al.
1981; Alpi and Beevers 1983) and barnyard grass (Rumpho et al. 1984). Presumably, anaerobic
respiration enables these seeds to germinate in the absence of oxygen.
Although most species germinate best in oxygen concentrations of air, some species
actually germinate better at gaseous concentrations different from that in air. Cattail (Typha
latifolia) and Bermuda grass (Cynodon dactylon) germination is aided by oxygen concentrations
below that of air (Morinaga 1926). Since cattail is an aquatic plant, its response might be
expected; however, the reaction of Bermuda grass is somewhat surprising. Lettuce and onion

Table 5.3. Effects ofCO/Oz Ratios on the Germination of

Oat Seeds (From Forward 1958).

Gas Mixture
Percent CO 2 Percent 02 Percent Germination
0.0 20.9 100
16.9 17.4 93
30.0 14.7 50
35.0 13.6 31
36.S 13.2 10
38.7 12.8 1
78 Seed Germination
seeds have been reported to require less oxygen for germination than most agronomic crop seeds
(Siegel and Roson 1962). On the other hand, seeds of carrot, curly dock, sunflower, cocklebur,
and various cereals germinate better under oxygen concentrations above that of air (Albaum et
al. 1942; Barton 1941; Morinaga 1926). The influence of carbon dioxide on seed germination
is usually opposite that of oxygen. Most seeds fail to germinate if the CO2 partial pressure is
increased over the 0.03% of air; however, a decrease usually does not hinder germination.
Several seeds reportedly have a minimum CO2 requirement, notably seeds oflettuce and timothy
(Mayer and Poljakoff-Mayber 1989; Thornton 1936). It should also be emphasized that these
relationships can be attributed to seed dormancy. Some studies have shown that oxygen is able
to decrease endogenous levels of inhibitors (Simpson 1990). When this occurs, germination
responses which are not directly attributable to increased levels of respiration will be enhanced.
Temperature. Seed germination is a complex process involving many individual reactions
and phases, each of which is affected by temperature. The effect on germination can be
expressed in tenus of cardinal temperatures; that is minimum, optimum, and maximum
temperatures at which genuination will occur. The minimum temperature is sometimes difficult
to defme since germination may actually be proceeding but at such a slow rate that
determination of germination is often made before actual germination is completed. The
optimum temperature may be defined as the temperature giving the greatest percentage of
germination within the shortest time. The maximum temperature is governed by the temperature
at which denaturation of proteins essential for germination occurs. Not only does germination
have cardinal temperatures, but each stage has its own cardinal temperature; therefore, the
temperature response may change throughout the germination period because ofthe complexity
of the germination process.
The response to temperature depends on a number of factors, including the species, variety,
growing region, quality of the seed, and duration of time from harvest. As a general rule,
temperate--region seeds require lower temperatures than do tropical-region seeds, and wild
species have lower temperature requirements than do domesticated plants. High-quality seeds
are able to germinate under wider temperature ranges than low-quality seeds.
The optimum temperature for most seeds is between 15 and 30°C. The maximum
temperature for most species is between 30 and 40°C. Some species will germinate at
temperatures approaching the freezing point. Certain flower, alpine, and rock garden species are
notable for their low-temperature germination. Seeds of Russian pigweed are reportedly able to
germinate in frozen soil or even on ice (Aamodt 1935). Figure 5.3 shows two groups of
seeds-one with low optimum germination temperatures and the other with higher optimum
temperature requirements.
Germination at Alternating Temperatures. Seeds of many species require daily
fluctuating temperatures for optimum germination. Such diurnal periodicity is common and
seems to be more prevalent in species that have not had a long history of domestication. For
example, seeds of many tree and native grass species germinate best under alternating
temperature conditions (Figure 5.4). The need for fluctuating temperatures during germination
seems to be associated with dormancy, but alternating temperatures may accelerate germination
of nondormant seeds as well. There is an adaptive advantage to the requirement for alternating
temperatures. For example, vegetation insulates the soil surface against large diurnal
temperature fluctuations and, in open areas, the soil serves as an insulator with deeper portions
Seed Germination 79

80
60
40
20

z 60
o
I- 40
~ 20
::I:

!~~~
w =--
d

"~
U
0::
W 40
0..
20 C

"G
100
eo
40 60

20
a 40 a
i
20
3.5° 7° 10° 13° 20. 25· 30·
( 2-5°) 20 0 250 30 0 35 0 40 0 45°

A GERMINATION TEMPERATURE ·C B GERMINATION TEMPERATURE °C

Figure 5.3. Maximum germination ofsome arable weed seeds at constant temperatures: (A) species
in which the non-after-ripened seeds have maximum germination at high temperatures; (a)
Chenopodium rubrum, (b) Chenopodium filicifolium, (c) Datura stramonium, (d) Polygonum
persicaria, (e) Gnaphalium uliginosum; (B) species which the non-after-ripened seeds have maximum
germination at low temperatures; (a) Juncus bufonius, (b) Veronica hederifolia, (c) Polygonum
convolvulus, (d) Campanula rapunculoides, (e) Delphinium consolida, (f) Fumaria officinalis, (g)
Arenaria serpyllifolia (From Vegis 1963).

of the profile maintaining more constant temperatures. Thus, the necessity for alternating
temperatures ensures that most seeds germinate on or near the soil surface in the absence of
surrounding vegetation. Where alternating temperatures are needed, the range between high and
low (temperatures) seems to be more important than the actual temperatures (Murdoch et al.
1989). McDonald et al. (1994a) reported that this range was 10°C for germination of most cool-
season grass seeds.
The reason for the effects of alternating temperatures on germinating seeds is not known.
The most commonly cited effect has been their differential influence on the sequential steps of
80 Seed Germination

lOO IS° 20° 20~25° 15~30o 25".30 0 15".35 0 25~35° 35 0

TEMPERATURE °c
95

90
/ i'-.. V f'.. t-.....
85
,, , , t- _
t~V, --- --- , , , I'--.V 1\
"
I'
" I
I
,-'
80
\\,
, I

75
I

--- """,

...... ,
.
\

70 '.
Z
Q 65 . I
E-<
< \
I
~60
:
J
,
::E
c:: : . I

..
\

w 55 .....
.
t
U \
I
650
. -- ,..11 DAYS
..._--- 5 DAYS
I
I

.
E-< = \

~ 45 ............. .:. 3 DAYS \

u •
:
I
I
~40
. .
• I
Q.. : I

35
:
'
.: \

: .
\
\

: '
30 ,
... :
.,,
: I
25 : 1
'I
"I
20
~, "'
15

10
5

'- 0 0 0 0

Figure 5.4. Germination of Kentucky bluegrass under alternating versus constant temperature
conditions (From Harrington 1923).

germination. Evidence indicates that alternating temperatures cause a change in the structure
of components in the seed which, in the original form, prevents germination (Cohen 1958). It
has also been suggested that alternating temperatures may create a balance of the intermediate
products ofrespiration at the high-temperature cycle, which, though unfavorable for germination
at that temperature, may promote germination at a lower one (Toole et al. 1958). Perhaps the
most likely explanation is that alternating temperatures create a shift in the inhibitor-promoter
balance where the inhibitor is decreased during the low-temperature cycle and the promoter
increased during the high-temperature phase, which ultimately leads to germination.
Seed Germination 81
Stratification or Prechilling. The practice of preconditioning imbibed seeds in cool, moist
conditions to promote germination is well known (Figure 5.5). This process is called
stratification, a term adapted from the nursery industry in which propagating stocks have been
stored between layers of moist sand and sawdust prior to planting in the field the next spring.
Today, the term is used to denote any combination of moisture and low- (or sometimes high-)
temperature preconditioning~ this is a common practice in seed testing laboratories and is called
prechilling. Stratification is discussed more extensively under dormancy in Chapter 7.
Chilling Injury. Seeds of lima bean and cotton are subject to chilling injury during
imbibition if dry seeds are exposed to temperatures of 5-1S0C (Pollock and Toole 1966;
Christiansen 1968). Injury can be avoided if imbibition occurs above 20°C, followed by lower
temperatures. This type of injury also occurs in several other species. The basis of low-
temperature imbibition injury is not known, but the first measurable effect is that the seeds leach
out more organic nutrients than uninjured seeds.

45

PRECHILL - DARK
40

35

PRECHILL - LIGHT
.J

~ 25
a:;
o
z
L.L..
020
w
<.!:>
i=!
z
~ 15
a:
w
a.

10

NO PRECHILL - LIGHT

7 14 21
DURATION OF TEST (DAYS)

Figure 5.5. Average rate of germination of 41 green needlegrass accessions under four conditions
(From NifJenegger and Schneiter 1963).
82 Seed Germination
It has generally been assumed that low temperatures create a stress condition in cell walls
that causes increased cell leakage during imbibitional chilling. However, Cohn et al. (1979)
showed that imbibitional chilling injury was a consequence of stelar lesions in radicles of corn.
Willing and Leopold (1983) proposed that low-temperature injury during imbibition interfered
with membrane expansion, possibly by lowering elasticity and hindering incorporation of lipid
material into the expanding cell membrane. Other studies have shown that imbibitional chilling
injury is not a consequence of reduced energy metabolism in com (Cohn and Obendorf 1976,
1978) and soybean (Ashworth and Obendorf 1980). It has been shown, however, that seeds
experiencing imbibitional chilling injury leak more than those that do not (Leopold and
Musgrave 1979) and that there is a correlation between the degree of chilling injury and soybean
maturity class. Early-maturing cultivars are more susceptible to imbibitional chilling damage
than later-maturing cultivars (Bramlage et al.I979). The practical implications of overcoming
such injury, either by presoaking or by other means, could be important for earlier planting at
lower temperatures.
Light. While moisture, oxygen, and favorable temperature are essential for germination
of all seeds, certain species also require light. The influence oflight on germination of seeds has
long been recognized. The light response of seeds of several hundred species has been studied
to determine those whose germination was promoted by light, by darkness, or were indifferent
to light. Almost half of the species investigated responded to light.
The mechanism of light control in seed germination is similar to that controlling floral
induction, stem elongation, pigment formation in certain fruits and leaves, radicle development
of certain seedlings, and unfolding ofthe epicotyl of bean seedlings. Both light intensity (lux or
candlepower) and light quality (color or wavelength) influence germination. The nature of light
quality can be illustrated by visible light, which is composed of different wavelengths of
different colors (Figure 5.6). By the use of a prism, the visible light that appears colorless to the
naked eye can be broken down into its several colors. A rainbow is such a phenomenon, with
the water in the clouds acting as a prism and exposing the component parts of the spectrum as
separate bands. Wavelengths shorter than 400 nm (in the ultraviolet region) and those longer
than 700 nm (in the infrared or far-red region) cannot be seen by the naked eye.

-<I)
-;:I
0_
'S:
<I)

.D
c::
<1)-
<1)-
...
0lJ>'
~
0
<I.>
~
C
"l
.....
0
-0
....
<I)

I I I I
8'<t 0
0
V)
0
0
\0
0
0
r--

Ultraviolet <I.> Radio

Cosmic ~
I
'"
';;: Infrared
rays x-rays
I
0.01 0.1 10 100 1,000 tO,OOO 100,000

Figure 5,6, The spectrum of radiant energy plotted on a logarithmic wavelength scale (in
nanometers).
Seed Germination 83
Light Intensity. The influence of light intensity on different species varies greatly.
Germination of some seeds that require light has been reported to be promoted by moonlight,
representing 100 lux at the most, whereas lettuce seeds need much higher light intensities. Light
intensities of 1080-2160 lux (l00-200 foot-candles) from indirect light in the average seed
testing laboratory are probably adequate for germination of most species. Most germinators
used for light-requiring seeds are equipped with supplemental light providing several thousand
lux. In comparison, a bright, sunny day provides up to 108,000 lux (10,000 foot-candles) and
a cloudy overcast day about 16,200 lux (1500 foot-candles). Figure 5.7 shows the effect of light
intensity on seed of New Zealand browntop.
Light Quality. Figure 5.8 shows the influence of light quality on seed germination. The
greatest promotion of germination occurs in the red area (660-700 nm) with a peak at 660 run,
followed by an inhibition zone in the far-red area above 700 run. Wavelengths below 290 nm
inhibit germination, with a second inhibition zone in the blue region (440 run), whereas between
290 and 400 run a clear-cut effect is lacking.

Germ % Germ %
100

90
~.l
80

70
.
/
--
• _02

60

50
./
/
/
/ •

; :

/
/
/
0

-- -d
I
40 /
/ ./
30 /
20
'/ /
/
-'-
<............•......
.-.-~-.
--3

'·-4 ."..---~-.---.-. 3
10 'I /:.<.
O~
'.'____+-____~____~

JO 20 30 days 10 20 30 days
I _ _ Direct sunshine I _ _ Stroflg light
2. _ _ Strong diffuse light 2 _ _ Rather strong light
J. _ _ Weak light .~ _ _ Weak light
4 Very weak light

Figure 5.7. The influence of light intensity on germination of New Zealand browntop seeds (From
Gadd 1955).
84 Seed Germination
100r-------------------------------~~~~~----------------~

5
~e
n.

I
c:
o
:;:::;
:c
:c
E

O~ ___________ L_ _ _ __ L_ _ _ _L __ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ ~~~~L-------~

480 530

WAVELENGTH nm
Figure 5.8. The action of radiation of specific wavelengths (nm) in relation to the germination of
light-sensitive lettuce seed The percentages ofgermination in the spectrum (immediately following
an exposure to red light sufficient to effect 50% germination) are indicated as ordinates, the
wavelengths of the spectral light being indicated as abscissae (From Flint and McAlister 1937).

In 1952, a research group at the U.S. Department of Agriculture in Beltsville, MD, fIrst
reported the photoreversibility of lettuce seed germination. By alternately exposing imbibed
seeds to red and far-red light, seed germination could be alternately promoted and inhibited-the
ultimate effect which was independent of temperature depended on the last wavelength given.
Table 5.4 illustrates that reversibility. Since 1952, this phenomenon has been discovered in
many other species such as tobacco, birch, peppergrass, pine, elm, and shepherd's purse.

Table 5.4. Reversal of Light Effect by Repeated Alternations of Red (R) and Infrared (I)
Irradiations on Grand Rapids Lettuce Seed (From Toole et al. 1953).

Seeds germinating when

exposures were made at
27' C 7' C
Irradiation Percent Percent
None (dark controls) ........................................ 14 11
R ......................................................... 70 72
R +I ............ ....... .......................... ........ 6 13
R +I+ R ................................................ 74 74
R+I+R+I............................................. 6 8
R + I + R + I + R ........................................ 76 75
R+I+R+I+R+I.................................... 7 11
R + I + R + I + R + I + R ................................ 81 77
R+I+R+I+R+I+R+I............................ 7 12
Seed Germination 85
Photoreversible germination in seeds appears to be due to a single coupled photoreversible
chemical reaction. The chemical reaction is controlled by the wavelength of light absorbed in
plant cells. The photoreversible substance was first isolated from corn seedlings grown in the
dark (Butler et al. 1964). It is a protein pigment known as phytochrome. In its most
concentrated form, it appears blue, is faded by red light, and reintensified by far-red light. The
far-red absorbing form (induced by exposure to red light) is believed to be the biologically active
form that functions by inducing the synthesis of enzymes essential for germination.
Day Length. Seeds of several species exhibit a photoperiodically controlled germination
response. The mechanism seems to be controlled by the activation of phytochrome similar to that
associated with floral induction. As was the case for floral induction, there are long-day species,
e.g., Begonia evansiana (Nagao et a1. 1959) and Betula (Black and Wareing 1955) and short-
day species, e.g., Amaranthus retroflexus (Kadman-Zahavi 1960) and A triplex dimorphostegia
(Koller 1971). Other categories are listed (Koller 1972) according to their response to
photoperiod, such as seeds inhibited by prolonged irradiation, e.g., Raphanus (Isikawa 1962)
which germinate in the total absence of light but are inhibited by continuous light. In some cases,
there may be an interaction of a high-energy light reaction with phytochrome, which determines
the photoperiodic effect (Ellis et aJ. 1986). It is further pointed out that the photoperiodic
response, especially that of long-day species, is temperature-dependent (Koller 1972). For
example, long-day species are especially sensitive to day length and are almost absolute in their
long-day requirement. The need for long days decreases progressively as temperatures increase
and, at high temperatures, germination can proceed in the absence of light.
Factors that Influence Light Sensitivity. Light sensitivity during seed germination
depends on the species and variety, as well as on environmental factors before and during
germination.
Age o/Seed Light influence is strongest immediately after harvest and diminishes with age
of the seed and eventually disappears (Toole et a1. 1957; Fujii and Yokohama 1965; Hagon
1976). This may be one reason why differing accounts of light requirements for seeds from the
same species exist in the literature.
Period 0/ Imbibition. Light sensitivity of Grand Rapids lettuce seeds was shown to
decrease during the imbibition period to 10 hr, remain constant for another 10 hr, and again
increase sharply (Borthwick et a1. 1954). A similar response has been observed for peppergrass
and tobacco seeds (Table 5.5). Tobacco seeds that ordinarily require high light intensities are
able to germinate under lower intensities after four days of imbibition (Kincaid 1935). The
sensitivity ofloblolly pine (Pinus taeda) seeds to light has also been shown to increase during
storage (Toole et a1. 1958b). Other studies have shown that light promoted the germination of
Avena jatua and Grand Rapids lettuce seeds when seed moisture content was high but was
inhibitory if seed moisture content was low (Hsiao and Simpson 1971). Holm and Miller (1972)
demonstrated that restriction of water with mannitol from the seed resulted in a reduction of
germination which could be overcome by a red light treatment. One general conclusion is that
less light is needed to stimulate seed germination with increasing time of imbibition.
Imbibition Temperature. Lepidium seeds, which had imbibed water at 20°C, germinated
only 31 % in response to light; however, when the seeds imbibed at 35°C, germination increased
to 98% (Toole et at. 1955b). In addition to the fact that increased temperature during imbibition
hastens metabolic events associated with germination, it also enhances water uptake, resulting
86 Seed Germination
Table 5.5. Germination of Seeds of Tobacco (Nicotiana) and Peppergrass (Lepidium) Seeds
Exposed to Red Light for Various Periods After 4 and 23 Hours Imbibition in Dark
(From Toole et al. 1953).
Germination percentage l after indicated hours of imbibition
4-hour imbibition 23-hour imbibition
Period of exposure to
red light (minutes) Nicotiana Lepidium Nicotiana Lepidium
0 4 0 6 11
1f4 6 14 22 62
1 6 46 37 65
4 14 61 58 64
16 29 66 88 76
64 69 64 95 76
'Nicotiana germinated at 25"C. and Lepidiurn at 20' to 3D' C.

in more rapid hydration and increased sensitivity to light.

Stratijlcation. Seeds of loblolly pine (P. taeda) and Eastern white pine (P. strobus)
became more responsive to light following stratification, and the increased sensitivity was
roughly proportional to the duration of stratification (Toole et al. 1962).
Germination Temperature. Many data exist showing the interaction of light and
temperature on germination; however, it is not possible to reduce these to a unified picture. It
is clear that the response to each can sometimes be increased, decreased, or changed
qualitatively by the other, while in other cases it cannot. Germination of peppergrass (Lepidium)
(Toole et al. 1955a) and Virginia pine (P. virginiana) (Toole et al. 1961) seeds is increased
when red irradiation is preceded by a low temperature and followed by a high one. Low-
temperature treatment could substitute for the red light requirement for germination of dock
(Rumex) seeds; and weak red light, while insufficient to promote germination alone, always
promoted it in combination with a high temperature treatment (lsikawa and Fujii 1961).
In some instances, the dependence of seed germination on light is reduced when seeds
experience alternating temperatures. For example, Agrostis tenuis seeds have their light
requirement either completely or partially removed when exposed to alternating temperatures
(Toole and Koch 1977). Light can influence germination of downy brome the least at optimum
temperatures and most at less suitable germination temperatures (Thill et aI.1984). In most
cases, total germination is enhanced when seeds are exposed to both alternating temperatures
and light (McDonald et al. 1994a).
The temperature treatment that increases the sensitivity to red light reduces the sensitivity
to far-red light (Toole et al. 1956). Virginia pine (P. virginiana) seeds germinated well when
exposed to light at 25°C but germinated weakly at either 20 or 30°C (Toole et aJ. 1961).
Peppergrass (Lepidium) and tobacco (Nicotiana) seeds that failed to germinate at 20°C became
responsive to light when temperatures were alternated between 15 and 25°C (Toole et al. 1957).
These few examples illustrate the complex nature of the light-temperature interaction on
germination. However, the exact mechanism of the light and temperature interaction remains
unknown.
Phytochrome. The action spectrum of light-sensitive lettuce seed was discovered in the
mid-1930s (Flint 1934; Flint and McAlister 1935, 1937) when it was shown that the greatest
Seed Germination 87
promotive response from a given irradiance occurs in the red region (600-700 run) and the far-
red (720-760 om) portion of the spectrum causes the greatest inhibition (Figure 5.8).
The light and dark response of phytochrome is illustrated by the scheme in Figure 5.9. Red
light (660 run) exposure converts phytochrome to the physiologically active, far-red absorbing
form, and germination proceeds. Exposure to far-red light (730 run) reconverts phytochrome to
the physiologically inactive red-absorbing form, and germination is blocked. Only a brief flash
of either red or far-red light is necessary to convert phytochrome to the opposite form. In seeds
of princess tree (Paulownia tomentosa). a prolonged dark period also permits conversion ofPFR
to PR (Toole et a1. 1958a). This conversion can also be promoted by prolonged or repeated brief
exposures to high temperatures or far-red light.
We now know that all of the PR is never completely converted to PFR by red irradiation
because of the overlap of the action spectra of PFR and PR (Butler et al. 1964). In far-red
irradiation, phytochrome is driven strongly toward PFR with about 2% remaining in the active
PR form (Figure 5.l0).
Light-requiring seeds need a certain number of PFR molecules to genninate and this may
vary among species. In some cases, the PFR requirement for germination may be so low that the
seeds germinate irrespective ofthe light treatment. Such seeds are referred to as light-insensitive.
Despite more than 50 years of intense research into phytochrome action, we still do not
know precisely how it functions. Recent advances in molecular and genetic techniques, however,
are beginning to increase our understanding of phytochrome action. It is now known that
phytochrome is synthesized in the seed as PR which is the biologically inactive form and most
seeds therefore require a light stimulus to convert the PR to PFR, the biologically active fonn that
leads to germination. The location of phytochrome in seeds has also been a matter of debate. It
is generally assumed that it is associated with cell membranes. But is there a specific part ofthe
seed that contains the phytochrome? It has been reported that light sensitivity is confmed to only
one portion ofthe seed in certain species. For example, in Phacelia seeds, light sensitivity is
localized at the chalazal and micropylar ends. In lettuce seeds, only the micropylar end is light
sensitive. This, of course, is the thinnest portion ofthe seed coats and may simply allow more
light to penetrate to the embryo. Evidence exists (Bewley and Black 1978) that the phytochrome
pigment is actually located in the embryonic axis. In addition, there now appears to be at least
five differing isofonns of phytochrome based on molecular studies of phytochrome mutants
(Viestra 1993). What role these have, if any, in mediating seed germination must still be
detennined'

Dormancy .... - - PR

Inactive Form
Red, 660 nm
..:_;::============~~ PFR
Far-Red, 730 nm
.. --.Germination

Active Form
+
1_ _ _ _ _ _ Dark _ _ _ _ _ _I
High Temperature

Figure 5.9. Photoreversible phytochrome response.

88 Seed Germination

l5~----------------------------------------~

& 1.0 RED IRRAQ.

"./
z
<
m
a:
o
en
m " \
< 0.5 \
\
\
\
\
\
\
,
'-
300 400 500 600 700 aoo
WAVELENGTH-nm

Figure 5.10. Absorption spectra ofphytochrome measured spectra. The solution was diluted tenfold
for measurements below 300 nm (From Butler et al. 1964).

While the physiological role of PFR is still unclear, the following four mechanisms have
been proposed:

1. PFR influences gibberellin synthesis. The observation that gibberellins can substitute for
light in breaking the dormancy of many light-sensitive seeds (Chen 1970) has led to the
suggestion that PPR increases gibberellin synthesis. It has been shown that inhibitors of
GA biosynthesis delay the light-induced germination of a number of species, suggesting
that gibberellins play some part in the germination of light-sensitive lettuce seeds (Jones
and Stoddart 1977). An alternative hypothesis is that PFR stimulates the release of
endogenous gibberel1ins from a bound form.
2. PPR selectively activates specific genes. Following activation of PFR, a number of
hydrolytic enzymes are known to increase (Chen and Varner 1973). It has been suggested
that PFR selectively exposes portions of the genome, which promotes the rapid synthesis
of enzymes essential for germination. For example, cloning of phytochrome genes from
moss plants has shown these to have kinase enzymatic activity (Algarra et al. 1993). The
same may be true for germinating seeds.
3. PPR alters membrane permeability (Bewley and Black 1978). Phytochrome is localized in
cell membranes. It has been suggested, therefore, that it may have a specific influence on
cell membrane permeability. If, for example, gibberellins bound to sugars are hydrolyzed
and released during imbibition, they may not exhibit a physiological function until they
are isolated from the sequestered form. This release may occur following a photoinductive
reaction, causing PFR to alter membrane permeability, resulting in the release and transfer
of gibberellins to their site of action (such as the aleurone layer).
Seed Germination 89
4. Kinase activity has been reported to increase (Algarra et al. 1993), which may cause a
change in the balance between the pentose phosphate shunt and the glycolytic pathway,
thus altering the capacity to germinate (Taylorson and Hendricks 1976).

The phytochrome-mediated control of germination has immense ecological significance.

This system allows buried seeds to remain dormant (although fully imbibed) until they are
exposed to light. It has also been demonstrated that light passing through a leafed canopy
possesses a high percentage of far-red radiation. This retards the germination of seeds under
heavy tree canopies where the light intensity is not adequate for seedling establishment. Thus,
phytochrome has evolved as a survival mechanism that promotes seed germination only under
conditions in which seedling establishment is most likely to succeed.

PATTERN OF SEED GERMINATION

Most seeds undergo a specific sequence of events during germination. The major events
are imbibition, enzyme activation, initiation of embryo growth, rupture of the seed coat, and
emergence of the seedling.

Imbibition

The early stages of imbibition or water uptake into a dry seed represent a crucial period
for seed germination. Seeds are sensitive to rapid imbibition, chilling, and anoxia; common
events that occur with the increasing emphasis on early planting and conservation tillage.
Imbibition is an essential process initiating seed germination. It is the first key event that moves
the seed from a dry, quiescent, dormant organism to the resumption of embryo growth.
Consequently, an orderly transition of increased hydration, enzyme activation, storage product
breakdown, and resumption of embryo growth must occur. Imbibition is not merely an
uncontrolled physical event: it is now recognized that chemical conformation events, seed coat
effects, and seed quality factors govern the directed flow of water into the seed (Leopold and
Vertucci 1989). Thus, any consideration of seed germination physiology and its resultant impact
on stand establishment should initially focus on water uptake. The extent to which water
imbibition occurs is dependent on three factors: (1) composition of the seed, (2) seed coat
permeability, and (3) water availability.
Composition of the Seed. Seeds typically possess extremely low water potentials
attributed to their osmotic and matric characteristics. These potentials may be as low as - 400
MPa in the dry seed of rape, wheat, and com (Shakeywich 1973). The low water potentials are
a consequence of the relationship of water with components of the seed. Water in a soybean
seed, for example, basically exists in three forms (Figure 5.11) dependent on its hydrational
status (Leopold and Vertucci 1986). Below 8% moisture (region 1), water is chemisorbed to
macromolecules through ionic bonding, has limited mobility, and acts as a ligand rather than a
solvent. Between 9% and 24% moisture (region 2), water is weakly bound to macromolecules
and begins to have solvent properties, and diffusion gradually becomes evident as water takes
on the properties of a bulk solution. Above 24% moisture (region 3), water is bound with
negligible energy, and its properties are similar to bulk water. The macromolecular surface of
90 Seed Germination

0.30 ,-------------------------r--r----,

0.25

I-
z
UJ 0.20
I-
z~
o
0"0
~

0:':-
UJ Cl
0.15
~ q"
3:J:
UJ..2!
:;,
en 0.10
en
i=

0.05 •
0.00 " - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - '
o 10 20 30 40 50 60 70 80 90 100
RELATIVE HUMIDITY (%)

Figure 5.11. Water sorption isotherms ofground soybean pellets at 5° and 2(f'C. showing the reverse
sigmoidal curve typically foundfor seeds. Relative humidity was controlled by saturated salt solutions
(From Leopold and Vertucci 1986).

the seed is considered fully wetted at 35% moisture (Leopold and Vertucci 1986).
The QIO value of imbibition is 1.5-1.8, indicating that imbibition is a physical process not
dependent on metabolic energy and related to the properties of the colloids present in seed
tissues. This is supported by the observation that imbibition occurs equaIJy in both dead and live
seeds (Figure 5.12). The principal component of seeds that is responsible for the imbibition of
water is protein. Proteins are zwitterions that exhibit both negative and positive charges that
attract the highly charged polar water molecules. The difference in imbibition of protein-
containing seeds compared with those containing starch is demonstrated by soybeans and com.
Soybean seeds typically imbibe two to five times their dry weight in water, while com seeds
imbibe 1.5 to 2.0 times their dry weight. Similarly, the observation that the embryo of cereal
seeds can absorb about twice as much water as the endosperm can be explained by the greater
proportion of protein present in the embryonic tissues (Chung and Pfost 1967). Other studies
on five legume species have shown that the embryonic axis characteristically had higher water-
binding properties than the cotyledons (Vertucci and Leopold 1987). McDonald et al. (1988b)
showed that the soybean embryonic axis hydrated more rapidly and completely than any other
seed part due to its higher protein composition.
Seed Germination 91

200

:c00
ii
J

-
0

.
'0
II
I~O
0
~

.c:
u

x - x 2° 6,-6, 26'
0--0 36° 0 - 0 ISo

Figure 5.12. Imbibition of heat-killed peas at different temperatures (From Mayer and Poljakoff-
Mayber 1989).

Other chemical constituents of seeds also contribute to imbibition. The mucilages of

various seeds increase imbibition as do the cellulose and pectins located in the cell walls. In
contrast, starch molecules have little impact on imbibition-even when large quantities of starch
are present, as in seeds of many grasses. Starch, because of its uncharged structure, only attracts
water at very acid pH or after high-temperature treatment--conditions that do not occur in
nature.
Seed Coat Permeability. Entry of water into seeds is greatly influenced by the nature of
the seed coat (or pericarp). Water permeability is usually greatest at the micropylar area where
the seed coat is ordinarily quite thin. The hilum of many seeds also permits easy water entry.
The same appears to be true in many grass seeds which possess a pericarp that completely
surrounds the seed except at the pedicel end. This open, porous structure results in a more rapid
hydration of the embryo that progressively moves from the radicle to the coleoptile end
(McDonald et al. 1994b). A slower, more progressive wetting front simultaneously moves
through the seed coat and into the endosperm (F igure 5.13).
Seeds of certain species have special tissues around these natural openings that prevent
water entry and contribute to hard seed coat (impermeable to water) dormancy (see Chapter 7).
This hard-seededness has been attributed to small elongated pores and a high density of waxy
material embedded in the testa epidermis (Calero et al. 1981; Tully et al. 1981; Yaklich et al.
1986; Mugnisjah et af. 1987). In other instances, hardseededness has been attributed to the
92 Seed Germination
presence of lipids, tannins, and pectic substances in the seed coat (Denny 1917). The incidence
ofhard-seededness is both genetically and environmentally controlled and is greatest when seed
maturation occurs under high temperature, high humidity (Dassou and Kueneman 1984; Potts
et al. 1978), and dry (Hill et al. 1986) conditions. McDonald et al. (l988a) showed that the
soybean seed coat delayed water uptake during the first eight hours of soaking. It also directed
water movement both tangentially and radially into the embryonic axis. The radial movement
of water was attributed to the presence of a radicle pocket that possessed a high incidence of
hourglass cells (Figure 5.14). These cells may increase the water storage capacity around the
radicle tip, ensuring a ready source of water for turgor pressure essential for germination.
The seed coat acts as a semipermeab Ie membrane, permitting the entry of water and certain
solutes while restricting others. For example, the leakage of the inositol pinitol from imbibing
soybean seeds is substantially greater than for sugars such as sucrose/raffinose, stachyose,
fructose, and glucose (Nordin 1984). Other studies have shown soybean seed leachate to contain
high concentrations of K+ and protein (Seneratna and McKersie 1983) and enzymes such as
malate dehydrogenase (Duke et al. 1983). This leakage is more pronounced if the seeds are
deteriorated (Duke et al. 1986; Seneratna and McKersie 1983; Schoettle and Leopold 1984).
This differential permeability may be the result of the ionization of the acidic and basic groups
of the membrane lipids (Weatherby 1943). Such membranes repel ions of similar charge while
attracting those of opposite charge. Thus, un-ionized molecules of liquids or bases do not
permeate as readily as ionized molecules. The cuticle of wheat seeds has been observed to carry
an electric charge that influences the permeability to various solutes (Brown 1932).
Water Availability. The environmental forces that determine the rate of water imbibition
by seeds are complex. The ability to imbibe water is dependent on cell water potential and is a
resu It of three forces:

1. Cell wall matric forces. Cell walls and intracellular inclusions such as mitochondria,
ribosomes, and spherosomes are characterized by the presence of membranes. These
membranes possess charges that attract water molecules and contribute to the total cell
water potential.
2. Cell osmotic concentration. The greater the concentration of soluble compounds, the
greater the attraction for water.
3. Cell turgor pressure. As water enters a cell, it exerts a swelling force on the cell wall
called turgor pressure. Unlike the cell wall matric forces and osmotic concentration that
attract water molecules into a cell, turgor pressure, which is a result of the restraining
force of the cell wall, tends to retard water absorption.

The soils in which seeds are planted also exhibit their own water potentials. The physical
properties of soils determine the retention and conductivity of water. For example, it is well
known that soils heavy in clays are able to absorb water more vigorously and retain it longer
than those possessing high quantities of sand. In effect, the seed water potential must compete
with the soil water potential for imbibition to occur. Initially, the difference between seed and
soil water potential is quite large. However, as imbibition continues, this difference is reduced
in the immediate vicinity of the seed. If it were not for the conductive ability of soils, imbibition
Seed Germination 93

Figure 5. 13. Movement of water through the seed coat and embryo of a corn seed The top four
photographs illustrate the movement of iodine into the corn seed endosperm after 0, 6, 24, and 48
hours. The bottom six photographs illustrate staining of the corn seed embryo with nitroblue
tetrazolium chloride after 0, 3, 6, 15, 24, and 48 hours of soaking in water (From McDonald et al.
1994).
94 Seed Germination

Figure 5.14. Scanning electron micrograph of (A) Cotyledon (C), radicle (R), radicle pocket (RP),
hourglass cells (H), and palisade cells (P); (B) Enlarged view of radicle (R), radicle pocket (RP),
hourglass cells (H), and palisade cells (P); (C) The seed coat opposite the hilum (note the absence of
hourglass cells showing the palisade cells (P), parenchyma (PA), and cotyledons (C). Bars represent
1.0 mm (From McDonald et at. J988b).
Seed Germination 95

would be quickly halted. However, most soils exhibit a high degree of hydraulic conductivity
that replenishes the available water surrounding the seed as it continues the process of
imbibition. This is important since seeds are sessile and a continuous flow of water is essential
for maximum imbibition. Seeds rarely attract water from beyond 10 mm in most soils.
Associated with seed and soil water potential is the degree of seed-soil contact. The
greater the intimate contact of the seed with the soil, the greater the amount of water imbibed.
At least three mechanisms have evolved to improve seed-soil contact. Some seeds possess
mucilage which is extruded from the epidermal cells of the seed as it imbibes water. The
mucilage serves to increase the contact ofthe seed with the soil by increasing the number of
pathways through which water may be absorbed by the seed. Another mechanism to enhance
seed-soil contact is to increase the amount of seed contact with a specific volume of soil. This
can be accomplished by altering seed coat configuration. Seeds possessing textured seed coats
are more likely to have a greater seed-soil contact than smooth-coated seeds, and thus they will
imbibe water more rapidly. A fmal factor is seed size. Small seeds possess a greater surface area
to volume ratio than large seeds. This greater surface area permits them to have access to a
greater amount of water than large seeds, which means that they will hydrate more rapidly.

Enzyme Activation

Dry seeds are characterized by a remarkably low rate of metabolism that is undoubtedly
attributable to their low moisture content (which may be as low as 5 to 10% in unimbibed
seeds), thus are said to be in a state of quiescence. As soon as the seed becomes imbibed,
however, marked changes in its metabolism occur. A triphasic pattern of water uptake has been
demonstrated during the germination of most seeds (Figure 5.15). Enzyme activation begins
during Phases I and II of imbibition. During Phase II, the seed undergoes many processes
essential for germination. Increased respiration and leakage of nutrients from the imbibed seed
leads to loss of dry weight. Finally, in Phase III, root elongation is observed. The root becomes
functional during this phase and is responsible for the increased water uptake noted in Phase III.
The process of enzyme activation during Phase II of water imbibition serves to break
down stored tissue, aid in the transfer of nutrients from storage areas in the cotyledons or
endosperm to the growing points, and trigger chemical reactions that use breakdown products
for the synthesis of new materials.
In monocots, gibberellins are released from the scutellum and move through the
endosperm of the aleurone layer. There they trigger the synthesis of hydrolytic enzymes
including a-amylase, ribonuclease, endo-~-glucanase, and phosphatase. The release of these
hydrolytic enzymes results in a degradation of the endosperm and endosperm cell walls. This
process begins near the scutellum, with subsequent hydrolysis occurring to the sides and
upwards through the endosperm. The mechanism for this degradation is not fully understood,
but scientists have offered two hypotheses. One suggestion is that since enzymes are released
from the scutellum, they arrive at the aleurone layer closest to the scutellum before they reach
those aleurone cells farther removed. This enables the aleurone cells closest to the scutellum to
initiate hydrolytic enzyme synthesis first. The other postulate is that a symmetrical release of
enzymes from the aleurone occurs. However, the scutellum, which is also rich in other enzymes,
simply provides an additional source of hydrolytic enzymes to digest the endosperm tissue.
96 Seed Germination

I'phase I" Phase II

,-..

~
.~
..c::
'"
<l)

<.!::
.S
<l)

'"'"
t
Start of visible
....
<1)

u germination
c::
c
-
....
<l)

'"
~L-

'0
________ ~ __________________

Time ----+-
<l)
.:.:
~
0..
~

Figure 5.15. Triphasic pattern of water uptake by germinating seeds (From Bewley and Black 1978).

Regardless of the mechanism, the germinating seed now possesses many products that are a
consequence of the activity of hydrolytic enzymes released from the aleurone and scutellum.
Trigger Chemical Reactions Used in the Synthesis of New Materials. It has already
been demonstrated that the early events that occur during the enzyme activation phase include
the synthesis of storage product enzymes such as a-amylase, ribonuclease, and phosphatase.
These events are mediated by the hormone gibberellic acid. However, during this lag phase many
other hydrolytic enzymes are formed. This synthesis is preceded by an increase in the
endoplasmic reticulum, ribosomes, and ribosomal RNAs--essential components of the enzyme-
synthesizing machinery. Such enzymes as ATPase, phytase, protease, lipase, and peroxidase all
increase during enzyme activation. In many cases, these enzymes further break down storage
compounds or take the hydrolyzed products and resynthesize them into molecules essential for
new growth. In other instances, the enzymes are indirectly involved in the synthesis of new
materials by ensuring that the energy molecules essential for these reactions are present in the
cytoplasm in adequate amounts. Clearly, the enzyme activation phase (which appears to be a
metabolically inactive period based on water uptake) is one of the most essential and dynamic
phases in preparation of the seed for embryonic axis elongation.

Breakdown of Storage Tissues

Generally, enzymes that break down carbohydrates, lipids, proteins, and phosphorous-
containing compounds are the first to be activated during Phase II of water uptake by seeds. The
controlling mechanism for directing this storage tissue degradation has not yet been precisely
elucidated.
Since the embryonic axis requires energy for growth, storage compounds must be
hydrolyzed to soluble forms, translocated from the endosperm to the embryo, and transformed
Seed Germination 97
to energy molecules that can be immediately utilized by the embryonic axis. The endosperm
initially becomes rich in soluble products such as glucose and maltose. These are then absorbed
by the scutellum. In the scutellum, glucose and maltose are transformed by a series of in situ
enzyme reactions to form sucrose. (Sucrose itself is not hydrolyzed within the scutellum because
the essential enzymes are not present.) The sucrose molecule is then transported to the adjacent
embryonic axis as the principal energy molecule for growth. This is illustrated in Figure 5.16.
In dicots, the hormonal regulation of storage product degradation is not as clear as in monocots.
This may be due to the absence of an aleurone-like tissue that synthesizes hydrolytic enzymes.
Additionally, the role of hormones in dicot seed germination has been debated. In some

ALEURONE LAYER

ENDOSPERM
stareb

lCD
1@
maItos.

MAIZE
Figure 5.16. Breadown of storage compounds and mobilization of their products during the
germination of corn seeds (From McDonald 1994).
98 Seed Germination

SOYBEAN
Figure 5.17. Breakdown of storage compounds and mobilization of their products during the
germination of soybean seeds (From McDonald 1994).

instances, gibberellins are known to trigger hydrolytic enzyme synthesis, but the degree of
activation is never as great as that noted in cereals. Some investigators believe that dicot seed
germination is mediated by the growing embryonic axis. As the axis continues to grow, it
incorporates breakdown products into the synthesis of new compounds. This reduces the
concentration of compounds in the cotyledons, which in turn stimulates the hydrolysis of other
storage reserves for use by the embryonic axis. Should this stimulation prove to be too great,
and hydrolyzed storage products begin to accumulate, a feedback mechanism may be operative
that retards further storage reserve hydrolysis. This is illustrated in Figure 5.17.
The mobilization and transfer of nutrients in most dicot seeds is through the conductive
tissue of the cotyledons to the growing embryonic axis. In bean seeds, for example, the
hydrolysis of protein bodies occurs at the center of the cotyledons and then moves toward the
outside of the cotyledons. Like grass seeds, storage compounds must initially be hydrolyzed to
a soluble form before they can be translocated.
Carbohydrate Metabolism. Amylopectin and amylose are hydrolyzed by u- and ~
amylase enzymes. These enzymes split either starch structure yielding the disaccharide maltose,
which is then split into two monosaccharide glucose units. Some glucose units are converted into
the highly mobile disaccharide sucrose for translocation to other sites, after which it is
reconverted to glucose or used directly in synthesis.
Seed Germination 99

Breakdown A-P-P A+B

products

Expenditure
of energy

Carbohydra te,
Fat, or Protein A-P-P-P
C
Adenosine triphosphate (AIP)

Figure 5. lB. In the breakdown of storage substances by the cells, energy is stored in the form of
special "energy-rich" phosphate bonds in the molecule of adenosine tripho~phate (ATP). The ATP
is then available for energy to transport or to drive reactions as A + B ~ C during which ATP loses
some energy and phosphate and is changed to ADP.

Glucose may be further broken down by respiration. The first step is known as glycolysis,
which yields two pyruvic acid molecules. These, then, are completely broken down into CO2
and water by a series of reactions known as the tricarboxylic acid (Krebs) cycle. The reactions
of glycolysis occur in the cytoplasm, while those ofthe Krebs cycle occur in the mitochondria.
Both processes yield energy as ATP (Figure 5.18).
As early as 1890, Haberlandt, in his study of rye germination, suggested that events
occurring in the growing points of the embryo initiate starch hydrolysis by the amylases. The
responsible substance, however, was not identifit;xi. It is now believed that gibberellins are
produced in the growing root-shoot axis and scutellum and migrate to the aleurone layers
surrounding the endosperm, where they stimulate synthesis of amylase and other hydrolytic
enzymes (Jones and Armstrong 1971; Paleg 1960; Jacobsen and Varner 1967; Briggs 1963;
Radley 1967). These enzymes hydrolyze the starch and other storage products of the endosperm
into solutes that are transported through the scutellum to the growing points, where they nourish
the growing seedling. Limited work with noncereal species indicates that similar systems occur
in other kinds of seeds.
Lipid Metabolism. For oil-bearing seeds, the first step in utilization of reserve storage
materials also involves a hydrolytic reaction using the enzyme lipase to cleave the ester bonds
and yield free fatty acids and glycerol. The free fatty acids are further degraded by one of two
processes-either u- or p-oxidation (Figure 5.19).
a-oxidation. This form of free-fatty acid breakdown plays a minor role in germinating
seeds, but has been observed in peanuts and sunflower. It involves the successive loss of one
carbon atom and CO2 bv the aid of fatty-acid peroxidase and aldehyde dehydrogenase enzymes.
p-oxidation. The major means of fatty acid breakdown during germination is by p-
oxidation, with the aid of p-oxidase, yielding acetyl coenzyme A and energy in the form of ATP.
Acetyl coenzyme A may then enter the Krebs cycle for complete oxidation to CO2, H20, and
energy (ATP), or it may enter the glyoxylate cycle for conversion by a complicated series of
reactions to yield sucrose. Sucrose is then available for translocation to growing sites and for
use in biosynthesis.
100 Seed Germination

A. Fat hydrolysis
CIS H31 COOH
C1s H31 COOH +
C1s H31 COOH

Palmitin Three molecules One molecule

of palmitic acid of glycerol
(fatty acid)

NAD+

Aldehyde
Dehydrogenase
NADH

RCHzCHO

C. {3-oxidation
( l
ATP • P.G.A - Triose-Hexose-Sucrose
t
DPN DPNH P.E.P.
t
Fatty Acid --~~--(,"-.. Oxalacetate
Acetyl Co A
1
Malate

Figure 5.19. Pathways offat degradation: (A) fat hydrolysis, (B) a-oxidation, (C) fJ-oxidation.

Protein Metabolism. Relatively little is known about the exact nature of reserve protein
breakdown during seed germination. However, proteinases, the proteolytic enzymes
(endopeptidases, carboxypeptidases, aminopeptidases) (Bond and Bowles 1983) are involved
in cleaving the peptide bonds ofthe protein and releasing the amino acids. Proteinases have been
observed in many seeds and increase rapidly during germination (Ryan 1973). Proteolytic
enzymes differ in their specificity in attacking certain peptide linkages. The specificity seems
to be determined by adjacent end groups on the protein molecule, the type of amino acid side
Seed Germination 101
chains, and the relationship between the size of the molecule and the number of free end groups.
Hydrolysis of protein bodies evidently occurs differently in cereals and legumes. Tn soybeans,
protein bodies are hydrolyzed by internal digestion (Wilson 1987) while in com the hydrolysis
begins at the surface of the protein bodies (Torrent et al. 1989).
As proteins are broken down during seed germination, there is a concomitant rise in amino
acids and amides in the cotyledon, followed by protein synthesis in the growing parts of the
embryo. Initial evidence of such changes has been observed in germinating lettuce seed as a
decrease in the ratio of protein to soluble nitrogen (Klein 1955).
After free amino acids are released from their protein complexes, they may be further
broken down by any ofthree processes: (l) deamination to give ammonia and a carbon skeleton
that subsequently enters various metabolic processes, (2) transamination enzymes to yield
ketoacids, which enter the Krebs cycle for further breakdown to CO 2, HP, and energy (ATP),
or (3) direct utilization for synthesis of new proteins in other parts of the germinating seed.
Regardless of the pathway followed, the breakdown products are eventually available for use
by the developing seed.
New proteins are synthesized on the surface of ribosomes in the cell cytoplasm. Here,
messages are received from the DNA of the nucleus by means of messenger RNA. These
messages determine the kind and sequence of amino acids to be combined into new proteins.
Phosphorus-Containing Compounds. About 80% of the phosphorus in seeds is stored
as calcium, magnesium, or manganese salts of inositol hexaphosphate, or phytin. The other 20%
is in organic compounds such as nucleotides, nucleic acids, phospholipids, phosphorylated
sugars, phosphoproteins, and a trace of inorganic phosphate. During seed germination, phytin
is broken down, releasing inorganic phosphorus for synthesis of other phosphorus-containing
compounds. Its breakdown is catalyzed by phytase, a phosphatase enzyme.
Nucleotides, such as ADP and ATP, are complex phosphorus-sugar compounds that have
the ability to store and release chemical energy in their phosphate bonds. During germination,
these are converted back and forth as energy is released and used again. Each time that energy
is released, inorganic phosphorus is also released.
Although the phospholipids are not a major food reserve in the seed, they are present, and
are broken down during germination. Their breakdown scheme is similar to that for other lipids,
and the catalyzing enzyme is a phospholipase.

Initiation of Embryo Growth

Studies have been conducted on the developmental changes that occur in seeds as they
initiate embryo growth. Because monocot and dicot seeds are different in their morphological
structure, it is not surprising that the alterations these seeds exhibit are unique. Monocot seeds
generally display a germination pattern similar to that exhibited by com. During the first 120
hours of germination, there is a marked decrease in the dry weight of the endosperm with a
concomitant increase in the dry weight ofthe embryonic axis (Figure 5.20). These changes are,
in part, a reflection of decreases in total nitrogen and insoluble protein that occur in the
endosperm, and the subsequent translocation of these compounds to the emerging axis. Similar
changes would be anticipated following endosperm starch hydrolyzation to maltose and then to
glucose, which is then enzymatically altered to sucrose and translocated to the axis. Figure 5.20
illustrates many of the metabolic changes that occur during the early stages of germination.
102 Seed Germination

5 TGrAL NITROGEN INSOLUBLE

250 4 DRY WEIGJ.lT PROTEIN
5 (a s N)
-, whol~ st!t!dtmg -~hot~ SC"t!dIi1l9
2000.....:---~ 4 - __ -'oj

~'50
~E
~
~Ildospum
15

100 10

aXI5 ..

SO scut~ltum A

8 TOTAL AMINO ACIDS 9 FAT

12

t!ndosp~rm

O~~~ __~~~___ ~"L-~~~~~--L- I-~~/~"~'~Lj

10 NUCLEIC ACIDS 11 SOLUBLE NUCLEOTIOES 12 SOL.CARBOHYDRATES
30

10 10
....
~ 08 08

--E 06
0..
06

04

o 24 48
HOURS

Figure 5.20. Changes in content of various components in different parts of Zea mays during
germination and growth (From Ingle et al. 1964).

Similar transitions are also apparent in dicot seeds. In Vigna sesquipeda/is (cowpea), the
major storage tissues, the cotyledons, undergo a decrease in dry weight as the hypocotyl and
subsequently the epicotyl, show increases (Figure 5.21). Like corn, soluble carbohydrates,
soluble nitrogen, and nucleic acid phosphorous levels decrease in the cotyledons and are found
in the emerging embryonic organs of the hypocotyl, roots, epicotyl, and plumule. These events
show that the storage tissues function primarily as reservoirs from which the emerging axis can
draw nutrients for rapid germination and emergence.

Protrusion of the Radicle

The actual protrusion of the radicle, which signals that the germination process is
complete, can be accomplished through either cell elongation or cell division. In general, cell
elongation precedes cell division. This is true in lettuce, corn, barley, beans, and peas (Berlyn
Seed Germination 103

Soluble Carbohydrates

Days o 5 6
Days
Soluble Nitrogen Nucleic Acid Phosphorous
40 70
04 60
30 50
0-3
y y
\-0 20

10
10

o 6
Days Days
Figure 5.21. Changes in content of dry weight, soluble carbohydrates, soluble nitrogen, and nucleic
acid phosphorus of Vigna sesquipedalis during germination (From Oola et al. /953).

1972; Brown 1932; Foard and Haber 1966; Haber and Luippold 1960). In pine seeds, both cell
division and elongation occur simultaneously (Berlyn 1972), while in cherry seeds, cell division
precedes cell elongation (Pollock and Olney 1959). Thus, the protrusion of the radicle through
the seed coat is initiated by cell elongation, followed by cell division in most seeds. The cell
elongation process reportedly occurs in two stages in seeds of peas, barley, and vetch (Rogan
and Simon 1975). In the first stage, a slow elongation of the radicle occurs without any increase
in dry weight and only a small increase in fresh weight. This stage may represent active cell wall
preparation for the synthesis of new wall materials during later elongation. The second phase
is a rapid elongation of the radicle with marked increases in both fresh and dry weight
accompanied by rapid mobilization of nutrients into the radicle. These events lead to protrusion
of the radicle and the seed's change from an autotrophic to a heterotrophic organism.

Seedling Establishment

Seedling roots, of course, grow down and shoots grow up and these gravitropic responses
are mediated by auxin (indole-3~acetic acid). Auxin is synthesized in the com root apical
104 Seed Germination
meristem (Feldman 1981). moves forward through the stele to the root cap. and is preferentially
redistributed toward the lower side of the cap. At that point, the redistributed auxin moves
through the root cortical cells to the zone of elongation. There, the auxin on the lower side of the
root inhibits cell growth and causes a downward curvature (Evans et al. 1986). Thus, while the
meristem is the site of auxin synthesis, the root cap functions in redistributing auxin
asymmetrically so that roots grow downward (Young et al.1990). With respect to corn shoot
growth, corn seedlings must rely upon the elongation of the mesocotyl and coleoptile before the
leaves can unfold above the soil surface. While both tissues elongate, it is the mesocotyl that is
primarily responsible for the greatest elongation. Auxin is believed to be synthesized in the
coleoptile tip and moves basipetally to the mesocotyl where it has its greatest effect (Bandurski
et aI. 1984; Parker and Briggs 1990).
In legume seeds, rapid growth of the seedling hypocotyl occurs immediately following
radicle protrusion. Cavalieri and Boyer (1982) demonstrated that water potentials decreased
from the root to the hypocotyl crook and radially from the stele to the cortex. They also showed
that water potential in the hypocotyl elongation zone was not uniform and was most negative
immediately below the hypocotyl crook. The control of gravitropic responses for elongating
seedling hypocotyls is not yet as clear as it appears to be in corn. Traditional concepts suggest
that there is a redistribution of auxin causing the lower tissues of a horizontal hypocotyl to
elongate more in response to a higher auxin gradient than upper tissues. However, Rorabaugh
and Salisbury (1989) found that differential growth in soybean hypocotyls was due not to
redistribution but differences in tissue sensitivity to auxin concentration. Other studies have
indicated that hypocotyl elongation rates are reliant on a balance between gibbereHins and
abscisic acid concentrations (Bensen et a1.1990) or ethylene evolution, which inhibits hypocotyl
elongation (Seyedin et al. 1982).
The seedling starts to establish itself when it begins water uptake and photosynthesis.
Initially, it undergoes a transition stage during which it begins to produce some of its own food,
but is still dependent on food breakdown from reserve storage tissue. As the seedling becomes
fIrmly established in the soil, begins water uptake, and manufactures most of its own food, it
gradually becomes independent of the exhausted storage tissues. Then the germination process
is complete.

A BIOCHEMICAL MECHANISM OF SEED GERMINATION

In 1968, Amen presented the f]fst model of the sequence of biochemical events that occur
during seed germination. At the outset, he recognized that this model must be a generalized
scheme because every plant species is unique; some require light while others need cold
temperatures to induce germination. His attempt to compile the literature of that time into a
workable model was noteworthy and provided the impetus and stimulus for further research to
test the model for its validity. Surprisingly, little new or additional information has been gained
to test Amen's hypothetical scheme. His model, however, highlights many ofthe events that must
occur during germination and is, therefore, important to consider in a discussion of seed
germination.
According to Amen, germination is governed by a balance between inhibitors and
promotors. When inhibitors are present in physiologically greater concentrations than
promotors, dormancy ensues. A trigger agent such as light or temperature is necessary to either
 V::i
&
~
;:sO
a
0'
• inhibitor inactivation " ::s

Phytochrome
I .. gibberellIn
. .if . proteolytIC enzymes "(activation)
i i
(trigger agent) (germination agent) (releasing enzymes) ~
germination /
inhibitor II I germination
ce u ase proteinase activation inhibitor
/ (brypain)

/hYdrOlYSiS /
glutenin bound
GERMINATION OIl structural protein hydrolysis polymerase
,~ ,
Starch hydrolysis ~ ~. feleasring
enzyme derepressing
(actlvatlOn)
.. r ~ ~
. protemases
. ' enzyme
n"cle" .cod \rolYS,"
RNA
repressed
mRNA ",polymerase
Ii-amylase ~-amylase ... \
DNA .....
!
--lr--:-- DNA

histone
~(s:;;:y;.:.n;.:t;.:he:.:s;:is:.s:.:t:.:im~u;;,;;la;:ti:..;;o:.:n",,)_ _..J..._ _ _ _ _ _... ribonuclease actinomycin-D

Figure 5.22. Possible biochemical pathways in the initiation ofseed germination (From Amen 1968).

ov.
-
106 Seed Germination
inactivate or decrease the level of inhibitors in seeds. When this occurs, a germination stimulant
such as gibberellic acid can exert its promotive influence and the process of germination is
initiated. Figure 5.22 shows that gibberellins increase the de novo synthesis of proteolytic
enzymes, a-amylase and ribonuclease, while activating the release of~-amylase. The amylases
hydrolyze starches, providing the essential sugars for germination. Ribonucleases are essential
for nucleic acid hydrolysis, which subsequently can be used to recode new RNA species for later
germination processes. The proteolytic enzymes serve in conjunction with cellulases to degrade
cell walls, which is an essential first step in the loosening of the seed coat prior to radicle
protrusion. Other studies have indicated that gibberellins may derepress portions of the DNA
molecule, allowing transcription ofmRNA which then can be translated to form many of the de
novo enzymes responsible for germination. Many of these events have been shown in
germinating seeds, and this model serves to integrate these independent processes into a unified
concept of germination.

OVERALL CHANGES DURING GERMINATION

It is useful to consider the overall changes that occur during seed germination. Because
of the great number of species available and their diversity, innumerable examples could be
cited. However, certain patterns of development occur that are common to all seeds.

Dry Weight

During the first few days, the germinating seedling undergoes a net loss in dry weight
(Figure 5.23) due to the high respiration rate and some exudation and leakage through the seed
coat. As the epicotyl and radicle begin to grow, they gain weight rapidly at the expense of the
cotyledon or endosperm, which undergoes a rapid weight loss as the developing hypocotyl uses
most of the initial breakdown products for rapid synthesis and gowth. After the third to fifth
days, its growth diminishes, and the remaining breakdown products are utilized for synthesis
in the epicotyl and developing root system.

V>
OJ)
.5
'0
0)
0)
CIJ
0
0

'0100
c
.... 80
.9 Adventitious
I::: 60 Root
.~
I:::
40 ~~p.J'~ S:ZZ~~ Sem inal
0
Vl 20 Root
t:LL...~L...:::::"::~~~Li.L.u..:,Lu..::.L~ZZ::::24:2::Z::~! Caryopsis
Vl
0
...J 18 II
OJ) " 14
E Days afte r sowing

Figure 5.23. Changes in weight distribution in perennial ryegrass seed through 21 days after planting
(From Anslo 1962).
Seed Germination 107
The storage carbohydrates, fats, and proteins decrease rapidly in the cotyledon and
endosperm during germination. Their degradation products are translocated to the growing
points, where they accumulate prior to utilization in further synthesis.
The increase in nucleic acids, especially DNA, closely parallels their increase during cell
division. Each cell division results in a doubling ofthe: nuclear material. Early hypocotyl growth
is largely from cell division; therefore, DNA increases rapidly. After a few days, further growth
is primarily from elongation of previously formed cells; therefore, DNA formation decreases.
Growth in the plumule and radicle results equally from cell division and cell elongation;
consequently, DNA increases steadily in these organs as germination proceeds.
RNA is located in both the nucleus and the cytoplasm; so its occurrence is less closely
associated with nuclear division. Therefore, the RNA changes reflect the overall effect of both
cell division and cell elongation.

CHEMICAL PROMOTION OF SEED GERMINATION

Gibberellins

Since about 1955, gibberellins have been known to promote seed germination in a great
variety of species. Like thiourea, the gibberellins are not used extensively in routine germination
testing, but may be useful in certain situations. A number of gibberellins promote germination,
but the form most often used is gibberellic acid (GA3)'
Like thiourea, gibberellins can substitute tor light and temperature in promoting
germination. They can also promote germination of seeds not having these requirements.
Formerly, the action of gibberellin and red light on imbibed seeds was assumed to be identical.
However, scientists now believe that their mode of action is only partially similar ([kuma and
Thimann 1960; Fujii et al. 1960). Gibberellins are believed to be important in controlling the
germination of seeds in nature (Phiney and West 1960; Koller et al. 1962). There is evidence
that natural gibberellin-like substances appear during successive stages of after-ripening and
germination. Gibberellin-like substances have also been isolated from seeds of beans, lettuce,
and many other species. It is now well established that gibberellins play an important role in the
regulation of seed germination.

Cytokinins

Another group of endogenous hormones that promote seed germination in some species are
the cytokinins. Kinetin (6-furfurylaminopurine) is the best known ofthese. While cytokinins can
break primary dormancy of some seeds, they appear to be more effective in overcoming
secondary dormancy (Tilsner and Upadhyaya 1985). The mechanism of cytokinin regulation of
seed germination is not known, but three possibilities have been suggested:
Transcriptional Mediation. The binding of cytokinins to ribosomal preparations has
been demonstrated in wheat seed embryos (Fox and Erion 1975). This finding may suggest that
cytokinins can regulate specific gene expression.
Translational Mediation. Cytokinins have been found in association with wheat embryo
tRNA and always with tRNA species which recognize uridine as their initial codon letter
(Burrows et al. 1970). It is suggested that the ribosome assumes a specific configuration
108 Seed Germination
in response to the uridine codon which controls which tRNA species are permitted access to the
codon, thus mediating the proteins to be synthesized (Burrows 1975). In this way, cytokinins
can regulate some of the initial processes of germination.
Membrane Permeability Mediation. Thomas (1977) showed that cytokinins influence
many phytochrome-controlled processes. Since phytochrome is located in cell membranes and
can alter membrane permeability through its reorientation, cytokinins may conceivably mediate
seed germination through their effect on membrane permeability. Such a system would permit
the release of gibberellins from the scutellum to the aleurone during the first stages of
germination.
The exact role of cytokinins in promoting seed germination has yet to be resolved.
However, cytokinins certainly can break dormancy in seeds with a chilling requirement such as
sugar maple, Leucadendron daphnoides, and Protea compacta, or with a light requirement such
as Rumex obtusifolius, lettuce, and celery.

Ethylene

Ethylene (C 2H4) is known to stimulate seed germination of many species, in addition to its
influence on fruit ripening, bud dormancy, leaf abscission, and other growth processes.
Apparently, it is involved in the regulation of seed dormancy, although its effect is not limited
to dormant seed. It has been shown to enhance the germination rate of aged as well as immature
seed. Ethylene is thought to be involved in regulating auxin levels in dormant seed, and is known
to be released during seed germination of several species. It can also act synergistically with
gibberellin and red light in the germination of lettuce seeds. There has also been a suggestion
that ethylene and nitrate interact in stimulating seed germination (Saint and Spencer 1986; Egley
1984). Ethylene is now used in seed testing to break the dormancy of peanut and sunflower
seeds.

Hydrogen Peroxide

The stimulating effect of hydrogen peroxide (H20 2) on seed germination and subsequent
seedling vigor has been observed in a number of species, including many conifers, legumes,
tomatoes, and barley. This chemical acts as a respiration stimulant that accelerates the
breakdown of reserve food substances, thus providing a more rapid supply of energy and
materials for synthesis in the growing points. Hydrogen peroxide also has disinfectant properties
and can be used to disinfect seed to prevent mold growth on the seed and germination media. A
laboratory test has been developed using the hydrogen peroxide treatment as a rapid viability
check of conifer seed.

Auxins

As noted in Chapter 3, auxins and other growth regulators are universal components of
plants and a common constituents in seeds. In view of their presence in seeds and their role as
plant growth regulators, it is not surprising that they can influence germination as well. The
best-known auxin, indoleacetic acid (IAA), has been shown to increase lettuce seed germination
and to be temperature dependent. A definite relationship between IAA concentration and its
Seed Germination 109
action has not yet been established, although high concentrations inhibit germination, while low
concentrations are generally promotive or ineffective. There is also evidence that IAA interacts
with light in influencing seed germination. Most studies have shown that auxins have little
promotive effect on seed germination.

Potassium Nitrate

Potassium nitrate (KN0 3) is the most widely used chemical for promoting seed
germination. Solutions of 0.1 to 0.2% KN0 3 are common in routine germination testing and are
recommended by the Association of Official Seed Analysts and the International Seed Testing
Association for germination tests of many species. Figure 5.24 shows the effect ofKN0 3 on
New Zealand browntop seed germination.
Most seeds that are sensitive to KN0 3 are also sensitive to light. At one time it was
assumed that KN0 3 substituted for Jight, but now it is believed that the light sensitivity is only
increased. On the other hand, KN0 3 can complet<::~ly counteract light inhibition of rice grass
(Oryzopsis) seed germination.

100

90
_.4
80 .' ...:..:;.;.:...,3
-1'-'-' ----
';/0--
2
E-< .. j
Z 70 .-:f
f.I.l
u .-:1
0:::
f.I.l
60
..~.:/
-

./
~
50
z
0 :/
E=:
<:
40 :i
~ :1
30
~ ./ _ _ ·1
f.I.l j • ______ e-

1/
0 20

10
0 ~1

10 20 30
Days of germination

1. _ _ Substrate moistened with tap water

2. - - - Substrate moistened with 0.1% KN0 3
3. _. _. Substrate moistened with 0.2% KN0 3
4 ......•.. Substrate moistened with 0.5% KN0 3

Figure 5.24. The effects ofdifferent potassium nitrate concentrations on germination afNe»> Zealand
browntop seed (From Gadd /955).
110 Seed Germination
KN0 3 may interact with temperature in influencing seed germination in some species. This
has been demonstrated in a comprehensive study of several native range grasses (Toole 1938).
It is also known to act synergistically, or cooperatively, with gibberellic acid (Hashimoto 1958)
and kinetin (Ogawara and Ono 1961) in inducing germination of tobacco seed.
However, KN0 3 may be detrimental to the germination of some seeds. Reported inhibition
of lettuce seed germination by KN03 is somewhat surprising in view of the light and low
temperature requirements of this species (AOSA 1991). Other studies have shown that the
stimulatory effects ofKN03 affect the respiratory system directly (Adkins et al. 1984) and that
these are more pronounced in light than in darkness (Hilton 1984). Still others have proposed
that nitrate acts to stimulate oxygen uptake (Hilton and Thomas 1986) or serves as a cofactor
of phytochrome (Hilborst 1990). These studies demonstrate that the precise role of KN03 in
stimulating seed germination remains to be determined.

Thiourea

Although not used in routine germination tests like KN0 3, thiourea does promote
germination of many seeds. Unlike KN0 3, thiourea is able to substitute for the light and
temperature requirements of germination, perhaps replacing the light and temperature
requirements for the physiological processes occurring naturally during after-ripening. Thiourea
has been reported to replace the growth promotor that develops naturally during stratification
(Villiers and Wareing 1960).

Other Chemicals

Other chemicals may also promote seed germination under natural conditions. For example,
the host plants of two parasitic species, witchweed (Striga) and mistletoe (Orobanche), secrete
a substance that stimulates the germination of nearby seeds of these parasitic species.
Scopoletin, a phenolic compound, better known as a growth inhibitor, has been observed to
promote germination of aged white mustard (Sinapsis alba) seed (Libbert and Lubke 1957). The
phenomenon of allelopathy suggests that many plants produce chemicals that either delay or
prevent seed germination (Karssen and Hilhorst 1992).

OTHER FACTORS THAT AFFECT GERMINATION

Osmotic Pressure

High osmotic pressures of the germination solution make imbibition more difficult and
retard germination (Rodger et al. 1957). The ability of seeds to germinate under high osmotic
pressure differs with variety as well as with species, but all are affected. Some halophyte
species, however, actually have seeds that germinate better in salinities ranging from 0.2 to 1.7
moll·l (Ungar 1987). Mannitol solutions at varying concentrations have long been used to obtain
different osmotic pressures for studying germination responses (Dotzenko and Dean 1959).
More recently, other research studies have employed polyethylene glycol (PEG) to subject seeds
to varying osmotic pressures. These osmoticums have been used to minimize the effects of salt
toxicity (Bliss et a1. 1984).
Seed Germination III
Hydrogen-Ion Concentration (pH)

Germination can proceed over a wide range of hydrogen-ion concentrations. The

germination of almost all species occurs readily between pH values of 4.0 and 7.6 (Justice and
Reece 1954).

Presoaking

Presoaking seeds in water can be used to speed up germination, and has been used to
accelerated germination of several grass species (Chippendale 1934; Johnston 1964). Another
approach has been to expose seeds to an osmoticum so that their moisture content is increased
but remains below the moisture level needed to initiate germination--a process called
osmoconditioning (see Chapter 13). In both cases, the seeds are usually redried prior to
germination. The basis for the acceleration is uncertain; however, it is likely that hydrolytic
processes begin during presoaking, and the resulting simple sugars that are released can be
utilized for synthesis immediately upon germination. It is also possible that membrane repair
may occur by enzymes activated during the hydration process.
Another well-known benefit from presoaking at moderate temperatures (20°C) is the
protection it provides from chilling injury during the subsequent germination at lower
temperatures (5° to 15°C) (Pollock and Toole 1966). In such cases, the seed is not redried but
is held moist until germination.
Conversely, soaking seeds may be detrimental to germination capacity and should not be
used unless definitely needed. Kidd and West (1918) reported reduction of germination and
seedling growth as a result of soaking. Prolonged soaking has been found to cause injury to
seeds of many species (Tilford et a1. 1924). Presoaking injury to soybeans has been attributed
to low oxygen concentration inside the seed (Resuhr 1941), although more often to leaching or
to exosmosis of enzymes and nutrients from the seeds. The latter view is supported by leachate
conductivity experiments.

Effect of Low Temperature

Frost and cold nights may cause considerable seed injury prior to harvest. After extensive
study of low-temperature injury to com seed, Rossman (1949a) concluded that the extent of
i11iury depended on: (1) temperature level, (2) duration of exposure (3) moisture content of the
seed, (4) physiological maturity of the seed, (5) husk protection, and (6) variety. Of these, seed
moisture was most crucial.
If conditions are otherwise favorable, seeds can withstand extremely low temperatures. Air-
dried (10-12% moisture) seeds of wheat, barley, and several other species have even withstood
temperatures of liquid nitrogen (-192°C) for 24 to 110 hours (Brown and Escombe 1898;
Adams 1905) and temperatures of liquid hydrogen (-252°C) for 6 hours (Thiselton-Dyer 1899)
without injury. Com seed at 20% moisture survived temperatures of -7°C, but at 25% moisture
considerable injury occurred (Rossman 1949a).
Rossman (l949b) reported several additional responses to freezing in com seed. These
include: (1) viability of soaked seed was decreased more than in freshly harvested seed with
comparable moisture content, (2) seedlings were killed outright, (3) repeated freezing and
112 Seed Germination
thawing cycles were less injurious than continuous freezing when the total times were equal, (4)
the frrst freeze-thaw was more injurious than subsequent ones, (5) rate of thawing had no effect
on the viability of frozen seeds, and (6) slow drying of frozen seeds was less injurious than fast
drying, presumably because it gives the protoplasm time to recover (Rossman 1949b).
As seed matures in the field, it becomes increasingly hardy. The cold resistance of com
seed with 15 to 20% moisture was adequate to avoid injury from ordinary autumn freezing
(Kiesselbach and Ratcliff 1920). When maturing oat seeds were collected from fields before and
after frosts, seeds in the milk to soft dough stage were easily injured, but resistance increased
rapidly thereafter (Fryer 1919). .
Mature soybean seeds are surprisingly resistant to freezing, and can be stored at 18%
moisture up to sixteen months at -10°C without significant injury (Robbing and Porter 1946).
Soybean seeds with 30 to 32% moisture have been stored at -29°C for extended periods of time
without injury, whereas sorghum seeds at 16 to 19% moisture did not survive under the same
conditions.
Judd et al. (1982) reported a curvilinear relationship between soybean seed moisture and
the temperature at which freezing occurred. Immature seeds (in green pods) at 65% moisture
content were not injured by freezing temperatures of -2°C; seeds at physiological maturity (in
yellow pods) at 55% moisture showed significant reductions in germination following an eight-
hour exposure at -7°C; while germination of seed in brown pods at 35% moisture was reduced.
The mechanism of low-temperature injury is not completely understood. It is generally
attributed to formation of ice crystals in the inter- and intra-cellular components, which then
expand and rupture the physical and functional integrity of the cell membranes. Extreme cell
desiccation and precipitation of the protoplasm (when water is withdrawn by intercellular ice
formation) may also contribute to freezing injury (Luyet and Gehenio 1938; Stanwood 1986).

Radiation

Exposure to gamma radiation usually retards seed germination. Its effect differs in varieties
as well as species, and is more pronounced at higher temperatures and higher seed moisture
contents. Germination losses up to 38.1 % have been observed in bluegrass, peanuts, and onions
at 10 Krad radiation; wheat and sorghum at 20 to 40 Krad; com at 40 to 80 Krad; and crimson
clover and radish at 80 Krad (Justice and Kulik 1970). Gamma radiation up to 80 Krad caused
retardation of shoot and root growth up to one-half to two-thirds that of the untreated seed
(Justice 1967).
The studies cited above reflect the effect of gamma radiation above 10 Krad radiation.
Sahid and Soemartono (1974) reported results of gamma radiation studies on rice seeds showing
that the speed of germination was actually increased by exposures to radiation below 10 Krad.
Like other investigators, however, they found that stronger exposures depressed germination.

Mechanical Damage

Mechanical damage during harvesting, conditioning, and handling is a major problem in

the seed industry, especially for fragile, large-seeded species (e.g., edible legumes). Injury
symptoms may be of several different forms: (I) gross damage to the seed coat that is easily
visible, such as "splits" and seed coat cracks; (2) internal damage, visible only after seed
germination; (3) microscopic breaks, especially of the seed coat, that reduce performance and
Seed Germination 113
increase susceptibility to microorganisms; and (4) cryptic (hidden, or internal) injury, which is
probably physiological in nature and reduces the germination vigor, lengthens the time to
maturity, and reduces yield (McDonald 1985).
Sources of Mechanical Injury. Injury may occ;ur from any kind of physical abuse. While
threshing abuse probably is the most serious, seed conditioning and handling may also contribute
to seed injury. The effect of all individual impacts the seed receives appears to be cumulative
in total seed damage; thus, it is essential always to minimize physical impact to assure the best
germination.
Influence of Moisture Content. Susceptibility to mechanical injury increases as the
moisture content of the seed decreases; however, safe moisture contents vary among species.
Large-seeded legumes are particularly vulnerable, and excessive injury begins to occur at
moisture contents below 15%. Germination of navy beans was raised from 36 to 76% by
increasing the moisture before threshing from 11 to 16% (Dexter 1966). Small-seeded grasses
and legumes can safely be threshed at moisture levels as low as 8 to 10%. The cereals are more
susceptible, and injury occurs at moisture levels below 11 to 12%.
Influence of Genotype. The genetic control of susceptibility to mechanical injury in some
crop species has been clearly established (Atkin 1958; Davis 1964). Twelve percent moisture
is the reported critical moisture for distinguishing between tolerant and susceptible navy bean
lines (Dorrel and Adams 1969).

Questions

1. What similarity do you see between seed germination and seed deterioration?
2. What type of germination is characteristic of the greater number of species-hypogeal or
epigeal?
3. What environmental factors are required for seed germination? Which do you consider to
be of the greatest importance?
4. How can rice seed germinate in the complete absence of air?
5. How would you defme the optimum temperature for seed germination of a given species?
6. What is the origin and meaning of the term stratification?
7. Do you think that the average light-supplied seed germinator can deliver optimum light
intensity for lettuce seed germination?
8. What factors influence the sensitivity oflight-r~uiring seeds to light intensity? How is the
light action on seed germination similar to that on floral induction?
9. What groups of chemicals can promote seed germination? Which are used to promote
germination in routine seed testing?
10. Why are soybeans more resistant to low-temp(:rature injury than most seeds?
11. What is imbibition pressure?
12. How does most water enter the seed during imbibition?
13. What is the importance of mitochondria during seed germination?
14. What is the importance ofnucleotides (ADP alfld ATP) during germination?
15. What are the breakdown pathways for starch, proteins, and lipids during germination?
16. What is the relationship between gibberellic add and amylase during barley seed germi-
nation?
17. Cite three potential mechanisms to explain the mode of action of phytochrome in promoting
seed germination.
114 Seed Germination
18. List the factors that influence the availability of water necessary for seed gennination.
19. Cite at least three processes that occur during the outwardly tranquil enzyme activation
stage of seed germination.
20. Contrast and provide examples of trigger and germination agents.

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Thill, D. C., K. G. Beck, and R. H. Callihan. 1984. The biology of downy brome (Bromus
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Thiselton-Dyer, Sir William. 1899. On the influence of temperature of liquid hydrogen on the
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Thomas, T. H. 1977. Cytokinins, cytokinin-active compounds and seed germination. In: The
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Seed Germination 123
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6
Seed Viability
and Viability
Testing
Although the concept of seed viability is well known, there may sometimes be dis-
agreement and confusion as to its precise meaning. To most seed technologists and most
people in the seed industry, viability means that a seed is capable of germinating and
producing a "normal" seedling. Therefore, it is used synonymously with germination capacity.
In this sense, a given seed is either viable or nonviable, depending on its ability to germinate
and produce a normal seedling; thus, only seed lots representing populations of seeds may
exhibit levels of viability.
In another sense, viability denotes the degree to which a seed is alive, metabolically
active, and possesses enzymes capable of catalyzing metabolic reactions needed for
germination and seedling growth. In this context, a given seed may contain both live and dead
tissues, and mayor may not be capable of germination. This meanil1g deals with tissue
viability as well as viability of the entire seed.
In either context, seed viability is probably highest at the time of physiological maturity,
though its environment on the parent plant may not permit germination. After physiological
maturity, the viability of seeds gradually declines. Their longevity depends on the
environmental conditions to which they are exposed.
Numerous tests exist for determining seed viability; these are discussed on the following
pages.

GERMINATION TEST

The germination test is most commonly used to determine seed viability. It has become
so universally accepted that seed germination and viability are probably considered one and
the same by most people. Regardless of its acceptance, the germination test is still an estimate
and has certain limitations as a universal estimate of seed quality. However, if these
limitations are recognized, the germination test is a useful viability index.

L. O. Copeland et al., Principles of Seed Science and Technology

© Kluwer Academic Publishers 2001
Seed Viability 125
Germination Terminology

Why test seeds for germination? The intuitive answer to this question is "Because we
need to know how the seeds will perform in the field." However, there are more reasons than
this. For example, we know that a seed lot consists of a population of seeds, each with its
own distinct capability to produce a plant. A germination test is an analytical procedure to
evaluate seed germination under standardized, favorable conditions that are seldom, if ever,
encountered in the field. Consequently, germination test results often overestimate field
performance of a seed lot. It is now increasingly recognized that seed vigor tests can more
accurately reflect the ability of a seed to perform in the field (see Chapter 8). However, the
use of favorable conditions leads to a maximum germination percentage and this result can be
consistently reproduced. This permits different analysts from different laboratories to obtain
similar germination percentages for the same seed lot. This allows verification of germination
results on seed labels, which permits shipment of seed lots of known value to move through
interstate and global commerce. A germination test allows a seed producer to determine and
compare the quality of a seed lot before it is planted. This information can be used to
determine the planting value of a seed lot and its storage potential to satisfY labeling laws and
provide for standardized marketing of seed.
To the seed analyst, germination has a definite and standardized meaning, which must
lead to uniform interpretations made by analysts in different laboratories. In the Rules for
Testing Seeds of the Association of Official Seed Analysts, seed germination is described as
"the emergence and development from the seed embryo of those essential structures which.
for the kind of seed in question, are indicative of the ability to produce a normal plant under
favorable conditions" (AOSA 2000). The seed analyst looks for the emergence of an
embryonic plant that must consist of a complete root and shoot axis that has the capacity of
normal growth under favorable conditions.
Seed testing associations (AOSA and ISTA-see Chapter 15) have developed sets of
standardized conditions (specifications) for seed germination of different species. These
conditions are published in their official rules, which specifY the optimum germination
conditions for hundreds of kinds of agricultural, vegetable, flower, and tree seeds. The rules
provide optimum germination conditions for each kind of seed. The rules also provide
additional measures that may aid in germination, particularly of dormant seeds. An excerpt of
the rules of the Association of Official Seed Analysts (AOSA) is given in Table 6,1.

Rules for Germination Testing Procedures

The AOSA Rules for Testing Seeds are divided into four groupings: agricultural,
vegetable and herb, flower, and tree and shrub seeds. Each kind of seed is listed according to
its recognized scientific nomenclature as well as common name. Following the name of the
seed is the substrate (germination media) recommended [e.g., petri dishes (P), blotters (B),
sand or soil (S), paper towels (T), or kimpac (C)]. Substrata should (1) be non-toxic to
germinating seedlings, (2) be free of molds, microorganisms, and their spores, and (3) provide
adequate aeration and moisture for germination. Timothy seed should be germinated on any
substratum suspected of being toxic to seedlings since its roots will not develop normally in
the presence of toxic substances.
 N
0\
-

Table 6.1. Some Methods for Testing for Germination.

First Final Specific

count count requirements and Fresh and
Kind of seed Substrata Temperature ·C days days photo numbers dormant seed
Eragrostis trichodes P 20-30 5 14 Light; KN0 3 Pre chill at 5° or 10°C
sand love grass for 6 weeks
Erodium cieutarium B,T 20-30 3 14 Clip seeds
alfileria
Fagopyrum eseulentum B,T 20-30 3 6
buckwheat
Festuea arundinacea P 15-25; 20-30 5 14 Light and KN0 3 Prechill at 5° or 10°C
tall fescue optional for 5 days and extend
test to 21 days
Festuea capillata P 10-25 10 28 KN0 3
hair fescue
Festuea elatior P 15-25; 20-30 5 14 Light and KN0 3
meadow fescue optional
Festuea ovina P 15-25 7 21
sheep fescue
alternate method P 20-30 7 28 Light
Festuea ovina var duriuseula P 15-25 7 21 Light and KN0 3
hard fescue optional
Data from AOSA {1978}.

V:l
~
iSS
~
~
~.
Seed Viability 127
Recommended temperatures (in degrees Celsius) for conducting germination tests are
followed by the fIrst and fmal evaluation times (counts). Temperature recommendations are
based on research that has shown optimum temperatures for each species of seed. The
Association of Official Seed Analysts permits only a 1°C variation from the recommended
temperature. One number in the temperature column indicates a constant temperature
throughout the testing period. Two numbers separated by a dash indicate an alternating
temperature during a 24-hour period-the fIrst temperature maintained for 16 hours followed
by the second temperature for 8 hours. Two temperature recommendations separated by a
semicolon are considered alternate recommendations. In such cases, both temperatures are
permissible and a laboratory may use the temperature that is most convenient. The time of the
fIrst count is considered to be approximate and a deviation of one to three days is permitted.
At this time, the analyst counts and discards germinated seedlings and inspects the test for
potential problems such as lack of moisture or pathogen infestation. The fmal count must be
on the day specifIed since it provides sufficient time for even weak seedlings to germinate.
Thus, the test should be terminated by the fmal count unless dormancy is suspected.
Seeds of certain species require special treatment for maximum germination or breaking
dormancy. Such requirements are listed in the two columns, "Special requirements" (Table
6.1) and include germination-promoting environments such as light and the addition of KN03.
Light is required for the germination of most grasses, many tree and shrub seeds, and some
vegetable seeds. The Association of Official Seed Analysts recommends that light be evenly
distributed with an intensity range of 810 to 1620 lux (75 to 150 foot-candles). For most seed
germination tests, seeds should be subjected to light for only a part of the test period. Eight
hours of light per day is usually sufficient. Potassium nitrate solution (0.2%) is used to
promote germination of some dormant seeds.
Special treatments for overcoming seed dormancy are listed under "Fresh and dormant
seed." In most instances, directions for prechilling are provided (Le., placing the seed on a
moist substratum at low temperatures-generally 10°C-for a specifIed period). Then the
seed is transferred to optimum temperature conditions for the duration of the test. Although
prechilling treatment (stratifIcation) prolongs a germination test substantially, it is essential to
break the dormancy of many species.
Although moisture and aeration are not specifIed in the germination methods, enough
moisture should be provided so that the seeds can imbibe the water before it is evaporated
from the substratum. Too much water creates an anaerobic environment in which essential
oxygen is not available to seeds. A formula determining optimum moisture for seeds
germinating in sand is provided in the "Rules for Testing Seeds" (AOSA 2000). Regardless of
the kind of seed being tested, all germination tests should be examined daily to ensure that the
moisture content of the substratum is optimal. The Association of Official Seed Analysts
further recommends that germination tests be conducted in germinators or germinator boxes
that maintain a relative humidity of 95% or higher to minimize moisture loss from the
germination substrata.
The rules for germination testing specify only environmental conditions and dormancy-
breaking procedures that are of proven effectiveness in promoting germination and lead to
standardized interpretations in routine seed testing. Too frequently, they do not reflect new
knowledge that is available through scientifIc research. For example, it is widely known that
many growth regulators are effective in breaking dormancy, yielding a more accurate
estimate of viability; yet only one inorganic chemical (potassium nitrate) is recognized by the
128 Seed Viability

AOSA "Rules for Testing Seed" (AOSA 2000) as an aid in performing germination tests. Of
the plant hormones, ethylene and gibberellic acid are approved (e.g., green needlegrass) for
helping to break dormancy. Ethylene is used for sunflower.

TETRAZOLIUM TEST

The tetrazolium test is widely recognized as an accurate means of estimating seed

viability. This method was developed at Hohenheim University in Germany in the early 1940s
by Professor Georg Lakon (1928) (see Figure 6.1) who had been trying to distinguish
between live and dead seeds by exposing them to selenium salts. He then tried tetrazolium
salts and found them more effective. Today, the test is used throughout the world as a highly
regarded method of estimating seed viability and is a routine test in many seed testing
laboratories. It is often referred to as a "quick test," since it can be completed in only a few
hours (compared to regular germination tests that require as long as two months for some
species). Tetrazolium test results can be extremely valuable for providing labeling
information for immediate shipment of
seed lots without waiting for completion
of germination tests. It is also a valuable
research technique for estimating seed
viability and determining reasons for
poor germination.

Principle

The tetrazolium test distinguishes

between viable and dead tissues of the
embryo on the basis of their relative
respiration rate in the hydrated state.
Although many enzymes are active
during respiration, the test utilizes the
activity of dehydrogenase enzymes as
an index to the respiration rate and seed
viability. Dehydrogenase enzymes react
with substrates and release hydrogen
ions to the oxidized, colorless,
tetrazolium salt solution, which is
changed into red formazan as it is
reduced by hydrogen ions (Figure 6.2).
Seed viability is interpreted according to
the topographical staining pattern of the
embryo and the intensity of the
coloration.

Figure 6.1. Professor George Lakon (Courtesy

of Prof A. M Steiner).
Seed Viability 129

N - N - CsHs

CSH5 - C
/ + 2 e + 2H+
) C6H5 - C \
\
N = N - CSH5

2,3,5-triphenyl tetrazolium formazan

chloride
(colorless) (red)

Figure 6.2. The chemical reaction that changes the colorless tetrazolium solution into jormazan.

Procedure

Seeds are first imbibed on a wet substratum to allow complete hydration of all tissues.
For many species, the tetrazolium solution can be added to the intact seed. Other seeds must
be prepared by cutting and puncturing in various ways (Figure 6.3) to permit access of the
tetrazolium solution to all parts of the seed. After hydration, the seeds are placed in a
tetrazolium salt solution and held in a warming oven at about 35°C for complete coloration.
Two hours is usually adequate for seeds that are bisected through the embryo. but others
require longer periods of staining. If seeds are held too long in contact with the tetrazolium
solution, they tend to become overstained, making interpretation difficult. Handbooks of
instructions for performing tetrazolium tests and interpretation instructions have been pub
lished by the Association of Official Seed Analysts (AOSA 2000) and International Seed
Testing Association (1985).

Evaluation

Although the living tissues of seeds stain red, interpreting these results to estimate
viability requires considerable skill and experience. Sound embryo tissues absorb tetrazolium
slowly and tend to develop a lighter color than embryos that are bruised, aged, frozen, or
disturbed in other ways. The experienced analyst learns to distinguish between seeds with the
capacity to produce normal seedlings and those that stain abnormally (Figure 6.4).
The tetrazolium test is often called the topographical tetrazolium test because the
pattern, or topography of staining is an important aspect of its interpretation. Many seeds are
neither completely dead nor completely alive. The staining pattern reveals the live and dead
areas of the embryo and enables the analyst to determine if seeds have the capacity to produce
normal seedlings. The cell division areas of the embryo are most critical during germination,
and if they are unstained or abnormally stained, a seed's germination potential is weakened.
The analyst must be familiar with crucial cell division areas of the embryo and learn to
interpret their staining pattern in terms of seed germinability.
130 Seed Viability

Small-seeded grasses

A B c
I

--4f--~- 4t lor
I>--
./
~
~
./ (side view}

Large-seeded grasses

-
.
-€?-.
.~-
. -
..
_ - .....::.---
... -
(top view)

- -

(side view)

Figure 6.3. Preparation procedures for small- and large-seeded grasses for the tetrazolium test.
Small seeded species (above) with a rather soft embryo and endosperm, such as orchardgrass, are
prepared by piercing with a needle near the embryo to allow access by the 1Z solution. Others are
prepared by cutting across or near the embryo with a razor blade. Larger seeded species (below)
are cut (after presoaking) to expose a cross-section ofthe embryo and endosperm (Slightly mod?fied
from AOSA 2000).

Other factors must be carefully observed when interpreting a tetrazolium test. Among
these are flaccid tissues (lack of adequate turgor) and critically located fractures, bruises, and
insect cavities. Any of these factors, when present in a vital position, may cause an otherwise
sound seed to be nongerminable.

Advantages and Disadvantages

The rapidity of the test is its most obvious advantage and may justify its use when speed
is important. Another advantage is its usefulness for dormant as well as for nondormant
seeds, although its nondetection of dormancy (other than hard seed) is sometimes cited as a
disadvantage. Actually, when used in combination with a germination test, it can be useful for
Seed Viability 131

2 3

5 6

7 8 9

No. I GERMINABLE. Seed completely stained.

Nos.2-4 GERMINABLE. Minor unstained areas on cotyledons.
No.5 GERMINABLE. Minor unstained area on upper portion of radiclehypocotyl
axis.
No.6 GERMINABLE. Extreme tip of radicle unstained; minor unstained spots on
cotyledons.
No.7 NON-GERMINABLE. More than extreme tip of radicle unstained.
No.8 NON-GERMINABLE. Unstained area at juncture of cotyledons and radicle-
hypocotyl axis.
No.9 NON-GERMINABLE. Unstained area near point of attachment of cotyledons
and radicle-hypocotyl axis over location where plumule develops.
No. 10 NON-GERMINABLE. Radicle-hypocotyl axis bisected by unstained area.
No. 11 NON-GERMINABLE. Unstained areas on radicle-hypocotyl axis and at point
of attachment of cotyledons to axis.
No.12 NON-GERMINABLE. More than one-half of cotyledonary tissue unstained.
No. 13 NON-GERMINABLE. Radicle-hypocotyl axis unstained.
No. 14 NON-GERMINABLE. Seed stained off color. grayish-red, orange-red or glassy
or transparent red color.
No. 15 NON-GERMINABLE. Seed completely unstained.

Figure 6.4. Tetrazolium staining patterns and their interpretation for clover seeds. Black areas
represent stained, living tissue; white areas represent unstained, dead tissue. Pairs representy both
sides of the seed (From AOSA 1970).
132 Seed Viability

testing dormant seed. The germination test tells the percentage of immediate germination; the
tetrazolium test tells the percentage of live seed; and the difference between the tetrazolium
and germination tests represents the percentage of dormant seed.
Perhaps the greatest disadvantage of the tetrazolium test is its difficulty and the
experience required to interpret it correctly. Another disadvantage has been the lack of
acceptance of tetrazolium test results by seed law enforcement agencies, the seed trade, and
certification agencies; however, this prejudice is gradually disappearing, as tetrazolium
testing procedures become standardized and more analysts are trained to use them.

OTHER BIOCHEMICAL TESTS FOR SEED VIABILITY

The tetrazolium method has largely supplanted other biochemical viability tests for
routine testing of seed quality. An excellent review of rapid biochemical viability tests that
have been attempted (with varying success) has been prepared by Gadd (1950) of the
Swedish seed testing station. The following information was taken largely from his review.

Vital Coloring Methods

The principle of the vital coloring methods is the differential coloration of live versus
dead tissues when exposed to certain dyes. An early method used sulfuric acid. Subsequently,
indigo carmine and other aniline dyes were used. This dye stains dead tissue blue, but it is
incapable of penetrating live tissue, which remains unstained. Gadd states that it is
particularly useful for determining viability of tree seeds.

Enzyme Activity Methods

These methods measure enzyme activity of imbibed seeds as an indication of their

viability. Some enzymes that have been measured are the hydrolyzing enzymes, which are
capable of splitting high-molecular organic compounds of proteins, starch, and fats into less
complicated soluble substances. Examples of such enzymes are lipase, diastase and amylase.
Another group of enzymes are the desmolases, which may be divided into two groups, the
oxidases (catalase and peroxidase), or oxidizing agents, and the dehydrogenases. They are
directly involved in the respiration process and, as a group, are closely correlated with seed
viability.
Oxidase Method i-Catalase. Some investigators have used the catalase content of
seeds as an estimate of seed viability (Leggatt 1929), while others have doubted its usefulness
because of its lack of absolute correlation with seed viability. Catalase activity varies over the
course of germination, and the catalase content of immature seeds has been reported to be
higher than that of ripe seeds. Although some workers have reported good correlations
between viability and catalase content, others have not. The probability of errors in the
detailed and complex method of measuring catalase activity makes it an unreliable measure.
Oxidase Method 2-Peroxidase. The peroxidase content appears to be more closely
correlated with viability than the catalase content. McHargue (1920) used a technique
involving guaiacol which, in the presence of H20 2, is transformed to blue tetraguaiaco-
quinone. By pretreating the seed sample with H20 2 and then grinding the seed, followed by
colorimetric determinations, he achieved results that closely agreed with germination tests.
Brucher (1948) used a similar method without the disadvantage of grinding the seed. He
Seed Viability 133
soaked the seeds for 12 hours in a mixture of guaiacol and benzidine in 10% dilutions of
saturated alcohol solutions and then treated them with the reagent. Unlike McHargue's
technique, this method permitted evaluation of each individual seed, and close correlation was
found between its results and those of regular germination tests. Brucher considered embryos
that were slightly stained or well stained to be capable of germinating. The disadvantage of
these methods is that the color disappears rapidly.
Other Oxidases. Gadd reported that other oxidases generally disappeared gradually
from mature seeds and were unsuitable for viability tests. Phenolase has been tested but with
unreliable results.
Dehydrogenase Activity Tests. These chemical tests are based on the color changes of
certain substances, depending on whether they are in an oxidized or a reduced state. The
tetrazolium test, described earlier, is the best example of a successful dehydrogenase activity
test. Another method is based on the change of methylene blue to colorless methylene white;
however, the seed must be ground and the determination made in a vacuum since exposure to
air reoxidizes the methylene blue. This process is also complicated by the presence of
microorganisms that cause a similar color change. Another test utilizes the dehydrogenase
reduction of dinitrobenzene to a red nitrophenylhydroxylamine compound in the presence of
ammonia. Gadd reported that the color reaction was rapid, and at 40°C takes place in one to
two hours; however, the color quickly disappears and the substance itself is poisonous.
The Selenite Method. The selenite test was a biochemical test of the 1930s based on
the reduction of colorless selenium salts to red elementary selenium by dehydrogenase activity
of living cells. Gadd states that the selenite principle was used as early as 1900 with bacteria
cultures; its potential in seed viability testing was discussed by Hasegawa (1935) and was
popularized by Eidmann (1938) in the 1930s. Eidmann worked primarily with tree seeds,
which he prepared by halving the embryo, piercing the seed coat or excising the embryo. The
usual method, according to Gadd, is to soak the seeds for about 24 to 48 hours at 20 to 30°C,
at which time they are washed in water and the color reaction determined. As in the
tetrazolium test, both the distribution and intensity of coloration are important in interpreting
seed viability. Gadd suggested the following classification of selenite results: (1) completely
and intensively red colored embryos (germinable), (2) slightly colored to intensively colored
embryos with pale spots (germinable), and (3) very slightly colored to entirely uncolored
embryos and those with larger uncolored spots amounting to more than one-third of the
surface (nongerminable). Gadd and Kjaer (1940) reported results of testing cereals with a
combination indigo-carmine-biselenite method in which viability estimates were improved. A
distinct disadvantage of the selenite method is that a poisonous gas slowly develops, making it
dangerous for large-scale testing.

OTHER VIABILITY TESTS

Free Fatty Acid Tests

The breakdown of fats during seed germination was discussed in Chapter 5. Similar
degradative reactions may occur in seeds as deterioration progresses, especially under high
moisture levels, high temperatures, and microorganism infestation, resulting in the formation
of free fatty acids. Consequently, under such conditions, the level of free fatty acids has been
134 Seed Viability
suggested as an index of viability. At best, the test provides only a broad quantitative estimate
of the general level of viability of a seed lot. It has never attained status as a recognized
viability test although it is reported to be useful for cotton seed.

Hydrogen Peroxide (H202) Test

In early seed germination tests, particularly those with tree seeds, hydrogen peroxide
was used as a seed treatment to minimize the effects of fungi. It was soon recognized,
however, that the compound stimulated germination. The reason(s) for this stimulation
remain unknown, although it has been suggested that the hydrogen peroxide degrades to water
and one-half molecule of oxygen, which enhances the immediate oxygen environment
surrounding the seeds and thus stimulates germination. The test is conducted by cutting the
seed coat at the radicle end allowing a 1% solution of hydrogen peroxide to permeate the
interior of the seed. This treatment results in more rapid root protrusion compared to the
standard germination test.
There are a number of other seed viability tests that focus on the integrity of the seed
coat which can have an influence on imbibition damage, seed leakage, and susceptibility to
invasion by pathogens. These include ferric chloride, indoxyl acetate, fast green, and sodium
hypochlorite tests.

Ferric Chloride Test

Mechanically injured legume seeds tum black when placed in a solution of ferric
chloride (Hardin 1980). This is a very fast and useful test that provides a rapid estimate of
the percentage of abnormal seedlings expected from a crop. The seed is placed in a 20%
solution of FeCl2 for 15 minutes, at which time all black staining seeds are separated.
Because of the rapidity of this test, a seed conditioner can examine seeds immediately after
conditioning and make adjustments to equipment as needed to reduce mechanical damage.

Indoxyl Acetate Test

Any damage to seed coats, particularly in soybeans and other large~seeded legumes, is
an indication of mechanical abuse during harvesting and conditioning and serves as an entry
site for pathogen infestation. The indoxyl acetate test (French et a1. 1962) is a rapid
laboratory test that reveals seed coat damage in soybeans and other light colored legume
seeds. Seeds are soaked for 10 seconds in a 0.1 % solution of indoxyl acetate prepared in 95%
ammonia for 10 seconds and allowed to air dry. Lesions in the seed coats that are difficult to
detect become visible because they turn purplish~green against the yellow or white seed
cotyledon background.

Fast Green Test

The fast green test reveals physical fractures in the seed coat of light-colored seeds such
as corn. Seeds are soaked in a 0.1 % fast green solution for only 15-30 seconds. During this
period, the fast green penetrates any area of the seed coat which has been fractured and stains
the endosperm green. After the soak period, the seeds are washed and the fractures then
become apparent in the seed coat.
Seed Viability 135
Sodium Hypochlorite Test

The sodium hypochlorite test is used to identifY soybean seeds with seed coat damage.
Seeds are immersed in a dilute solution of sodium hypochlorite for 10 minutes. Seeds with
seed coat cracks rapidly absorb the sodium hypochlorite and swell to three times their original
size. This enables identification of seeds with seed coat cracks from those without seed coat
deformations.
Other seed viability tests focus on seed membrane and embryo integrity such as the
conductivity, excised embryo, and x-ray tests.

Conductivity Test

Seed producers have long dreamed of a simple method for determining seed viability by
merely subjecting them to an electrical current and noting the different responses of live and
dead seeds. According to Gadd, this concept has been used with varying degrees of success
beginning with Waller in 1901 who showed that live and dead seeds gave different 'blaze
currents" that could be measured with a galvanometer. A more reliable electrical method was
suggested by Fick and Hibbard (1925) who found that after soaking a seed sample in water
for a few hours under controlled temperature conditions, the conductivity of the solution
reflected the general level of viability of the seed sample.
Conductivity tests are based on the premise that as seed deterioration progresses, the cell
membranes become less rigid and more water-permeable, allowing the cell contents to escape
into solution with the water and increasing its electrical conductivity. In the past, the
conductivity test suffered from the disadvantage of being a bulk test where 50-100 seeds were
soaked simultaneously and a mean conductivity value obtained for all seeds. However, a
commercial instrument (Figure 6.5), has been developed and possesses the capability of
monitoring individual seed conductivity values. The instrument provides a rapid indication of
seed viability for seed lots (McDonald and Wilson 1979) and brings a sophistication to the
concept of conductivity tests by improving the value and reliability of test results. However,
there is some doubt about the reliability of test results in the medium quality range.

Excised Emhryo Tests

The excised embryo test (Figure 6.6) provides a unique way of assessing seed viability
that can greatly reduce the time required for viability estimates of dormant seed. It is
particularly useful for seeds of woody species for which the time for viability estimates 9~n
be dramatically reduced. Most of the original research on this technique was performed \ly
Flemion (1936, 1938, 1941, 1948) at the Boyce Thompson Institute. She observed that if
such embryos of dormant seeds were carefully removed without injury and placed on a moist
blotter or filter paper under favorable conditions, they would readily tum green and begin to
grow. When embryo dormancy occurred, growth was slow, although much more rapid than
in the intact seed. Where extreme embryo dormancy existed, no growth occurred. Even in the
latter case, dormancy was more easily overcome when the embryos were excised than when in
the intact seed.
136 Seed Viability

Figure 6.5. This machine has the capability of simultaneously measuring the conductivity of
leachate from 100 individual seeds sequentially (Courtesy of Wavefront, Inc., Ann Arbor, MI).

The excised embryo method is especially valuable with seeds of trees and shrubs where
dormancy is a major problem in evaluating seed viability, and germination sometimes takes
as long as six months. Laboratories that test considerable quantities of tree and shrub seeqs
routinely conduct excised embryo tests; however, a standard germination test may be
performed for comparative results.
One disadvantage of the excised embryo test is the high degree of skill and time required
to prepare the embryos for the test. Considerable caution must be taken to avoid injuring the
embryo during removal. Another disadvantage is that the test does not reveal damage to the
embryo other than in the root-shoot axis that might prevent normal germination of the intact
seed

X-Ray Tests

Although the x-ray test is not a viability test, it does provide information that can help
assess viability. It can reveal morphological deficiencies that indicate the structural potential
for viability. However, by using different metallic salts with different absorption capacity into
live and dead tissue, it can be used to distinguish viable from nonviable seeds.
The most beneficial use of x-rays in seed testing is to obtain a quick indication of
abnormal morphology or mechanical damage that might impair germination capacity.
Perhaps most x-ray work has been performed on seeds of sugar beets and tree seeds. In both
cases, it is particularly useful, since it reveals the inner seed structure within the hard seed
coat and shows developmental deficiencies (Figure 6.7) as well as mechanical fractures. The
Association of Official Seed Analysts has developed a handbook for x-ray testing of seeds
(AOSA 1979).
Seed Viability 137

Figure 6.6. Excised embryo test (Courtesy ofAOSA, Public Service Kit).

Questions

1. What do you consider to be the most useful seed viability test and why?
2. Do you believe the germination test will ever be replaced as the most common seed
viability test?
3. Does the seed analyst's concept of germination differ from that ofthe layperson? How?
4. What is the principle of the tetrazolium test?
5. Do any of the other biochemical viability tests have use in routine seed analysis?
6. List the advantages of other viability tests, including those for free fatty acid, indoxyl
acetate, conductivity, excised embryo, and x-ray tests.
7. List the substrate, temperature, and fmal count days recommended for germination of
tall fescue seeds according to the AOSA "Rules for Testing Seeds."

General References

Association of Official Seed Analysts. 1970. Tetrazolium testing handbook for agricultural
seeds. Association of Official Seed Analysrs Handbook No. 29, ed. D. F. Grabe.
Association of Official Seed Analysts. 2000. Tetrazolium testing handbook. AOSA
Publication.
Association of Official Seed Analysts. 2000. Rules for Testing Seeds. Proceedings of the
Association of Official Seed Analysts 60(2):39.
138 Seed Viability

Normal Empty

Damage-(mechanical) Damage-( deterioration)

Figure 6.7. Radiograph of longleaf pine seed showing normal embryo development, empty seed,
damaged (mechanical) and damaged (deterioration) (From "Radiographic Analysis ofAgricultural
and Forest Tree Seed, " AOSA Handbook No. 31).

Association of Official Seed Analysts. 1979. Radiographic AnalYSis of Agricultural and

Forest Tree Seed: Handbook No. 31, by E. Belcher and J. Bozzo.
Brucher, H. 1948. Eine Schnellme node zur Bestimmung der Keimfahigkeit von Samen.
Physiologia Plan/arum 1:343-358.
Eidmann, F. E. 1938. Eine neue biochemische Methode zur Erkennung des Aussaatwertes
von Samen. Proceedings of the International Seed Testing Association 10:203-211.
Fick, G. L., and R. P. Hibbard. 1925. A method for determining seed viability by electrical
conductivity measurements. Michigan Academy of Science, Arts, and Letters 5 :95-103.
Flemion, F. 1936. A rapid method for determining the germinative power of peach seeds.
Contributions from Boyce Thompson Institute 8:2154-293.
-,--_ _ . 1938. A rapid method for determining the viability of dormant seeds. Contributions
from Boyce Thompson Institute 9:339-351.
Seed Viability 139

--::-_-:-:-.1941. Further studies on the rapid determination of the germinative capacity of seeds.
Contributions from Boyce Thompson Institute 11 :455-464.
___ .1948. Reliability of the excised embryo method as a rapid test for determining the
germinative capacity of dormant seeds. Contributions from Boyce Thompson Institute 15:
229-241.
French, R. c., J. A. Thompson, and C. H. Kingsolver. 1962. Indoxyl acetate as an indicator
of cracked seed coats of white beans and other light colored legume seeds. Proceedings of the
American Society for Horticultural Science 80:377-386.
Gadd, I. 1950. Biochemical tests for seed germination. Proceedings of the International Seed
Testing Association 16:235-253.
Gadd, I., and A. Kjaer. 1940. Uber die Yerwendbarkeit der Selenund Indigokarmin-methoden
bei der Prufung von frost-und fusariumgeschadigten Getreide. Proceedings of the
International Seed Testing Association 12: 140-149.
Hardin, E. E. 1980. Personal communication.
Hasegawa, K. 1935. On the determination of vitality in seed by reagents. Proceedings of the
International Seed Testing Association 7: 148-153.
International Seed Testing Association. 1985. Handbook on tetrazolium testing. Zurich:
ISTA. 72 pp.
Lakon, G. 1928. 1st die Bestirnmung der Keimfahigkeit der Samen ohne Keimversuch
moglich. Angewandte Botanik (Zeitschrift der Vereinigungfor angewandte Botanik) 10:470.
Leggatt, C. W. 1929-1930. Catalase activity as a measure of seed viability. Scientific
Agriculture (Ottawa) 10:73-110.
McDonald, M. B., Jr., and D. O. Wilson. 1979. An assessment of the standardization and
ability of the ASA-610 to rapidly predict potential soybean germination. Journal of Seed
Technology 4(2): 1-12.
McHa.rgue, J. S. 1920. The significance of the peroxidase reaction with reference to the
viability of seeds. Journal of the American Chemical Sacuety 42:612-615.
7
Seed
Dormancy
The ability of seeds to delay their germination until the time and place are right is an
important survival mechanism in plants. Seed dormancy may be a complex and puzzling
challenge to the seed analyst and the seed researcher, but it is the method through which plants
are able to survive and adapt to their environment.
Seed dormancy is a genetically inherited trait (Naylor 1983) whose intensity is modified
by the environment during seed development. Plants with a long history of domestication
generally show less seed dormancy than wild or more recently domesticated species. When
domesticated species exhibit dormancy, they become a problem to seed producers, their
customers, and seed analysts. However, a degree of dormancy in certain crops (e.g., winter
cereals) is desirable since it prevents preharvest sprouting and helps maintain seed quality.
However, dormancy may cause seeds of numerous species to remain ungerminated in the soil
for many years. This explains the presence of unwanted crop plants or weeds in fields that are
cultivated regularly. Considerable attention is now given to the composition of species in soil
seed banks (Fenner 1992; Leck et at. 1989).
A common misconception of seed dormancy is that it is merely a resting state in the
absence of suitable germination conditions. This state is often called quiescence. However, true
dormancy is defined as a state in which seeds are prevented from germinating even under
environmental conditions normally favorable for germination. Several physical and physiological
mechanisms of dormancy, including both primary and secondary dormancy, occur in seeds.

PRIMARY DORMANCY

Primary dormancy is the most common form of dormancy and takes two forms: exogenous
and endogenous dormancy.

Exogenous Dormancy

Exogenous dormancy is a condition in which the essential germination components (e.g.,

water, light, and temperature) are not available to the seed and thus it fails to germinate. This
type of dormancy is generally related to physical properties of the seed coat. However, proper
light conditions and other environmental stimuli favorable for germination may also be absent.
Thus, this form of dormancy is under exogenous control.

L. O. Copeland et al., Principles of Seed Science and Technology

© Kluwer Academic Publishers 2001
Seed Dormancy 141
Factors Responsible for Exogenous Dormancy

There are three factors responsible for exogenous dormancy: water, gases, and mechanical
restriction.
Water. The impermeability of seed coats to water is typical of many species in a number
of families (e.g., Fabaceae, Malvaceae, Chenopodiaceae, Liliaceae). Seeds that exhibit water
impermeability are known as hard seeds. Water impermeability is caused by both genetic and
environmental factors.
The effect of genotype (variety), or inheritance, on seed coat impermeability is variable.
Dexter (1955) reported only small differences in seed impermeability of different alfalfa
varieties. However, most workers have found this trait to be highly heritable. One study showed
that 80% of 388 bean varieties studied had impermeable seed coats with a range of 1-79%
(Gloyer 1932). Genetic control of seed impermeability was demonstrated in another study in
which crimson clover breeding lines were selected for water impermeability for nine generations
during which the impermeable seed content was increased from 1% to 63 % (Bennett 1959). The
result ofthis breeding program was the crimson clover variety "Chief" with superior reseeding
characteristics. Two genes control hardseededness in cotton seeds (Lee 1975). Similarly, two
or more genes control the semihard condition in garden bean seeds (Dickson and Boettger 1982)
and a single recessive gene modifies the expression of hard seeds in lentil (Ladizinsky 1985).
The environment also influences seed impermeability, although little is known about the
nature of this controL Weather and soil conditions during the final stages of seed maturation are
especially influentiaL Lee (1975) showed that the two genes which control hardseededness in
cotton were expressed more in dry conditions during seed maturation. Aitken (1939) reported
that subterranean clover seeds produced on plants under moisture stress were more prone to
hardseededness than seeds produced on plants without moisture stress. Hill et al. (1986)
concluded that high soil moisture availability during soybean seed fill reduced seed coat
impermeability. Thicker and more impermeable seed coats occurred on soybean seeds
developing under mineral nutrient-deficient conditions (Nooden et al. 1985).
While genetics and the environment modify the expression of hard seededness, the cause of
this trait is a subject of considerable debate. What is known is that impermeability continues to
increase with decreases in seed moisture content (Hyde 1954; Quinlivan 1971; Standifer et at.
1989) and seeds do not become impermeable until their moisture content decreases to about 14
%. Hyde (1954) demonstrated that the dry-down process in hardseeded white clover seeds was
regulated by the hilum which acted as a one-way valve, closing when relative humidities were
high and opening when relative humidities were low. This results in a continual loss of moisture
content in hard seeds (Figure 7.1). A similar mechanism has been proposed for species in the
Caesalpinaceae and Mimosaceae (Werker 1980/1981).
The actual cause of hardseededness has been attributed to both physical and chemical
attributes of the seed coat (Tran and Cavanagh 1984; Eg 1ey 1989). The impermeability to water
may be due to the presence of a cuticle and a well-developed layer of palisade cells or both.
Heavy deposits of suberin, lignin, or cutin are common in the integuments of many legume seeds
(Comer 1976) as well as those of other hard-coated species. The cuticle of bean seeds can
effectively limit water penetration, leaving only the micropyle for imbibition (Pammel 1899).
It has been demonstrated that Great Northern bean seeds with the micropyle sealed off gain only
0.25% of their weight while control seeds gain 79%. According to Kyle (1955), the raphe of
142 Seed Dormancy

70
16

....
5 12
.r--:,
,:
.
I
u
L. >(

ttl
I
t.! I
~ g I I
c •• I

8 • I
I
Z! 6 I·
I
~ -<I- Hard seeds t•
£. ,
4

2
••>C •• ScariFed ~eeds
\ ,
~ •••J!

°o~~------~----~----~-----L----~----~
4 11 18 25 32 39 46
Storsge time - da~s

Figure 7.1. Changes in moisture content in white clover (Trifolium repens) hard or scarified seeds
transferred once a week successively to chambers ofdifferent relative humidity (70%, 30%, O%) (From
Hyde 1954).

Great Northern bean seed contributes more to water entry during the fIrst 24 hours than either
the micropyle or seed coat. Cutin deposits have been reported in the nucellar layers of
watermelon seeds (Thornton 1968).
In other seeds, the impermeability to water may be related to the structure of the hilum
(Gutterman 1993). A mucilaginous layer beneath the cuticle which is hard and thick has been
observed in legume seeds (Werker et al. 1979). In cotton (Figure 7.2), the palisade layer
becomes discontinuous at the chalazal region and the cap is blocked by a plug of closely packed
parenchyma and mesophyll cells that are high in tannin content. In those cotton seeds that are
hard, this palisade layer is tightly adherent to the chalazaI cap and its cells are heavily lignifIed
(Werker 1980/1981). In other seeds, the strophiolar cleft, which is a small opening in the seed
coat near the hilum, is thought to control water uptake. The strophiolar cleft is fIlled with a
corklike substance of suberin called the strophiolar plug (Hagon and Ballard 1970; Dell 1980;
Kelly and Van Staden 1987). This plug must be removed or loosened before water uptake can
occur. Vigorous shaking of seeds possessing this mechanism can often either loosen or disrupt
the strophiolar plug and pennit water entry. This process is called impaction and may be used
to break the dormancy of sweetclover (Melitotus alba). Other water-impermeable seeds do not
possess a strophiolar plug and the seed coat must be abraded or punctured to eliminate
dormancy. Thus, different parts of the seed are important in the control of water entry during
the initial stages of imbibition.
Seed Dormancy 143

Chalazal cap
Epidermis
Outer
pigment
laye,
Colourless Brow"
layer seed
POhsade
coot
layer
Inner
pll~ment
layer

Fringe
celis Whitt
} "membrane"
HMH-- Living
cells

--Embryo

~~SL--- Micropyle

Figure 7.2. A schematic drawing of cotton seed coat (From Marchaim et al. 1974).

Gases. The impenneability of gases through the seed coat has been described as a
mechanism ofdonnancy governed by the seed coat. This is somewhat surprising since water and
oxygen represent small molecules of similar molecular weight, yet the seed coat is able to be
selectively penneable to one but not the other. In addition, the mechanism by which gases are
restricted from the embryo is difficult to detennine because of the volatile nature of the gases
and the small seed tissues through which the gases flow. Still, the nucellar membrane of
cucumbers (Brown 1940), the pericarp of Fraxinus excelsior (Villiers and Wareing 1964), and
the endocarp of coffee "seeds" (Huxley 1965) are known to restrict the entry of oxygen. The
general mechanism of action is thought to be related to seed coat permeability to oxygen:
Dormant seeds are less permeable to oxygen, thus retarding the aerobic metabolism required for
germination. This appears to be the case in a number of grasses (Major and Roberts 1968;
Landgraff and Juntilla 1979; Probert et at. 1985).
The causes of gas impermeability in seeds have been attributed to physical and biochemical
barriers. From a physical perspective, it may be that the mere process of imbibition replaces
pore spaces in the seed coat with water that hinders the ready movement of gases to the embryo.
Many cruciferous seeds also have epidermal cells in the seed coat that are mucilaginous and
swell when wetted (Vaughan and Whitehouse 1971). This makes the path of oxygen diffusion
longer before it actually reaches the embryo. From a biochemical perspective, chemical
compounds in the seed coat may consume oxygen as it permeates the testa, thus reducing the
amount of oxygen to the embryo. For example, apple seeds have restricted oxygen permeability
144 Seed Dormancy
at 20°C compared to 4°C (Come 1968) indicating a temperature-oxygen permeability
interaction. Later studies suggested that the high level of phenolic compounds found in the seed
coats were oxidized to quinones by enzymatic reactions which were more rapid at the higher
temperatures, thus reducing the rate of oxygen diffusion to the embryo (Come and Tissaoui
1973). Peroxidases found in seed coats also have been reported to reduce the availability of
oxygen to the embryo (Renard and Capelle 1976). Other biotic factors also influence seed coat
permeability to gases. The decomposition of plant material in the soil produces saponins which
can alter oxygen permeability of alfalfa seeds (Marchaim et al. 1972) and possibly other seeds
as well.
Other seeds have been shown to be differentially permeable to oxygen and carbon dioxide.
The nucellar membrane of cucumber seeds is one example in which the inner membrane is more
permeable to CO2 (15.5 ml/cm2/hr) than to oxygen (4.3 mVcm2/hr) (Brown 1940). Perhaps the
best~known example of seed coat impermeability to oxygen is Xanthium, or cocklebur. For
many years, we have known that the dimorphic seeds present in the bur differ in their
germination capacity. The upper, smaller seed requires pure oxygen for 100% germination,
while the lower seed needs only 6% oxygen for complete germination. The germination of these
dimorphic seeds under varying oxygen levels was used as the classical example to illustrate the
selective gas permeability in some seeds. However, other data now indicate that the small, upper
seed of Xanthium contains a much greater quantity of inhibitor than the larger, lower seed
(Wareing and Foda 1957). It has been suggested that the small seed requires more oxygen to
oxidize and inactivate the inhibitor before 100% germination is achieved (Porter and Wareing
1974). Thus, this classical example of Xanthium demonstrating seed coat impermeability to
oxygen demonstrates the complexity of seed coat effects on impermeability of gases as a cause
of seed dormancy.
Mechanical Restriction. Dormancy has also been attributed to the physical restraint by
the seed coats on an enlarging embryo. This assumes that the thrust developed during imbibition
and growth is inadequate to rupture the seed coat and permit germination. Most evidence to
support this contention is based on the observation that removal of a thick-walled seed coat
produces germination. According to Koller (1955), this type of dormancy has been described
in seeds of water plantain (Alisma plantago), pigweed (Amaranthus retroflexus), raspberry
(Rubus idaeus), peach (Prunus persica), and cherry (Prunus carasus). However, it should be
noted that seed coats are often the source of inhibiting substances which are also eliminated
during seed coat removal. Additionally, seed coats may interfere with the leaching of inhibitors
or restriction of water flow, accounting for a myriad of changes that occur with the removal of
seed coats. To date, no experimental evidence unequivocably demonstrates that seed coats act
as a mechanical obstruction to the germinating embryo. More studies are needed to measure the
mechanical resistance ofthe seed coats as well as the force produced by a germinating embryo.
Only one study has attempted to provide this information. Esashi and Leopold (1968) concluded
that dormant seeds of Xanthium required slightly more force to rupture the seed coats than
nondormant seeds. More importantly, they showed that the nondormant embryo developed twice
the thrust of a dormant embryo, clearly indicating that the physical restraint ofthe seed coat was
not the major dormancy-imposing mechanism for these seeds (Table 7.1). Further studies of this
nature are needed to establish whether seed coat restriction is a causal factor in seed dormancy.
Seed Dormancy 145
Methods of Breaking Exogenous Donnancy

Under natural conditions, exogenous dormancy is overcome by the freezing-thawing of the

soil, ingestion by animals, microorganism activity, forest fires, natural soil acidity, and other
factors. All of these factors affect the integrity ofthe seed coat in some way. For example,
temperature fluctuations cause a gradual expansion and contraction of cells that eventually
disrupt the testa at weak points such as the strophiole (Quinlivan 1966; Taylor 1981; Egley
1989) and leads to germination (Figure 7.3). Such processes may take many years to complete
and thus serve to expand the range of time over which seeds germinate. While this may represent
an advantage to some species, in many cases we need to ensure that crop seeds will germinate
more rapidly and uniformly. Overcoming this dormancy is accomplished by the mechanical and
chemical removal of the seed coat, a process called scarification.
Mechanical Scarification. Grinding seeds with abrasives or sand or shaking them
(impaction) are techniques that are often used to scarify seed coats. Other techniques such as
heating, chilling, drastic temperature shifts, brief immersion in boiling water, piercing the seed
coat with a needle (Table 7.2), or exposure to certain radio frequencies alter seed coat integrity,
permitting penetration of both water and gases. However, the duration of these treatments is
critical, since prolonged treatment may result in seed damage while brief treatments may not be
sufficient to break dormancy.
Chemical Scarification. Seeds may also be treated with chemicals to cause degradation
of the seed coat. Sulfuric acid has been used most widely and is effective in industrial and
concentrated forms. Other compounds such as sodium hypochlorite and hydrogen peroxide have
also been reported to scarify seeds (Hsiao and Quick 1984). However, chemical scarification
has not been commercially popular because the materials are hazardous to handle, the seeds
must be thoroughly washed and dried after treatment, and reduction of germination may occur
from even slight overscarification.
Recent techniques include the use of selective seed coat enzymes such as cellulase and
pectinase to degrade seed coats (Brant et al. 1971; Lester 1985). Since many seed coats contain
water-insoluble compounds that retard water entry into the seed, organic solvents such as
alcohol and acetone have been used to dissolve and remove these insoluble constituents and
permit imbibition (Rolston 1978). It should be emphasized that while scarification enhances
germination, it invariably leads to seed injury due to the disruption of essential cells. This
enhances fungal invasion and mechanical injury.

Table 7.1. The Amount of Embryonic

Thrust Developed and Force Required to Rupture
the Seed Coats of Nondormant and Dormant Cocklebur Seeds.
Force
Physiological State Thrust Developed Required to Rupture
Nondormant 84 67
Dormant 41 56
From Esashi and Leopold (1968).
146 Seed Dormancy

60

50
~
t- -0-- 1S·30·C
Z
w 40
t- '" '. ". ..'l!i ............ 6
Z .. .. tJ: ...
0 1S-IS·C
U
0 30
w --G-
w
(J)
lS·60·C

0
a:
<{ 20 -e- 1S·7S·C
I

10

O+----r----.---,,---.----~--_r--~
o 30 60 90 120 150 180 210

TIME INTERVAL (DAYS)

Figure 7.3. The rates ofsoftening of Geraldton subterranean clover (Trifolium subterranean) seeds
during 210 days of dry storage with increasing amplitudes of the temperature fluctuations (From
Quinlivan 1966).

Table 7.2. The Effects of Piercing and Hull

Removal on the Germination Percentage of Dormant Oat Seeds.
Hulls pierced with Hulls removed,
Tested at needle, then tested at then tested at
Test No. 20· C for 10 days 20· C for 10 days 20· C for 10 days
1 43 94 98
2 36 90 79
3 47 95 91
4 10 84 87
5 77 96 96
6 74 90 92
7 75 91 84
8 71 88 93
9 75 88 93
10 89 95 97

Average 59.7 91.1 91

From Forward (1958).
Seed Dormancy 147
Endogenous Dormancy

Endogenous dormancy is the most prevalent dormancy found in seeds and is due to the
inherent properties of the seed. For example, the seed may possess an excess of inhibitors that
must be removed or reduced prior to germination. This form of dormancy, therefore, is under
endogenous control.
Causes of Endogenous Dormancy. Environmental conditions during seed development
and maturation influence the duration of endogenous dormancy. Among these factors are day
length, moisture status, position of the seed in the fruit or inflorescence, age ofthe mother plant,
and temperature during seed maturation.
The day length experienced by the mother plant influences the dormancy of developing
seeds. This effect is greatest in the final stages of seed maturation (Gutterman 1978). Plants
differ in their response to photoperiod and their expression of seed dormancy (Gutterman 1993).
For example, longer days promote the germination of Polypogon monspeliensis (Table 7.3),
Carrichtera annua, and Cucumis prophetarum seeds. Short days enhance the germination in
Chenopodium album, Portulaca oleracea, and Lycopersicon esculentum. The reasons for the
day-length effect on seed dormancy are still not known. In some cases, long days result in
increased seed coat thickness (Dome 1981; Pourrat and Jacques 1975). In other cases, fruits
matured under long days evolve more ethylene gas (Gutterman 1978).
The moisture status of the mother plant or developing seed also influences the degree of
seed dormancy. Water deficits increase barley seed dormancy when they occur close to
flowering but decrease dormancy at the final stages of seed maturation (Aspinall 1965). In
Avena jatua, water stress on the parent plant during seed maturation reduces the level of
dormancy (Simpson 1990). Desiccation ofthe developing seed decreases dormancy in wheat and
barley (Nicholls 1986) and exposure of developing cereal seeds to high relative humidities
causes preharvest sprouting (Black et al. 1987). It has been proposed that water stress during
seed maturation causes the seed to switch from a developmental to a germinative system
(Kermode et al. 1986).

Table 7.3. The Influence of Different Day Lengths Under Greenhouse and Outdoor
Conditions on Po/ypogon monspeliensis During Growth and Seed Maturation and
on Seed Germinability.

Day length during Seeds from greenhouse Seeds from outdoor

growth of mother plants Germination (%) plants Germination (%)
plant and seed
maturation (h) 3 days 7 days 3 days 7 days
9.0 0.0 0.0 1.5 10.0
11.0 0.0 0.0 9.5 17.0
12.0 17.5 19.0 64.0 86.5
13.5 38.0 44.5 98.5 98.5
15.0 60.5 66.0 97.0 98.5
18.0 90.5 91.0 98.0 99.0
Control natural 93.0 96.0 91.5 93.0
daylength
From Gutterman (1982).
148 Seed Dormancy

Position of the seed on the mother plant also influences dormancy. These effects are
primarily attributed to differences in seed maturation which are often expressed by seed weight.
The classic example is with members of the Apiaceae (carrot) family where the inflorescence
is an umbel which is produced in sequential order from primary to secondary, etc. Table 7.4
shows that seeds from primary umbels are heavier, more mature, and more dormant than those
produced elsewhere on the plant (Thomas et al. 1979). These differences in dormancy serve as
an effective mechanism for maintaining viable but dormant seeds in the soil seed bank
(Gutterman 1992).
Age of the mother plant at the time of flower induction can also affect seed dormancy. For
example, dormancy of seeds from Amaranthus retroflexus (Kigel et al. 1979) and Oldenlandia
corymbosa (DoCao et al. 1978) plants increased as the age of the plant increased. Temperature
during seed maturation also influences the expression of dormancy. In general, higher
temperatures produce less dormant seeds. For example, Datta et al. (1972) showed thatAegi/ops
ovata plants exposed to 28/22°C produced more germinable seeds than when exposed to
15/10°C.
Environmental conditions during seed development and maturation also influence the
duration of endogenous dormancy. The level of dormancy in seeds oftouch-me-not (Impatiens
balsam ina) depends on water supply and mineral nutrition, especially nitrogen, during seed
formation and development. Seeds from plants that had been adequately watered and well
supplied with nitrogen had less dormancy than those from plants deficient in these factors
(Junges and Ludwig 1963). Lettuce (Lactuca sativa) seed germination requirements have been
shown to differ when grown under different environments (Koller 1962). Stage of maturity has
little effect on the duration of endogenous dormancy in Kentucky bluegrass seeds following
harvest, but the level of dormancy was directly associated with the seed moisture content at the
time of harvest: The higher the moisture content, the greater the degree of dormancy (Delouche
1958). Table 7.5 shows the effect of maturity on endogenous seed dormancy in barley.

Table 7.4. Seed Position of Umbel, Weight (mg) and Germination (%) After 21 Days at 18°C
in light in Three Celery (Apium graveolens) Cultivars. LSD at 5% in Parenthesis.

Umbel Mean seed

Cultivars position weight (mg) Germination (%)
Green snap Primary 0.590 51
Secondary 0.440 85
Tertiary 0.386 94
Quaternary 0.382 80
(0.069) (9.8)
Lathom Blanching Primary 0.474 50
Secondary 0.438 72
Tertiary 0.380 94
Quaternary 0.348 82
(0.069) (9.2)
Ely White Primary 0.590 59
Secondary 0.468 62
Tertiary 0.490 80
Quaternary 0.520 87
(0.086) (7.3)
From Thomas et a!. (1979).
Seed Dormancy 149
Unlike exogenous dormancy, which requires a physical alteration of the seed coat, only
physiological changes such as rudimentary embryo maturation, response to growth regulators,
changes in temperature, exposure to light, and endogenous rhythms are able to relieve
endogenous dormancy in seeds.

Rudimentary Embryo Dormancy

Seeds of some species are shed before they are morphologically mature. This results in
dormancy because the immature embryo is unab Ie to germinate. Rudimentary embryo dormancy
occurs in Ranunculus, Plantago, Fraxinus, Viburnum, flex, and Pinus. Further embryo
maturation occurs following seed dispersal and may take a few days or several months. The
embryo of holly (flex opaca) is an undifferentiated mass of cells when the seed is shed, but
during subsequent maturation the cells become a well-defined structure (Ives 1923). Cherry
seed embryos increase in size, weight, length of leaf primordium, and oxygen uptake of the
embryonic axis after being shed from the tree (Pollock and Olney 1959). Similar changes occur
in other species (Zagaja 1962; Scott and Waugh 1941; Zagaja and Czapski 1962). In L'iOpyrum
biternatum, seed dispersal occurs in late May but embryo growth is delayed by high
temperatures until October (Baskin and Baskin 1986). In contrast, Apium graviolens seeds
require light before embryo growth continues (Jacobsen and Pressman 1979). The changes that
occur in dry, dormant seeds after dispersal is known as after-ripening. This process will be
more fully discussed later in this chapter.

Physiological Dormancy

Seed dormancy in higher plants is generally believed to be regulated by a balance of

endogenous growth inhibitors and promoters (Amen 1968). Thus, dormancy may be considered
a result of the presence of growth inhibitors, the absence of growth promoters, or a combination

Table 7.5. Timetable of BarJey Seed Dormancy During Seed Development

(Germination Tests at 15°C).

1-4 weeks Condition of

after anthesis 4-6 weeks 6-8 weeks Rain in 9th week ripe seed sample
No germination Rise of Germination Reduction of Majority
germination percentage germ inability nondormant
percentage over 90% of 14%
Moisture content Moisture content Moisture content Rise of moisture Some still in
over 80% decreasing ca. 16% content to ca. 30% primary dormancy
Isolated embryo Some still in
germinated secondary
dormancy
Covering layers Covering layers Collapse of cells
green turn yellow of covering layers,
disorganization
of their contents
Primary Dormancy No dormancy Secondary
dormancy disappearing dormancy
From Evenari (1965).
150 Seed Dormancy

of both. The levels of these endogenous compounds are controlled by certain environmental
stimuli such as light and temperature. Many substances that are present in seeds help determine
whether a seed will be dormant. The goal of research workers is to identity these componds and
determine their metabolic function in order to better understand their role in the regulation of
seed dormancy. A large number of compounds have been isolated that can induce dormancy
through their influence on metabolic and osmotic inhibition.
Metabolic Inhibition. Certain compounds that are present in seeds inhibit specific
metabolic pathways. An example of this type of inhibitor is cyanide, which is found in apple and
peach seeds. Such compounds act by suppressing germination through their effect on
respiration. However, at very low concentrations, cyanide stimulates germination (Taylors on
1988; Tilsner and Upadhyaya 1987) and this effect is pH-dependent (Cohn and Hughes 1987).
Phenolic compounds also inhibit germination, and because oftheir widespread occurrence,
have been regarded as natural germination inhibitors. Compounds such as substituted phenols
and cresols have been shown to inhibit germination, although they are not true dormancy-
inducing compounds {picman and Picman 1984; Rice 1983}. The first dormancy-inducing
inhibitor found was coumarin, which is characterized by an aromatic ring and represents an
unsaturated lactone structure. Coumarin is widely distributed and rapidly metabolized in seeds
and is considered a natural germination inhibitor. Additionally, coumarin derivatives such as the
glycosides of the lactone or substituted coumarins have also been found in many fruits,
supporting the role of coumarin as a natural seed inhibitor. The exact mechanism of coumarin
inhibition has yet to be elucidated, although it is suspected that it interferes with respiration and
oxidative phosphorylation and indirectly with the availability of energy by its effect on
phosphorous metabolism.
Another extremely active inhibitor of seed germination was discovered in 1966 (Cornforth
et al. 1966). Because of its dormancy-inducing properties, the compound was initially named
dormin. Later, chemical characterization revealed that dormin had the identical chemical
structure of abscisic acid - a compound that was being investigated for its promotion of the
abscission of young cotton fruit and its ability to induce bud dormancy in trees. By general
agreement, the name of abscisic acid (ABA) was given to dorm in in 1967.
Since these initial reports, many studies have documented that ABA is present in a wide
range of seeds and is a naturally occurring endogenous hormone that is active at very low
concentrations. Further, reports have shown that the levels of endogenous ABA are reduced
following stratification of dormant ash, rose, and other seeds (Sondseimer et al. 1968; William,
et al. 1973; Webb et al. 1973).
ABA has also been shown to be localized in the testa of apple seeds (Lewak and Rudnicki
1977) and is translocated to the embryo during imbibition to induce dormancy. The exact
mechanism of ABA inhibition of germination has not been clearly defmed. It is known to inhibit
the synthesis of enzymes that are important in the early stages of germination (Varty et al.
1983), perhaps by inhibiting their translation from mRNA (Ho and Varner 1976).
Due to the ubiquitous occurrence and reported dormancy inducing capability of ABA,
attempts have been made to determine whether promotive hormones such as gibberellins could
interact with ABA in the recognized inhibitor-promotor hypothesis.
Many studies have, in fact, shown that ABA can completely or partially reverse the pro-
motive action of either the gibberellins or cytokinins (Rudnicki et al. 1972). Additionally, it has
been demonstrated that as ABA levels decrease, gibberellin and cytokinin levels increase during
stratification in seeds such as sugar maple (Van Staden et al. 1972; Webb et al. 1973). Thus,
Seed Dormancy 151

an inverse relationship between inhibitors and promotors occurs during the dormancy-breaking
(after-ripening) process. However, it should be emphasized that many of these reports are still
preliminary and many were conducted using bioassay techniques. Correlations between
inhibitor levels and dormancy in seeds do not demonstrate direct physiological roles until these
are determined. More refined analyses are necessary before a general role in the induction of
dormancy can be attributed to ABA. We should also emphasize that ABA is not the only natural
germination inhibitor in seeds. Other compounds such as coumarin may serve a similar function.
Osmotic Inhibition. Many substances possessing high osmotic pressures can inhibit the
germination of seeds. Compounds such as sugars or salts in sufficient concentration may
compete so successfully for water that the seed never becomes fully imbibed and thus remains
ungerminated (quiescent). However, such seeds readily germinate when removed from the
osmotic-inhibiting environment. Many such substances are located in the fruits or fruit walls
that surround the seeds. One study concluded that the osmotic pressure exerted by inorganic
substances in the fruit ball of sugar beet is responsible for the inhibition of germination (Duym
et al. 1947). In another study, the concentration of electrolytes from sugar beet fruit ball extracts
retarded the growth of wheat seedlings (Snyder et al. 1965). Demineralized extracts of sugar
beet fruits are also known to inhibit germination of peppergrass and other species (Frorchell
1957); the cause is attributed to the presence of a specific organic substance or substances
rather than osmotic inhibition. Ferulic and caffeic acids occur in tomato fruit (Ackerman and
Ve1dstra 1949); parasorbic acid in fruits of European mountain ash (Sorbus aucuparia) (Kuhn
et al. 1943); and a mixture of organic acids exists in fruits oflemons, strawberries, and apricots
(Varga 1957). These substances are known to be capable of inhibiting germination. Tomato
juice completely inhibits the germination of garden cress (Lepidium sarivum) seeds even when
diluted in a proportion of 1:25 (Meyer et al. 1960). It is well known that seed germination is
delayed in most fleshy fruits while the seed is still embedded in the fruit itself. Thus, the high
osmotic pressure of such fruit juices undoubtedly contributes to seed dormancy.
Physiological studies of imbibition have frequently utilized the concept of osmotic
inhibition to allow partial intake of water into the seed while preventing it from germinating.
Initially, many of these investigations were conducted with salts until it was revealed that the
seeds often experience deleterious effects due to ionic toxicity. Subsequently, such studies began
to employ mannitol as the osmotic-inhibiting compound. However, it was again demonstrated
that mannitol was imbibed into the seed and altered later metabolic functions. More recent
studies have utilized polyethylene glycol, which is a long-chain polymer incapable of being taken
into the intact seed but still water soluble and, therefore, osmotically active.
Methods of Breaking Physiological Dormancy. There are various methods of alleviating
physiological dormancy. Seeds that are dormant due to osmotic inhibition can be germinated
after removing the seed from the influence of the inhibitor or diluting the inhibitor from around
the seed, a process called leaching. Leaching typically requires exposing the seeds to an excess
of water that dilutes or removes the inhibitor from the seed. An example of seeds that are
typically leached to remove an inhibitor from the fruit wall is sugar beet seeds, compared to
tomato seed that can be germinated only after removing from the juice which otherwise causes
osmotic inhibition.
Seeds that possess metabolic inhibitors located in the seed coat can be relieved of dormancy
by removal of the seed coat by mechanical or chemical scarification. Conventional scarification
to break down otherwise hard, impermeable seed coats to allow water penetration is usually
accomplished by various procedures including the use of abrasives or acids, or by simply
152 Seed Dormancy

piercing the seed coat. However, scarification may also be effective in degradation of the seed
coat, alleviating physiological dormancy and permitting germination by one of two processes.
First, seed coats have often been shown to be the source of inhibitors, and when they are
removed, the inhibitor is also removed. Second, seed coats can function as differentially
permeable membranes, permitting the entry of water but retarding the loss of inhibitors.
Removing the seed coats eliminates the barrier to inhibitors, resulting in leaching of the
inhibitors from their site of action, and thus permitting germination. An example ofthis process
is provided by Glory cabbage seed which has an inhibitor in the seed coats (Cox et al. 1945).
Soaking these seeds in dilute sulfuric acid for several minutes results in rapid germination.

Temperature Requirement

Seeds with a specific temperature requirement for germination often contain both inhibitors
and promotors. Evidence now exists to support the view that dormancy of this type is controlled
by an inhibitor-promotor balance that is altered by exposing seeds to low temperatures under
imbibed conditions (stratification) or under higher temperatures while unimbibed.
Stratification. It is now well documented that both physical and physiological changes may
occur in imbibed seeds exposed to low temperatures. Embryo growth ,occurs in stratified apple
embryos (Table 7.6) which will otherwise not germinate. Similarly, the embryonic axis of
stratified cherry seeds increases in cell number, dry weight, and total length (Olney and Pollock
1960). On a cellular basis, increased oxygen uptake and energy supply to the embryonic axis
of stratified seeds has been observed. Increases in catalase, phosphatase, alkaline lipase, and
peroxidase enzymes have also been detected (Zarska-Maciejewska and Lewak 1976). Thus, it
appears that many phases of embryonic development and metabolism are influenced by
stratification. All of these may be considered as after-ripening processes.
Shifts in hormonal levels also occur in stratified seeds. For example, the level of ABA
drops during stratification of apple, ash, walnut, and hazelnut seeds. However, the addition of
exogenous gibberellins can substitute for the stratification requirement in some seeds (Winfield
1968), implicating this hormone as a promotive agent. Increases in the levels of endogenous
gibberellins have been observed during stratification, supporting this premise (Frankland and
Table 7.6. Development of Apple Embryos Isolated After Different Times of Stratification
and Subsequently Cultured at 25°C for 9 Days.

Embryos with Embryos with only

Weeks of both greening one greening Mean length of
stratification Germination (%) cotyledons (%) cotyledon (%) main root (mm)
a 9 7 42 1-9
2 12 22 27 2.2
4 24 73 5 3.7
6 39 97 2 9.1
8 51 100 a 9.9
10 59 100 0 10.9
12 65 100 a 11.9
14 79 100 0 21.4
From Lewak and Rudnicki (1977).
Seed Dormancy 153
Wareing 1966). Using gibberellin-deficient mutants, Karssen et al. (1989) showed that low-
temperature exposure of imbibed seeds increased their sensitivity to gibberellins. Thus, beyond
the observed increases in growth and metabolic activity of stratified seeds, shifts in the levels
of inhibitors and promotors have also been detected that further contribute to after-ripening and
the release from dormancy.
The Rules for Testing Seeds of both the Association of Official Seed Analysts and the
International Seed Testing Association (see Chapter 15) list stratification as an aid for
germinating seeds of many species. Since the analyst is seldom aware of the history of the seed
lot being tested, stratification is ordinarily performed as a routine part of the seed testing
procedure. The effect ofprechilling (stratification) is shown in Table 7.7.
The germination rate of many seeds may also be increased if exposed to daily alternating
temperature cycles. Germination of Chinese red pine (Pinus densiflora) and Japanese black
pine (P. thunbergii) seeds was improved under such conditions. Seeds oftigertail spruce (Picea
poUta), which needed light to germinate at a constant temperature, germinated in the dark under
alternating temperatures (Asakawa 1959). The need for alternating temperatures is apparently
associated with endogenous dormancy and characterizes many agricultural species.
Stratified seeds are usually preconditioned at temperatures between 3 and 10°C, although
the specific temperatures and duration of exposure may vary. For some species, low-temperature
stratification is an absolute requirement for germination; for others, it may only hasten
germination and increase the speed of growth. For some species, stratification may decrease the
sensitivity to external conditions, thus increasing the range of temperatures under which
germination can proceed. Such is the case for sugar pine (Pinus lambertiana) seeds that
germinated only at temperatures above 25°C, but after stratification were able to germinate at
lower temperatures (Stone 1957). The length oftime required for stratification varies depending
on the species. Seeds of wild rose (Rosa multiflora) require a two-month stratification period
(Crocker and Barton 1931), however swamp persicaria (Polygonum coccineum) and mild water
Table 7.7. The Effect of Prechilling (vs. Control) on the Gennination of
Oat Seed.

Pre chilled at 10' C

Tested for 5 days, then tested
Test No. at 20' C for 10 days at 20' C for 5 days
1 43 99
2 36 B5
3 47 97
4 10 91
5 77 97
6 74 97
7 75 9B
B 71 96
9 75 9B
10 B9 100
Average 59,7 95.8
From Forward (1958).
154 Seed Dormancy

pepper (P. hydropiperoides) may require an 8 II2-month stratification period (Justice] 944).
The seeds of more than 60 species, many of them woody types, have been listed as requiring
low-temperature stratification for germination (Crocker and Barton 1957).
The stratification requirement of a particular seed lot also depends on the intensity of the
temperature exposure and seed age. Meyer et al. (1990) showed that the depth of dormancy for
a range of ecotypes of Artemisia tridentata correlated with the mean January temperature:
colder temperatures required longer stratification periods. Freshly harvested seeds of Chinese
maple (Acer truncatum), which require a two-month stratification, germinated well without
pretreatment after a year of storage (Ackerman 1957). Decrease in the persistence of dormancy
with increasing age of seeds is a universal characteristic of endogenous dormancy, but the speed
at which dormancy is lost varies among species.
After-Ripening. The disappearance of dormancy in dry seeds over time at room
temperature is widespread among many seeds and almost universal in cereals. For most cereals,
storage for 1 to 2 months at 15 to 20°C suffices to allow maximum germination. In Ph/eum
arenarium seeds, longer periods of dry seed after-ripening not only result in increased
germination but also increased germination over a broader range oftemperatures (Figure 7.4).

Light Requirement

The light quantity and quality perceived by a seed depend on its position in the soil, the
vegetative covering, and the light absorption characteristics of the seed coat or comparable
structure(s). Measurable quantities of light do not penetrate the soil to greater depths than a few
millimeters or centimeters depending on soil type (Tester and Morris 1987). Sandy soil has the
greatest light transmission which is reduced with increasing levels of humus. In addition, the
100

80

60
c
.51
;;
's.
c
40
,~
.....
*- 20

o 10 2S 30
Temperature (0c)
Figure 7.4. Dry after-ripening in seeds of Phleum arenarium. Changes in the germination response
to temperature during storage at 15°C and 15%R.H. Germination was tested on a thermogradient bar
after 1, (.. ); 6 (II); and 13 months (~storage. The seeds were incubated on wettedfilter paper and
received an 8-h photoperiod/day (From Probert 1992).
Seed Dormancy 155
quality ofthe light is also modified by soils. Shorter wavelengths oflight are absorbed more by
soils than longer wavelengths (Bliss and Smith 1985). The presence of a leafed canopy over the
soil also reduces the photosynthetically active (400-700 run) portion of the light spectrum. Thus,
canopy shade is richer in far-red and poorer in red portions of the spectrum. The light-absorbing
phytochrome pigment which controls germination in seeds is believed to be located in the
embryo. As a result, seed coats that are thick and/or intensely pigmented can alter both the
quantity and quality of the light perceived by the embryo (Widell and Vogelmann 1988).
Light intensity, wavelength, and photoperiod are all known to affect the germination of
seeds that have physiological dormancy. Three well-known species whose dormancy is broken
by exposure to red light (670 run) are lettuce, birch, and Virginia pine.
Duration of light exposure (photoperiod) can also affect seed germination. The seeds of
some species respond to short days, others to long days, while still others are unaffected by
daylength.
Continuous light may inhibit germination of some seeds, and thus impose a different kind
of dormancy. Continuous light is reported to inhibit both onion and leek seeds (Lovato and
Amaducci 1965). Although light may initially promote the radicle growth of Douglas fir, an
extended period of light may become inhibitory, especially for seeds with large radicle size
(Villiers 1961).
For a more complete discussion of the influence of light on seed germination, refer to
Chapter 5.

Circadian Rhythms

Plants have long been observed to follow an orderly sequence of growth and developmental
processes. There is evidence that they are somehow able to measure time independently of the
outside environment, and thus they are able to regulate certain growth and developmental
processes (Cummings and Wagner 1968). This orderly sequence of growth and development is
referred to as circadian rhythms.
Circadian rhythms also seem to influence the pattern of seed germination. They have been
classified as those of a single yearly cycle and those with more than one cycle per year
(Kummerow 1965). Several hundred species have been reported to show yearly oscillations
(Banning 1965). A periodicity of pigweed (Amaranlhus relrofiexus) seed germination has been
reported when the seed was held moist at 20°C for 78 months (Crocker and Barton 1957); two
maxima occurred, one at 8-10 months and the other at 20-22 months. Maguire (1969) reported
circadian-controlled rhythm patterns in bluegrass seed germination (Figure 7.5). In a study of
over 300 species, Baskin and Baskin (1988) showed that seasonal temperature changes
influenced the breaking of dormancy in a number of species.

Interaction of Primary Dormancy Mechanisms

The types of dormancy described are by no means mutually exclusive, and more than one
mechanism for the imposition of dormancy may be possessed by seeds. Such an example is
provided by Indian rice grass (Oryzopsis hymenoides) seeds which possess both seed coat
(exogenous) and physiological (endogenous) dormancy (McDonald and Khan 1977). In this
species, seed coat dormancy can be effectively removed by scarification with sulfuric acid.
However, addition of exogenous GA3 further enhances germination to 70% for freshly harvested
156 Seed Dormancy

100
90

80
-- ...., " ~;
;--------............ ..... ...... --
70 I';
c COUGAR
.g 60
o::l
c
'E... 50
dJ
<.:> 40
~
30
20
10
0
I 2 3 4 5 6 7 8
A - 1964 Seed Months after harvest

100
90
80 COUGAR
/ . . ,1
,
70
c 60 I '"
,2
'(;:l
c 50 /'
E
...
dJ 40
<.:>
~ 30
20
10
01
1 2 3 4 5 6 7 8
B - 1965 Seed Months after harvest

Figure 7,5. Endogenous germination rhythms observed in Cougar and Newport varieties ofKentucky
bluegrass seed stored at 5°C (From Maguire 1969).

seeds (Table 7.8). Dry seeds that have been after-ripened by natural storage are not as
responsive to exogenous GA3, presumably because the seed has had adequate time to synthesize
this compound.
Seed Dormancy 157

Table 7.8. Influence of Various Exogenous Hormonal Treatments on the Germination of 1- and 2-
Year-Old Intact and Scarified Indian Ricegrass Seeds After 72 Hours Soaking.

1 year 2 years
Treatment Intact Scarified Intact Scarified
H2O 0 31 5 67
10).lM GA3 0 65 5 76
10 ).lM Kinetin 0 54 6 72
10).lM ABA 0 1 2 2
From McDonald and Khan (1977).

Embryo Excision

The complexity of dormancy and its exact cause can often be shown by removing the
embryo and growing it independently of the seed. Flemion (1936) was the first to report that
the embryos of dormant seed could be germinated if they were excised and grown separately.
Other studies have shown that birch seeds, which normally require light, can be germinated in
the dark if excised (Redmond and Robinson 1954). Excised embryos of Canadian hemlock
(Tsuga canadensis) can be germinated in the dark, whereas intact seeds or those with only the
integuments removed required light for germination (Steams and Olson 1958). The light
requirement of Grand Rapids lettuce seed can be eliminated by removing or puncturing the
endosperm surrounding the embryo.
It is possible to force germination of peach seeds normally requiring stratification by
embryo excision or by seed coat removal, but such forced seeds usually produce dwarfed or
deformed seedlings (Mayer and Poljakoff-Mayber 1989).
The excised embryo test for seed viability was discussed in Chapter 6.

SECONDARY DORMANCY

Sometimes nondormant seeds encounter conditions that subsequently cause them to become
dormant. This may be caused by exposure of the seed to conditions that favor germination in
all respects except one. A study of spring wheat and winter barley reported that secondary
dormancy could be imposed by: (a) exposure of dry barley seed to temperatures between 50 and
90°C, (b) seven-day storage of winter barley at high moisture contents at 20°C, (c) one-day
storage of spring wheat at high moisture contents in airtight containers at 50°C, and (d)
placement of the seed under water and in darkness for one to three days at 20°C (Frischknicht
et al. 1961). Exposures of dry seeds to 50°C required four days for imposition of secondary
dormancy; to 70°C required four hours; and to 90°C only one hour. Induction of secondary
dormancy was possible one and one-half months after seeds reached physiological maturity,
although the persistence of dormancy obtained in this way decreased almost continuously as the
time between physiological maturity and treatments increased. For example, compared to
primary dormancy, which was partly broken, secondary dormancy in spring wheat could not be
broken by two weeks of storage at 40°C. However, it was completely broken by treatments of
0.1 % gibberellic acid, 0.5 to 1.0% ethanol, low-temperature stratification, removal of the
158 Seed Dormancy

pericarp above the embryo, and storage at 20°C.

We have discussed secondary dormancy as being either thermo- (temperature), photo-
(light), or skoto- (darkness) imposed (Evenari 1965), though other causes such as imposition
by excess or adverse amounts of water, chemicals, and gases might also be involved. Two
suggestions have been made to explain the mechanism of secondary dormancy: (1) the
imposition of a block at crucial points in the metabolic sequence leading to germination, or (2)
an unfavorable balance of growth-promoting versus growth-inhibiting substances.

Questions

I. Is it correct to describe dormancy as merely a resting state? Explain.

2. What is the difference between hard seed and firm seed? What is reseeding?
3. How can preharvest factors influence hard-seed coat dormancy? Many students confuse
scarification with stratification. What is the difference?
4. What is after-ripening?
5. Name at least two species whose seeds have each type of dormancy.
6. What is the argument, if any, against prechilling or stratifYing seeds during laboratory
germination testing?
7. Do you think that more than one inhibitor is responsible for chemical inhibition of
germination in most seeds?
8. Name several chemicals with germination-inhibiting ability.
9. How can secondary dormancy be induced in nondormant seeds?
10. Do you believe there is adequate evidence for endogenous rhythms in seed germination?
II. Distinguish between primary and secondary dormancy.
12. Name at least three ways that seed coats can regulate seed dormancy and cite two
procedures used to overcome this dormancy.
13. Describe the inhibitor-promoter concept for the regulation of seed dormancy and suggest
potential hormones that might be involved in this interaction.
14. Describe endogenous dormancy and provide at least four factors that can relieve this con-
dition.
15. Differentiate between stratification and after-ripening.
16. What is the difference, if any, between dormancy and quiescence?

General References

Ackerman, A. M., and H. Veldstra. 1949. The chemical nature of Kockerman's blastocholine
from Lycopersicon esculentum Mill. Recueil des Travaux Chimiques des Pays-bas et de la
Belgique 66:411-412 (Biological Abstracts, 1949.23:12727).
Ackerman, W. L. 1957. After-ripening requirements for germination ofAcer !runcatum Bunge.
Proceedings of the American Society for Horticultural Science 69:570-573.
Aitken, Y. 1939. The probl~m of hard seeds in subterranean clover. Proceedings of the Royal
Society of Victoria 51: 187-192.
Amen, R. D. 1968. A model of seed dormancy. Botanical Review 34:1-31.
Asakawa S. 1959. Germination behavior of several coniferous seeds. Journal ofthe Japanese
Forestry Society 41 :430-435.
Seed Dormancy 159

Aspinall, D. J. 1965. Effects ofsoi! moisture stress on the growth of barley. III. Germination
of grain from plants subjected to water stress. Journal of Institute of Brewing. 72:174-176.
Baskin, C. c., and 1. M. Baskin. 1988. Germination ecophysiology of herbaceous plant species
in a temperate region. American Journal of Botany 75:286-305.
Baskin, J. M., and C. C. Baskin. 1986. Germination ecophysiology of the mesic deciduous
forest herb Isopyrum biternaturm. Botanical Gazette 147:152-155.
Bennett, H. W. 1959. The effectiveness of selection for the hard-seeded character in crimson
clover. Agronomy Journal 51: 15-16.
Black, M., J. Butler, and M. Hughes. 1987. Control and development of dormancy in cereal. In:
4th International Symposium, Preharvest Sprouting in Cereals, ed. D. J. Mares, pp. 379-392.
Boulder, Colo.: Westview Press.
Bliss, D., and H. Smith. 1985. Penetration of light into soil and its role in the control of seed
germination. Plant Cell Environment 8:475-483.
Brant, R. E., G. W. McKee, and R. W. Cleveland. 1971. Effect of chemical and physical
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160 Seed Dormancy

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Seed Dormancy 163

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 8
Seed Vigor and
Vigor Testing

Seeds, as reproductive units, are expected to produce plants in the field. However,
farmers and seed producers have long recognized that the labeled germination often
overestimates the actual field emergence of seed lots. This occurs because by defmition,
germination is the "emergence and development from the seed embryo of those essential
structures which, for the kind of seed in question, are indicative of the ability to produce a
normal plant under favorable conditions (AOSA 2000)." As a result, the standard
germination test may fail to provide accurate information concerning a seed lot's field
performance potential for at least four reasons. These include the following.

1. The defmition of seed germination emphasizes that the seed analyst must focus on
essential structures which lead to the production of a normal plant. But, this
emphasis on seedling morphology may have little relationship with rapidity of
growth; a prime element in the potential for successful stand establishment.
2. Methodology for the conduct of a germination test is standardized so that test results
are reproducible within and among seed testing laboratories. This process means
that favorable conditions are utilized as described in the definition to ensure greater
uniformity in test results. Tests must be conducted on artificial, standardized,
essentially sterile media in humidified, temperature controlled chambers; conditions
that are so synthetic that they seldom relate to field conditions that seeds are likely
to encounter. In essence, because the standard germination test is conducted under
favorable conditions, it basically establishes the maximum plant-producing ability
of the seed lot. When field conditions are optimum, the standard germination test
may correctly predict field performance of the seed lot. For the most part, however,
standard germination values overestimate actual field emergence. We know, for
example, that when the standard germination test result is 80%, actual emergence
under field conditions seldom reaches 80%. In most instances, the field emergence is
considerably less.

L. O. Copeland et al., Principles of Seed Science and Technology

© Kluwer Academic Publishers 2001
166 Seed Vigor
3. The standard germination test is designed to provide for a first and fmal count. The
first count has a purpose of basically removing most of the strong seedlings that
have already germinated. The fmal count is designed to provide a sufficiently long
period that even weak seeds are coaxed or provided every opportunity to be
considered germinable. The germination percentage, therefore, is the sum of strong
and weak seedlings. The difficulty with such a process is that weak seedlings seldom
perform adequately when provided environmental stresses associated with field
emergence.
4. By definition, germination is scaleless. A seed is considered either germinable or it
is not. There are no distinctions provided for strong or weak seedlings. Those
considered germinable may vary from weak to semi-lame to robust in field
performance. This inability to document the quality of the seed fails to take into
account the progressive nature of seed deterioration, which has a major impact on
stand establishment.

These deficiencies have led to a continuously disquieting murmur for years that not all
facets of seed quality were being properly identified by the standard germination test. As a
result, it is useful to review the history and development of seed vigor testing.

mSTORY OF SEED VIGOR TESTING

In 1876, Fredrich Nobbe first distinguished the concept of seed vigor from that of
germination. He introduced the term triebkrajt, which means driving force or shooting
strength to convey the idea that, in addition to germination, speed and uniformity of
emergence were important parameters of seed quality. However, it was not until the 1950
International Seed Testing Congress that renewed interest was focused on seed vigor. At that
time, European and American laboratories were expressing concern that germination test
results were not standardized. In an attempt to explain these disparities, Franck (1950)
pointed out the differing concepts of germination testing between European and American
laboratories. He noted that in Europe, germination tests were made under optimum,
reproducible conditions, to assure that seed lots could be sold across national boundaries,
while special tests, such as the brick grit and soil tests were developed to evaluate "seedling
vigor." The American concept of germination, which was based on soil test results, was to
determine the plant-producing ability of a seed lot. Franck (1950) contended that both groups
needed to come to grips with these differing philosophies. To start the debate, he proposed
that germination testing should be conducted under favorable conditions in order that uniform
test results be obtained. The plant-producing ability in the field of a seed lot was to be defmed
by a new term: vigor.

Definition of Seed Vigor

The development of a satisfactory defmition of seed vigor has been a central theme in
the development of vigor tests. Without a defmition, the ability to measure or test this
undefined entity becomes difficult, ifnot impossible. Fortunately, many definitions have been
proposed and a study of their evolution portrays the initially confusing and changing status in
the expectations for seed vigor. As an example, in 1957, Isely defmed seed vigor as "the sum
Seed Vigor 167
total of all seed attributes which favor stand establishment under favorable conditions."
Building on this defmition, Delouche and Caldwell (1960) stated that "seed vigor is the sum
of all seed attributes which favor rapid and uniform stand establishment." Note the subtle
differences from Isely's definition. Delouche and Caldwell clarified stand establishment to
emphasize rapid and uniform performance and they also deleted the reference to favorable
conditions. It was clear at this point that rapid and uniform field performance was acceptable
parameters of seed vigor. However, the reference to" ... sum total of all seed attributes ... "
still left unresolved what the factors were that determined seed vigor. To address this issue,
Woodstock (1965) proposed that seed vigor was "that condition of good health and natural
robustness in seed, which, upon planting, permits germination to proceed rapidly and to
completion under a wide range of environmental conditions." Perry (1973) identified seed
vigor as the "physiological property determined by the genotype and modified by the
environment which governs the ability of a seed to produce a seedling rapidly in soil and the
extent to which the seed tolerates a range of environmental factors." He clearly emphasized
that seed vigor was determined by both genetic and environmental components. By this time,
consensus was rapidly emerging on a definition of seed vigor. In 1977, the International Seed
Testing Association (ISTA) defined vigor as "the sum total of those properties of the seed
which determines the potential level of activity and performance of the seed or seed lot during
germination and seedling emergence" (Perry 1978). Among the aspects of performance are:
(1) biochemical processes and reactions during germination such as enzyme reactions and
respiration activity, (2) rate and uniformity of seed germination and seedling growth, (3) rate
and uniformity of seedling emergence and growth in the field, and (4) emergence ability of
seedlings under unfavorable environmental conditions. Factors that influence the level of seed
vigor include the genetic constitution of the seed; environment and nutrition of the mother
plant; stage of maturity at harvest; seed size, weight, and specific gravity; mechanical
integrity; deterioration and aging; and pathogens. This defmition is considered an academic
definition because it discusses, identifies, and describes seed vigor (i.e., it attempts to relay
what seed vigor is).
In 1979, the Association of Official Seed Analyst's Vigor Committee defmed seed vigor
as "those seed properties which determine the potential for rapid, uniform emergence and
development of normal seedlings under a wide range offield conditions" (McDonald 1980b).
This definition quantifies vigor in terms of rapid uniform emergence and development of
normal seedlings. Thus, it focuses on what seed vigor does and is considered to be an
operational defmition.

Progress in Seed Vigor Testing

Initially, progress in seed vigor testing was slow in the United States. While the 1950
IS T A Congress and the perceptive comments of Franck on the need for vigor testing were
emphasized, the United States still considered this issue primarily a European concern. It
wasn't until the publication of two articles on seed vigor testing (Isely 1957; Delouche and
Caldwell 1960) that the key stimulus was provided in the United States to refocus and
redirect the development of the concept of seed vigor. In 1961, the first AOSA vigor test
committee was formed and was chaired by Dr. R. P. Moore. The principal objectives of that
committee were to bring into focus the advantages and disadvantages of direct vs. indirect
vigor tests as well as outlining various concepts of seed vigor. It seemed at this point that the
168 Seed Vigor
challenge of vigor testing was straightforward and the solutions imminent. But a review of the
history associated with vigor test development (McDonald 1994) demonstrates how naive this
notion was. By 1966, Moore noted that "Since quite diverse points of interest are involved,
the progress of the Committee could no doubt be promoted by restriction of the assignment to
measurement of vigor which commonly conveys rate and magnitude of growth." Clearly, the
more the topic was studied, the more challenging it became.
The next major advance occurred in 1974 when the increasing attention given to seed
vigor and its potential for advancing the ability to better estimate field performance resulted
in a clamor for standardized vigor tests. As a result, the Association of American Seed
Control Officials formally resolved that AOSA develop standardized seed vigor test
procedures. This resolution prompted the AOSA vigor test committee to even greater activity.
Then Chairman Lowell Woodstock wrote "there has been more real movement towards
consensus in seed vigor testing and more real progress by the AOSA Vigor Testing
Committee in meeting its responsibilities for developing, evaluating, codirying, and
standardizing vigor testing procedures during the past nine months than during any recent
period." Under Dr. Woodstock!s leadership, the "Seed Vigor Testing Progress Report" was
published as a special edition of the AOSA Newsletter in 1976. This document was a
significant milestone because it provided specific guidelines for the conduct of eight proposed
vigor tests. These tests could be evaluated using a referee format.
The AOSA vigor test committee set out to determine the standardization capability of
each of these tests. Corn and soybeans served as the crops receiving the greatest emphasis.
Concurrently, the committee was deriving a satisfactory definition of seed vigor. By 1980, a
seed vigor definition had been approved by every major organization involved with the
commerce and testing of seeds. As referee and research results were studied, the committee
became convinced that seven useful vigor tests were available. The procedures for these tests
were published in the AOSA Vigor Testing Handbook in 1983. The tests were placed into a
suggested vigor test section because of the remaining concern that the tests needed to stand
the scrutiny of routine use before they could be declared standardized. This handbook also
contained a historical perspective of vigor test development, the types of vigor tests available,
and the applications of vigor testing information. By 1987, the accelerated aging test was
significantly revised and improved and became the first vigor test for soybeans to be moved
from the suggested to recommended vigor test section. With increasing reliability and
standardization of the tests, the committee set out to evaluate acceptable tolerances for vigor
test results. This first evaluation was completed in 1991.
Clearly, the AOSA vigor testing committee in conjunction with its ISTA counterpart has
provided effective leadership in this important seed quality testing area. Its success can be
followed by monitoring the number of laboratories routinely using seed vigor tests. In 1978,
52% of seed-testing laboratories were conducting vigor tests (McDonald 1994). In 1983, the
number was 60% and by 1990, 75% of the seed-testing laboratories were using one or more
vigor tests. These data indicate that these committees not only provided useful vigor testing
protocols but have also educated the users so that the value and limitations of vigor testing
are fully understood. Useful reviews on the many facets of seed vigor have been provided by
Heydecker (1972, 1977), Pollock and Roos (1972), McDonald (I975, 1999), Cantliffe
(1981), HaImer and Bewley (1984), and TeKrony and Egli (1991). To better understand the
importance of seed vigor, we must first identiry the factors that influence this important seed
quality attribute.
Seed Vigor 169
FACTORS INFLUENCING SEED VIGOR

The development of a seed encompasses a series of important ontogenetic stages from

fertilization, to accumulation of nutrients, to seed dry down, to dormancy. Each of these
stages represents a change in morphological and physiological ontogeny that can alter seed
performance potential. The point at which the seed reaches its maximum dry weight is called
physiological maturity. At this point, it has its greatest potential for maximum germination
and vigor (Delouche 1974). However, since seeds generally achieve physiological maturity at
high moisture levels unsafe for storage, seed is typically not harvested until it attains harvest
maturity, which is low enough for safe storage, but high enough to minimize mechanical
injury. Between physiological maturity and harvest maturity, the seed is essentially stored on
the plant where it may be exposed to severe environmental conditions that adversely affect
seed quality.
Among the factors that influence seed vigor are genetic constitution, environment during
seed development, and seed storage environment.

Genetic Constitution

For many years, plant breeders have inadvertently selected for increased seed vigor. In
their attempts to increase yields, they have also imprOVed such seed characteristics as
mechanical integrity (hard-seededness), resistance to disease, protein content, and seed size.
These factors lead to better field emergence and often result in enhanced yields. In addition to
such physical manifestations of seed vigor, plant breeders have introduced hybrid vigor.
Thus, factors that are under genetic control such as hybrid vigor, hardseededness,
susceptibility to seed damage, and the seed chemical composition influence the expression of
seed quality.
Hybrid Vigor. Hybrid vigor is a component of heterosis and represents the measurable
superiority of the hybrid progeny over its inbred parents. The superiority of the hybrid is
often greater under conditions of stress than under optimal conditions. For example, seeds of
hybrid com and barley germinate faster and grow more rapidly than their inbred parents
(Whaley 1950; McDaniel 1969). This increased growth potential has been attributed to more
efficient mitochondria and extra enzyme systems for carbon assimilation (McDaniel and
Sarkissian 1968). The production of hybrid com seed is discussed in more detail in Chapter
10.
Hard Seed. For the most part, hard-seededness is an undesirable genetic trait because it
results in extreme variations in germination and hence, stand establishment. However, recent
efforts have been made to reintroduce hard-seededness into some cultivars to help protect the
seed from aging and to protect against leakage of nutrients during imbibition (Potts et al.
1978). Hard-seededness can be eliminated from cultivars relatively easily. Lebedeff (1947)
showed that, in crosses between soft lines and lines with varying degrees of hard-seededness,
the F] was intermediate or approached the soft-seeded parent. The F2 seed showed all possible
degrees of permeability between the parental extremes. The data indicated that hard-
seededness was controlled by several genes.
Kyle and Randall (1963) examined the nature of hard seed in the great northern and red
Mexican dry bean classes and reported that the micropyle was the site of water imbibition in
great northern, while in red Mexican it was the raphe and hilum. The impermeability of the
170 Seed Vigor
hilum and raphe was due to a simple recessive gene and was closely associated with the p
gene for white seed. Gloyer (1932), however, showed in a cross of red and white kidney beans
that it was easy to select for soft seed in whiteseeded segregants.
Susceptibility to Mechanical Damage. Susceptibility to mechanical damage, whether
induced by harvesting or conditioning equipment, has been shown to be under genetic control.
F or example, Barriga (1961) demonstrated that 41 strains of navy beans possessed differing
tolerances to mechanical abuse. Atkin (1958) and Wester (1970) reported that colored snap
bean cultivars were more resistant to mechanical damage than white-seeded cultivars (Figure
8.1). Kannenberg and Allard (1964) showed that seed thickness in lima beans directly
afffected the extent of seed coat cracking. Green and Pinnell (1968) studied visible seed
defects in soybeans and found them inherited quantitatively. Along with Walters and
Caviness (1973), they were the first to suggest the need for enhanced seed quality through
improved breeding lines.
Seed Chemical Composition. Breeding for improved nutritional quality such as high-
lysine corn has often resulted in increased seed quality problems. For example, this process
often produces small, shrunken, low-vigor seeds. Plant breeders are now attempting to find
gene systems that control nutritional quality but do not result in reduced seed vigor. Nass and
Crane (1970) found that various genes for endosperm expression influenced seed germination
at 15,20, and 25°C. Seeds with the Al gene produced more vigorous seeds than those without
this gene. Ullrich and Eslick (1978) also compared the effect of different shrunken endosperm
genes on kernel weight for barley. They found that mutant kernel weights ranged from 38 to
95% of normal and suggested that seed quality could be improved by selecting those
segregations with the greatest kernel weight.

Environment During Seed Development

The concentration of seed production (Chapter 10) for some crops in specific areas is
persuasive testimony to the environmental influence on seed development and quality
(Delouche 1980). For example, a major portion of the seed of temperate climate forage and
lawn or turf grasses is produced in the Pacific Northwest, especially in Oregon where the
climate is favorable for high-quality seed production. In these locales, the seeds complete
maturation and dry down, and can normally be harvested with little risk from rain and
adverse humidity. Other examples are the arid irrigated regions of California, Idaho, and
Arizona where high-quality cotton, vegetable, flower, and forage legume seeds are produced
because of the low humidity, minimal rainfall, and favorable temperatures that occur during
seed maturation. These environmental conditions reduce the spread of seed-borne diseases as
well as the risks associated with inclement weather during late harvesting periods. However,
most of the seed used in crop production is produced in the same area where the commercial
crop is grown. Consequently, environmental conditions may range from bad to good for high-
quality seed production. Among the factors that affect seed quality are soil moisture and
fertility, seed maturity, and the postmaturation preharvest environment (Delouche 1980).
Soil Moisture and Fertility. Moisture stress during seed development often results in
light, shriveled seed which, in turn, results in poor-vigor seed. However, it is generally agreed
that when soil fertility becomes limiting, plants respond by producing less seed. Thus, the
fewer seeds produced under marginal fertility conditions are usually as viable and vigorous as
are the greater number of seeds produced under favorable conditions. Exceptions do occur,
Seed Vigor 171

Figure 8.1. (Fop) Faster germination and more vigorous seedlings from green (nonbleached) than
from white (bleached) bean seed (Bottom) Field view showing much better stand from green-sorted
(nonbleached) than from white (bleached) seed, especially when the green-sorted seed is sound
(From Wester 1970).
172 Seed Vigor
however. Legatt (1948) demonstrated that boron-deficient pea seeds produced abnormal
seedlings and that such abnormalities could be corrected only with the addition of borax. In
contrast, soybean seeds produced in areas with high molybdenum soil concentrations,
possessed such high levels of molybdenum that they did not require sodium molybdate seed
treatment for planting in molybdenum-deficient soils (Harris et al. 1965). Peanuts are
particularly susceptible to soil mineral deficiencies. Soils low in boron and calcium produce
peanut seeds that exhibit a discoloration of the cotyledons associated with boron deficiency,
and a watery hypocotyl and physiological root breakdown associated with calcium deficiency
(Cox and Reid 1964; Sullivan 1973).
In other instances, a direct relationship between soil fertility and seed vigor has been
reported. For example, nitrogen fertilization of wheat is known to cause increased protein
levels in seed (Fernandez and Laird 1959; McNeal et al. 1971). Other studies have shown
that high-protein wheat seed results in increased germination (Fox and Albrecht 1957), seed
vigor (Lowe et al. 1972; Lowe and Ries 1973), and subsequent crop yield (Ries et at. 1970).
Seed Maturity. Abundant information demonstrates an association between parameters
of maturity such as seed size (and weight) and seed vigor (Austin 1972; Heydecker 1972;
McDonald 1975). Consequently, the environment during seed maturation has an indirect
effect on its potential vigor. Large soybean seeds have been shown to be superior to small
seeds in germination and vigor, as well as crop yield potential (Burris et al. 1971,1973;
Fontes and Ohlrogge 1972). Other studies have reported no difference in performance among
soybean seeds of varying size (Singh et al. 1972; Johnson and Leudders 1974), while others
(Aguiar 1974) have found that medium-sized soybean seeds were superior in vigor to both
large and small seeds. In hybrid com, the influence of seed size is reported to have little or no
effect on com performance in terms of emergence rate (Hunter and Kannenberg 1972), date
of tasseling (Hicks et al. 1976), fmal leaf number (Hawkins and Cooper 1979), and grain
yield (Hicks et al. 1976; Hawkins and Cooper 1979). Similarly, Shieh and McDonald (1982)
reported that seed size of two com inbreds had no effect on seed quality, although flat seeds
were shown to be superior to round seeds.
Postmaturation-Preharvest Environment. Deterioration of seed during the
postmaturation-preharvest environment is a serious seed production problem in the eastern
half of the United States because of high humidity, frequent rainfalls, and warm
temperatures-conditions that produce a rapid loss in seed viability and vigor. For example,
Simpson and Stone (1935) reported a 20 to 30% loss in cotton seed viability after only one
week's exposure to rainy conditions. Caldwell (1972) and Woodruff et al. (1967) showed that
cottonseed from lower bolls that opened first and were exposed longer to the field
environment before harvest were consistently lower in seed vigor than seed from bolls in the
upper half of the plant.
In soybeans, delayed harvest of seed because of inclement weather results in loss of
viability (TeKrony et a1.1980b) and an increase in mechanical damage during harvest (Green
et al. 1966). In a study on the effect of planting and maturity dates on soybean seed quality,
Green et al. (1965) found that soybeans from early planting dates that matured during hot,
dry weather produced lower-quality seeds compared to those from later plantings. It has also
been shown that early-maturing soybean cultivars are more susceptible to seed deterioration,
not because they are inherently predisposed to deterioration, but because the seed matures in
the warmer temperatures oflate September and early October compared to cultivars maturing
in late October (Delouche 1980). Preharvest loss of seed quality is known to be increased by
Seed Vigor 173
fungal invasion, which increases during wann and humid conditions (Nicholson et al. 1972;
Roncadori et al. 1972; TeKrony et al. 1980a).

Seed Storage

Seldom are seeds harvested and immediately planted without undergoing at least a brief
storage period. Consequently, the time of storage, type of seed stored, and storage
environment (temperature, relative humidity, and oxygen levels) influence seed vigor. A more
detailed discussion of these factors is provided in Chapter 9.

SEED VIGOR TESTS

The challenge of vigor testing has been to identify one or more quantifiable parameters
that are common to seed deterioration. Although not all changes that occur during seed
deterioration are understood, we can speculate on the probable sequence of events. A
hypothetical model (Figure 8.2) has been developed by Delouche and Baskin (1973) that
outlines some ofthe major parameters used in measuring seed vigor.
Because a vigor test is a more sensitive index of seed quality than the standard
gennination test, any of the events that precede loss of germination could serve as a basis for
vigor tests. The earlier the parameter can be measured during the loss of gennination, the
more sensitive the index of seed vigor. Thus, since the onset of membrane degradation
precedes loss of germination (Figure 8.2), the most sensitive vigor test should be one that
monitors membrane integrity.
Considerable experimental evidence supports this contention. Membranes are essential
for many metabolic events occurring in the seed, including respiration (cristae in
mitochondria), which provides the seed with the energy required for subsequent growth. The
endoplasmic reticulum is also a membraneous organelle on which many enzymes are fonned
as ribonucleic acid is translated. Thus, any impairment of membrane function can decrease
the amount of ATP fonned as an energy source, as well as retard the synthesis of specific
enzymes essential for growth. Koostra and Harrington (1969) showed that membrane
degradation had occurred in deteriorated seeds, and this observation has been supported by
others (Streeter 1965; Villiers 1973; Spencer et al. 1973; Simon 1974; McDonald 1975;
Harman and Mattick 1976; Stewart and Bewley 1980), though at least one report challenges
this concept (Priestley and Leopold 1979). Beyond these studies, ATP (Ching 1973), RNA
(Van Onckelen et al. 1973), and respiration (Woodstock and Grabe 1967) levels have been
shown to decrease following storage in accelerated or natural aging conditions. Subsequent to
the loss in respiration and biosynthetic capacity, the rate of germination declines, culminating
in a loss of seed lot uniformity. Other associated events that occur during deterioration are
loss in storability and ability to resist disease infection. Deteriorated seeds that are subjected
to biological and environmental stresses also exhibit reduced field emergence which may
(Johnson and Wax 1978) or may not (TeKrony and Egli 1977; Yaklich and Kulik 1979;
Burris 1976) be related to final yields of certain crops. Eventually, these subtle
manifestations of loss in seed quality are expressed by an increasing incidence of abnonnal
seedings-a component of the germination test. The final parameter of seed deterioration (and
the one most often employed) is seed gennination, underscoring the need for seed vigor tests
to supplement routine standard laboratory gennination tests.
174 Seed Vigor

PLANT RESISTANCE> /
" ~--------~~~'-
YIELD>

EMERGENCE (field) > ABNORMAL

'~~_______
SE_'E~'D~L~IN~G~S~<~

----0----
LOSS OF GERMINABILITY

Figure 8.2. Probable sequence of changes in seed during deterioration (From Delouche and
Baskin 1973).

As a result of the biochemical and physiological changes known to occur during seed
deterioration, most vigor tests have focused on measuring one or more of these parameters.
These vigor tests may be separated into various categories based on the parameters
monitored. For example, Isley (1957) divided vigor tests into direct and indirect tests. Direct
tests imitate the field environment in some way and measure the ability of seeds to emerge
under simulated field stress conditions. The cold test is an example of a direct test because it
subjects seeds to adverse conditions by placing them in cold, wet soil under direct stress from
temperature, moisture, and microorganisms. However, direct tests have been criticized
because they may not detect differences in quality when seeds are exposed to favorable soil
conditions.
Indirect tests measure specific physiological components of seeds. For example, the
conductivity test is an indirect test that monitors cell leakage. However, indirect tests fail to
evaluate all the physical and physiological factors that determine field establishment. In the
case of the conductivity test, improved performance due to seed treatments, the degree of
morphological damage, the influence of soil microorganism attack, and other factors are often
not adequately assessed.
Seed Vigor 175
Vigor tests may also be classified on the basis of the component of seed vigor measured.
For example, Woodstock (1973) separated vigor tests into physiological and biochemical
tests. Physiological tests measure some aspect of germination or seedling growth while
biochemical tests evaluate a specific chemical reaction or reactions (e.g., enzymatic activity
or respiration) related to the expression of germination (and hence, vigor). McDonald (1975)
added one additional grouping, a physical category that included seed size, shape, and
density, factors long associated with seed vigor because oftheir relation to seed maturity.
Other investigators have classified vigor tests into stress and quick test categories
(Pollock and Roos 1972). Stress tests consist of subjecting seeds to one or more of the
environmental stresses that might be encountered under soil conditions. These tests usually
involve measurement of some aspect of germination (e.g., hypocotyl length) while only the
conditions of stress are varied. Stress conditions may include high temperatures and relative
humidity as in the accelerated aging test, low temperatures with or without soil as in the cold
test or cool germination test, or osmotic stress imposed by using solutions such as
polyethylene glycol. Quick tests are tests in which some chemical reaction associated with
seed vigor is monitored; these usually require much less time than stress tests. Examples of
quick tests include the tetrazolium test, conductivity test, and various tests associated with
enzymatic activity.

Characteristics of a Seed Vigor Test

A vigor test should possess certain essential characteristics that can make it useful to the
seed producer and consumer. These characteristics have been described by McDonald
(1980a) as follows:
Inexpensive. Due to limited budgets for seed testing, it is important that a vigor test be
reasonably priced and require a minimum investment in labor, equipment, and supplies.
Rapid. Every seed laboratory has periods of peak activity, thus it is important that the
vigor test be conducted rapidly to minimize analyst time and germinator space. Furthermore,
seed producers desire a quick turn-around time for samples submitted for vigor tests since
such quick information on seed quality can provide them with a competitive marketing
advantage.
Uncomplicated. Where possible, vigor test procedures should be simple so that they can
be performed in seed laboratories without requiring additional staff with special backgrounds
and training.
Objective. For a vigor test to be easily standardized, a quantitative or numerical index
of quality that avoids subjective interpretations by analysts should be utilized.
Reproducible. The success of any test depends on its reproducibility. If these results
cannot be repeated because of intricate procedures or subjectivity of interpretation, then
comparison of results among laboratories becomes meaningless.
Correlated with Field Performance. Most defmitions of seed vigor emphasize the
relationship between seed vigor and field performance, and many studies have demonstrated
that this association exists. Consequently, the ultimate value of any vigor test may be its
ability to predict field performance.
176 Seed Vigor

Types of Seed Vigor Tests

The standard germination test is conducted under optimum conditions for seed
germination. Consequently, when field conditions at planting are near optimum, the results
usually correlate well with field emergence (Perry 1977; Egli and TeKrony 1979; Luedders
and Burris 1979). However, under suboptimal field conditions, standard germination results
usually overestimate field emergence (Sherf 1953; Delouche and Caldwell 1960; TeKrony
and Egli 1977; Johnson and Wax 1978; Tao 1978a; Yaklich and Kulik 1979). Therefore,
additional tests are needed to better predict seedling emergence under a wide range of field
conditions. Many vigor tests have been suggested; however, only a few have attained
acceptance by seed analysts and seed testing organizations (AOSA 1983; Perry 1981). These
are discussed below.
Cold Test. The cold test is one ofthe oldest methods of stressing seeds and is most often
employed for evaluating seed vigor in com and soybeans. Seeds are placed in soil or paper
towels lined with soil and exposed to cold for a specified period, during which stress from
imbibition, temperature, and microorganisms occurs (Figure 8.3). Following the cold
treatment, the seeds are placed under favorable growth conditions and allowed to germinate.
The greatest difficulty with the cold test is the lack of uniformity in field soil. Soils differ in
moisture, pH, particle composition, and pathogen levels, all of which contribute to divergent
results. Use of vermiculite, a more uniform medium, has been suggested as a possible
solution to the variability of soil conditions. However, it is widely believed that a cold test
requires field soil to be successful. Regardless ofthese difficulties, cold test vigor rankings of
seed lots tend to remain consistent within laboratories, which support the usefulness of this
test, especially for in-house purposes.
Accelerated Aging Test. This test incorporates many of the important traits desired in a
vigor test. Initially proposed as a method to evaluate seed storability, the accelerated aging
test subjects unimbibed seeds to conditions of high temperature (41°C) and relative humidity
(around 100%) for short periods (3 to 4 days). The seeds are then removed from the stress
conditions and placed under optimum germination conditions.
The accelerated aging test is rapid, inexpensive, simple and useful for all species; it can
be used for individual seed evaluation and requires no additional training for correct
evaluation. Modifications of the aging chamber have proven beneficial (Baskin 1977;
McDonald and Phaneendranath 1978) and resulted in this vigor test being the first to be
standardized. Another study has shown that differences in initial seed moisture should be
considered when interpreting this test (McDonald 1977b).
Conductivity Test. Low-vigor seeds have been shown to possess decreased membrane
integrity as a result of storage deterioration and mechanical injury. During imbibition, seeds
having poor membrane structure release cytoplasmic solutes into the imbibing medium. These
solutes with electrolytic properties carry an electrical charge that can be detected by a
conductivity meter.
Measurement of the conductivity of leachates from seeds is a rapid, precise, inex-
pensive, and simple procedure. However, initial seed moisture (Simon and Wiebe 1975) and
seed size (Tao 1978b) can affect the rate of solute leakage. Additionally, treatment of seeds
with antibiotics may influence conductivity measurements, necessitating their removal before
determinations are made.
Seed Vigor 177

Figure 8.3. The rolled towel cold test procedure: (AJ Planting seeds on cold wet towels and
covering with a sand-soil mixture; (BJ Emergence of corn seedlings following the rolled towel cold
test procedure.
178 Seed Vigor

One limitation of the present conductivity test is that it expresses results as an average
conductivity evaluation for 25 seeds. Such an expression presumes that all seeds are equally
deteriorated and will provide the same quantity of electrolyte leakage. A seed lot, however, is
composed of a population of individual seed-each with its own unique potential to perform
in the field. Conductivity test results, therefore, would better reflect the vigor capability of a
seed lot if they were presented on an individual seed basis. A commercial instrument is now
available to monitor the electrolyte leakage of individual seeds. Recent studies suggest that
this instrument provides a more accurate appraisal of seed vigor in soybeans (McDonald and
Wilson 1979, 1980; Miles and Copeland 1980), cotton (Hopper and Hinton 1980), cowpea
(Beighley and Hopper 1981), navy bean (Suryatmana et al. 1980) and corn (100 et al. 1980)
than does the present conductivity method.
Cool Germination Test. Unlike the cold test, the cool germination test is conducted
under standard laboratory conditions at low temperatures (18°C) and does not rely on the
activity of microorganisms to stress the germinating seeds. It has been demonstrated that low
vigor seeds from warm-season crops, such as cotton, have decreased growth rate and lower
germination under these conditions. The major advantage of this test is that it is similar to the
standard germination test and the same criteria for interpretation of normal seedlings are
employed. Its principal limitation is that it is currently limited to use in cotton.
Seedling Growth Rate Test. Vigorous seeds are able to efficiently synthesize new
materials and rapidly transfer these new products to the emerging embryonic axis, resulting in
increased dry weight accumulation. The seedling growth rate test is based on this concept and
vigor results are expressed as mg dry weight of germinable seedlings. This test is generally
conducted according to the standards for the routine germination test. After evaluations are
made, the growing segments of the embryos from normal seedlings are excised from the
storage organs (cotyledons or endosperm), dried in beakers at 80°C for 24 hours, and
weighed to determine their increase in dry weight. Since seedling growth rate is correlated
with vegetative development in the field (Burris 1976; Pinthus and Kimel 1979), this test
offers substantial promise. However, certain standardization problems still need to be
addressed. For example, small differences in moisture and light intensity can have significant
effects on the rate of seedling growth. Also, the test may require standardization for specific
cultivars since rate of seedling growth can be genetically controlled (Burris 1975).
Seedling Vigor Classification Test. This vigor test is an expansion of the routine
germination test, requiring the seed analyst to further classifY "normal" seedlings into
"strong" and "weak" categories (Figure 8.4). The test requires no additional equipment and
employs concepts and terms familiar to seed analysts; thus it is particularly attractive to seed
analysts. Despite its advantages, it has one serious difficulty. To further separate "normal"
seedlings into two additional categories is a subtle task and can introduce additional
variability. For this reason, seed testing organizations maintain active referee programs to
help laboratories identifY their interpretation difficulties.
Tetrazolium (TZ) Test. The TZ test is one of the most valuable techniques for
analyzing seed quality. It relies on the action ofthe TZ molecule to react with hydrogen atoms
released as a result of dehydrogenase enzyme activity in living tissue. This results in the
formation of a water-insoluble red pigment called formazan which a trained seed analyst
evaluates for staining pattern and color intensity. The analyst then subjectively places the
seeds into vigor categories ranging from strong to weak (Figure 8.5). Though the results of
this test correlate well with seed vigor when interpreted by a qualified analyst, it is still
Seed Vigor 179

Figure 8.4. Classification ofpeanut seedlings at the final count in the seedling vigor classification
test. Groups of seedlings from left to right are normal strong, normal weak, and abnormal.
Deficiencies of normal weak seedlings are: (a) partial decay of epicotyl, (b) no primary root, (c)
primary root cracked, (eI) one primary leaf missing, (e) split primary root, (f) curled hypocotyl.
Deficiencies of abnormal seedlings are: (g) shortened hypocoty/, (h) stubby primary root with no
secondary roots, (i) no epicotyl, 0) hypocotyl and roots missing, (k) poorly developed hypocotyl and
roots.

subject to certain standardization difficulties: fIrst, the ability of the analyst to ascertain
whether a seed is vigorous; second, the failure of the test to detect seed treatment
phytotoxicity and reveal seed dormancy.
Speed of Germination. Speed of germination is one of the oldest seed vigor concepts.
Seed lots with similar total germination often vary in their rate of germination and growth.
Many methods for determining germination rate have been employed (Nichols and Heydecker
1968; Tucker and Wright 1965; Timson 1965). The number of days a lot requires to reach
90% germination was used by Belcher and Miller (1974) as an index of vigorous seed. For
lower-quality lots, another percentage value (e.g., 50%) could be used. A different approach
was proposed by Maguire (1962) who suggested the following formula:

x = number of normal seedlings + ... + number of normal seedlings

days of fIrst count days of fInal count

Similar germination indices were suggested by Czabator (1962) and Djavanshir and Pourbeik
(1976) for tree seeds based on the following formula:
 Seed Vigor
180

Figure 8.5. Examples of (A) high, (B) medium, (C) low vigor and (D) ungerminable soybean seeds
following staining with tetrazolium chloride.
Seed Vigor 181
Germination value = peak value of cumulative number of normal seedlings
days of germination counts

x total number of normal seedlings

days of final count

Normal seedlings ranging from those with radicles emerged and small hypocotyls visible
to those with fully developed seedlings with seed coats completely shed, were classified by
Wang (1973) into six different classes depending on the stage of seedling growth. He tested
red pine seeds from seven sources and found that the combined percentages of the top three
classes were significantly correlated with the nursery emergence.
In the standard germination test, a preliminary and [mal count are routinely performed.
The percentage of normal seedlings recorded in the first count represents fast germinating
seeds and can be used as an index of vigor. (This index is similar to that from the seedling
vigor classification test except that the strong normal seedlings in the second count are
excluded.) In soybean studies, the first count (four days) was found to provide a good
estimate of seedling vigor (Burris et al. 1969) and has been used as a vigor index component
(TeKronyand Egli 1977). More recently, it was reported that foliage development, dry matter
accumulation, and crop yield from rapidly germinating soybean seeds exceeded those from
slowly germinating seeds (Pinthus and Kimel 1979).
The advantage of the speed of germination test is that little additional work is required
compared to the standard germination test. However, variations in temperature or moisture in
the test chamber and substrate may affect test results.
Brick Grit Test. The brick grit test is also known as the Hiltner test. It was originally
developed by Hiltner and Ihssen (1911) for detecting seed-borne Fusarium infection in
cereals. Results of further studies indicated that the test also detected seed weaknesses other
than those caused by fungi. For example, it revealed cereal injury caused by frost, preharvest
sprouting (Schoorel 1960), and hot-water treatment (Tempe 1963) which makes it useful as a
vigor test (SchooreI1960; Tempe 1963).
In the Hiltner test, seeds are planted on damp brick grit or in a container of sand and
covered with 3 cm of damp brick grit, then germinated in darkness at room temperature for a
specific time. Seeds weakened by pathogenic fungi, mechanical injury, or storage
deterioration are unable to penetrate the brick grit layers. The percentage of normal seedlings
from this test is considered to be an indication of the vigor level.
The Hiltner test has not been popular in the United States. Comparative trials between
germination in sand and ground brick (similar to brick grit) have shown that it fails to provide
any more information about vigor than does the standard germination test (Fritz 1965). The
test also has several disadvantages including high cost, large space requirement, and
variability in test results, as well as difficulties in obtaining, washing, and drying of brick grit
(Perry 1978).
Osmotic Stress. When seeds are sown in the field, they are often subjected to drought
stress which results in poor emergence. Such drought conditions can be simulated in a
laboratory test by use of soil, soil solution, and other solution systems (Parmar and Moore
1968; McWilliams and Phillips 1970; Sharma 1973). Since standardization of soil conditions
is difficult to achieve, a solution system is preferred. Seeds are germinated in solutions such
as sodium chloride, glycerol, sucrose, polyethylene glycol (PEG), and mannitol (Parmar and
182 Seed Vigor
Moore 1968; Sharma 1973) with specific osmotic potentials. There is evidence, however, that
some low molecular weight osmotic substances (sucrose, sodium chloride, glycerol, and
mannitol) enter germinating seeds and cause toxicity. High molecular weight PEG (4000 or
more) is a satisfactory compound for simulating true drought (Manohar 1966; Parmar and
Moore 1968) without causing toxic side effects. The osmotic potentials of PEG 6000
solutions at various concentrations and temperatures have been determined (Michel and
Kaufmann 1973). The rate of germination under such conditions is markedly reduced, and
emergence of the plumule is generally more affected than that of the radicle (EI-Sharkawi and
SpringueI 1977). Since vigorous seeds can tolerate greater osmotic stress, this method has
been suggested as a vigor test (Hades 1977).
The advantage of the osmotic test is that no special equipment or training is required.
However, small com seeds reportedly germinate better than large seeds under such conditions
because of their lower water requirement (Muchena and Grogan 1977). A significant
interaction of osmotic stress and temperature of germination has been reported (EI-Sharkawi
and SpringuelI977).
Respiration. Seed germination and seedling growth require the use of metabolic energy
acquired from respiration. Thus, a decrease in the rate of respiration of germinating seeds has
been shown to precede a decline in the rate of seedling growth (Woodstock 1968).
Respiration rate, measured during the first 18 hours of germination, can be used to detect
injury from gamma radiation in com, sorghum, wheat, and radish (Woodstock and Combs
1965; Woodstock 1968) and chilling injury in lima bean (Woodstock and Pollock 1965) and
cacao (Woodstock et a1. 1967). Positive correlations have been reported between rate of
oxygen uptake during imbibition and seedling growth (Woodstock and Grabe 1967).
However, this relationship has not been confrrmed by other studies (Abdul-Baki 1969;
Anderson 1970; Byrd and Delouche 1971; Bonner 1974).
Respiration tests are rapid and quantitative, but require a respirometer and trained
personnel. Furthermore, mechanical injury (which lowers seed vigor) may increase respiration
rates (Woodstock 1969), thus producing confusing results.

Standardization of Vigor Tests

For any seed quality test to be useful, it must provide reproducible results. To evaluate
reproducibility of various vigor test results, both the AOSA and ISTA have conducted
extensive referees (McDonald 1977a, 1980b; McDonald et a1. 1978; Perry 1978; Tao 1978c,
1980a, 1980b). Generally, results of these referees have shown that seed testing laboratories
can reproduce their own results on the same sample of seed within acceptable confidence
limits. However, when the same tests are conducted by different laboratories, the amount of
variability is often unacceptable.
Many possibilities exist to explain the lack of standardization among laboratories
(AOSA 1983). For example, most vigor tests used to date require a degree of subjectivity.
The seedling vigor classification and tetrazolium vigor tests possibly involve the most
subjective interpretations because seeds and seedlings or both are separated into categories
based on characteristics that are difficult to precisely describe. Even the cold test and
accelerated aging test require the classification of seedlings as normal or abnormal.
Variations in temperature, moisture, and other environmental conditions are more
critical for tests in which rate of growth or rate of a biochemical process is measured than for
Seed Vigor 183
those such as the standard germination test which measures the completion of a process. For
example, a 1°C difference in temperature during the course of the standard laboratory
germination test probably has little effect on the final percent germination, but a 1°C
temperature difference may have a considerable effect on results in the seedling growth rate
test or seed deterioration in the accelerated aging test, as would minor variations in the
moisture content of germination substrata or the relative humidity of the air. Consequently,
conditions and equipment that are suitable for the standard laboratory germination test may
not be suitable for vigor tests.
The cold test would appear to be difficult to standardize because it exposes seeds to soil
microorganisms to measure their ability to resist attack under cold, wet conditions. Thus, the
standardization of the cold test procedure in its present state still requires standardization of
the substrate (soil) microflora, a task that would be difficult, if not impossible. The use of
sterile media inoculated with specific microorganisms has been suggested, an approach that is
probably too simplistic, since it is difficult to culture microorganisms and maintain a constant
level of pathogenicity. Also, pathogens behave differently in soil, where there is a complex
population of microorganisms, from how they behave in the absence of such an interacting
popUlation.
A recent study by Byrum and Copeland (1995) indicated that the cold test for com in the
U.S. corn belt is as repeatable as the standard germination test. This study was conducted on
four corn seed lots with varying levels of quality and each sample was tested by 10 different
laboratories throughout the Midwest and Midsouth representing official, commercial, and
crop improvement associations. This study illustrates the present state of the art of cold
testing in the United States and demonstrates the potential for the eventual standardization of
all vigor tests.
It may be important to determine whether such factors as dormancy affect vigor test
results. Many species (e.g., legumes) have seeds that are impermeable or only slowly
permeable to water. This can affect the percentage of seeds that germinate as well as the
length of seedling growth in stress tests and can cause a biased index of vigor in a seedling
growth rate test. It can also affect the leaching of electrolytes from seeds in a conductivity
test.
Dormancy can also confound the interpretation of vigor test results. For example,
dormancy can cause lettuce and cereal seeds to germinate poorly in tests conducted at high
temperatures. This difficulty is often overcome in the brick grit test by exposing cereal seeds
to low temperatures in the moist brick grit prior to the germination period. If dormancy is
suspected of affecting vigor results, the seed should be checked by a standard warm
germination test or a tetrazolium test.
Clearly, vigor testing for all species and regions has not yet achieved the same level of
standardization possessed by the standard germination test. However, both seed testing
organizations and the seed trade are focusing on improving the reproducibility of vigor test
results. Considering the important role that vigor tests can have in plant breeding, seed
production, quality control, and marketing programs, standardization is badly needed.
Although further research and testing are required before vigor testing becomes a routine
phase of seed testing, the promise of vigor testing in the future is bright.
184 Seed Vigor
Use of Seed Vigor Tests

The AOSA Seed Vigor Testing Handbook (1983) has included a consideration of the
many possible uses of seed vigor tests. For example, many farmers know from experience
that seed lots with equal germination levels may emerge from the soil quite differently,
resulting in erratic field stands, replanting (on some occasions), or both. These same farmers
recognize that this problem is usually more severe when adverse environmental conditions
occur at or immediately after planting. Thus, if farmers know that they will be planting seed
in adverse soil conditions or can predict an adverse field environment following planting, high
vigor seeds should provide higher field emergence than low vigor seeds. If this prevents
replanting, and the associated delayed maturity or yield reductions due to poor stands,
additional money spent on high vigor seeds may be worthwhile. It cannot be assumed,
however, that high vigor seeds will produce excellent emergence and stands in any soil
environment. It should improve the chances for satisfactory emergence. but it will not
guarantee it. Conditions prevailing after planting may be so severe that even the most
vigorous seeds cannot perform satisfactorily.
Many seed companies in the United States conduct a series of vigor tests on each
soybean seed lot and combine this information into a seed vigor index. They establish an
acceptable vigor level for a specific production season and seed lots are evaluated (in-house)
prior to conditioning, treatment, and marketing. In a few cases, those seed lots that exceed a
specified vigor level after conditioning are advertised and promoted in genera] terms as vigor-
proven, vigor-rated, or high-vigor seed. If the seed is sold with strong emphasis on vigor, a
buyer should ask the dealer for specific information on how many vigor tests were conducted
and the criteria used to assess vigor potential. One should not assume that the statement
"vigor-tested" automatically means a sound vigor testing program or implies higher vigor
than other seed which is not promoted as vigor tested
Does high vigor seed mean higher yields? This question is asked by many farmers
following implications concerning yield by some seed producers. Unfortunately, there have
been fewer comparisons of seed vigor to final yield than to field emergence. As with other
investigations relating to seed vigor, the yield comparisons are variable depending on the crop
planted, the stand achieved, and the original vigor level of the seed. Many factors that may
not relate to the vigor of the seed can influence yield during the growing season. Also, certain
crops and genotypes have the ability to compensate for minor stand differences following
emergence, and this may result in little difference in final yield. As would be expected, if
adequate field stands resulted from high vigor seeds, higher yields should occur. However,
less evidence supports improved yields using high vigor seeds than supports equal stands
achieved from planting medium or low vigor seeds. To date, insufficient evidence is available
to show a strong relationship between yield and vigor in the absence of stand differences.
Future research on plant growth and development or the physiological or biochemical basis
for seed vigor may improve our understanding of this relationship.
Seed Vigor 185
Questions

1. What are some evidences of genetic influence on seedling vigor?

2. How much loss of vigor do you feel results from harvesting before complete seed
maturity? Which crops are more likely to be affected?
3. Which preharvest environmental factors are most influential in affecting seed vigor?
4. The seed corn industry traditionally has not emphasized the different performance of the
different seed grades. Is this consistent with the documented effect of seed size on
seedling vigor?
5. Mechanical damage to seed is more serious in certain crops. Name several species in
which damage is most likely.
6. Should all carryover seed by tested for vigor?
7. List some advantages which might accrue if it were agronomically feasible to presoak
seed prior to planting to avoid chilling injury.
8. Name several ways of seed stimulation. Which are agronomically feasible?
9. Name several seed vigor tests. Which are most valuable for measuring seed quality?
10. Do you think cold tests would be valuable for measuring seed vigor in crops other than
corn and soybeans?
11. Do you believe physical growth rate tests or physiological measurements most truthfully
measure seed quality?

General References

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Aguiar, P. A. A. 1974. Some relationships between seed diameter and quality in soybeans.
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Anderson, J. D. 1970. Physiological and biochemical difference in deteriorating barley seed.
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Atkin, J. D. 1958. Relative susceptibility of snap bean varieties to mechanical injury of seed.
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Barriga, C. 1961. Effects of mechanical abuse of Navy bean seed at various moisture levels.
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Beighley, D. H., and N. W. Hopper. 1981. The relationship of chemical composition and
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Bonner, F. T. 1974. Tests for vigor in cherrybark oat acorns. Proceedings of the Association
of Official Seed Analysts 64: 109-114.
186 Seed Vigor
Burris, J. S. 1975. Seedling vigor and its effect on field production of com. Proceedings of
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. 1976. Seed/seedling vigor and field performance. Journal of Seed Technology 1:58-74.
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Hadas, A. 1977. A suggested method for testing seed vigor under water stress in simulated
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188 Seed Vigor
Kannenberg, L. W., and R. W. Allard. 1964. An association between pigment and lignin
fonnation in the seed coat oflima bean. Crop Science 4:621-622.
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vulgaris L. and its use in breeding and inheritance studies. Proceedings of the American
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Seed Vigor 189
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Streeter, J. G. 1965. Possible mechanisms in the loss of seed viability. Association of Official
Seed Analysts Newsletter 39(3):27-35.
Sullivan, G. A. 1973. Effects of dolomitic limestone, gypsum, and potassium on planting seed
quality, yield, and grade of peanuts, Arachis hypogea L. Ph.D. Dissertation, North Carolina
State University, Raleigh.
Suryatmana, G., L. O. Copeland, and D. F. Miles. 1980. Comparison oflaboratory indices of
seed vigor with field performance of Navy bean (Phaseolus vulgaris L.). Agronomy
Abstracts 1980: 113.
Tao, K. J. 1978a. Effect of soil water holding capacity on the cold test for soybeans. Crop
Science 8:979-982.
__ . 1978b. Factors causing variations in the conductivity test for soybean seeds. Journal
of Seed Technology 3 (1): 10-18.
__ . 1978c. The 1978 "referee" test for soybean and com. Association of Official Seed
Analysts Newsletter 52(4):43-66.
_ _. 1980a. The 1979 Vigor "referee" test for soybean and com. Association of Official
Seed Analysts Newsletter 54(1):40-58.
_ _.1980b. The 1980 vigor "referee" test for soybean and com seed. Association of
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Seed Vigor 191
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Butterworth and Co., Ltd.
TeKrony, D. M., A. D. Phillips, and D. B. Egli. 1980b. The effect of field weathering on
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Woodstock, L. W., B. Reiss, and M. F. Combs. 1967. Inhibition of respiration and seedling
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Yaklich, R. W., and M. M. Kulik, 1979. Evaluation of vigor tests in soybean seeds.
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 9
Seed Storage and
Deterioration

Seeds are uniquely equipped to survive as viable regenerative organisms until the time and
place are right for the beginning of a new generation; however, like any other fonn of life, they
cannot retain their viability indefmitely and eventually deteriorate and die. Fortunately, neither
nature nor agricultural practice ordinarily requires seeds to survive longer than the next growing
season, though seeds of most species are able to survive much longer under the proper
conditions.

THE LIFE SPAN OF SEEDS

Long~Lived Seeds

Museum botanists in Canada reported the germination oflupin seeds that had been buried
deep in a Canadian peat bog for an estimated 10,000 years (Porsild and Harrington 1967). This
is the longest known record for safe seed storage even with a literature that abounds with
evidences of long seed storage (Justice and Bann 1978). Germinating Indian lotus seeds from
a Manchurian lake bed were first estimated to be 120 to 400 years old (Ohga 1926), but were
later found, using radiocarbon dating, to be over 1000 years old (Libby 1951). At the National
Museum of Paris, viable seeds of several species that had been collected 100 to 160 years
previously were found (BecquereI1934). Frequently, one hears of reports of gennination ofthe
so-called "mummy" seed from Egyptian tombs; however, these reports have not been
substantiated.
Many long-tenn storage studies are in progress in the United States. These have already
provided considerable data on the seed longevity of many species. The two oldest and best-
known studies are those founded by Beal in 1879 at Michigan State University and Duvel of the
United States Department of Agriculture in 1902. In both studies, seeds buried in soil continue
to germinate demonstrating the remarkable life span of many seeds (Figure 9.1).

L. O. Copeland et al., Principles of Seed Science and Technology

© Kluwer Academic Publishers 2001
 Limit of longevity (years) ~
10 20 30 40 50 60 70 80 90 100 ~
~
r ________ --Jill ~
Polygonaceae:
Polygonum ,hydroplper
~
Rumex crispus
-- - - - - - -- -- ~
Portulacaceae:
Portulaca oleracea ---_..::::::=- ~
Caryophyllaceae:
~
Agroslemma gilhago ~
Siellaria media ~.
Amaranthaceae:
Amaranlhus retroliexus g'
-----.:::~
Brassica nigra --
Cruciferae:
Capsella bursa'paslorls ~:-:-:::::::::::::i::
Lepidium virgmicum _
Leguminosae:
Trifolium repens
••••
Euphorbiaceae.
Euphorbia maculata
Malvaceae:
Malva pusilia
Onagraceae:
-
..:.:::::=-=:::::::::::::::::::::::::::::.:::::::.:.:.::::::::=--
Oenolhera blennis
Scrophulariaceae:
Verbascum spp.
Plantaginaceae:
Plantago major
. -::: :::::=-:::.:..-----------
c::::=-: :::::.::::::::..
Compositae:
Ambrosia arlemisilfolia r::::::::::::::::::~
Anthemis cotula
Erechtiles hieracifolla
Gramineae:
Bromus secalinus
Setaria pumlla I
Figure 9.1. Limits of longevity for seeds in Beal's burial experiment. Germination tests were performed every five years for the first 40 years and
every 10 years thereafter. Dashed Jines indicate test periods when no positive germinations were noted "Verbascum spp." indicates V. thapsus
and/or V. blattaria (From Priestley 1986). ..-
\0
W
194 Seed Storage and Longevity
Short-Lived Seeds
In contrast to evidence on long-lived seeds, some species have seeds that are remarkably
short-lived, particularly when stored in open air. To distinguish between these types oflong- and
short-lived seeds. Roberts (1973) proposed the terms orthodox and recalcitrant seeds.
Orthodox seeds are long-lived seeds. They can be successfully dried to moisture contents as low
as 5% without injury and are able to tolerate freezing temperatures. Recalcitrant seeds are those
seeds which cannot be dried to moisture contents below 30% without injury and are unable to
tolerate freezing. As a result, recalcitrant seeds live for only short durations and are difficult to
successfully store because their high moisture content encourages microbial contamination and
results in more rapid seed deterioration. Furthermore, storage of recalcitrant seeds at subzero
temperatures causes the formation of ice crystals which disrupts cell membranes and causes
freezing injury.
Recalcitrant and orthodox seeds differ greatly in their ecology and seed morphology (Chin
et a1. 1989). Recalcitrant seeds are primarily from perennial trees in the moist tropics such as
coconut, coffee, and cacao. In some cases, they also come from temperate region trees such as
citrus. Other species with remarkably short life spans that lose viability after a few months in
the open air are willow, poplar, cottonwood, and elm. Seeds of river maple (Acer saccharinum)
reportedly last only a few days under natural conditions (Jones 1920). Survival in nature
apparently depends on the ability of the seeds to germinate and become established quickly after
they have fallen from the tree. Seeds of wild rice (Zizania aquatica) also lose viability rapidly
in open-air storage and must be kept in water at low temperatures for longer survival (Duvel
1905). Most orthodox seeds come from annual temperate species adapted to open fields.
Recalcitrant seeds also differ from orthodox seeds in their seed morphology. Most
recalcitrant seeds mature and exist in their fruits and are covered with fleshy or juicy arriloid
layers and impermeable testa (Chin et al. 1989). At physiological maturity, they are generally
much higher (50-70%) in moisture content than orthodox seeds (30-50%). At the same time,
they are larger than most orthodox seeds, even though their embryos are only about 15% the size
of an orthodox seed embryo (Table 9.1). These differences have made successful storage of
recalcitrant seeds difficult and an active area for research. It is generally believed that
recalcitrant seeds never go into dormancy but, instead, continue their development and progress
toward germination (Berjak et a1. 1990). Most attempts at storing these seeds have focused on
using endogenous seed inhibitors such as abscisic acid (Goldbach 1979) or replacing the high
water content with other substances such as sugars or ethylene glycol to permit successful
storage even under low temperatures without inducing ice-crystal formation and subsequent seed
damage (Crowe and Crowe 1986). A more detailed consideration of recalcitrant seeds is
provided by Chin and Roberts (1980). It should be noted that some seeds now appear to fit
neither an orthodox nor recalcitrant category and, for these seeds, an intermediate category has
been suggested (Ellis 1991). Citrus and coffee seeds may fit this intermediate classification.

CONCEPTS OF SEED DETERIORATION

Seed deterioration is a major problem in agricultural production. It has been estimated that
about 25% of the annual value of seeds in inventory are lost because of poor seed quality
(McDonald and Nelson 1986).
Seed Storage and Deterioration 195

Table 9.1. Seed Size, lOOO-seed wt., and Moisture Content of Typical Recalcitrant and
Orthodox Seeds.

Seed size, 1000 Moisture

Crop species length x width Seed wI. content
mm g gHzO g-lfw
Recalcitrant
Nephelium lappaceum L. 28 x 16 3,555 0049
Arlocarpus heterophyllus Lam. 35 x 24 8,520 0.52
Artocarpus champeden (Lour.) Spreng. 30 x 20 5,814 0.71
Lansium domesticum Carr. 17 x 13 2,335 0.52
Bouea ganadaria 22 x 15 3,530 0046
Durio zibethinus Murr. 42 x 25 14,783 0.50
Theobroma cacao L. 25 x 25 1,995 0.36
Orthodox
Hibiscus esculentus L. 6 x 4 146 0.18
Vigna sesquipedalis (L.) Fruw. 12 x 5 192 0.16
From Chin et al. (1989).

and Nelson 1986). Worldwide, these losses are even greater, particularly in lesser-developed
countries and in geographic regions where high temperatures and high relative humidities prevail
during seed maturation and storage. Although the significance ofthese losses is readily apparent,
the overall importance of seed deterioration becomes even more manifest when it is realized that
even greater economic loss occurs annually due to deterioration and as a consequence of
breakage and microorganism spoilage during production, storage, and shipping of feed grains
(Salunkhe et al. 1985).
Seed deterioration can be characterized by the following three general concepts (Delouche
1973):

1. Seed deterioration is an inexorable process. All living things must eventually

deteriorate and die. Although death still remains an inevitable result of life, it is possible
to retard the rate of deterioration through optimum storage practices.
2. Seed deterioration is an irreversible process. Once seed deterioration has occurred,
this anabolic process cannot be reversed. Simply stated, low-quality seeds cannot be made
into high-quality seeds. Although some mechanisms for preconditioning or treating seeds
with fungicides improve field emergence, these treatments only allow the optimum
expression of seed potential; they do not alter the basic physiological quality of the seed.
3. Seed deterioration varies among seed populations. It is now well established that
certain varieties exhibit less deterioration than others. Even within a variety, the storage
potential of individual lots varies, and even within a seed lot, individual seeds have
differing storage potential.
196 Seed Storage and Longevity

FACTORS INFLUENCING THE LIFE SPAN OF SEEDS

Internal Factors

The physical condition and physiological state of seeds greatly influence their life span.
Seeds that have been broken, cracked, or even bruised deteriorate more rapidly than undamaged
seeds (McDonald 1985; Priestley 1986). Even in the absence of physical symptoms, seeds may
be physiologically impaired and become susceptible to rapid deterioration. Several kinds of
environmental stresses during seed development, and prior to physiological maturity, can reduce
the longevity of seeds-for example, deficiency of minerals (N, K, Ca) (Harrington 1960b),
water (Haferkamp et al. 1953), and temperature extremes (Justice and Bass 1978). Immature
and small seeds within a seed lot do not store as well as mature and large seeds within a seed
lot (Wien and Kueneman 1981; Minor and Paschal 1982). Hard-seededness also extends seed
longevity (Flood 1978; Patil and Andrews 1985).

Relative Humidity and Temperature

The two most important factors that influence the life span of seeds are relative humidity
and temperature. The effects of relative humidity (and its subsequent effect on seed moisture)
and temperature of the storage environment are highly interdependent. Most crop seeds lose their
viability at relative humidities approaching 80% and temperatures of 25 to 30°C, but can be
kept 10 years or longer at relative humidities of 50% or less and a temperature of 5°C or lower
(Toole 1950). According to Harrington (1973), because of this interdependency, the sum of the
percentage of relative humidity plus the temperature in degrees Farenheit should not exceed 100
for safe storage. Another report indicates that for safe storage from one to three years, this
combined total may be as high as 120 as long as the temperature contributes no more than half
the total (Bass 1967). It has been suggested that the relative humidity should be no higher than
60% for seeds at 21°C and no higher than 70% for seeds at 4 to lOoC; however, at 5°C and 45
to 50% relative humidity, many crop seeds can be safely stored for 10 years or longer (Toole
1957).
These data reflect the intimate association among seed moisture, storage temperature, and
seed longevity. This interaction was recognized by Harrington (1972) when he suggested the
following two "rules ofthumb" regarding optimum seed storage: (1) each 1% reduction in seed
moisture doubles the life of the seed, and (2) each 5°C reduction in seed temperature doubles
the life of the seed. Let's examine each of these important variables critically.

Seed Moisture

Harrington (1972) recognized that there must be some qualifications to the above rules of
thumb. First, the rule regarding seed moisture does not apply above 14 or below 5% seed
moisture. Seeds stored at moisture contents above 14% begin to exhibit increased respiration,
heating, and fungal invasion that destroy seed viability more rapidly than that indicated by the
first rule of thumb. Below 5% seed moisture, a breakdown of membrane structure hastens seed
deterioration (This is probably a consequence of reorientation of hydrophyllic cell membranes
due to the loss of the water molecules necessary to retain their configuration). Thus, storage of
most seeds between 5 and 6% seed moisture appears to be ideal for maximum longevity.
Seed Storage and Deterioration 197

Water in seeds is a complex system and is only partially understood (Leopold 1986;
Stanwood and McDonald 1989). The probability of various structural features existing have
been derived through statistical thermodynamical approaches (Nementhy and Sheraga 1962;
Rahman and Stillinger 1971). Empirical physical properties of water such as viscosity, density,
and thermodynamical properties were compiled by the United States Bureau of Statistics in the
1920s and are valid for bulk water but not that associated with plant substances. The way in
which water interacts with plant substances is more complex (Kavanau 1964). In much of the
literature, we fmd references to bound water, adsorbed water, and free water which have been
characterized into three phases or zones (Vertucci and Leopold 1986) depending on the way it
is held by the plant substances. Bound water is tightly held to ionic groups such as amino or
carboxyl groups and exists as a monolayer around macromolecules ofthe seed. Adsorbed water
is considered to exist in multilayers, loosely held by bonding to hydroxyl and amide groups
above the monolayer of bound water. Free water is considered as capillary or solution water held
only by capillary forces to the seed tissues. However, these concepts about seed moisture may
be oversimplifications.
Even though much is not known about water-plant substance relationships, it is known that
water is associated with the seed system in several patterns. In some cases, it is actually part of
the chemical structure of other molecules of the seed tissue, held by hydrogen bonding
(vectorized polar bonds), and does not exist as discrete water molecules. In other cases, water
is held as discrete molecules in bonding interactions with seed tissue molecules, though the
arrangement and stability of this type of water is highly variable. These interactions may extend
into the surrounding liquid, forming gradient patterns of structure in a dynamic state ofturnover.
There may be water, but always in association with other systems, and at which point water is
bound and free is difficult to ascertain.
Moisture Equilibrium. The hygroscopic nature of seeds allows them to maintain
equilibrium moisture content with any given relative humidity (Table 9.2). Equilibrium is
attained when the seed has no further tendency to absorb or lose moisture. Hygroscopic
equilibrium curves, also called absorption isotherms, are graphic expressions of the relationship
between the moisture content of seeds and their ambient relative humidity at constant
temperatures. They are established by measuring the absorption or desorption at successive
relative humidities and can be used to predict seed moisture content at any given relative
humidity.
The hygroscopic moisture equilibrium curve in Figure 9.2 is a sigmoid like curve, with three
rather distinct phases representing different stages of water absorption or desorption. Phase one
represents very tightly held water that may actually be a part of the chemical structure of the
seed. This kind of water cannot be removed without destruction of the seed tissue. This phase
may also include some water held as discrete molecules in bonding interactions with the seed
tissue molecules.
Phase two of the moisture equilibrium curve represents water that is more loosely held than
that of phase one. For most seeds, this portion of the moisture equilibrium is represented by a
straight-line relationship between relative humidity and moisture content. Water represented by
the upper portion of phase two is easily removed by drying; however, the lower portion
representing strong bonding is difficult to remove. Water in the upper portion of phase two
contributes significantly to seed deterioration during storage.
 \C
00
-
Table 9.2. Approximate Moisture Content of Vegetable and Field Crop Seeds in Equilibrium with Air at Different Relative Humidities at Room
Temperature (Approximately 77° F) (Moisture Content Wet Basis, in Percentages).

Vegetable seeds Field crop seeds

% Relative Humidity % Relative Humidity
Species 10 20 30 45 60 75 Species 15 30 45 60 75 90 100
Bean, Broad 4.2 5.8 7.2 9.3 11.1 14.5 Barley 6.0 8.4 10.0 12.1 14.4 19.5 26.8
Bean, Lima 4.6 6.6 7.7 9.2 11.0 13.8 Buckwheat 6.7 9.1 10.8 12.7 15.0 19.1 24.5
Bean, Snap 3.0 4.8 6.8 9.4 12.0 15.0 Shelled Corn,
Beet, Garden 2.1 4.0 5.8 7.6 9.4 11.2 yd 6.4 8.4 10.5 12.9 14.8 19.1 23.8
Cabbage 3.2 4.6 5.4 6.4 7.6 9.6 Shelled Corn.
Cabbage. Chinese 2.4 3.4 4.6 6.3 7.8 9.4 wd 6.6 8.4 10.4 12.9 14.7 18.9 24.6
Carrot 4.5 5.9 6.8 7.9 9.2 11.6 Shelled Corn.
Celery 5.8 7.0 7.8 9.0 10.4 12.4 pop 6.8 8.5 9.8 12.2 13.6 18.3 23.0
Corn. Sweet 3.8 5.8 7.0 9.0 10.6 12.8 Flaxseed 4.4 5.6 6.3 7.9 10.0 15.2 21.4
Cucumber 2.6 4.3 5.6 7.1 8.4 10.1 Oats 5.7 8.0 9.6 11.8 13.8 18.5 24.1
Eggplant 3.1 4.9 6.3 8.0 9.8 11.9 Peanut 2.6 4.2 5.6 7.2 9.8 13.0
Lettuce 2.8 4.2 5.1 5.9 7.1 9.6 Rice. Milled 6.8 9.0 10.7 12.6 14.4 18.1 23.6
Mustard, Leaf 1.8 3.2 4.6 6.3 7.8 9.4 Rye 7.0 8.7 10.5 12.2 14.8 20.6 26.7
Okra 3.8 7.2 8.3 10.0 11.2 13.1 Sorghum 6.4 8.6 10.5 12.0 15.2 18.8 21.9
Onion 4.6 6.8 8.0 9.5 11.2 13.4 Soybeans 4.3 6.5 7.4 9.3 13.1 18.8
Onion Welsh 3.4 5.1 6.9 9.4 11.8 14.0 Wheat.
Parsnip 5.0 6.1 7.0 8.2 9.5 11.2 White 6.7 8.6 9.9 11.8 15.0 19.7 26.3
Pea 5.4 7.3 8.6 10.1 11.9 15.0 Wheat.
Pepper 2.8 4.5 6.0 7.8 9.2 11.0 Durum 6.6 8.5 10.0 11.5 14.1 19.3 26.6 f(l
Radish 2.6 3.8 5.1 6.8 8.3 10.2 Wheat. Soft ~
!:l..
Spinach 4.6 6.5 7.8 9.5 11.1 13.2 Red Winter 6.3 8.6 10.6 11.9 14.6 19.7 25.6
5.6 ~
Squash. Winter 3.0 4.3 7.4 9.0 10.8 Wheat. Hard Cl
'-;
Tomato 3.2 5.0 6.3 7.8 9.2 11.1 Red Winter 6.4 8.5 10.5 12.5 14.6 19.7 25.9
Turnip 2.6 5.1
~
~
4.0 6.3 7.4 9.0 Wheat. Hard
Watermelon 3.0 6.1 t:l
4.8 7.6 8.8 10.4 Red Spring 6.8 8.5 10.1 11.8 14.8 19.7 25.0 ::s
!:l..
Data from Harrington (1960a). ~
Cl
~
~
~
-.
~
Seed Storage and Deterioration 199

Moisture
Equilibrium
Curve

Relative Humidity (%)

Figure 9.2. An absorption isotherm showing relationship of seed moisture content with relative
humidity of air at a given temperature.

Water in phase three represents water loosely held by very weak bonding and free water
in the intercellular and intertissue spaces. It is easily eliminated during drying; but if not
eliminated, contributes to rapid seed deterioration.
Effect of Temperature. The moisture equilibrium curves are only slightly influenced by
temperature; an increase in temperature causes a slight reduction in moisture content at a fixed
relative humidity (Haynes 1961). Most equilibrium moisture curves are established at 77°P;
however, a formula for predicting the effect of temperature at any relative humidity has been
devised (Hibbard and Miller 1928).
The Hysteresis Phenomenon. The desorption equilibrium curve will usually be slightly
higher than the adsorption curve. The difference in the equilibrium moisture content during
adsorption and desorption is known as hysteresis. The higher desorption isotherm has been
attributed to the appearance of additional points of attachment (polar sites) for bound water as
a result of tissue swelling (Abdul-Baki and Anderson 1970). On desorption, it was suggested
that the disappearance of these polar sites was delayed by their tendency to hold and keep the
bound water that intervenes to block the collapse of the seed tissue. The study concluded that
hysteresis could only occur in absorptive substances with a high degree of structural rigidity.
200 Seed Storage and Longevity

Temperature

At temperatures of O°C, formation of intracellular ice crystals can disrupt membrane

integrity and contribute to seed deterioration. Seeds with moisture levels below 14% do not form
ice crystals. It should be noted, however, that at 14% initial moisture, seeds stored in cold rooms
below O°C will likely gain moisture. Most cold rooms have a high relative humidity, and seeds
achieve equilibrium with that relative humidity after a brief period of storage. Thus, seeds stored
at low temperatures must be in conditions in which the relative humidity is controlled or placed
in moisture-proof containers to avoid increases in moisture content and increased deterioration.

Association Between Seed Moisture Content and Temperature

The axiom that the sum of the percentage of relative humidity and temperature in degrees
Farenheit should not exceed 100 for safe storage implies an equivalence of the effects of
temperature and humidity on seed longevity. We now know that this is not true. Both parameters
influence seed metabolism. High relative humidities increase seed moisture content, which
results in biochemical events such as increased hydrolytic enzyme activity, enhanced respiration,
and increases in free fatty acids. High temperatures serve to enhance the rate at which many
enzymatic and metabolic reactions occur, causing a more rapid rate of deterioration. However,
seed moisture content is considered to be the most critical factor in maintaining seed longevity.
Although seed moisture content and high temperatures are interrelated, high temperatures hasten
the deterioration of high-moisture seeds by increasing the metabolic activity of hydrolyzed
substrates and enzymes. High temperatures exert only a minimal deteriorative effect on low-
moisture seeds. It has been shown that low-moisture seeds store well at temperatures up to 25°C
but high-moisture seeds will store well only if the temperature is reduced to lOoC or less. Thus,
although temperature and relative humidity interact to determine seed longevity, the control of
relative humidity and its subsequent effect on seed moisture content is more critical than storage
temperature in achieving optimum storage conditions.

Genetic Factors

Seeds of some species are genetically and chemically equipped for longer storability than
others under similar conditions. Most long-lived seeds belong to species possessing hard,
impermeable seed coats. Seeds of Canna (S ivori et al. 1968), Lotus (Wester 1973), and Lupinus
(Porsild and Harrington 1967) have been reported to be viable even after 500 years. Other hard-
seeded genera reported by Harrington (1972) to be germinable after 100 years include Albizia,
Cassia, Goodia, and Trifolium.
Seeds of other species are characteristically short-lived. These include vegetables such as
lettuce, onion, and parsnip and agronomic crops such as rye (Table 9.3). Wheat exhibits
intermediate storability compared to alsike clover which has excellent storability (Table 9.3).
Generally, seed species possessing high oil content do not store as well as those with low oil
content. However, such total compositional analyses may be misleading. What is more important
is the quantity of oil present in the portion of the seed responsible for germination. For example,
whole wheat seeds contain only about 3% oil, but their embryo, has about 27% oil. Seeds of
different species may also be chemically similar but have greatly different storability due to
differences in genetic potential. For example, Chewings fescue and annual ryegrass seeds are
Seed Storage and Deterioration 201

similar in appearance and chemical composition; however, ryegrass seeds have much better
storability under comparable conditions. These genetic factors affect seed storability and have
led to the development of tables in which species differences are divided into three broad
categories according to their relative storability (Table 9.3).
Genetic differences in storage potential are not limited to seeds of different species.
Differences in seed storability may also occur among cultivars. For example, the bean cultivar
Black Valentine stores better than Brittle Wax (Toole and Toole 1953). Similarly, some inbred
lines of com have been shown to germinate 90% after 12 years of storage while others were
completely dead following the same storage period (Lindstrom 1942). Kueneman (1983)
demonstrated by means of reciprocal soybean crosses that the maternal plant was responsible
for increased storability of F1 seeds perhaps by influencing the permeability of the seed coat.
Thus, inheritance clearly exerts a marked effect on seed longevity and can be a focus of breeding
programs. It should not be forgotten, however, that the environment strongly alters the genetic
potential for seed longevity.
Table 9.3. Relative Storability Index (1 = 50% of the Seeds are Expected to Germinate
After 1 to 2 Years Storage, 2 = 50% of the Seeds are Expected to Germinate
After 3 to 5 Years Storage, 3 = 50% of the Seeds are Expected to Germinate
After 5 or More Years) of Important Crop Seeds (from Justice and Bass 1978).
Relative Storability Relative Storability
Crop Index Crop Index
Alfalfa 3 Bahiagrass 2
Barley 2 Field bean 2
Sugarbeet 3 Creeping bentgrass 3
Velvet bentgrass 3 Bermuda grass 1
Kentucky bluegrass 2 Big bluestern 2
Mountain brome 2 Smooth brome 2
Buckwheat 2 Buffalograss 3
Reed canarygrass 1 Carpetgrass 3
Alsike clover 3 Berseem clover 2
Crimson clover 2 Subterraneum clover 2
Field corn 1 Cotton 1
Cowpea 1 Dal1isgrass 1
Chewings fescue 2 Tall fescue 2
Flax 2 Blue grama 2
Hardinggrass 1 Korean lespedeza 1
Yellow lupine 1 Foxtail millet 1
Pearl millet 1 Oats 2
Orchardgrass 1 Peanut 1
Rape 2 Redtop 1
Rice 2 Rye 1
Perennial ryegrass 2 Sorghum 1
Soybean 1 Sunflower 1
White sweetclover 3 Timothy 2
Birdsfoot trefoil 2 Hairy vetch 3
Wheat 2 Broccoli 2
Cabbage 2 Carrot 2
Sweet corn 2 Cucumber 2
Lettuce 1 Onion 1
Parsley 1 Parsnip 1
Pumpkin 2 New Zealand spinach 2
Tomato 3 Turnip 2
202 Seed Storage and Longevity

Presence of MicroOora

Two types of fungi invade seeds: field fungi and storage fungi. Field fungi infect seeds that
are developing on the mother plant and typically require high relative humidity (90 to 95%) or
high seed moisture content (30 to 35%). Since these conditions occur only during seed
maturation or imbibition, field fungi seldom contribute to seed deterioration during storage.
In contrast, storage fungi have the capacity to grow without free water. In general, they
grow at seed moisture contents in equilibrium with relative humidities from 65 to 90%. The
optimum temperature for growth of storage fungi is about 30 to 33°C, with a maximum at 55°C
and a minimum ofO°C. Most storage fungi belong to one of two principal genera, Aspergillus
and Penicillium. Species from these genera are saprophytes and survive on dead tissue. Most
storage fungi ultimately invade the embryo of the seed. They cause seed deterioration not only
through invasion, but also by producing toxic metabolites that destroy cells that provide the dead
tissue on which they subsist. An excellent treatment of the importance of microflora and has
been prepared by Christensen and Meromick (1986) and been the subject of symposia (West
1986; APS 1983). Interestingly, bacteria do not substantially reduce the germination of stored
seeds since they require free water for growth.

Mechanical Damage

Seed production practices such as harvesting, cleaning, and handling inevitably lead to
mechanical damage. Although the immediate effect of such damage on seed quality is generally
not serious, the delayed effects of mechanical damage on seed longevity are much more
troublesome and of much greater economic significance (McDonald 1985). Seed deterioration
is inexorable and progressive; thus, small mechanically damaged areas that initially have little
impact on seed performance may later increase in size and cause deterioration of vital embryonic
tissues, resulting in poor seed quality (Moore 1972). Direct injuries to embryonic tissues are
much more detrimental to seed longevity than are large injuries to nonembryonic tissues.
Mechanical damage also promotes invasion by storage fungi, which can enter the seed through
cracks in the seed coat (Beattie and Boswell 1939; Mamiepic and Caldwell 1963).
Another important production process that permits the germination of many hard seeds is
scarification. However, this mechanical or chemical treatment invariably causes some
mechanical damage. Studies have also shown that scarified seeds do not store as well as
nonscarified seeds (Battle 1948; Brett 1952). Delinting of cotton seeds also reduces storability
(Simpson 1946) as does hulling of cool-season grasses (Canode 1972).

Seed Maturity

Factors such as temperature, moisture, variety, and nutrient status influence seed maturity,
which, in tum, influence seed storability. The greatest storage potential is attained at the time
of physiological maturity, or maximum dry weight of the seed. Many plants (e.g., carrot,
grasses) have an indeterminate flowering pattern; that is, the most mature flowers grow at the
base of the inflorescence with more immature flowers formed on the newer branches. Thus,
varying stages of seed maturity, from recently fertilized ovules to mature seeds, occur on the
same plant. Harvested seeds from such plants show varying degrees of maturity and different
storage potential.
Seed Storage and Deterioration 203

Although immature (or small) seeds have been shown in a number of studies to be inferior
to mature seeds in viability and vigor (Ries 1971; Lopez and Grabe 1973; Clark and Peck 1968;
Austin and Longden 1967; Fontes and Ohlrogge 1972), other factors such as nutrient status can
also influence seed longevity. Harrington (l960b) demonstrated that carrot and pepper plants
grown in nutrient solutions deficient in nitrogen, potassium, or calcium produced seeds that did
not store well over an eight year period. Seeds deficient in phosphorus, however, did not exhibit
decreased storage potential. A similar nutrient deficiency symptom that affects seed quality is
"marsh spot" of pea. These seeds have a magnesium deficiency that expresses itself as a brown
necrotic area on the adaxial surface ofthe cotyledons and results in low quality pea seeds.

PREDICTING SEED DETERIORATION

The ability to predict or forecast seed deterioration would be extremely valuable to seed
companies and germplasm repositories since the loss of seed quality could be anticipated and
seed stocks replenished in orderly fashion. Since we know that the type of seed, its initial
germinability and moisture content, and the temperature and relative humidity of the storage
environment greatly affect the degree of seed deterioration, it has been suggested that seed
deterioration can be modeled by mathematical equations (Roberts 1973, 1986). The general
principle is that low-moisture, high-quality seeds stored under cool, dry conditions maintain seed
quality better than high-moisture, low-quality seeds under hot, humid conditions. To utilize this
principle, Roberts (1986) developed the following equation:

KE - Cwlog m - CHt - CQf

v = ~ - plIO

where v equals the probit of percent germinability after a storage period of p days; K, is the
probit of initial germinability of the seed lot; KE, Cw, CH, and CQ are species constants; m is seed
moisture on a fresh weight basis; and t is the storage temperature in degrees celsius. This
assumes that loss in seed germination is normally distributed in time under a constant
environment. This is useful for two reasons. First, the sigmoidal curve typical of the decline in
germination during storage can be made a straight line using the probit transformation (Figure
9.3). Thus, the curve is completely described by two parameters, slope and intercept. Second,
germination following long-term storage may be anticipated from short-term results by
extrapolation ofthe line. This is the method by which the four species constants are determined.
Considerable research is needed to validate this mathematical test of seed deterioration rate.
Its limitations are associated with precisely characterizing the quality of the seed being stored
and the storage conditions. Bewley and Black (1982) identified these factors as (1) cultivar and
harvest variability, (2) pre- and post-harvest conditions, (3) oxygen pressure effects during
storage, and (4) fluctuating environmental conditions. Wilson and McDonald (1989) tested the
predictions using Phmeolus vulgaris seeds and found that further modification of the equation
to account for seed moisture influence on deterioration would be useful. Even so, this attempt
at mathematically describing seed deterioration has been useful to physiologists as they attempt
to better understand the factors that cause loss in seed quality during storage.
204 Seed Storage and Longevity

MAINTAINING SEEDS IN STORAGE

The purpose of seed storage is to preserve planting stocks from one season to the next. In
some cases (e.g., seed companies), the objective of seed storage is to maintain seed quality for
the longest duration possible. This approach creates a greater diversity in seed inventory and
provides a guarantee of seed supply in years when acceptable seed quality and production is
low. In addition, seed storage enables the maintenance of germplasm over time for improved
plant breeding programs. Four principal approaches are recommended for storing seeds:
conditioned, cryogenic, hermetic, and containerized storage.

"iil100
III " ,,
", ,,
~ 90 ,, (al ... (b)
"iil ;) \
,,
U
,
\ \
ill
c: 80 \

,
~

III \ \

;) :;: 70 \

,,
\ \
U 0
\

,,
\
~ \
III
0- III 60 ,, \
,,
0- E ,,
:J 50 \
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, \
,
0~ c 40 '.,
\
,,
:>: ;)
'- 30 ,, \
III , \
\
,
::aro 0-
III 20
\
\
\
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'.
..
1:; \
> ~ 10 , ------~ ...... ~:~...
III
0 0
.,
~,,-+-a-

4 99.99
3
99
......"....
...
,,
(c)
, ,
>- 2
.. , ,
~ ",
90
Q

.a
ro
:>:
.::: 70 , '"
.. . ,
">
::a0 -
0
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:0
ro
:>
50
30
10
'-a
. ,
, ... , ,
ct - 2
,,
, ., ,
,
,
-3 ,,

4 001
Storage period, P

Figure 9.3. Graphs (a) and (b) show typical seed survival curves, which are cumulative normal
distributions of negative slope, under conditions when the temperature and moisture content remain
constant during storage. The frequency distribution of lijespans which give rise to these survival
curves are also shown. The standard deviation, (a), is indicated below each curve. Graphs (c) and
(d) show the same survival curves as (a) and (b), respectively, when percentage viability is transformed
to probit values; these curves are described by Eq. (1) and the slope, lIa, is indicated on one of the
curves in (c). Graphs (a) and (c) show two survival curves representing two different seed lots of the
same species stored under identical conditions which, therefore, have identical slopes but where the
seed lot constant, K" of one seed lot (---) has a greater value than the other seed lot (-).
Graphs (b) and (d) show two survival curves of the same seed lot (therefore the same K,-
value, stored in two different environments so that the slope, lin, in the less deleterious
environment (---) has half the value of the other environment) (After Roberts and Ellis
1982).
Seed Storage and Deterioration 205
Conditioned Storage

Seeds of most species may be safely stored for several years by careful control of
temperature and relative humidity. Although such conditions are too costly for most agricultural
seed lots, they may be extremely valuable for preserving germplasm and certain high-value seed
stocks. In some parts of the world, especially in the tropics, conditioned storage is necessary in
order to maintain high seed viability of some crop species from harvest to planting (Harrington
1973). At least four factors should be considered when evaluating the economics of seed storage
(Welch and Delouche 1974).
Type of Seed To Be Stored. Seed storage requires a large investment in construction,
maintenance, and power. However, the relatively high cost of seed, particularly that which must
be carried over, may justify such an investment. Factors that should be considered in making
conditioned storage decisions include: (a) the differential between seed and grain prices, (b) the
prevailing price of seed to be stored and its storage potential, (c) the weight per volume of seed
(costs of construction, conditioning, and storage capacity will reflect storage volume rather than
seed weight), (d) the extent ofloss resulting from disposing the seed in alternative markets (e.g.,
selling as grain, or dumping in the case of treated seed), and (e) the projected seed inventory
(volume). Although all seeds benefit from conditioned storage, it becomes practical only when
income derived from seed storage exceeds the costs involved.
Length of Storage. Costs of installation and maintenance of a conditioned storeroom can
be substantial over time and will vary with geographical location. The costs of operating a
conditioned storeroom will be less in the Great Lakes area where the prevailing environment is
generally cool and dry compared to hot and humid areas of the Gulf of Mexico. In most
instances, it is only essential to maintain conditioned storage for seeds from one growing season
through the following summer to the next spring planting, a period of 18 to 20 months.
Quality of Seed Stored. Many people believe that conditioned storage is more beneficial
for low-quality than high-quality seeds. Figure 9.4 demonstrates that low quality garden bean
and sorghum seeds store poorly compared to high-quality seeds even though all lots have the
same initial germination. Seed quality cannot be improved by seed storage. Even the best storage
conditions can only maintain the quality of the seed placed in storage, and low-quality seeds
deteriorate more rapidly under conditioned storage than high-quality seeds.
Loss of Seed Weight During Conditioned Storage. High-moisture seeds that are placed
in a conditioned storeroom at 40% relative humidity will gradually decrease to 10% moisture
content as moisture equilibrium is attained. Thus, the seed will weigh less following conditioned
storage as a result of water loss. Consequently, conditioned storage will eventually result in
higher density seed lots with more seed per unit volume and may become an important economic
consideration for seed producers.
Requirements of Conditioned Storage. Conditioned storage involves placing seeds in a
dry and cool environment for extended periods. But, how dry and how cool? Seeds of most grain
crops (e.g., corn, wheat, barley, sorghum, and oats) will maintain satisfactory germination and
vigor for about one year at moisture contents of 12 to 13% under normal warehouse
temperatures. When longer storage is needed, seed moisture content should be less than 11 % and
the temperature should not exceed 20°C. Similarly, seeds of most temperate zone grass and
legume crops can be stored safely for one year at moisture contents of 10 to 11 % under normal
warehouse temperatures. When longer storage is needed, 10% seed moisture content and
temperatures of 20°C or less are recommended.
206 Seed Storage and Longevity

Seed of some species (e.g.~ soybean and peanut) may lose viability even over one year if
kept at 11 to 12% moisture content at temperatures of 20 0 e or less. Although carryover of
soybean seed lots is generally not recommended, they may be kept safely for 18 to 20 months
in conditioned storage facilities of 20°C and 50% relative humidity. While most orthodox seeds
can be successfully stored under conditioned storage facilities, operation of the facilities is costly
and mechanical breakdowns of the cooling units are common. As a result, more reliable
approaches to long-term preservation of seeds have been sought.
To achieve these storage conditions, conditioned seed storage rooms typically contain
refrigeration and dehumidification equipment. The refrigeration is necessary to lower the
temperature of the seed storage room and dehumidification is required because lower storage
air temperatures hold less water causing the relative humidity of the air to increase. In general,
the temperature and relative humidity conditions of the storage environment are determined to
meet satisfactory storage conditions identified by Harrington (1972) while satisfying the
objectives of the seed storage manager.
Storage of seed for maintaining germplasm requires conditions that are not feasible for seed
industry quality control programs. For this purpose, seeds are often stored at temperatures
ranging from -15° to -20 0 e.

Cryogenic Storage

While most orthodox seeds can be successfully stored in conditioned storage facilities,
routine operation of such facilities is costly and mechanical breakdowns are common. Another
approach to minimize these difficulties is cryogenic storage. This storage method places seeds
into liquid nitrogen at a temperature of -196°e (seeds are actually placed into the gaseous phase
ofthe liquid nitrogen, at approximately -150°C, to facilitate handling and safety. The differential
in seed temperature is not considered significant in influencing seed longevity).
The advantage of this approach is that seeds are placed at a temperature where little
detrimental physiological activity occurs, thereby prolonging storage life. From a practical
perspective, the cost of the liquid nitrogen is minimal compared to the cost of maintaining
conditioned storerooms. In addition~ no working parts are necessary so repair of equipment is
not required. Liquid nitrogen is also an inert gas that volatilizes easily. Therefore, it remains
relatively safe, although air circulation in the storage room is necessary to guard against the
room being filled with nitrogen gas and to prevent asphyxiation. Finally, this seed storage
method is limited in capacity to the amount of storage space available in the cryogenic tanks.
It is not practical for most commercial seed, although it may be useful for maintaining valuable
seed germplasm over prolonged periods.
Many studies are now under way on the ability to safely store a variety of seeds using
cryogenic storage (Stanwood 1985). The general conclusion is that most agronomic crops can
be successfully stored in liquid nitrogen. It seems clear that under such circumstances, there is
little question of metabolism or even conventional chemical activity ensuring successful seed
storage for prolonged periods. Yet, not all seeds are able to tolerate liquid nitrogen storage. The
reasons for this are still unknown although seed moisture content is certainly involved. In seeds,
there is a distinct moisture content below which freezable water which kills cells does not occur.
Sesame seeds, for example, are able to tolerate liquid nitrogen freezing up to 12% moisture, at
which point germination declines rapidly. A central theme of cryogenic storage has been to
Seed Storage and Deterioration 207

75
Z
0
1=
z......<
50
~~
0
~
GARDEN BEANS
25

MONTHS IN STORAGE

100

75
Z B
0
1=
z<
...... SO
~
~
0
t.~
SORGHUM
25

10 20 30
MONTHS IN STORAGE
Figure 9.4. Differences in longevity of three seed lots each ofgarden beans and sorghum under open
storage conditions (From Delouche 1973).
208 Seed Storage and Longevity

replace freezable water in seeds with protective cryogenic compounds such as sugars and
glycols. The objective is to replace the water in the seeds by infiltrating the protective
compounds into the seeds without inducing subsequent seed injury.

Hermetic Storage

In recent years, packaging seeds in moisture-resistant or hermetically sealed containers

for storage and marketing has been explored. The purpose of such containers is to maintain
seeds at safe storage moisture levels. Figures 9.5 and 9.6 show the effectiveness of several types
of packaging materials in preventing moisture uptake and maintaining seed viability. Ordinary
paper and cloth containers were least effective, while various laminate and polyethylene
materials were moderately effective. Metal cans were completely effective in maintaining seed
moisture at the initial 5% level. Such completely moisture-proof containers hermetically seal the
seed and are effective for long-term seed storage up to 10 years or more. The effectiveness of
other materials was directly associated with their ability to resist moisture.
The moisture content of seeds placed in hemetic storage must be lower (2 to 3%) than that
at which seeds are normally packaged in nonsealed storage. Most observations indicate that
starchy seeds above 12% and oily seeds above 9% deteriorate faster in sealed storage than in
nonsealed storage (Harrington 1973). It is believed that the atmosphere in a moisture-proof
container holding seed at 13% moisture will equilibrate at a relative humidity of about 65%.
Some seed storage fungi that are detrimental to seed quality multiply under such conditions. In
contrast, the relative humidity surrounding com seed packaged in porous containers such as
paper bags at 13% moisture will rise to nearly 100% and decrease below 65% at other times.
As a result, seeds placed in open containers will generally decrease in moisture during the winter
when relative humidities are the lowest and thus inhibit the growth of storage fungi.
One of the central benefits of hermetic seed storage is the ability to remove ambient air
from the seeds and replace it with specific gases known to prolong seed storage life. This is
necessary because the composition of the gaseous environment in hermetic storage changes with
time and the effect is more pronounced with high seed moisture content. For example, Roberts
and Abdalla (1968) demonstrated that pea seeds stored at 18.4% moisture content at 25°C in
ambient air had a decrease in oxygen from 21 to 1.4% and an increase in carbon dioxide from
0.03 to 12% after only 11 weeks. This decrease in oxygen concentration and/or increase in
carbon dioxide level may differentially affect the storage capability of the seeds.
The most extreme case of hermetic storage is to store seeds in vacuo at low moisture
contents. While some reports have indicated that this approach extends seed storage life
(Bockholt et al. 1969; Lougheed et al. 1976), the majority of reports suggest no benefit from
vacuum storage (Justice and Bass 1978; Bass and Stanwood 1978). Studies have also been
conducted which replace ambient air with a pure gas in sealed containers. Some advantage has
been reported for pure carbon dioxide (Bennici et a1. 1984; Harrison 1966) and the inert gases
nitrogen, argon, and helium (Harrison 1966; Quaglia et al. 1980), but most reports show little
or no advantage to this approach (Justice and Bass 1978). Interestingly, a consensus is emerging
that seeds stored in oxygen at low moisture contents deteriorate more rapidly (Harrison 1966;
Roberts and Abdalla 1968; Ohlrogge and Kernan 1982). This effect may be associated with the
physicaVphysiological mechanisms occurring during lipid peroxidation that will be described
later.
Seed Storage and Deterioration 209

100

Can

'\
".\
s:: " \
o
.... '. \
\
.S
"

60
...
E
o
<I)

~
40

20

o 3 6 9 12 18 24 36 42 48 60 72
Months Storage

Figure 9,5. The effect ofdifferent packaging materials on the germination ofcreeping redfescue seed
(From Grabe and Isley 1969).

25

20

15

10

--------------- Can

3 6 9 12 18
Months' Storage

Figure 9.6. The effect ofdifferent packaging materials on the moisture content ofcreeping redfescue
seed (From Grabe and lsely 1969).
210 Seed Storage and Longevity

Containerized Seed Storage

Absolute humidity control in seed storage areas requires considerable investment in

specially constructed rooms and dehumidifying equipment. Most seed stock organizations and
commercial seed companies have special facilities for storing germplasm and high-value seed
stocks where relative humidity and temperature are closely regulated. The National Seed Storage
Laboratory operated by the United States Department of Agriculture at Fort Collins, Colorado,
as a permanent repository for valuable germplasm of all kinds of plants, provides temperature
and relative humidity conditions satisfactory for long-term storage of all kinds of seeds.
Where elaborate facilities are not available, humidity can be closely regulated in a closed
container by use of chemical desiccants that have known moisture equilibrium values. Either
saturated salt solutions or acid solutions may be used. Sulfuric acid (H2S04) is a common acid
desiccant and is diluted with water to provide increasing relative humidity levels (Table 9.4).
Since H2S04 is very caustic and corrosive, it should be handled with extreme caution to prevent
injury to personnel as well as to the seeds. Saturated salt solutions are less hazardous to both
seed and personnel; however different salts must be used to attain various humidity levels (Table
9.5). Complete saturation is attained when a given quantity of water dissolves its full capacity
of salt crystals. To maintain comp lete saturation, some additional salt should be added so that
the relative humidity is held constant. Care must be used to keep the seed separate. This can
be accomplished by using metal platforms, cheesecloth, or wire containers for holding the seed.
Depending on the size of the container, small fans may be used to keep the relative humidity
distributed uniformly throughout. Exposure of seed to uniform relative humidity can also be
aided by use of small seed containers or appropriate use of wire screens throughout the seed
mass.
A common seed desiccant is silica gel treated with cobalt chloride which serves as an
indicator dye and turns from blue to pink when the relative humidity exceeds 45%. The silica
gel-cobalt chloride granules are placed in a metal box with the seeds in a proportion of 1 kg
desiccant per 10 kg of seeds. Seeds can thus be stored safely for several years because the
relative humidity within the box is kept below 45%. The advantages of this desiccant storage
system are: (I) construction of conditioned storage facilities is not necessary, (2) maintenance
and operation costs are minimal, (3) the metal storage boxes are insect-, rodent-, and moisture-
proof, and (4) the seeds are not damaged by storage fungi since they are maintained at 45%
relative humidity. The only care required for desiccant seed storage is frequent inspection ofthe
silica gel to insure that the indicator silica gel remains blue.

SYMPTOMS OF SEED DETERIORATION

The process of seed deterioration is complex; therefore, the conclusions from seed
deterioration studies are often difficult to critically evaluate. There are at least two reasons for
this. First, most seed deterioration studies have focused on whole seed germination response or
enzyme analyses without considering the fact that seed deterioration probably does not occur
uniformly throughout a seed. For example, young vigorous embryos that are excised and
transplanted onto deteriorated endosperms do not grow as well as those planted onto
nondeteriorated endosperms. This suggests that endosperm deterioration markedly alters the
performance of a vigorous embryo (Floris 1970; MandaI and Basu 1981). Similarly, Bulat
(1963) showed that unique seed deterioration patterns occurred in 41 species. In many cases,
Seed Storage and Deterioration 211
the necrosis was first detected in the embryo and usually around the radicle tip. Thus, it should
be recognized that seed parts vary in their chemistry and susceptibility to deterioration. Studies
examining this differential susceptibility of seed parts which collectively influence the whole
seed response should be useful in understanding the mechanisms of seed deterioration.
Second, all seed lots are composed of individual seeds, each possessing its own unique
capability to perform in the field. A hypothetical distribution of loss in seed quality over time,
therefore, can be represented by Figure 9.7. Two observations can be made regarding this
deterioration (McDonald and Wilson 1980). First, the proportion of high-quality seeds within
a population decreases with increasing storage time. Second, the curves shift from high to low
seed quality and the range becomes wider with increased storage time. As seeds age, overall
quality level of the seed lot declines. Since some seeds are inherently stronger than others, a
remnant of high quality seeds remains even after a three-year storage period. The point of this
example is that seeds in storage do not die simultaneously, but age at differing rates. Yet, most
studies of seed deterioration are expressed as an average of some subsample greater than one
seed (e.g., conductivity of 20 seeds), or as some average value of a subsample that represents
the entire population of seeds (e.g., percentage germination). Thus, total population studies of
seed deterioration do not represent what is occurring at the individual seed level.

Table 9.4. Relative Humidity in Equilibrium with Different

Concentrations of Aqueous Acid Solutions at Various Temperatures.

Acid by weight. percent

Acid Temperature OF 20 40 60 80
H2S04 (Sulfuric) 0 87.3 55.7 15.0 3.14
50 87.4 56.6 15.8 3.88
68 87.7 56.7 16.3 4.76
86 87.5 56.6 17.0 5.75
104 87.6 57.5 17.8 6.88
112 88.8 58.2 18.8 8.2
Acid by weight, percent
20 30 40 50
HN0 3 (Nitric) 0 89.2 78.4 65.3 45.7
50 86.7 77.0 63.0 45.6
68 86.6 75.2 61.5
86 86.6 74.9 61.3
104 85.9 74.1 60.5
112 86.5 74.6
140 86.9 75.6
Acid by weight, percent
10 20 30 40
Hel (Hydrochloric) 0 83.5 56.0 27.4 8.9
50 83.5
68 83.2
86 84.2

Calculated by.£. from basic data.

Po
From Hall (1957).
212 Seed Storage and Longevity

Table 9.5. Relative Humidity in Equilibrium with Saturated Salt Solutions

at Different Temperatures.

Temp. RH Temp. RH
Salt (OF) (Percent) Salt (OF) (Percent)
BaCI 2 ·2Hp 77 91.2 LiCI·Hp 50 13.3
(Bari urn chloride) 86 90.8 (Lithium chloride) 68 12.4
95 90.2 86 11.8
104 89.7 104 11.6
CaCI 2 20 44 MgCl 2 73 32.9
(Cald urn chloride) 32 41 (Magnesium chloride) 86 32.4
50 40 100 31.9
70 35
Ca(NO J )2 73 51.8 Mg(NO J )2 73 53.5
(Calcium nitrate) 86 46.6 (Magnesium nitrate) 86 51.4
100 38.9 100 49.0
CuCI 2'2H 0 68 68.7 Na 2 Cr 2 0 7 ·2H 2 O 50 57.9
(Cupric chloride) 77 68.7 (Sodium dichromate) 68 55.2
86 68.3 86 52.5
104 67.4 104 49.8
KCI 68 89.2 NH 4 CI 20 82
(Potassium chloride) 77 87.2 (Ammoni urn chloride) 32 83
86 85.3 50 81
95 83.8 70 75
KN0 2 68 49.0 NaC 2H 30 2·3Hp 68 76.0
(Potassium nitrite) 77 48.2 (Sodium acetate) 77 73.7
86 47.2 86 71.3
100 45.9 95 68.8
KN0 3 32 97.6 NaN0 2 68 65.3
(Potassium nitrate) 50 95.5 (Sodium nitrate) 77 64.3
68 93.2 86 63.3
86 90.7 100 61.8
104 87.9
K2 Cr0 4 68 86.6 NaCI 32 74.9
(Potassium chromate) 77 86.5 (Sodium chloride) 50 75.2
86 86.3 68 75.5
100 85.6 86 75.6
104 75.4
122 74.7
From Hall (1957).

Despite these difficulties, the most visib Ie symptoms of seed deterioration are observed first
at the whole seed morphological level and then during germination and seedling growth.
However, these are preceded by numerous ultrastructural and physiological changes whose
symptoms are not as readily apparent but can be detected by sophisticated monitoring techniques
that attempt to identify changes in the deteriorating seed at the physiological level (McDonald
1999).
Seed Storage and Deterioration 213
Seed Symptoms

Morphological Changes. Seed coat color often provides an indication of seed

deterioration, particularly for legumes. Darkening of the seed coat in deteriorating clover
(Vaughan and Delouche 1968), peanut (Marzke et al. 1976), and soybean (Saio et al. 1980)
seeds has been reported. Such color changes are presumably due to oxidative reactions in the
seed coat which are accelerated under conditions of high temperature and relative humidity.
(Hughes and Sandsted 1975). Beyond these seed coat effects, other morphological changes in
deteriorating seeds have been reported. Lettuce seeds develop red necrotic lesions in the
cotyledons known as cotyledonary necrosis (Bass 1970) and lentil seeds characteristically tum
yellow after prolonged storage (Nozzolillo and DeBezada 1984).
Ultrastructural Changes. Deteriorated dry seeds have been examined for ultrastructural
changes using electron microscopy and two general patterns of coalescence of lipid bodies and
plasmalemma withdrawal associated with deterioration have been observed. Coalescence oflipid
bodies in the embryo has been found in a broad group of species, including wheat (Anderson et
a1. 1970), peas (Herman and Granett 1972), and pine (Fernandez Gracia de Castro and
Martinez-Honduvilla 1984). In lettuce, this coalescence of lipid bodies has been detected in the
embryonic axis but not the cotyledons (Smith 1983). Withdrawal of the plasmalemma has also
been detected in these species (above) as wen as in rye (Hallam et al. 1973). It is significant that
both of these events influence cell membrane integrity.
Cell Membranes. One common facet of deteriorating seeds is their inability to retain
cellular constituents which leak out during imbibition. This has three important seed quality

200

--1 year

150
Ul
"0
(I)
(I)
Ul
15 100
, ...,,...--2 years
,,,
(jj
.0
\
E \
:J
Z
I
50 I
I

High Mediunn Low

Seed Quality

Figure 9.7. Hypothetical distribution ofloss in soybean seed quality over a three-year period (From
McDonald and Wilson 1980).
214 Seed Storage and Longevity
implications: (1) many of these cellular constituents are essential for normal, vigorous
germination, (2) some of the exuded compounds are necessary for maintenance of internal
osmotic potential which is responsible for normal water uptake and provides the turgor pressure
required for radicle protrusion, and (3) the external leakage of these substances encourages the
growth of pathogenic microflora.
The reasons for this increased leakage have been attributed to cell membrane disruptions
associated with the loss of membrane phospholipids. Phospholipid decrease has been reported
in deteriorating cucumber (Koostra and Harrington 1969), peanut (Pearce and Abdel-Samad
1980), pea (Powell and Matthews 1981), soybean (Priestley and Leopold 1983), tomato
(Francis and Coolbear 1984), and sunflower (Halder et al. 1983) seeds. The loss of
phospholipids in deteriorating seeds is generally considered to be due to either phospholipase
enzyme activity or lipid peroxidation. Interestingly, some (Koostra and Harrington 1969;
Petruzzelli and Taranto 1984) have suggested that the decline in phospholipids occurs only
under conditions of high relative humidity (as encountered in an accelerated aging test) but not
under long-term dry storage. Since phospholipases are hydrolytic enzymes, they function most
actively under conditions of high seed moisture content. Thus, the decline in phospholipids
reported in the literature under accelerated aging conditions may be attributed to these enzymes
and could be one reason that the rate of seed deterioration is accelerated compared to that in dry
seed storage. In the latter case, lipid peroxidation may be more responsible for the observed
declines in membrane integrity.
Loss of Enzyme Activity. The most sensitive tests for measuring incipient seed
deterioration are those that measure activity of certain enzymes associated with breakdown of
food reserves or biosynthesis of new tissue during germination. Examples include amylases
(Saxena and Maheshwari 1980), proteinases (Nowak and Mierzwinski 1978), cytochrome c
oxidase (Ching 1972), and glyceraldehyde phosphate dehydrogenase (Herman et a1. 1976).
Among the biochemical tests that have been used to measure loss of enzyme activity are
the tetrazolium test (dehydrogenase) (see Chapter 6) and the glutamic acid decarboxylase
(GADA) activity test. Other enzymes that have been correlated with seed deterioration are the
oxidases such as catalase (Crocker and Harrington 1918), peroxidase (McHargue 1920),
amylase (Anderson 1970), and cytochrome oxidase (Throneberry and Smith 1954). Although
these positive associations have been reported, other studies have shown that high levels of
peroxidase activity existed in old wheat seeds (Brocq-Rosseu and Gain 1908) and high levels
of dehydrogenases have been found in heat damaged barley seeds (MacLeod 1952). Changes
in the levels of specific enzymes may not always provide an accurate indication of seed
deterioration. Many of these studies were conducted under accelerated aging conditions which
increase seed moisture content and contamination by microflora-both conditions which are
seldom encountered in long-term dry seed storage.
Reduced Respiration. Respiration is a composite expression of activity of a large group
of enzymes that react together in breaking down food reserves. As seeds deteriorate, respiration
becomes progressively weaker, and ultimately leads to loss of germination. However, prior to
loss of germ inability, the respiration level during the early stages of germination has been
correlated with subsequent seedling vigor (Woodstock and Feeley 1965).
Changes in respiration have been studied under natural aging (Woodstock and Grabe 1967;
Kittock and Law 1968; Abdul-Baki 1969; Anderson 1970), accelerated aging (Woodstock and
Feeley 1965; Abdul-Baki 1969), chilling injury (Throneberry and Smith 1954; Woodstock and
Pollock 1965, Woodstock and Feeley 1965), and irradiation (Woodstock and Combs 1965;
Seed Storage and Deterioration 215
Woodstock and Justice 1967) conditions. Although the association between oxygen utilization
and seed deterioration has been demonstrated, a more sensitive index of deterioration may be the
respiratory quotient (RQ), which represents the CO2 evolved divided by the oxygen utilized.
High RQ values (1.5 or higher) have been reported in deteriorated seeds (Woodstock and Grabe
1967; Anderson 1970; Woodstock et al. 1984) and may be due to increases in CO2 evolution,
reductions in oxygen uptake, or both. Regardless of the cause, it appears that reductions in the
rate of respiration are closely associated with seed deterioration. This may be causally related
to a breakdown in membrane structure, particularly in the mitochondrial cristae, which would
reduce total respiration.
Increases in Seed Leachates. A frequently observed symptom of deteriorated seeds is their
increased leachate content when soaked in water. The degree of deterioration is associated with
the concentration of seed exudates that may be found in the steep solution. These exudates are
a reflection of the amount of membrane degradation that has occurred. The leachate
concentration has been measured by electrical conductance methods (Hibbard and Miller 1928),
and also by determining the soluble sugar content of the leachate (Abdul-Baki and Anderson
1970).
Increase in Free Fatty Acid Content. The hydrolysis of phospholipids leads to the release
of glycerol and fatty acids, and this reaction accelerates with increasing seed moisture content
(Harrington 1973). The continual accumulation of free fatty acids culminates in a reduction in
cellular pH and is detrimental to normal cellular metabolism (Earnshaw et a1. 1970).
Furthermore, it denatures enzymes, resulting in a loss of activity (Tortora et a!. 1978).
Individual cotton seeds containing 1% or more of free fatty acids usually will not germinate
(Hoffpauir et a1. 1947).
There is debate as to whether the increase in free fatty acids is due to production of lipases
by microflora or by the seed itself. Hummel et al. (1954) concluded that the increase in free fatty
acids in wheat was entirely attributable to mold. Fungal invasion is also thought to be a major
cause ofthe breakdown oflipids to fatty acids (Christensen et al. 1949). Others (Priestley 1986)
have indicated that lipase activity can be detected in seeds which are at moisture contents too
low for fungal growth.

Performance Symptoms

Eventually the deterioration of seeds is observable in their lowered performance during

germination. De1ayed seedling emergence is among the fIrst noticeable symptoms, followed by
a slower rate of seedling growth and development and decreased germination. As seeds
deteriorate, the environmental conditions under which they will germinate become narrower
(Heydecker 1969); this symptom also occurs early in seed deterioration. Loss of fIeld emergence
potential is another frequently observed symptom of deterioration (Grabe 1965).
Another symptom of deteriorated seeds is decreased resistance to environmental stresses
during germination and early seedling growth (Isely 1957; Woodstock and Pol1ock 1965). Still
another is reduced yield potential. This may occur even in the absence of the more obvious
symptoms accompanying germination and seedling e:stablishment.
As some areas of the seed lose their viability, the seed may still be able to produce a
seedling, although it may be morphologically abnormal due to malfunction of the deteriorated
areas. Such a condition in lettuce has been called red cotyledon malfunction, causing lack of
hypocotyl elongation and stunting of the radicle, resulting in a stunted seedling incapable of
216 Seed Storage and Longevity

survival (Dempsey and Harrington 1951). Aged onion seeds do not develop the cotyledonary
knee necessary for emergence through the soil, and aged seeds of cucurbits do not develop the
hypocotyl peg that normally cracks open the seed coat, allowing the cotyledons to emerge
(Harrington 1973).
The ultimate performance symptom of seed deterioration is the complete loss of
germinability and death of the seed.

PossmLE CAUSES OF SEED DETERIORATION

An understanding of the fundamental factors that induce aging is essential in a study of

seed deterioration. The following discussion cites several theories that have been suggested as
basic causes of deterioratio~ some of which are entirely speculative. It is fairly certain that seed
deterioration occurs from a combination of several causes.

Lipid Peroxidation

Of all the models presented to explain seed deterioration, the lipid peroxidation model has
stimulated the greatest interest (Wilson and McDonald 1986b; Bewley 1986; McDonald 1999).
A free radical is an atom or a group of atoms with an unpaired electron. They can be produced
either through autoxidation or enzymatically by lipoxygenase which is present in many seeds.
The autoxidation mechanism is often initiated by oxygen around unsaturated or polyunsaturated
fatty acids such as oleic and linoleic acids which are most common in seed membranes (Figure
9.8). The result is the release of a free radical, often hydrogen (H-) from a methylene group of
the fatty acid that is adjacent to a double bond. In other cases, the free radical hydrogen may
combine with other free radicals from carboxyl groups (ROOH) leaving a peroxyfree radical
(ROO-). Once these free radicals are initiated, they create profound damage to membranes,
particularly those where electron transport is most frequent, and continue to propagate other free
radicals until they combine with free radicals which terminates the reaction. The result is the loss
of membrane integrity in the case of phospholipids.
It has been noted that lipid autoxidation occurs in all cells; but in fully imbibed cells, water
acts as a buffer between the reactive compounds and the macromolecules, thus preventing
enzyme inactivation (Harrington 1973). Lipid autoxidation is accelerated at high temperatures
and increased oxygen concentration. Harrington (1973) considered this to be a cause of seed
deterioration only at moisture contents below 6%, since moisture contents from 6% to 12%
maintain seed viability, and above 12% other factors are responsible for deterioration.
Lipoxygenase enzymes also generate free radicals. However, their activity is greatest when
the seed moisture content exceeds 14%, while autoxidation is believed to occur primarily at
lower seed moisture contents. Thus, the mechanism of lipid peroxidation may be different under
accelerated aging (lipoxygenase) compared to long-term aging (autoxidation) conditions. It
should also be noted that oxygen is deleterious to seed storage based on this proposal, which is
consistent with the success of hermetic seed storage and that lipid peroxidation causes loss of
membrane integrity.
Free radicals also attack compounds other than fatty acids. Changes in protein structure
of seeds have also been observed and attributed to free radicals. Free radicals oflipid peroxides
damage cytochrome c by changing its physical and catalytic properties, suggesting that free
radicals attach themselves to the protein by covalent linkages (Tapper 1962). Sulfhydryl levels
Seed Storage and Deterioration 217

Initiation

RH + Oz ---------..
- free radicals (R-)

ROOH - - - - - -... free radicals (ROO.)

Propagation

R' + O z - - - - -.....- ROz"

RO z• + RH -----t.._ R· + ROOH (hydroperoxide)

Termination
R· + R.--_ _ _ _ ~

~ stobIe end products

R- + RO·----
Figure 9.8. Free radical chain reaction resulting in autoxidation (From Wilson and McDonald, 1986).

decrease in wheat flour with increasing oxygen content (Yoneyama et a1. 1970). The "sick wheat
syndrome" which results in a discoloration of the embryo with increasing storage time has been
attributed to a condensation reaction between lysine or methionine protein residues and reducing
sugars (Feeney and Whitaker 1982). Even more importantly, free radicals are also suspected of
assault on chromosomal DNA. As seeds age, the propensity for genetic mutations increases.
Many of these mutations can be detected as chromosomal aberrations (Ghosal and Mondal
1978; Murata et a1. 1981) which delay the onset of mitosis necessary for germination (Murata
et al. 1980). While these chromosomal aberrations delay seedling growth, continued
development of the seedling results in fewer cells with these chromosomal irregularities,
presumably because abnormal cells are not able to c:ompete with normal ones (Murata et al.
1984). As a result, Roos (1982) has argued that mitotic lesions are unlikely to affect the genetic
integrity of stored germplasm.
Noteworthy products of peroxidation include the volatile aldehydes. Harman et al. (1978)
reported that deteriorated seeds produced 20 times more volatile aldehydes during imbibition
than did fresh seeds. They further suggested that these compounds served to stimulate fungal
spore germination and provided directive signals for growth (Harman et a1. 1982). Wilson and
McDonald (1986a) simplified the analysis of volatile aldehydes from imbibing seeds and
proposed that their levels could be used as an index of seed vigor.
Four approaches have been described to minimize lipid peroxidation in seeds (Wilson and
McDonald 1986b): (1) Lipid modification-breeders have focussed on changing the proportion
of saturated/unsaturated fatty acids to favor an increase in saturated fatty acids which are less
prone to lipid peroxidation. This has been accomplished in certain oil seeds (Knowles 1969;
Wilson et al. 1981). Lipoxygenase-deficient soybean lines have also been reported (Hildebrand
and Hymowitz 1982). (2) Regulation of oxygen pressure-reducing the quantity of oxygen
218 Seed Storage and Longevity

surrounding the seeds might also decrease the initiation of free radicals. This may be one of the
reasons for the success of storing seeds for longer periods in hermetically sealed containers. (3)
Antioxidant treatments-Antioxidants such as vitamin E or a-tocopherol are known to be free
radical scavengers. It has been estimated that one tocopherol molecule can afford antioxidant
protection to several thousand fatty acid molecules (Bewley 1986). Tocopherol is known to
occur in seed oils and a correlation between the degree of unsaturation in the oil and the
tocopherol content has been reported (Hove and Harris 1951). To date, however, attempts to
eliminate lipid peroxidation using antioxidants have not been successful (Yang and Yu 1982;
Parrish and Bahler 1983). Part of the difficulty may be the infusion of these compounds into the
seed since they are water insoluble. Woodstock et al. (1983) showed that freeze-drying of
tocopherol impregnated seeds prolonged the storage life of parsley and onion seeds. (4)
Hydration/dehydration treatments-hydration/dehydration treatments, also known as priming
(Chapter 13), do not appear to extend seed storage life but do lead to improved seedling
performance during germination. One possible explanation of this observation has been that
repair of free radical damage to membranes and perhaps other organelles occurs during the
hydration phase (Ward and Powell 1983). Studies supporting this notion have shown that
deteriorated seeds perform better following imbibition (Goldsworthy et al. 1982), placing them
in high humidity (Sanchez and de Miguel 1983), or placing seeds in an osmoticum followed by
a drying treatment (Brocklehurst and Dearman 1983; Burgass and Powell 1984). This practical
approach to repair of lipid peroxidation-induced membrane damage has been implemented in
many commercial seed quality assurance programs.

Degradation of Functional Structures

As seed deterioration progresses, cellular membranes lose their selective permeability,

permitting the cytoplasmic metabolites to leach into the intercellular spaces. Membrane
degradation occurs from both hydrolysis of phospholipids by phospholipase and phospholipid
autoxidation; therefore, in the strictest sense, it might be considered a result of aging rather than
a cause.
Mitochondrial degradation and functional changes appear to playa major role in seed
deterioration. A comprehensive review of the literature on aging in both plants and animals
shows that many mitochondrial changes undoubtedly playa role in seed deterioration (Abu-
Shakra 1963; Priestley 1986). Such changes decrease in number as deterioration proceeds;
mitochondria become permanently swollen and lose their natural swelling-contracting ability
(Cowdry 1956). Later they become pigmented and fragmented (Payne 1946; Weiss and Lansing
1953; Andrews 1956). Degradation of the mitochondrial membranes also occurs, leading to loss
of function and eventual fragmentation (Abu-Shakra 1963). Two important aspects of
mitochondrial deterioration are an increase in ATPase (Kiel1ey and Kielley 1951; Sacktor 1953)
and a decline in oxidative phosphorylation ability necessary to complete its respiratory function
(Lehninger and Remmert 1959; Wojtczak and Wojtczak 1960; Weinbach et al. 1963). Since
ATPase catalyzes the breakdown of ATP and ADP, it depletes energy available in the
mitochondria. Levels of ATP are lower in aged seeds of a number of crops (Luon and Madsen
1981; Banerjee et al. 1981). A TP also helps restore contraction ability to aged, swollen
mitochondria; thus, the loss of ATP accelerates mitochondrial disintegration (Weinbach et al.
1963).
Seed Storage and Deterioration 219

Functional changes in mitochondria may be repaired partly or in whole by the addition of

unsaturated fatty acids, phospholipids, chelating agents, and fatty acid binding compounds
(albumin) (Chefurka 1963; Wojtczak and Wojtczak 1960; Rossi et al. ] 964). It is believed that
certain growth regulators contribute to protection of mitochondria and other cytoplasmic
membranes and thus help maintain their integrity and selective permeability.

Inability of Ribosomes to Dissociate

Associated with the degradation of function in deteriorating seeds is the dissociation of

ribosomes. Evidence indicates that the dissociation of polyribosomes must occur before
attachment of preformed mRNA occurs, leading to protein synthesis in germinating seedlings
(App et al. 1971). In nonviable seeds, the ribosomes fail to dissociate (Bray and Chow 1976)
and protein synthesis is retarded. Such declines in protein synthesis are a measurable symptom
of aging. There is also evidence that longlived mRNA is lost during extended seed storage
(Osborne 1983; Ghosh and Choudhuri 1984). It has also been suggested that aging depresses
the synthesis of newly formed mRNA (Osborne et al. 1977; Weidner and Zalewski 1982).

Enzyme Degradation and Inactivation

Decreased activity of enzymes such as catalase, dehydrogenase, and glutamic acid

decarboxylase in deteriorating seeds is well documented. The general decrease in enzyme
activity in the seed lowers its respiratory potential, which in turn lowers both the energy (ATP)
and food supply to the germinating seed. Several changes in the enzyme macromolecular
structure may contribute to their lowered effectiveness (Walter 1963). They may undergo
compositional changes by losing or gaining certain functional groups, by oxidation of sulfhydryl
groups, or by conversion of amino acids within the protein structure. The enzymes may also
undergo configurational changes such as: (I) partial folding or unfolding of ultrastructure, (2)
condensation to form polymers, or (3) degradation to subunits.

Formation and Activation of Hydrolytic Enzymes

As seed moisture content approaches levels necessary for germination, hydrolytic enzymes
are activated. If the seed moisture content remains high or reaches higher levels, normal
germination may occur; however, if moisture levels for germination are not attained, the seed
deteriorates because of energy expenditure or accumulation of breakdown products. The
increase in free fatty acids, a symptom of deterioration caused by activation of lipase enzymes
in oil-containing seeds has already been discussed. A related group of enzymes, the
phospholipases, hydrolyzes the phospholipids and thus destroys the membrane structure of the
seed. Activation of phosphatase enzymes converts ATP to ADP, resulting in an energy loss
accompanied by increased phosphate acidity. Other hydrolytic enzymes activated by high
moisture levels are amylases and proteolases. This kind of deterioration is rapid and is important
only over short periods at moisture levels around 20% and above. Below 20% moisture, other
kinds of deterioration predominate.
220 Seed Storage and Longevity

Breakdown in Mechanisms for Triggering Germination

Harrington (1973) has made a strong case for the idea that the breakdown of various
triggering mechanisms also causes seed deterioration. Evidence has been cited on the role of
gibberellins (Paleg 1960; Penner and Ashton 1965) and cytokinins in triggering enzyme activity
leading to germination. Further evidence for this theory is the improved germination in aging
seeds after exposure to growth hormones (Harrington 1973). For example, the exposure of
partially aged rape seed to ethylene gas enabled them to produce normal seedlings (Takayanagi
and Harrington 1971). It has been noted that gibberellic acid improved the germination and vigor
of partially aged celery seeds (Harrington 1973) and can protect seeds from age-induced damage
(Petruzzelli and Taranto, 1985), although other studies on aged barley seeds have shown no
effect by gibberellic acid (Huber and McDonald 1982).

Genetic Degradation

A great deal of indirect evidence supports the view that seed deterioration is associated with
random somatic mutations that impair the cellular functions of vital seed tissues. The increase
in chromosomal aberrations in deteriorating seeds as a result of somatic mutation has been
observed in many species. Fusion and fragmentation aberrations that were observed in both x-
rayed and aged barley seeds occurred at higher rates than in fresh seeds (Gustafsson 1937). Two
general observations have been made on work with barley seeds (James 1960): (1) the properties
of chromosomes change gradually with age, and (2) mutations in seeds are highly correlated
with seed age and decline in viability in aging seeds. The area of greatest sensitivity to x-ray
damage and mortality is in the central root nuclei. The following observations have been made
in support of the genetic theory: (1) extracts from aged seeds retard the germination of fresh
seeds, (2) mutations in seeds are highly correlated with seed age and decline in viability, (3)
spontaneous mutations arise in aging seeds and become evident by chromosome breakage in the
presplit phase, (4) differences in the shoot and root tips in aged seed closely parallel those in
irradiated seeds (D'Amato and Hoffinan-Ostenhof 1956). Earlier, D'Amato (1952) suggested
that mutations in aged seeds were induced by mutagens formed during the aging process and
probably were decomposition products resulting from seed metabolism.
One observer (Harrington 1973) discounts genetic degradation as a primary cause ofaging.
Harrington cites evidence that although more chromosomal aberrations occur with increasing
age, animal tissues rid themselves of the altered cells (Kohn 1963). This notion has been
supported by Roos (1982). Further, radiation-induced injury in bacterial cells is reduced by
quick repair of damaged DNA (Witkin 1966). If enough cells in the germinating seedlings are
damaged, slower growth and seedling abnormality can occur; however, ordinarily radiation-
killed cells are quickly crushed or replaced by normal tissue (Harrington 1973).

Depletion of Food Reserves

Depletion of food reserves is one of the oldest theories on deterioration; however, it has not
survived critical scrutiny. In fact, most seeds contain enough food materials to last thousands
of years (James 1960). Even 1000- to 2000-year-old wheat seeds that have been found in ancient
tombs sti1l retain most of their food reserves; thus, even nonviable seeds contain enough food
reserves for seedling growth and development (Barton 1961). We know that the biochemical
Seed Storage and Deterioration 221
degradation processes in dry seeds are almost imperceptibly small and could not account for
depleting the food reserves within the life span of most seeds.

Starvation of Meristematic Cells

The theory of starvation of meristematic cells was introduced at the USDA-ARS Seed
Quality Research Symposium in 1971 (Harrington 1973); however, in principle, it had been
implied earlier in an evaluation of the reserve food depletion theory (Harrington 1973). It was
noted that respiration may deplete the tissues involved in the transfer of nutrition from reserve
storage areas and thus prevent them from reaching the embryo. Another study elaborated on this,
noting that meristematic cells, even though only a few cells away from abundant reserves of
energy, may die from lack of food or from injury (Harrington 1973). It was speculated that
perhaps the meristematic cells exhausted their energy supply, with no way to convert ADP to
ATP.

Accumulation of Toxic Compounds

Under low-moisture storage, the reduced respiration and enzyme activity may be
responsible for accumulation of toxic substances that reduce seed viability (Harrington 1973).
When aged wheat embryos were transplanted onto young endosperms and young embryos onto
aged endosperms, a progressive decline in germination and vigor of both transplants has been
observed (Floris 1970), strongly indicating a gradual accumulation of toxic metabolites. It has
been suggested that the presence of abscisic acid, a germination inhibitor, in several seeds
supports this theory as a probable cause of aging (Harrington 1973; Ryugo 1969; Martin et at.
1969) as well as phenolic compounds (Sreeramulu 1983) and polyamines (Mukhopodhyay et
al. 1983).

Questions

1. Name some crop and weed species noted for their short-lived seed.
2. What internal and external factors influence seed longevity?
3. Why do some seeds have higher moisture equilibrium curves (isotherms) than others?
4. How do the relative amounts offats, carbohydrates, and proteins affect seed storability?
5. If it were possible to regulate temperature and relative humidity independently, which
control would be most important for preserving seed viability in storage? Why?
6. Name several symptoms of seed deterioration. List them in order of their relative
appearance in decline of germination in seed storage.
7. What are some possible causes of seed deterioration?
8. How would you realistically provide for adequate seed storage in the midwestern United
States? Alaska? the Andes mountains in South America? Indonesia?
9. Describe Harrington's "rules of thumb," emphasizing their relationship to seed moisture
and temperature during storage.
10. Explain why seeds placed in sealed storage at 13% moisture will deteriorate more
rapidly than seeds in open storage.
222 Seed Storage and Longevity
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Physiology 44:733-738.
Abdul-Baki, A A, and J. D. Anderson. 1970. Viability and leaching of sugars for germinating
barley. Crop Science lO:31-34.
Abu-Shakra, S. 1963. Biochemical study of aging in seeds. Ph.D. Dissertation. Oregon State
University, Corvallis.
American Phytopathological Society. 1983. Deterioration mechanisms in seeds. Phytopathology
73:313-339.
Anderson, 1. D. 1970. Physiological and biochemical differences in deteriorating barley seed.
Crop Science 10:36-39.
Anderson, 1. D., J. E. Baker, and E. K. Worthington. 1970. Ultrastructural changes of embryos
in wheat infected with storage fungi. Plant Physiology 46:857-859.
Andrews, W. 1956. The mitochondria of neuron. International Review ofCytology 5: 147-170.
App, A. A., M. G. Bulis, and W. J. McCarthy. 1971. Dissociation of ribosomes and seed
germination. Plant Physiology 47:81-86.
Austin, R. B., and P. C. Longden. 1967. Some effects of seed size and maturity on the yield of
carrot crops. Journal of Horticultural Science 42:339-353.
Banerjee, A, M. M. Choudhuri, andB. Ghosh. 1981. Changes in nucleotide content and histone
phosphorylation of ageing rice seeds. Zeitschrift fur Pjlanzenthysiologie 102:33-36.
Barton, L. V. 1961. Seed Preservation and Longevity. London: Leonard Hill.
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Bass, L. N. 1970. Prevention of physiological necrosis (red cotyledons) in lettuce seeds
(Lactuca sativa L.). Journal ofAmerican Society of Horticulture Science 95:550-553.
Bass, L. N., and P. C. Stanwood. 1978. Long-term preservation of sorghum seed as affected by
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Battle, W. R. 1948. Effect of scarification on longevity of alfalfa seed. Journal ofthe American
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Beattie, J. H., and V. R. Boswell. 1939. Longevity of onion seed in relation to storage
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Becquerel, P. 1934. La \ongevite des graines macrobiotiques. Comptes Rendus Academie des
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Bennic~ A, M. B. Bitonti, C. Floris, D. Gennai, and A M. Innocenti. 1984. Ageing in Triticum
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Berjak, P., 1. M. Farrant, D. J. Mycock, and N. W. Pammenter. 1990. Recalcitrant
(homoiohydrous) seeds: The enigma of their desiccation sensitivity. Seed Science and
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Bockholt, A J., 1. S. Rogers, and T. R. Richmond. 1969. Effects of various storage conditions
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Bray, C. M., and T. Y. Chow.1976. Lesions in ribosomes of nonviable pea (Pisum arvense)
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Brett, C. C. 1952. Factors affecting the viability of grass and legume seed in and during
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Brocq-Rosseu, D., and E. Gain. 1908. Sur la duree desperoxy distases des graines. Comptes
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Bulat, H. 1963. Das allmahliche, durch ungunstige Largerungsbedingungen beschleunigte
Absterben der Samen bzw. Ruckgang der Keimfahigkeit im Bilde des topografischen Tetrazoli-
umverfahrens. Proceedings of International Seed Testing Association 28:713-751.
Burgass, R. W., and A. A. Powell. 1984. Evidence for repair processes in the invigoration of
seeds by hydration. Annals ofBotany 53:753-757.
Canode, C. L. 1972. Germination of grass seed as influenced by storage conditions. Crop
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Chefurka, W. 1963. Comparative study ofthe dinitrophenol-induced ATPase activity in relation
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Chin, H. F., B. Krishnapillay, and P. C. Stanwood.l989. Seed moisture: Recalcitrant vs.
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pp. 15-22. Crop Science Society of America, Madison, WI.
Chin, H. F., and E. H. Roberts. 1980. Recalcitrant Crop Seeds. Kuala Lumpur, Malaysia:
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Seed Storage and Deterioration 225
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 10
Seed
Production

During the last 50 years, the availability of high-quality seed of improved crop varieties,
along with modern power equipment, improved fertilizers, and better methods of weed and
insect control, have revolutionized farming. The seed industry has played a vital role in this
modem revolution, with its expanding production capability, efficiency in rapid seed increase
of new varieties, and effective maintenance of genetic purity. The quantity of seed needed by
farmers each year is enormous. It is estimated that North American farmers alone use over 12
billion pounds annually offield crop, vegetable, flower, and tree seeds.
Prior to the era of modern agriculture, seeds were usually a by-product of grain or hay
production. Frequently, the poorest part of the crop was reserved as seed, which was often
gathered from haylofts, along with chaff, we<xi seeds, and other incidental material.
Occasionally, farmers would have enough excess seed to share with a neighbor, and a farm-
to-farm exchange resulted. Although seed was sometimes available on the commercial
market, the supply was sporadic, and often of questionable quality. Unscrupulous marketing
gimmicks were sometimes used to take advantage of the customers' inability to identifY seeds
and their quality. This led to skepticism about thc~ quality of seeds bought off the farm and
created a climate of distrust that acted against the development of a legitimate seed industry.

DEVELOPMENT OF THE SEED INDUSTRY

Several factors are responsible for the development and evolution of the seed industry in
North America: (1) an increased number of new, available varieties, (2) the development of
seed certification and seed law enforcement programs, (3) the development of cleaning and
conditioning technology, (4) a better knowledge of seed quality, and (5) the emergence of the
seed grower as a specialist, and more recently, (5) the growing sophistication of the seed
industry.
The Hatch Act of 1875 accelerated the development of the u.s. seed industry. This Act
established a network of experiment stations throughout the country and stimulated the
development of improved crop varieties. Seed certification was a direct outgrowth from the
experiment station network of the land grant colleges. Certification programs were estab-

L. O. Copeland et al., Principles of Seed Science and Technology

© Kluwer Academic Publishers 2001
232 Seed Production

lished in most states between 1915 and 1930, and most of the new "college-bred" field crop
varieties first became available as certified seed. During this period, many farmers became
accustomed to purchasing seed from recognized seed producers rather than planting their own
"bin-run" seed.
With the availability of new varieties, a strong seed industry was necessary with the
ability for rapid seed increase and distribution with adequate safeguards for varietal purity.
Otherwise, the impact of new improved varieties would have been lost. The National
Foundation Seed Project, another outgrowth of the experiment station network, assured the
success of many new varieties by providing for rapid increase of basic seed stocks; however,
it is no longer an active program.
Since the early 1900s, the technology of seed cleaning and conditioning has made
tremendous strides. Machinery has been designed that can condition the seed more efficiently
and improve the quality over bin-run seed by removing contaminating weed seeds, other crop
seeds, and inert material. Machines have also been developed to improve germination by
eliminating poor-quality crop seeds. Facilities have become available to chemically treat and
inoculate the seed and package it in attractive containers for merchandising appeal. Seed
quality testing has been developed as a service to both seed producers and farmers. Such
testing has provided new insights about seed quality and has done much to increase quality
consciousness among farmers and seed producers.
The development ofthe seed producer as a specialist and the growth of the seed industry
have been closely related. Since seed production, for the most part, is no longer a by-product
of grain or hay production, a large seed industry has developed in areas widely removed from
where the seed is used and a seed increase and merchandising network has developed to
market seed over wide geographical areas. Seed stocks of a new variety are sent to a seed
production area for increase and quantities of commercial seed are returned to the area of
adaptation for forage or turf production. Thus, seed production outside the area of use has
contributed in a major way to the development of the seed producer as a specialist and has
helped the growth of the seed industry. Seed production is a complex business involving many
different and integrated operations. Several reviews of these operations have been captured by
Hebblethwaite (1980), George (1985), Kelly (1988), and McDonald and Copeland (1995).

SEED PRODUCTION IN AREA OF USE

The seeds of many field crops are produced in the same area where they are planted for
commercial production. As a general rule, when adequate quantities of high-quality seed can
be produced in the area of use, it is best to do so. Crop varieties usually produce higher seed
yields in the area where they are best adapted. Also, transportation and marketing costs from
producers to consumers are minimized. This is particularly important for seed of cereals,
soybean, and certain other crops that involve larger seed volumes and greater planting rates
than small-seeded grasses and legumes. Other than small-seeded grasses and legumes and
certain specialty crops, seed of most field crops is produced in the area of principal
adaptation and use.
Seed Production 233
SEED PRODUCTION OUTSIDE THE AREA OF USE

A large part of the seed planted in the United States and Canada is produced outside its
major area of use, including almost all seeds of I::ertain grass and legume crops, and many
vegetable and ornamental crops. Most of this specialized seed production is in the western
regions of North America, where a unique combination of climate and cultural factors has
contributed to the specialized seed production capability. A closer look at this western seed
industry indicates why it has developed and flourished far removed from areas where most of
the seed is used.

Advantages-Climatic Factors

The probability of dry harvesting weather is the single greatest reason for the success of
the western seed production region. Weather records for these areas show that the probability
of rain during the harvest season is minimal. In the western valleys of the Pacific Northwest,
the relative assurance of favorable harvesting weather, combined with plentiful natural
precipitation during the spring and early summer, provides a unique competitive advantage in
turf and forage seed production as well as other specialty crops. In the drier western areas,
water is provided during the growing season from vast irrigation systems.
Nonirrigated Production. Although the inland valleys of the Pacific Northwest are
particularly well suited for non irrigated grass seed production, seeds of forage legumes, sugar
beet, garden vegetables, and certain flower species are also produced in this area. Few
geographical regions are more climatically suited for grass seed production. Rainfall is
plentiful and almost continuous throughout the mild winters and spring, providing ideal
conditions for plant development and seed head production. In early summer, the rainy season
ends, and the weather becomes dry with a low probability of rain from mid-June until
September. Weather records show many summers without precipitation for 30-60 days. Such
conditions favor maturation of seed heads and threshing at harvest, without the continuous
threat of rainy weather that often plagues seed harvest in other areas.
Irrigated Production. Irrigated seed production in the western United States is
principally for both large- and small-seeded legumes, although there are areas of highly
successful grass seed production as well. Although pockets of irrigated legume seed pro-
duction are found throughout the West, major production is centered in California, eastern
Oregon, and Washington eastward to Idaho and Utah. These areas have a low probability of
rainfall through the harvesting season and have adlequate water for irrigation. Another major
advantage in legume seed production is the relative abundance of natural insect pollinators.
Great success has been achieved in building up native bee pollinators and introducing various
types of wild bees from other areas. Dry weather during the flowering period and an
abundance of insect pollinators usually assures good pollination and seed set.
The warm, dry summer climate of the west<,rn United States is also ideally suited for
production of disease-free seed of vegetable and field crops that are highly susceptible to
seedborne diseases. Practically all of the seed of garden pea and snap bean and much of the
dry edible bean types are produced in Idaho, Washington, and California. The warm, dry
summer weather combined with stringent phytosanitary seed production regulations prevent
buildup of many of the bacterial, fungal, and viral diseases that are highly destructive in the
warm, humid summers of the central and eastern regions where much of the seed is planted.
234 Seed Production

Disadvantages

Transportation and marketing costs are major disadvantages of producing seeds outside
their area of use. However, this cost is minimal compared to the advantage provided.
Among the potential dangers of producing seed outside the major area of crop
adaptation is the likelihood of a genetic drift in the varieties caused by the different
environmental stresses (such as day length, temperature, soil type) to which the plants are
exposed during seed production. The potential for a drift in germplasm composition is
especially critical in cross-pollinated forage species; environmental stresses in the seed
production area may cause the failure of the individual plants in the varietal population to
survive or to produce their proportionate share of seed. As a result, the germplasm
represented by their segment of the population decreases, while other segments of the
population increase proportionately. Thus, a gradual drift in the genetic balance of
germplasm occurs, which may change the varietal characters. Although this drift may be
gradual and hardly noticeable in the region of seed production, after a few years of
production it may seriously affect the varietal performance in its area of adaptation.
Safeguards against a serious genetic drift are provided through a limited-germination
certification system, in which the number of generations of seed increase outside its area of
adaptation is carefully controlled. The certification system provides these limitations and
minimizes the likelihood of genetic drift.

GRASS SEED PRODUCTION

Seed production of grasses can be appropriately discussed in terms of cool and warm
season grasses. The latter includes many the Great Plains grasses as well as southern grasses
(see Table 10.1).

Cool Season Grasses

Cool season grasses are those adapted to the northern regions of North America. They
include most of the lawn and turfgrass species in the United States, except for those of the
most southerly regions. These grasses grow actively at cool temperatures during the spring
and fall. Depending on the severity of the temperatures, their winter response varies. During
the mild winters of the Pacific Northwest, their growth is minimal but they maintain a green
color. Under more severe winter conditions, cool season grasses go dormant and lose their
green color. Dormancy is followed by very rapid spring growth until flowering in early June.
Seed is mature about the first of July.
Most seed production of the cool season grasses is located in nonirrigated areas of the
Pacific Northwest; however, considerable seed is also produced under irrigation in the Pacific
intermountain region. Relatively small but important pockets of production exist in
midwestern and central states, particularly in Minnesota, South Dakota, Kentucky, and
Missouri.
Seed Production 235
Table 10.1. Classification of Grasses of the United States.

Cool Season Grasses Warm Season Grasses

Great Plains Grasses Southern Grasses
tall fescue gramagrass Bermudagrass
red fescue bluestems Dallisgrass
ryegrass buffalograss rescuegrass
tall oatgrass Indian ricegrass Bahiagrass
Kentucky bluegrass lovegrass Zoysia grass
smooth brome grass blue panicum napier grass
orchard grass switchgrass pangola grass
Reed canarygrass dropseeds centipedegrass
timothy buffelgrass Rhodegrass
bentgrass Texas wintergrass carpetgrass
Russian wild rye Indiangrass Vaseygrass
wheatgrasses vine mesquite St. Augustine grass
curly mesquite grass
thatchgrass

Seed fields may be established in rows or in solid plantings by using a grain drill. In
recent years, row spacings of 24 to 48 in. have become more popular, although much seed
production is still from solid seedings. In the Great Plains, Russian wild rye has been planted
in 84-in. rows. In addition to providing assurance of cleaner, weedfree fields, row seedings
offer several other advantages. Row plantings g4~erally produce higher yields than solid
seedings and require lower seeding rates. It is thOUght that row plantings offer more
economical use of fertilizer and prolong stand productivity. Solid stands, especially in the
sod-forming grasses, tend to become sod bound after a few years and their productivity
declines. This condition seems to be a result of ovt:rpopulation and excessive competition for
available nutrients, as well as the physical crowding caused by overpopulation. The adverse
effects of sod binding can be minimized by increased nitrogen applications and by postharvest
field burning.
Seeding rates lower than those used for forage or turf production have been found best
for grass seed production. Small-seeded grasses, such as bentgrass or bluegrass, are often
seeded at V4 to I Ib/ac, while larger-seeded grasses, such as tall fescue and Russian wild rye,
may be seeded at 4-8 lb/ac.
The ability of grass seed producers in the Pacific Northwest to increase sman quantities
of seeds of new forage and turf varieties for widespread availability in the United States and
in the world market has gained them international recognition. These seed producers use
techniques of stand establishment, weed control, culture, harvesting, and conditioning that are
geared for high varietal purity, freedom from weed and other crop seed and higher
gennination standards. Since much of their seed is used for establishment of quality lawns
and high-grade pastures, their attention to seed quality is important.
One method of stand establishment uses a special drill that places a charcoal barrier
directly over the seed immediately after planting (Figure 10.1). A nonselective herbicide (e.g.,
Kannex), is then applied to the entire field. The berbicide directly above the drilled seed is
absorbed by the charcoal layer, allowing seed germination without injury while all seedlings
236 Seed Production

IS

Figure J O. J. Establishing a grass seed field by use of a protective charcoal barrier: (AJ laying
down the barrier and planting the seed, (BJ results following seedling emergence (Courtesy of
Richard Bailey).
Seed Production 237
between the bands are eliminated. Once the crop is established, the use of selective herbicides
and hand roguing keeps weed contamination to a minimum.
Seed is harvested (Figure 10.2) by swathing the crop into windrows and allowing it to
dry a few days before threshing. Much ofthe seed is taken directly to farm cleaners where it
may be cleaned completely or given a rough scalping to remove most ofthe foreign material,
such as straw, stones, and other large and easily separated contaminants. If cleaned
completely, it is sold to a wholesaler on a pure seed basis. Alternatively, it may be sold "in
the rough" to a wholesaler who finishes the cleaning procedure. In the latter case, the grower
is paid on a pure seed basis. Much of the grass seed is grown on a contract basis, so
commitments are made in advance concerning quality of seed to be delivered, the degree of
cleaning, and the price to be paid.
Field Burning-A Disappearing Dilemma? Grass seed fields in the Pacific Northwest
traditionally have been burned after the seed is removed. This practice has been widely recog-
nized as a valuable cultural practice to the grass seed grower, since burning significantly
increases yields and controls destructive insects and diseases that overwinter in unburned
plant residues. Burning also aids in weed control, delays the development of sod binding, and
returns potash and other mineral nutrients to the soil.
Although the benefits are significant and conspicuous, so is the air pollution from the
burning fields. In recent years, ecologists and the nonagricultural population have become
increasingly critical of this practice and have influenced state legislatures to ban open-field
burning. Considerable research has been conducted on alternatives to open field burning,
including the use of portable burners using straw or propane as fuel. Non-burning options
include: (1) removal of all stubble and chaff from the field by mechanical methods and (2)
chemical control of diseases and insects. As a result of this research and continued public
opposition to open field burning, it now appears clear that grass seed production in the
Willamette Valley of Oregon will continue by using a combination of residue removal,
cultural practices, and chemical control of inse<:t and disease pests that in the past were
controlled by post-harvest burning.

Warm Season Grasses

Wann season grasses make their maximum growth during the summer. This growth is
aided by the adequate summer rainfall in their area of adaptation. These grasses start their
spring growth about three weeks later than cool season grasses and cease growing with the
first hard frost in the fall. In the winter they are completely dormant. Warm season grasses
predominate in the central and southern Great Plains area and throughout the South.
Great Plains Grasses. These are mostly native grasses of the high plains of western and
southern Texas and western Oklahoma, Kansas, Nebraska, and the surrounding areas. Seeds
of these grasses were originally harvested from native pastures for use in converting
croplands to grass and for improving rangelands. Even today, much of the seed of these
grasses is harvested from native stands. However, a specialized seed production industry is
developing due to at least two reasons: (1) seed production from native stands is sporadic and
undependable, occurring only when seasonable moisture conditions are unusually favorable,
and (2) as improved varieties of native grasses become available, the need for a program for
maintaining seed availability intensifies.
238 Seed Production

Figure /0.2. Grass seed production in Oregon: (A) general view of a fescue seed field, (B)
swathing (windrowing) a ryegrass seedfield, and (C) postharvest field burning (B and C, Courtesy
of Harold Youngberg).
Seed Production 239
Most seed fields of the Great Plains grasses are established in rows, fertilized as needed
for optimum seed yields, and perhaps irrigated during periods of drought. Harvesting is
usually performed by direct combining, although some of the grasses are harvested by a
stripper of the type previously used for harvesting native bluegrass stands. The harvested
seeds of many of the Great Plains grasses are extremely chaffy and are not free flowing. This
creates extreme difficulty in cleaning and handling the harvested seed and has even
discouraged production of certain species. However, handling techniques are becoming
available that will remove much of the chaff and increase the ease of conditioning. Thus, a
seed industry for these species is emerging.
Southern Grasses. The adaptation of a plant to its environment is reflected in its
development and seed yield. Consequently, seed of the warm season southern grasses (for
example, Bermuda grass) must be produced in southern regions of the United States rather
than in the Pacific Northwest. Almost all Bermuda grass seed production is limited to
Arizona and southern California. Bermuda grass seed fields may be established either
vegetatively by "spriggings" (planting stolons or rhizomes), or by seeding (rows or
broadcast). Regardless of the method used, the plants soon spread into solid stands. Zoysia
grass is another southern species in which seed fields are established by vegetative plantings.
Both Bermuda grass and Zoysia grass are harvested for turf purposes in vegetative plugs or
sprigs, from which lawns are established (though Bermuda grass seed may also be harvested).
Much of the southern forage grass seed production is harvested from pastures that have
been grazed during the summer, fall, and early spring. This dual use and early removal of
grass material appears to stimulate seed production relative to ungrazed stands. Recent
availability of improved varieties has stimulated more seed production in rows rather than as
a byproduct of pasture production. Harvesting may be done by combining directly from a
windrow, or occasionally by shocking and threshing.

LEGUME SEED PRODUCTION

Seed of the forage legumes is produced under two distinct types of management systems
and climatic regions. One kind is the specialized seed production industry that has developed
in the western United States and Canada. The second, often a byproduct offorage production,
is not centered in any geographic area, but is scath~red throughout North America.

Western Legume Seed Production

The greatest proportion of high~quality legume seed is produced in the western regions
of North America, particularly in western Canada, California, Oregon, Washington, and
eastward to Colorado and Oklahoma. This area has great natural advantages for legume seed
production, especially alfalfa seed. The dry climate, adapted soils, abundance of water for
irrigation, and availability of insect pollinators have raised seed production of alfalfa and
other legumes to levels unattainable in other areas of North America. Occasionally in this
region, alfalfa seed yields of 2000 lbs of seed per acre can be achieved.
Cultural Practices. Alfalfa seed fields are usually established in 21 ~ to 41-in. rows,
although some solid stands are used. Seeding rates vary from 2-4 Ib/ac for rows to as much
as 15 lb/ac for solid stands. Research has showll1 that within certain limits, lower seeding
rates tend to give the highest seed yields. Alfalfa stands respond to lower plant populations by
240 Seed Production
producing more seed per plant, thus increasing total production. Spacing the plants
individually or in hills within the row often increases seed yields, although it may also
increase weed problems. In the San Joaquin Valley of California, highest yields are obtained
in 40-in. rows planted at only llb/ac. Row planting also offers advantages over solid stands
through more efficient use of seed and fertilizer and in facilitating weed control.
One of the more serious weeds in alfalfa seed production is field dodder (Cuscuta spp.).
Dodder (see Figure 10.3) is a parasitic weed which grows particularly well on alfalfa, though
several other crops and weeds may also serve as hosts. It is particularly troublesome for
alfalfa seed production because its similar seed size makes it difficult to separate from alfalfa
seed lots. Although the seed is similar in size, its rough seed coat texture allows it to be
separated by means of a velvet roll machine (see Chapter 11). Dodder is controlled in alfalfa
seed fields by a combination of chemical and manual methods. Most states have added dodder
to their noxious weed lists to prevent incoming alfalfa and clover seed lots from becoming
sources of dodder contamination.
Insect Pollinators. The availability of insect pollinators gives the western seed
production area a great advantage for alfalfa and other insect-pollinated legumes. Several
important insect pollinators used include: (1) honeybees, (2) leaf-cutting bees, (3) alkali bees,
and (4) bumblebees. Both wild and domesticated honeybees are used, although neither are
very efficient in "tripping" (pollination/fertilization) the alfalfa flowers (see Figure 10.4).
When the honeybees visit the flowers for either nectar or pollen collection, they "trip" the
alfalfa flower by dislodging the sexual column (the stigmatic tissue), which is concealed by
the keel petals. The tripping exposes the stigma to the pollen carried by the bee from other
flowers. When the sexual column is released, it snaps upward with considerable force,
striking the bee on the underside. Honeybees soon learn how to visit flowers without tripping
the flower and thus become less effective pollinators. Some honeybees are primarily nectar
gatherers, while others gather polIen and are more effective as pollinators. In spite of their
rather low effectiveness, the large numbers of honeybees and the availability of domesticated
colonies make them the most important insect pollinator for alfalfa.
Leaf-cutting bees (Megachile spp.) are a wild species that are more effective pollinators
than honeybees, although they rarely occur naturally in sufficient numbers to pollinate
commercial seed fields alone. They are small, dark-colored bees (Figure 10.5) that build their
nests in colonies in small holes in wood. They may bore the holes themselves or use existing
holes made by other insects. Oblong leaf cuttings are used to make a cell into which the eggs
are laid. Each cell is provided with pollen moistened with nectar. After the eggs are laid, the
cell is sealed with circular leaf cuttings. If the hole is long enough, several cells may be
placed in tandem within the same hole.
In recent years, alfalfa seed growers have increased the number of leaf-cutting bees by
providing artificial egg-laying sites where colonies can be established. These colonies are
often portable and can be moved to different parts of the seed field as needed. One can
observe the effectiveness of leaf-cutting bees by the conspicuous circles of well-pollinated
plants (with wilted flowers) around their colonies. Leaf-cutting bees have two disadvantages
that lower their value and require specific management by the commercial seed grower: (1)
their range of activity is relatively short-usually only a few hundred feet around the colony
and (2) like most wild bees, their populations are sporadic and undependable.
Seed Production 241

c
Figure 10.3. Three views of dodder and its control in alfalfa seed production: (A) closeup of
dodder in the flowering stage growing on alfalfa, (B) a severe dodder infestation of alfalfa, and (C)
an aerial view of an alfalfa seed field in which dodder has been sprayed with a contact herbicide,
allowed to dry, and subsequently burned (this is a common control method) (Courtesy of Howard
Roylance).
242 Seed Pmd1Jrfinn

st.andard
petal

wing
petal

keel
petal

standard
petal

S\"Jeua!
column

keel
petal

Figure 10.4. The parts of an alfalfa flower important in insect pollination: above, untripped;
below, tripped (Courtesy of William P. Nye, Logan, Utah).
Seed Production 243

13

Figure 10.5. Leaf-cutting bees, their habitat and use in alfalfa seed production: (A) leaf-cutting
~~ro~~~~~W~ro~~~~~~~~~~
portable leafcutting bee colonies (A, Courtesy of William P. Nye; Band C, Courtesy of Howard
Roylance).
244 Seed Production
Alkali bees (Nomia melanderi) are highly effective pollinators that are native to many
western alfalfa seed production areas. Large colonies become established in bare, moist, salt
flats, where the female digs a pencil-sized tunnel 8-10 in. into the soil and hollows out several
egg-laying cells. She provides each cell with balls of pollen moistened with nectar on which
eggs are laid, and the cell is then plugged with soil. The eggs hatch in a few days, and the
larva feed on the pollen and nectar that has been provided. Depending on the time of year, the
larva may pupate and emerge as adults in about 30 days or wait until the next season. After
emerging, female adults reestablish the colony by laying eggs and starting the life cycle over
again. Alkali bees are shown in Figure 10.6.
Alkali bees have become important alfalfa pollinators in certain seed areas where they
occur naturally or have been introduced. They range much farther than leaf-cutting bees and
usually occur in much greater numbers. Many attempts have been made to introduce alkali
bees into new seed areas, but with moderate success. This is done by lifting large cores from
well-established nesting sites and inserting them into sites where a new bed is desired.
Considerable site preparation is necessary if the establishment is to be successful.
Bumblebees (Bomb us spp.) and melissodes bees (Melissodes spp.) are also effective
pollinators; however, they seldom occur in numbers sufficient to increase seed production
significantly. Bumblebees are limited in intensive agricultural areas because of destruction of
their nesting sites. Melissodes bees have shown remarkable ability to survive under
intensively cultivated conditions; however, they are not as gregarious as other wild types and
seldom occur in sufficient quantities for pollination of commercial seed fields.
Weather conditions influence the activity of all bee types. Temperatures between 75 and
100°F are most favorable. Bees do not work in the rain or while the flowers are wet. Wind
velocities over 5 mph also appreciably reduce bee activity.
The use of insecticides is a potential threat to all bee types, and many documented cases
exist where bee populations have been seriously reduced by misuse of insecticides.

Legume Seed Production in Areas of Use

Large volumes of alfalfa and clover seed are still produced in the midwest and
mideastern regions; however, it is usually a by-product of hay production and is subject to
large fluctuations due to weather conditions and availability of insect pollinators. Although
these regions have few of the advantages available for seed producers farther west,
considerable seed volumes are harvested annually from Midwest and Mideast hay fields.
Usually the first hay crop is cut or grazed, and the second is left for seed. Clipping of some
types of legumes, however, may be detrimental to seed production. For example, in Michigan,
seed production of medium red clover is stimulated by early clipping, while mammoth red
clover seed yields are reduced. Harvesting is done by combining directly or by windrowing
and allowing the crop to dry a few days before threshing. The probability of late summer
rains during the harvest season is a serious hazard to dependable seed production in this area.
Unlike seed production of the northern adapted species, seeds of southern legumes are
produced almost entirely in their area of adaptation and use. Although a specialized seed
production industry has developed, a first cutting of hay may still be taken to stimulate
better seed production by the second growth. Seed of southern legumes is harvested in the
Seed Production 245

A
feeding

mother mature diseased C

bee larva larva

Figure 10.6. Alkali bees: (A) alkali bee starting her nest excavation, (B) portion of an alkali bee
nesting site showing wind-blown entrance mounds among clumps of salt grass and saphire, (C)
horizontal section of an alkali bee nest at cell level, (D) an artificial (man-made) alkali bee nesting
site, (E) sign along an Idaho road in an alfalfa seed production area (A-D, Courtesy of William P.
Nye; E, Courtesy of Howard Roylance).
246 Seed Production

fall, usually in October. Harvesting methods vary from direct combining, shocking, and
threshing to combining from windrows.

HYBRID SEED PRODUCTION

Corn
Discovery of hybrid vigor in field crops after the beginning of the 20th century and
development of techniques for producing hybrid corn seed have probably contributed as much
to agriculture as any other single factor. The first announcement of potential advantages of
hybrid over open-polJinated varieties was made in 1908. In 1917, a method of double-cross
seed production was proposed that could be used to supply hybrid seed corn at prices farmers
could afford and in large enough quantities to assure an adequate and constant supply of
seed. Since these beginnings, hybrid corn has established an enviable standard for
performance and acceptance by farmers. In 1933, only 0.2% of the com acreage in the United
States corn belt was planted with hybrid seed. By 1944, its use had grown to 83%; today
virtually all corn is grown with hybrid seed.
A hybrid variety is produced by crossing inbred lines that have been developed by
inbreeding and selection for at least five successive generations. Inbreeding results in (I)
depression of vigor (plant height, yield, etc.), (2) increasing uniformity (homozygosity), and
(3) appearance of undesirable recessive gene effects that can be eliminated from the
population. The first-generation progeny of hybrid seed results in yield increases far above
either parent, and above that of nonhybrid populations. This yield increase is from heterosis,
or hybrid vigor, which is due to an accumulation of a large number of dominant favorable
growth factors (genes).
Modern corn production may be from seed representing double-cross, single-cross, or
three-way hybrids (see Figure 10.7). Until the early 1960s, double-cross hybrids were the
predominant types used for commercial seed production. They offer several advantages:

1. They are more variable than single or three-way crosses; they are not all alike
genetically. Thus, the plants may be buffered more against unfavorable conditions that
occur during the growing season.
2. A longer pollination period than other crosses may provide more complete filling ofthe
ear with seed and result in higher yields.
3. Lower seed cost is an obvious advantage when the yield of double crosses is equal to or
better than the best single-cross or three-way hybrids.
4. Double-cross hybrids generally have higher seed quality than single-cross hybrids and
may give more optimum stands when adverse conditions occur after planting.

Single-cross corn hybrids have become more prevalent in recent years. A corn field
planted from single-cross seed is impressive because the plants tend to be uniform. Plant
height, ear height, tasseling, silking, and pollen shedding are uniform, giving the field good
eye appeal. Also, since only two inbred parents are involved in a single-cross hybrid, a higher
level of resistance to diseases, insects, and other unfavorable situations may be incorporated
into them during the breeding process. Within a given set of inbred parents, the best single-
cross hybrid has a higher genetic yield potential than the best double-cross hybrid; however, a
particular double cross [(AxB)x(CxD)] may yield better than a particular single cross (ExF)
Seed Production 247

FIRST YEAR
OETASSElED
DE TASSELED

INBRED A INBRED

(61 A I, PRODUCED 1f'4

B
SINGI.E C!lOSSEO SEEo."""
t SECOND YEAR

8 INBRED C INBRED
'-SINGLE - CROSSED SI:: ED.
IC.OI, PRODUCED IN
AN ISOLATED FIELD
D

AN ISOLATED FIELD

I
SINGLE -CROSSED SINGLE -CROSSED
PLANT, (BIA) PLANT. (1':.0)

Figure 10.7. The method by which single-cross and double-cross hybrid corn seed is produced
(From Hughes and Metcalf, 1972).

that uses other inbred parents. The main disadvantages of single-cross hybrids are lower seed
yield and relatively lower quality of seed, since seed is produced on inbred parents. To
overcome these disadvantages in seed production, some single-cross hybrids are modified
single crosses. Two closely related sister inbred lines are crossed (A) x A2), to produce the
seed (female) parent. The pollen (male) parent may also be a cross of two different but
closely related sister inbreds (B l xB 2) or a full inbred (B). Seed yields are higher since sister
inbred (AI x A2) plants are somewhat more vigorous and produce better-quality seed than full
inbred plants.
Hybrid seed com is produced throughout th~: com belt of the United States by highly
specialized producers. Most hybrid com seed is produced under the control of large seed
companies, although a substantial portion is produced by many small companies located
throughout the corn belt. Regardless of the control, hybrid corn seed production techniques
are quite standardized. Double-cross seed production fields are usually planted in a pattern of
six seed (female) parent rows and two pollen (male) parent rows; single-cross production is
usually a pattern of two seed parent and one pollen parent, or four seed parent and two pollen
parent rows. Seed fields must be well isolated from other com fields that represent potential
contamination from outcrossing. The required isolation can be reduced if additional border
rows ofthe pollen parent are planted around the p(:rimeter of the seed fields.
Hybridization is achieved by allowing crossing of the desired male (pollen) and female
(seed) parents. Control of pollen may be achieved by detasseling the female parent or by use
of male-sterile female parent inbreds. The fITst hybrids were all a result of detasseling;
248 Seed Production
however, during the 1960s, use of male sterility seemed likely to completely eliminate the
need for detasseling. The situation was dramatically altered in 1971 by outbreaks of a new
race "T," of Southern Corn Leaf Blight and the recognition that the Texas source (T-
cytoplasm) cytoplasmic male sterility was much more susceptible to the new race "T." For a
few years after 1971, almost all seed was produced by detasseling, however, incorporation of
genetic steri Iity and other types of cytop lasmic sterility (other than the Texas source) heralded
another movement away from mechanical detasseling. When male sterility is used for hybrid
seed production, restorer mechanisms must be incorporated so the subsequent commercial
corn plants will produce pollen as well as seed.
Hybrid corn seed is usually harvested at about 30 to 40% moisture content and dried
while still on the ear. After drying down to about 12% moisture, it is cleaned and graded into
different size and shape classes (e.g., small rounds, large flats) to facilitate planting precision.
It is almost always treated with a fungicide (e.g., slurry or flowable) and often with an
insecticide before bagging and marketing.

Sorghum

Like corn, virtually all sorghum produced in the United States is from hybrid seed.
However, unlike corn, the sorghum plant is largely self-pollinated, so mechanical removal of
pollen-producing structures is not practical. Consequently, hybrid sorghum seed production
became practical only after discovery of cytoplasmic and genetic sterility mechanisms.
Restorer mechanisms are used for producing hybrid sorghum seed of grain sorghum; when
sorghums are to be used as forage and silage, restorer mechanisms are not needed.

Wheat

Commercial hybrid seed production of most self-pollinated crops is considered more

difficult than that for cross-pol1inated crops. Their flowers and pollen dispersal are not
structured for cross-pollination with other plants. Aside from the flower structure and pollen
dispersal pattern, male sterility and restorer mechanisms must be incorporated. A few hybrid
barley varieties have been released; however, hybrid wheat has often been the real goal
towards which large research investments have been made by both public and private
agencies.
The first obstacle to overcome in hybrid wheat was the development of suitable male
sterile lines. Now that male sterility has been found, the greatest obstacle to hybrid wheat
development is the lack of simple, effective mechanisms for restoration of the fertility of the
male line. Genetic restorer systems that can be incorporated into male parents do exist, but
they vary widely in effectiveness, are influenced by climate, are genetically complex, and
require a long development program.
Another method of producing hybrid wheat involves the use of chemical stamatacides on
the female parent. This allows fertilization by pollen from adjacent male rows and assures the
production of hybrid seed (assuming adequate wind for pollination). Male rows are harvested
for grain while female strips are harvested and sold for hybrid seed.
Seed Production 249
If hybrid wheat should prove successful, it could create a huge and specialized seed
production industry, since seed would need to be produced every year for planting the next
season's crop. The magnitude of such a program is difficult to imagine, compared to small-
grain seed production today. Not only would larg(! amounts be needed, but isolation from
commercial wheat fields would be necessary. Whether adequate isolation can be found within
the commercial wheat production region is questionable.
Figure 10.8 illustrates one method of hybrid wheat seed production.

Hybrid Alfalfa Seed

In 1971, a patent was granted to a commercial seed company for producing hybrid
alfalfa seed by a process using cytoplasmic male sterility. This is the first successful use of
the hybrid concept in the forage legumes, and the first patent awarded for using the hybrid
concept. Unlike hybrids of grain crops, a restorer mechanism is not needed for alfalfa, since
the vegetative material rather than the seed is commercially important.
Cytoplasmic male sterile plants are usually discovered by screening large plant
populations. These are then increased and maintained by crossing to "maintainer type plants,"
resulting in a seed population from which male stt~rile plants can be produced. Commercial
hybrid alfalfa seed is produced by growing male sterile plants in rows adjacent to those of
male fertile plants, allowing insect cross-pollination. The resulting single-cross hybrid seed is
then harvested from the male sterile rows. This seed could be marketed as a single cross but
is generally used as a female parent to produce a three-way hybrid. The alternate pollinator
rows in hybrid seed fields are also harvested and marketed as a byproduct of hybrid seed
production.

Female Parent Increase Male Parent Increase

Figure 10.B. Production of hybrid wheat seed (From Rogers and Lucken, 1973).
250 Seed Production

Questions

1. Why has the small-seeded grass and legume seed industry largely moved to the western
United States and Canada?
2. What are the advantages and disadvantages of field burning for grass seed production?
3. How can an optimum balance be maintained between insect control and pollinating
insects for cross-pollinated legume seed crops?
4. What are the advantages and disadvantages of contract seed production?
5. How much seed is sold directly to consumers from the farms on which it is produced?
How does this practice vary among various crops?
6. What is the future of hybrid seed production for crops other than corn, sorghum and
cotton? What about self-pollinating crops?
7. What is genetic drift in a variety? Explain its likelihood in self-pollinated versus cross-
pollinated crops?
8. Explain sod binding in grass seed production fields and how it can be avoided.
9. List the relative merits of solid versus row seedings for seed production of grasses and
legumes.
10. Explain the relationship between seeding rate and seed yields of grasses and legumes.
11. What do you consider to be the ideal seed moisture content for harvesting seed to avoid
mechanical damage? Do you consider mechanical damage to be a serious factor in seed?
Which species are most seriously affected?

General References

Bohart, G. E., and T. W. Koerber. 1972. Insects and seed production. In: Seed Biology, vol.
HI, ed. T. T. Kozlowski, pp. ]-54. New York: Academic Press.
Cowan, J. R. 1961. Producing high quality seed. In: Turfgrass Science, ed. A. A. Hanson and
F. V. Juska, pp. 424-441. Madison, Wisc.: American Society of Agronomy.
Douglas, 1. E., ed. 1980. Successful Seed Programs: A Planning and Management Guide.
Boulder, Colo.: Westview Press.
George, R. A. T. 1985. Vegetable Seed Production. New York: Longman Press.
Hebblethwaite, P. D., ed. 1980. Seed Production. London and Boston: Butterworth and
Company.
Hughes, H. D., and D. S. Metcalfe. 1972. Crop Production, 3rd ed., p. 185. New York:
MacMillan.
Kelly, A. F. 1988. Seed Production ofAgricultural Crops. New York: Longman Press.
McDonald, M. B., and L. O. Copeland. 1995. Principles and Practices of Seed Production.
New York: Chapman & Hall.
Rogers, K. J., and K. A. Lucken. 1973. Hybrid wheat seed production in North Dakota.
North Dakota Agricultural Experiment Station Report No. 806, Farm Research 30(6):4.
Thompson, J. R. 1979. An Introduction to Seed Technology. New York: John Wiley and
Sons.
Seed Production 251
U.S. Department of Agriculture. 1961. Seeds: The Yearbook of Agriculture. Washington,
D.C.: U.S. Government Printing Office.

1. Airy, J. M., L. A. Tatum, and 1. W. Sorenson, Jr. Producing seed ofhyhrid corn
and grain sorghum, pp. 145-153.
2. Bodger, H. The commercial production of seeds of flowers, pp. 216-220.
3. Bohart, G. E., and F. E. Todd, Pollination of seed crops by insects, pp. 240-246.
4. Cochran, L. c., W. C. Cooper, and E. E. Blodgett. Seeds for rootstocks of fruit
and nut trees, pp. 233-239.
5. Culbertson, J. 0., H. W. Johnson, and L. G. Schoenleber. Producing and
harvesting seeds of oilseed crops, pp. 192-199.
6. Graumann, H. O. Our sources of seeds of grasses and legumes, pp. 159-163.
7. Hanson, E. W., E. D. Hansing, and W. T. Schroeder. Seed treatments for control
of diseases, pp. 272-280.
8. Harmond, J. E., J. E. Smith, Jr., and 1. K. Park. Harvesting the seed of grasses and
legumes, pp. 181-188.
9. Hawthorn, L. R. Growing vegetable seeds for sale, pp. 208-215.
10. Hills, C. A., K. E. Gibson, and W. F. Rochow. Insects, viruses, and seed crops,
pp. 258-263.
11. Hoekstra, P. E., E. P. Merkel, and H. R Powers, Jr. Production of seeds offorest
trees, pp. 227-232.
12. Kreitlow, K. W., C. L. Lefebre, 1. T. Presley, and W. J. Zaumeyer. Diseases that
seeds can spread, pp. 265-272.
13. Lieberman, F. V., F. F. Kicke, and O. A. Hills. Some insect pests of important
seed crops, pp. 251-258.
14. McMurtey, J. E., Jr. Producing and harvesting tobacco seed, pp. 206-207.
15. Pedersen, M. W., L. G. Jones, and T. H. Rogers. Producing seeds of the legumes,
pp.171-181.
16. Ricker, P. I. The seeds of wild flowers, pp. 288-294.
17. Rogier, G. A., H. H. Rampton, and M. D. Atkins. The production of grass seeds,
pp.163-171.
18. Rudolf, P. O. Collecting and handling seeds of forest trees, pp. 221-226.
19. Shaw, W. C. and L. 1. Danielson. The control of weeds in seed crops, pp. 280-287.
20. Stevens, H., and 1. R. Goss. Seeds of oats, barley, wheat, and rice, pp. 153-159.
21. Stewart, D. New ways with seeds of sugar beets, pp. 199-205.
22. Todd, F. E., and S. E. McGregor. Insecticides and honeybees, pp. 247-250.
23. Waddle, B. M., and R. F. Colwick. Producing seeds of cotton and other fiber
crops, pp. 188-192.
Wheeler, W. A., and D. D. Hill. 1957. Grassland Seed'!. New York: D. Van Nostrand
Company.
 11
Seed Conditioning
and Handling

Seed as it comes from the field is almost never pure. It usually arrives at the cleaning plant
containing large quantities oftrash, leaves, weed segments, other crop seeds, and insects. If it
contains such materials as green leaves and other high-moisture materials, it cannot be safely
stored, efficiently handled, nor accurately cleaned until most of the foreign material has been
removed. The process of removing these unwanted materials from a seed lot, along with overall
improvement of seed quality, is known as seed conditioning.
Seed conditioning is a vital part of the total technology involved in making available high-
quality seed of improved varieties. It assures farmers of high-quality seed with minimum
adulteration. A good seed conditioning job can assure that previous efforts of plant breeders in
developing superior varieties, and of seed producers in growing them, can result in maximum-
quality seed. If seed is not conditioned and handled properly, all past efforts in varietal
development and seed production can be lost.
Figure 11.1 shows the steps involved in a typical seed conditioning plant.

PRINCIPLES OF SEED CONDITIONING

The seed conditioner has five objectives when cleaning seed: (1) complete separa-
tion-removal of all contamination, (2) minimization of seed loss-some good seeds
are removed along with contaminants in almost every conditioning operation and this
loss must be kept at a minimum, (3) upgrading-improvement of seed quality through
removal of decayed, cracked, broken, insect-damaged, or otherwise injured or low-
quality seed, (4) efficiency-the highest capacity consistent with effectiveness ofsepa-
ration, and (5) minimization of labor-labor is a direct operating cost and cannot be
recovered.
The quality of seed is improved during conditioning in two ways: (l) separation of
contaminating seeds of other crops, weeds, and inert matter, and (2) upgrading, or the
elimination of poor-quality seed. The ultimate goal of seed conditioning is to obtain the

L. O. Copeland et al., Principles of Seed Science and Technology

© Kluwer Academic Publishers 2001
Seed Conditioning and Handling 253

Receiving f----

~
Conditioning
and
Pre cleaning

/
f-- I Cleaning

~
r Separating
and
Upgrading

/
Treating
Bulk and
Storage Bagging

a-J LI Shipping I

Figure 11.1. Basic flow diagram showing essential steps in seed conditioning (Reprinted through the
courtesy o/the publisher, Vaughan and Delouche, 1967).

maximum percentage of pure crop seed with maximum germination potential. This
concept is reflected in terms of the pure live seed percentage. This is calculated by
multiplying the percent purity and the percent gt~rmination as in the following example:

Pure seed content (95%) x Germination (93%) = Pure live seed (88.35%)
(0.95 x 0.93 x 100 = 88.35%)

The pure live seed content provides a more realistic picture of the actual quality of a given
seed lot than either the purity or germination alone.
Seeds can be separated by mechanical means only if they differ in some physical
characteristic that can be detected by a mechanical or electrical process. Thus, the seed
conditioner uses differences in physical characteristics of the seed crop being cleaned and
those of seeds of other crops and weeds as well as inert matter. Inert matter such as chaff,
stems, and stones is usually the easiest to remove; other crop seeds and weed seeds may be
much more difficult, especially those similar in appearance and physical characteristics.
Physical characteristics that are used to separate seeds include: size, length, width, thickness,
shape, weight (specific gravity), surface texture, color, affinity for liquids, and electrical
properties.

SEED CLEANING EQUIPMENT

The seed conditioner should carefully analyze each seed lot as it comes from the field
to determine what the conditioning problem will be and which machine or machines will do
the best job. Since more than one machine must generally be used to completely condition
a given seed lot to maximum purity and germination, their sequence of use is also important.
254 Seed Conditioning and Handling

Frequently seed arriving at a conditioning plant contains excessive trash, which makes
it difficult to move through the elevators, thus interfering with proper conditioning by
lowering efficiency and slowing capacity. When this occurs, the seed may require one or more
precleaning operations to improve conditioning efficiency and separation precision, and prevent
loss of seed during subsequent conditioning.

Precleaning Equipment

Probably the most commonly used precleaning machine is a scalper. This machine is used
to rough-clean various kinds of trash from the seed lot. Although many different scalpers are
available, they usually consist of a vibrating or rotating screen (or sieve) through which the
small seed pass readily, while the larger seed and inert matter are "scalped off' and removed.
This separation is usually accompanied by an air flow that blows away the chaff, stems, and
other lighter contamination. To remove the lighter contamination from the seed without a size
separation, some scalpers utilize only the air flow of an aspirator, through which the seed falls.
Although scalping is the most important precleaning operation, other operations may also
be needed to increase the cleaning efficiency. For example, oats, barley, and many noncereal
grass seeds have awns, hairs, or other chaffY appendages that cause them to cling together, and
thus make them hard to condition. This is particularly true for the so-called chaffY grasses of
the Great Plains region of the United States. Seed of these species is often preconditioned by a
vigorous hammering, rubbing, or abrading action. Several kinds of machines are available;
however the debearder perhaps is used most frequently. It has a hammering or flailing action
that removes awns, beards, or lint from the seed and tends to break up seed clusters ofthe chaffY
grasses, as well as multiple seed units of nonchaffy types, permitting them to be conditioned
more efficiently. Another conditioning machine, the huller-scarifier, removes hulls or pods from
seeds, such as crownvetch, crambe, and lespedeza by an abrading or rubbing action. This
machine also reduces the hard seed content of many species, such as sweet clover, crownvetch
and alfalfa, whose seed coats are impermeable to water. It will do a good job of scarification,
but must be carefully adjusted to prevent seed injury.

Air Screen Machine

The air screen machine (Figure 11.2), sometimes called the fanning mill, is the basic seed-
cleaning machine in most conditioning plants. It uses a combination of airflow and perforated
metal or wire screens to separate seed on the basis of size, specific gravity, and resistance to
airflow. Many sizes of air screen cleaners exist- from the small, two-screen farm models to
large industrial cleaners with seven or eight screens and three or four air separations.
The air screen machine works in three different ways. Seed first enters the machine by
gravity from a feed hopper. In some machines, it falls directly onto a scalping screen, which
allows the good seed to fall through, while separating out the larger-seeded species and foreign
material. In other machines, the seed first falls through an aspirating airstream which blows off
the lighter material from the seed mass. The last operation grades the seed by allowing the good
seed to ride over screen perforations, while the smaller particles drop through. All air screen
machines use these three principles, though their sequence may differ.
Seed Conditioning and Handling 255

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Figure J1. 2. The air screen machine, with its parts labeled (Courtesy of Crippen Manufacturing
Company, St. Louis, MI).

Screens used in an air screen machine are constructed of either perforated sheet metal or
woven wire mesh on a wooden frame. Metal screen openings may be either round, oblong, or
triangular, while openings in wire mesh screens are either square or rectangular. The
perforated metal screens are measured in 64ths of an inch (hole size) and the wire mesh
screens are measured in number of openings per inch. Most air screen machines used in
conditioning seed use three or four screens. These screens can be easily changed, and almost
100 sizes are available.
The screens are kept clean during operation by tappers, or hammerlike screen knockers
(whichjar loose material wedged in the perforations), and brushes (which move back and
forth underneath the screen to dislodge and brush away material clinging to it). Some ofthe
newer machines have highly resilient rubber balls that bounce between the screens and help
dislodge such material.

Gravity Separator

The specific gravity separator (Figure 11.3) is perhaps the second most commonly used
piece of seed cleaning equipment. It can be used to separate undesirable seed and inert
contaminants that are so similar in size, shape, and seed coat characteristics to the crop seed
that they cannot be removed in any other way. In addition to its separation of other crop and
weed seed contamination, it is probably the best machine available for upgrading seed
quality. For example, deteriorated, moldy, or decayed seed, which are usually similar in size
and shape to good seed but have a lower specific gravity, can be removed by this machine.
Insect-damaged seed, empty seeds, off-color seeds and any other seeds that have defects that
decrease their specific gravity can also be remov~:d. Heavy nonseed particles such as mud
256 Seed Conditioning and Handling

Figure 11.3. A gravity separator (Courtesy of Forsbergs Inc., Thief River Falls, MN).

balls, soil particles, and stones can also be removed.

Although several types and styles of gravity machines exist, they all have basically the
same components and use the same principle of separation. They consist of a base (or
frame), one or more fans, a plenum chamber (air chest), a porous vibrating deck, a feed
hopper, and a seed discharge system. Seeds are introduced from the feed hopper onto the
porous metal or fabric deck, where the combination of shaking and airflow up through the
deck causes them to stratify according to specific gravity, producing a layering effect; the
heavier particles remain close to the deck surface, while the lighter components float on a
cushion of air above them. The deck can be tilted in two directions to facilitate seed
separation and discharge. The slight incline and vibration of the deck causes the heavy
particles in close contact with the deck surface to "walk" or be forced up toward the top of
the deck, where they ultimately fall off and are separated. The lighter particles tend to float
on an air cushion above the heavier seeds, following the path of least resistance, and drift
to the lower end of the deck where they fall off and are discharged. Seeds and other particles
in the medium specific gravity ranges, called middlings, fall from the deck near the middle
area; shields, or discharge aprons, can be mounted on the discharge side of the deck to
direct the middlings into one spout from which they are returned for another separation
on the deck.
Seed Conditioning and Handling 257
The gravity separator is widely used for cleaning and upgrading seed of a great
many species. However, it is normally used only after other machines (such as the air
screen machine) have been used to eliminate most chaff, stems, and off-sized
contamination. It is especially useful for upgrading seed offield beans, alfalfa, grasses,
as well as many other crops.
A modification of the specific gravity separator is represented by a machine called
the stoner. This machine separates seed on the same principle as conventional specific
gravity machines; however, it discharges only a1t each end. The desirable seeds flow to
the lower end and are discharged, while stones and heavier concreted earthen material
"walk" their way up to the upper end of the deck and are discharged. This machine is
useful in conditioning seed offield beans, sinct~ it eliminates most of the small stones
that commonly occur in bean seed lots.

Dimensional Sizing Equipment

Several types of seed conditioning machines separate or grade seeds on the basis
of width or thickness.
Length separators are specifically designed to separate seeds differing in length.
Two types are available, and both are commonly used for conditioning seed, especially
for cleaning smaller-seeded grasses and legumes. These are the indent disk and indent
Gylinder separators (Figures l1A and 11.5). The indent disk separator uses a series of
indented disks that are rotated inside a tilted cylinder through which the seed moves.
Disks may be used with indents that will lift OUit shorter contaminating seeds from the
longer crop seeds, or remove smaller crop seed and leave the large-seeded contamination
to flow on through the cylinder. The indent cylinder works in much the same way, but
the indentations are in the inside cylinder housiing, which lifts out undesirable seeds as
it rotates on an almost horizontal axis. The cylinder is tilted so that the seeds may flow
through it by gravity and discharge from the lower end after a separation is attained. Both
the indent cylinder and indent disk separators are best suited to free-flowing species.
Width and thickness separators are commonly called graders or sizers by the seed
industry and are primarily used for separating hybrid corn seed into different size and
shape grades. There are three types of graders used: (1) ribbed horizontal-flat screen
types, (2) vertical ribbed screen types, and (3) cylindrical screen types.

Surface Texture Separators

Three types of seed conditioning machines separate on the basis of differences in
surface texture. The roll machine is probably tht! best known and most widely used. It is
also known as the "velvet roll" or "velvet roll mill," but is most frequently called the
"dodder mill" because of its effectiveness in removing dodder from clover and alfalfa
seed.
Like gravity separators, roll mills are finishing machines and should be used only
for seed that has already been conditioned by other machines. This machine is effective
258 Seed Conditioning and Handlin/(

Figure 11.4. The indent disk separator. The illustration on the left shows how the disks revolve
through the seed mass, and the one on the right shows how the shorter seeds are liftedfrom the seed
mass, while the longer ones are rejected by the disk indents (Reprinted through the courtesy of the
Carter-Day Company, Minneapolis, MN).

in removing rough-textured, irregular, broken, cracked, and immature seed, or inert

matter from smooth-textured crop seeds. The machine consists oftwo slightly inclined
velvet-covered cylinders which roll in opposite directions. The seed to be cleaned is fed
through rolls at the upper end of the incline, slides down between them, and is
discharged from the lower end. Rough-textured seeds are separated by their tendency to
cling to the velvet-covered rolls and be lifted out. A metal shield covers each set of rolls
and directs the rejected seed and inert matter into the discharge spout. The machine can
be adjusted by varying the roll speed, cylinder tilt, and shield clearance. Its capacity may
be increased by using multiple sets of stacked rolls. Most commercial units use up to 10
sets of rolls.
The roll mill has done much to aid in separating seed of dodder from westem-
produced alfalfa seed and is to a large extent responsible for the elimination of this
noxious weed as a serious problem in many alfalfa seed consumption areas.
Two other machines that use differences in surface texture are magnetic separators
and inclined drapers. The magnetic separator depends on the affinity (especially when
wet) of rough-textured, undesirable seed for metal dust. When the seed is pretreated with
Seed Conditioning and Handling 259

Figure 11.5. Three views of the indent cylinder machine. The view at the lower left shows: (A)
materials being lifted by the cylinder, (B) material being deposited into the conveying trough, and (C)
the position of tbe separating edge (Reprinted through the courtesy of the Carter-Day Company,
Minneapolis, MN).

a combination of oil and water and exposed to a fine iron powder, the rough-textured
contaminants such as dodder attract the mt!tal dust and they are removed by a
magnetized metal drum over which the seed moves. The revolving drum is then brushed
or scraped clean on the lower tum, and the seed is eliminated. The inclined draper, a
special-purpose finishing machine that does not have wide use, separates seeds on the
260 Seed Conditioning and Handling
basis of their different tendency to roll or slide down an inclined surface. Crop seeds
containing contamination with different sliding or rolling tendencies can often be
cleaned very thoroughly by this method.

Spiral Separator

This device has no moving parts but permits separation of seeds that differ in their ability
to roll do~ an inclined spiral. Round seeds can be easily separated from flat or irregular seeds
since they tend to roll much easier. Thus, when seed mixtures are fed into the spiral, the round
seeds move at much faster speeds and are thro~ by centrifugal force from the top spiral into
a second spiral below, where they are discharged separately. The spiral separator is very useful
for removing nonround contamination from vetch seed. It is also effective in separating vetch
from small grain seeds.

Color Separators
Color separators make it possible to separate seeds that cannot be separated by any other
method. This machine requires only that contamination, or undesirable seed, be slightly different
in color than the good crop seed. This machine should be used only after the seed is cleaned and
graded by other processes. If properly used, it can be valuable in upgrading germination and
overall seed quality by eliminating off-color, poor-quality crop seeds. For example, it can
separate weathered, deteriorated, and off-color diseased seeds from high-quality seed of navy
beans. The use of color separators has completely eliminated long hours of tedious and
comparatively inefficient hand-sorting belts previously used in the bean industry.
Color separators contain a photoelectric cell that changes its electric characteristics in
relation to the amount of light, or radiant energy it receives. Phototubes can be built that are
sensitive to a particular color of light by varying the construction materials, and the light
reaching them can be controlled by placing various filters in front ofthem. Light is then reflected
into the cell from a variable background ofkno~ color. It is through this beam oflight that the
seed is dropped by gravity (one at a time). When an off-color seed appears between the preset
background and the photocell, a rejection system is triggered which ejects the seed into a
discharge area apart from the desirable seed.
Electrostatic Separators
Electrostatic separators separate seed by using differences in their natural or induced
electrical properties. Their effectiveness depends on the natural charges of various seeds in the
mixture and their relative ability to accept and retain an induced charge.
Seed is passed over a positively charged metal-embedded belt a few inches away from a
negatively charged electrode. Those seeds with positive electric charges are attracted toward the
electrode, lifted from the metal drum, and fall outside the normal path of seed discharge.
Negatively charged seeds are repelled from the electrode and are pinned to the belt and carried
underneath the belt where they are brushed off into a separate discharge spout. A third
separation effect is achieved by the difference in the ability of seeds to accept and retain an
electric charge. Those that accept a negative charge from the electrode (conductors) are attracted
to the belt, where they adhere until the charge is lost, causing them to fall at varying distances
past the location of normal seed discharge.
Seed Conditioning and Handling 261
Timothy Bumper Mill

The bumper mill is a specialized machine used only for cleaning timothy seed. It
utilizes a knocking (or bumping) action to remove weed seeds from timothy and
separates on the basis of differences in shape, surface texture, and weight of seeds. The
seed to be separated is fed onto a series of slightly inclined, identical, superimposed
decks. When the decks are sharply bumped by a knocking action, the timothy seeds tend
to roll back a considerable distance as they move uphill, while the contaminating seeds
and inert matter are moved a shorter distance uphill. By the time the seeds move from
the feeder to the discharge area, the timothy seeds are separated far enough from the
contamination so that they fall into a separate discharge spout.

Vibrator Separator
This separator has a coated vibrating deck onto which the seed is fed. It separates
seeds on the basis of their different reaction to the vibrating deck and materials that coat
it. The interaction of some seeds with the deck material creates friction causing the seed
to be "walked" upward before being discharged at the upper edge of the deck, while
others slide or roll downward and are discharged into a separate spout.
Single-vibrator separator decks are sometimes used to separate small samples in
seed-testing laboratories. In order to condition commercial seed lots, multiple decks
provide greater capacity. The vibrator separator removes curly dock from crimson clover
and dog fennel from timothy. As new deck materials are used, it has the potential for
various other separations.

Affinity for Liquids

One process uses affinity for liquids to effect a separation. It is used to separate
seeds of buckhorn plantain from crop seeds of similar size, shape, and density. Buckhorn
seeds are covered with a mucilaginous layer that becomes sticky when moistened. Seed
producers mix the seed with water followed by sawdust, which readily adheres to the
buckhorn seed coat, increasing its size, and changing its specific gravity, allowing it to
be separated by either the air screen machine or gravity separator. Later, the sawdust is
dried and reused.

SEED TREATMENT

Seed treatment is the process of applying c:hemical substances to seeds in order to

reduce, control, or repel seedbome, soilborne, or airborne organisms. Chemical
treatment is accepted as a sound agronomic practice for seeds of many field and garden
crops and is usually a routine part of seed conditioning. In the past 50 years, the
treatment of seeds with protective chemicals prior to planting has become a standard and
widely accepted practice.
262 Seed Conditioning and Handling
History of Seed Treatment
Although seed treatment has only recently become an important part of seed conditioning,
it is by no means a new concept. According to the 1966 DuPont Seed Treating manual, a
knowledge of the benefits of treating seed dates back to the 17th century, when saltwater was
accidentally discovered to reduce bunt and stinking smut infestation of wheat seed. Later, in
1755, Matthieu du Tillet, a French botanist, recommended the use oflye and lime for chemical
treatment of wheat seed. Fifty years later, Prevost, a Swiss botanist, introduced the use of
copper sulfate seed treatments. However, a new concept of treating seeds began in the 1920s
with the introduction of Ceresan and Semesan, the first of the organic mercurial compounds. In
recent years, the increasing recognition ofthe environmental implications of organic mercurials
has led to their disrepute and spurred the development of effective alternatives to mercurials for
seed treatment.

The Ideal Seed Treatment Chemical

The ideal seed-treatment chemical should be: (1) highly effective against pathogenic
organisms, (2) relatively nontoxic to plants, (3) harmless to humans and livestock, even if
misused, (4) stable for relatively longer periods oftime during seed storage, (5) easy to use, and
(6) economically competitive. Unfortunately, none of the chemicals currently available meets
all these requirements.

Treatment Formulations and Equipment

The types of pathogenic organisms controlled by seed treatment include: (I) fungi and
bacteria-----<.:ausing seed rots, seedling blights, and smuts. These pests may be seedborne,
soilborne, or airborne; (2) soil insects-such as seed corn maggot and wireworm, and (3)
storage insects-including weevils, moths, and beetles.
The activity of seed treatment chemicals falls into three principal categories: (1) seed
surface disinfestation-----<.:hemicals in this group cover the seed and kill or suppress the activity
of spores, and other disease agents on the seed surface; (2) seed protection- chemicals in this
group protect seed before and during germination from soilborne diseases and insects; and (3)
systemic protection-----<.:hemicals in this group penetrate the seed and kill or suppress the activity
of pests or pathogens. They may also extend into and protect the resulting plant.
Seed treatment chemicals are normally combined with other materials that enhance or
maintain their activity. Many formulations contain several inert ingredients in addition to the
active ingredients. Inert ingredients act as carriers, binders, wetting agents, sticking agents,
emulsifiers, suspending agents, and dyes. These materials do not have to be listed on the label
since they are added to the formulation to improve appearance, increase coverage and adherence,
prevent dusting off, or make the formulation easily recognizable.
Seed treatment chemicals may be applied as a slurry (flowable, paste, or a mixture of a
wettable powder and water), a liquid, or a dust. Other formulations are used, but the slurry or
film-coating methods are most common. Special equipment is available for applying each type
of formulation. If no commercial application equipment is available, the treatment may be
applied by home methods, such as in a revolving metal drum or even in a cement mixer. Special
care should be taken to see that the chemical is applied uniformly to the seed.
Seed Conditioning and Handling 263

Both state and federal agencies normally require licensing of all businesses that offer
pesticide application to the public, including those that custom-treat seed for a fee. If seed is
already treated and then sold, no pesticide applicator license is required. An individual in each
licensed facility must normally be certified in the Icategory of seed treatment, and licenses are
not issued until one person is certified. Seed treaters who do not offer their services to the public
will not need certification unless they use restricted-use pesticides.
Identifying Treated Seed
State and federal laws require that treated seed be identified in two ways: (1) by
incorporating a dye into the treatment that will give the seed a contrasting color, and (2)
by a statement indicating the seed has been treated and the name ofthe chemical(s) used
[either the common name, chemical (generic) name or abbreviated chemical name]. Seed
treated with highly toxic pesticides must bear a label with a skull and crossbones and a
precautionary statement. The skull and crossbones must be at least twice the size as the
type on the label and the precautionary statements must be in red letters on a contrasting
background. Seed treated with substances not listed as highly toxic must bear a label
with an appropriate precautionary statement. T his information may appear on a separate
tag or be printed conspicuously on the side or top ofthe seed container.

SEED HANHLING

Receiving, Elevating, and Conveying Equipment

Every seed conditioning plant must have adequate equipment for receiving seed and
conveying it throughout the plant. A well-equipped plant has a pit area where incoming
trucks can be unloaded quickly. From the pit an~a, conveying facilities must be available
to move the seed throughout the plant vertically, horizontally, or on an inclined plane
as needed. Conveyors can be classified as: (1) bucket elevators, (2) belt conveyors, (3)
vibrating conveyors, (4) pneumatic (air) conveyors, (5) screw conveyors, (6) chain
conveyors, or (7) lift trucks.
Bucket elevators are normally used to move seeds vertically within the conditioning
plant, especially from the pit area to overhead storage bins. They consist of an endless
belt or chain equipped with evenly spaced buckets. Several types are available, but the
most satisfactory ones have good capacity, are relatively quiet, require little mainte-
nance, and are self-cleaning. The space between the rear of the bucket and the belt
should be checked frequently because it is particularly troublesome as a seed hangup
area. Another trouble spot that must be checked and cleaned frequently is the boot area
at the lower end of the elevator leg.
Aside from the seldom-used screw and pne:umatic conveyors, most other conveyors
move the seed horizontally within the plant, especially between storage bins or condi-
tioning operations. Horizontal seed conveyors are suitable for bagged or unbagged seed.
264 Seed Conditioning and Handling

Scales
Seed is weighed at least twice in most seed conditioning operations: (1) when it is received,
and (2) when it is bagged. The first weighing requires heavy-duty platform scales for weighing
trucks, trailers, or wagons. The second scale needs to weigh up to I OO-Ib bulks for measuring
the amount of seed put into bags. These are usually placed at the bottom of a bagging bin to
permit a predetermined amount of seed to flow into the bag before it is stopped by a tripping
mechanism, which may be either manually or automatically operated. In addition to these types
of scales, most conditioning plants have small portable platform scales for weighing small
amounts of seed.

Miscellaneous Equipment
All seed conditioning plants must have considerable miscellaneous equipment to aid in
adequately conditioning seed and to prepare it for marketing. This includes sewing machines,
heat-sealing devices, air compressors, vacuum cleaners, and tag print equipment.

DESIGN OF SEED CONDITIONING PLANTS

The design and layout of seed conditioning plants should be carefully planned to ensure
that: (1) the seed receives the necessary conditioning in the proper sequence, (2) there are no
bottlenecks, (3) operating costs are kept to a minimum, (4) the seeds are not injured from
excessive handling, (5) facilities are completely cleanable, and (6) the chance of contamination
is kept to a minimum.
If possible, the seed should be elevated only once to overhead storage bins, where it can
be held until conditioning. From there it should be allowed to flow by gravity between the
various pieces of conditioning equipment. In order to accomplish this, an initial elevation of
about 40 to 60 ft is needed. Mistakes in design will require two or more vertical elevations and
increase the opportunity for seed injury and seed lot contamination, as well as require extra
maintenance.
Figure t 1.6 shows a suggested design for a seed conditioning layout that can handle most
crops, including small-seeded grasses and legumes. Figure 11.7 shows the sequence of
conditioning operations for cleaning seed of various species.

Questions

t. What is the single most important piece of equipment for cleaning seeds?
2. Can you explain the operation principle of each piece of seed cleaning equipment, as welt
as its advantages and disadvantages?
3. Do you believe that the proper use of seed cleaning equipment could eliminate almost any
quality problem in most seed lots? Can you think of poor-quality factors that could not be
eliminated by the proper selection and use of equipment?
4. What is the pure live seed concept?
5. Can proper selection and use of equipment compensate for a poor job of seed production?
6. Do you consider seed treatment to be a desirable practice? What do you consider to be the
future importance of seed treatment?
Seed Conditioning and Handling 265

STORAGE
BIN

STORAGE
rn [ rn
STORAGE
BIN
g
ELEV
STORAGE ~~ '"~ ~ Mtd 10 be plo.,d r---
E:IN '" DlMP f/H COIlY.yor Iltto hop,.,
~!~ a..

mEJ
~,u DO ELEV ::;;:
STORAGE 3~ <l:
tl:
BIN
~
z
STORAGE ,11 15

r---
<l:
0
BIN o ElE'V. (UNCL.EANED SEED) ...J
~, ' EL.E'b 0 SACt<

~"'O'6[cT
DUMP
---,-".--- pm
DUN.

UNLOADING
STORAGE
RAMP

A. - Floor plan for seed conditioning plant.

TRUCK DUMP

B. - Isometric drawing of flow plan illustrated above.

Figure 11.6. A suggested layout for a single-story seed conditioning plant (Reprinted through the
courtesy a/the publisher, Vaughan et al. (1967).
 N
0\
0\

~eceiving Hoppe~
d
,/ ,,-
'" /
'"
I scal~r I [ Sca~, I
---
[Debe,arder! Scarifier
,,

Air Screen
Cleaner "-
(fannmg mill)
r~:n" I C~L_~ /
" "- ....
~

," Gravity
If Separator
Indent "
Cylinder ~
"

~~"e< I ~
(\)
l:l..
g
·····~gi.ng!
r
1
Machme I
...~
c·
;::s
t ~.
Small-seeded Grasses Field Beans Small Grains Soybeans Small-seeded Legumes

Figure I J. 7. Suggested sequence for conditioning seeds of various kinds. Discontinuous lines indicate optional processes. ~
~
;::s
~
~.
Seed Conditioning and Handling 267
7. How does the widespread use of augers for c:onveying seed affect its quality? Name the
places where a seed lot is likely to encounter an auger throughout the production process.
8. Can you suggest principles of adjustment that might apply to almost any piece of seed
cleaning equipment? Can you suggest alternative methods that might be used in the place
of augers?
9. Do you believe most seed conditioners only attempt to condition to those quality levels that
will meet minimum acceptance levels of the seed trade?

General Refe~rences

Douglas, 1. E., ed. 1980. Successful Seed Programs: A Planning and Management Guide.
Boulder, CO: Westview Press.
Gregg, B. R., A. G. Law, S. S. Virdi, and 1. S. Balis. 1970. Seed Processing. New Delhi, India:
Mississippi State University, National Seeds Corporation, and United States Agency for
International Development, New Delhi.
Harmond, J. E., N. R. Brandenberg, and L. M. Klein. 1968. Mechanical Seed Cleaning and
Handling. Agricultural Handbook No. 354. Washington, D.C.: Agricultural Research Service,
U.S. Department of Agriculture in cooperation with Oregon Agricultural Experiment Station.
Klein, L. M., 1. Henderson, and A. D. Stoess. 1961. Equipment for cleaning seeds. In: Seeds:
The Yearbook of Agriculture, ed. Alfred Stefferud, pp. 307-321. Washington, D.C.: U.S.
Department of Agriculture.
Thompson, J. R. 1979. An Introduction to Seed Technology. New York: John Wiley and Sons.
Vaughan, C. E., B. R. Gregg, and 1. C. Delouche, eds. 1967. Seed Processing and Handling.
Handbook No.1. State College, Miss.: Seed Technology Laboratory, Mississippi State
University.
Wheeler, W. A., and D. D. Hill, 1957. Grassland Seeds. New York: D. Van Nostrand.
 12
Seed Dryingl

The drying and storage of seeds are often essential steps in the production and maintenance
of high quality seed. In the drying process, excess water is removed, either in the field by natural
means (i.e., sun and wind) or in a bin or dryer with the aid of a fan and heater. In storage, an
atmosphere is maintained in which the temperature and moisture content of the seed remain
constant, regardless of the length of the storage period.
The important quality factor to be maintained during the drying and storage of seed is
viability. Otherwise, seeds rapidly lose viability when dried with air at excessively high
temperatures, or when stored under excessive moisture contents. Thus, these conditions should
be avoided in the production of high-quality seed.
Sensitivity to high temperatures and high moisture varies among different species and
varieties or hybrids. Thus, the design and operation of a drying and/or storage system depends
on the kind of seed and location of production.

Drying Principles

Knowledge of the basic principles of drying is essential for understanding the process of
seed drying. In particular, it is important to have a rudimentary understanding of air properties,
equilibrium moisture content, and airflow rate (Brooker et al. 1992).
The medium in which seed is dried and stored is a mixture of dry air and water vapor. The
temperature of the moist air may refer to the dry-bulb temperature (i.e., measured with an
ordinary thermometer) or to the wet-bulb temperature (i.e., measured with a thermometer
covered with a wet wick). The amount of water vapor contained in the drying air can be
expressed in terms of vapor pressure (the partial pressure exerted by the water vapor molecules
in moist air) or the relative humidity (the ratio of the vapor pressure in the air to the saturated
vapor pressure at the relevant dry-bulb temperature).

lThis chapter was prepared by Professor Fred Bakkar-Arkema, Department ofBio-Systems Engineering,
Michigan State University.

L. O. Copeland et al., Principles of Seed Science and Technology

© Kluwer Academic Publishers 2001
Seed Drying 269
The equilibrium moisture content (EMC) is the moisture content of the seeds after they
have been exposed for a long period of time to ambient conditions and is a function of the kind
and variety (or hybrid) of seed and storage temperature.
The airflow rate in seed drying and storage systems is a function of the resistance to the
airflow by the seed, and depends on the characteristics of the fanes) used in a dryer or storage
bin. When air is forced through a layer of seed, resistance to the flow (the so-called static
pressure) occurs due to air friction and turbulence. The static pressure depends on the species,
the airflow rate, and the depth of the seed layer. The characteristics of a fan are expressed in
terms of the air flow rate at various static pressures, e.g., 500 m3/min at 5 cm of water (column).

Storage Principles

The objective of seed storage is to preserve seed quality (viability) throughout the storage
period. Although favorable storage conditions cannot improve seed viability, incorrect storage
can reduce its germination potential, usually due to physiological deterioration or mold
development (Sauer 1992).
Molds can be prevented from developing in stored seeds, thus maintaining their viability,
as long as the temperature and seed moisture content are controlled within narrow limits. In
general, most seed stores best at 12-13% moisture content and 10-13°C temperature. In tropical
regions, seed chillers must be used to attain these conditions. If a chiller is not available, seed
should be stored at 10% (or below) moisture conttmt in the tropics.
When bulk seed is stored in bins, intermittent aeration (i.e., cooling of the seed with
ambient air at a low airflow rate) may be necessary in order to off-set the non-uniform
heating/wetting of the seed due to natural convecticm currents occurring in bins due to changes
in the ambient conditions. Manual control of aeration fans is possible, but automatic control
is recommended (Brooker et al. 1992). If properly done, this will prevent moisture migration
and the development of wet, or "hot spots," which can otherwise lead to seed deterioration.

Drying Methods

Seed can be dried in various ways, including sun drying, bin drying, portable batch drying,
wagon bed drying, continuous-flow/crossflow dryilng, and rotary drying. In addition, seed com
is dried in specially designed ear-corn dryers. Each of these dryer types is explained in detail
in texts on grain drying (Brooker et al. 1992; Loewer et al. 1994).
In drying seed, it is important to use the maximum temperature at which the viability of
the seed is not impaired. The maximum temperature is a function of the moisture content and
seed type. For com at 24% moisture content, the maximum temperature for safe drying is 61°C;
at 18%, it is 67°C. It should be noted that the maximum seed temperature and not the maximum
drying-air temperature is specified since, in seed dryers, the the two temperatures are not
necessarily the same.
Figure 12.1 shows the influence of the drying temperature on the viability of seed com at
32% moisture content in a shallow-bed « 5 cm) bin dryer (Brooker et aI.1992).
270 Seed Drying
Sun Drying
Sun drying is the practice of spreading moist seeds in a thin 3-6 cm layer on the ground or
on a concrete floor and exposing them to ambient conditions. The practice is still commonly
used in the tropics and sub-tropics, especially in developing countries.
Sun drying is labor-intensive because it requires intermittent stirring ofthe layer to prevent
non-uniform drying and overheating of the layer of seed in direct contact with the heated air.
Also, during the night hours, and when it rains, the seed should be raked into a pile and covered.
The time required for sun drying depends on the ambient weather conditions, the thickness
of the seed layer, the initial moisture content, type of seed, and the frequency of raking or
stirring. For example, on Java (Indonesia) about three days are required during the non-rainy
season to dry rice seed from 20-22% down to 14%, assuming the 4 cm layer is raked twice daily
(Suhargo 1993).

Bin Batch-Dryer
In bin batch-dryers, the seed is placed in a (usually round) bin, and ambient or slightly-
heated air is blown through it by a fan. The maximum thickness of the seed layer in the bin
depends on the initial moisture content, the type of seed, the air temperature and relative
humidity, and fan horsepower. To obtain a uniform airflow through the seeds, a full perforated
floor is required.
Figure 12.2 shows a typical bin batch-dryer (Loewer et al. 1994). A 0.8-1.0 m layer of
seed at 20% moisture can be dried to 14% within 24 hours without affecting germination using
30-3 SOC and 50-65% relative humidity air at a rate of 5-8 m3 per minute per m3 of seed.
After the seed in a bin has reached the acceptable average moisture content, a moisture
gradient will remain from the top to the bottom of the seed. The surface layer will have a
moisture content above the average and the bottom layer of the bin will be lower than average.
Thus, proper mixing of the seeds is essential before further storage or packaging. This can be
addressed by installing one or more grain stirrers to mix the entire content of a bin for 3 to 12
hours (Figure 12.3).

Wagon Batch-Dryer
A grain-transport wagon can be transformed into a wagon batch-dryer by equipping it with
a plenum, a perforated floor, and a fan/heater unit coupled with a canvas transition to the wagon
(Figure 12.4). The drying principles ofa wagon batch-dryer and a bin batch-dryer are similar.
Wagon batch-dryers are most frequently used for drying fragile seeds such as large-seeded
legumes (e.g., field or garden beans and peanuts). The recommended airflow rate for the
ambient-air wagon drying of a 1.5 m layer of peanut seeds is 0.25 m3 of air per m2 of floor area
(Cabrera 1999).

Bin Layer-Dryer
In the process of bin batch-drying, a batch of seed is placed in a bin and the entire batch
is dried. In bin layer-drying, the seed is dried in layers; successive layers of moist seed are
Seed Drying 271

100

--
0~
80

--~
..J
m
60
Initial M.C. =32% w.h.
<t
>= 40
I:r-A 75.0·C
0
UJ 0-0 70.0·C
W
(fJ
20 ..... 50.0·C
A--.l 65.0·C

IHI 40.0·C

o L--L~~~~~~~~__~__L-~~
o 10 20 30 40 50
DRYING TIME (min)
Figure 12.1. Influence of drying temperature on the viability of corn seed at 32% moisture content
(w.b.) (Brooker, et al. 1992).

ROOF HATCH GRAIN SPREADER

L.P. GAS HEATER

UNLOADING AUGER

.... ~
..... r~

Figure 12.2. Batch-in-bin drying equipment (Loewer, et al. 1994).

272 Seed Drying

ROOF HATCH GRAIN SPREADER

..,.-- STIRRING DEVICE

UNLOADING AUGER

~~-L~~~~~~~--\~
~~ PERFORATED
.,,J
DRYING FLOOR

Figure 12.3. Layer drying equipment (Loewer, et al. 1994).

\ l' {
Peanut Seed

Plenum Chamber

Figure 12.4. Wagon batch-dryer (Cabrera 1999).

Seed Drying 273
added periodically to the drying bin after the previous layers have been partially dried. The final
seed depth in the layer-drying bin is much greater than in a bin batch-dryer.
Bin layer-drying requires a thorough understanding of the in-bin drying process, since the
depth of the successive layers of seed to be added to the drying bin depends on the type of seed
and its moisture content, the drying-air conditions, and the fan rating.
In drying seed corn from 20% down to 14% in a layer-drying system with air at 25-30°C
and 55% RH in 13 days, Loewer et al. (1994) recommended that 1.3 m, 1.1 m and 0.6 m of wet
seed must be added to the bin after, 3.9, 3.8 and 3.5 days (respectively) of drying while
maintaining airflows of7.l, 7.9 and 12.8 m2 per m2 of un dried seed.

Column Batch-Dryer

In a colunm batch dryer, drying air is forced perpendicularly from an air chamber through
the wet seed held in a relatively thin column (0.25-0.45 m). The airflow ranges from 15-30 m3
of slightly-heated air per minute per m2 of screen area. The temperature of the drying air should
not exceed 35-40°C. Figure 12.5 shows a colunm batch dryer (Loewer et al. 1994). After the
seeds in a colunm batch-dryer have been dried to near the desired average moisture content, the
seeds are cooled by ambient air after turning off the heating unit. The total time required for
drying and cooling 20% moisture seed in a colurrm batch-dryer to down to 14% is about 3.5
hours, depending on the seed type, the air temperature, and the airflow rate.
As in other batch-type dryers, the seeds in a colunm batch-dryer are not uniformly dried.
Thus, the seeds have to be well mixed prior to further storage or packaging.

Continuous-Flow Crossflow Dryer

The basic designs of the colurrm batch-dryer and the continuous-flow crossflow dryer are
similar. In both dryers, heated air is forced from a heated-air plenum through the moist seeds.
In the colurrm batch-dryer, the seed is stationary, while in the continuous-flow crossflow dryer,
the seed moves continuously, first through the drying section and next through the cooling
section. The thickness ofthe seed colunms in the two dryers is similar, as are the airflow rate,
the drying-air temperature, and the moisture gradient across the seed column. The residence time
in the continuous-flow crossflow dryer of seed dric~d from about 20% down to 14% at 40-45°C
is about 2-4 hours. Figure 12.6 shows a typical continuous-flow crossflow dryer (Loewer et al.
1994). Such dryers are usually designed to operate at 80-1 OO°C for the drying of seed corn.
Since such temperatures can be deleterious to seed viability, continuous-flow crossflow dryers
are seldom used for drying seeds.

Rotary Dryers

Most rotary dryers consist ofa slightly inclined long drum (shell) which slowly rotates (4-8
revolutions per minute). The moist seed and the drying air are introduced at one end of the shell,
and the dried seeds and exhaust air exit at the other end continuously. The inside of the shell
often has a set of flights which cascade the seeds through the shell. Figure 12.7 shows a
continuous-flow cascading rotary dryer for seeds (Brooker et al. 1992).
274 Seed Drying

weT GRAIN SUPPLY

GRAINSUDE

HEATED AIR
CHAMBER
DRYING COLUMNS
WITH PERFORATED
WALLS
CONVEYOR FOR
REMOVING DRIED
GRAIN

Figure 12.5. Typical column batch dryer (Brooker, et ai. 1992).

WET HOLDING I3IN

GRAIN COLUMN

METERING AUGER ---~~~~i!i~~

FOR COOL DRY GRAIN
Figure 12.6. Continuous flow dryer without heat recovery (Loewer, et al. 1994).
Seed Drying 275

I~---- Air out

Figure J2.7. Concurrent-flow cascading rotary seed dryer (Brooker, et al. 1992).

Rotary seed dryers are frequently batch-type units because of small lot sizes. Dryers often
consist of a perforated rotating drum in which the seeds are tumbled while being heated with
drying air which is injected perpendicularly to the drum axis. A batch-type rotary dryer is
particularly suitable for drying small lots of high moisture vegetable and/or flower seeds. Unlike
in sunlbinlwagonlportable-batch dryers, seeds are dried uniformly in rotary dryers. This is the
great advantage of this dryer type. However, it has relatively high fIxed and maintenance costs.

Ear-Corn Drying

Ear-com requires a totally different dryer design than those discussed thus far (Cabrera
1999). Not only must the com seed be dried, but the cob as well.
An ear-com dryer can be considered as a modified bin batch-dryer (see Figure 12.8). Wet
ear com at a moisture content of20-40% is placed in the dryer in a bin with a perforated 45°
angled floor; the drying depth is 2.0-3.0 ft and the seed is dried to 11-12%.
The drying air moves upwards through the ear-com at a rate of 0.25-0.35 in3 of air per
minute per in3 of product; the preferred conditions of the upward air are about 35°C and 65-
75% relative humidity. [Note that the air has alrt::ady passed (in an adjoining bin) through a
partially-dried layer of ear com.] The movement of the air is reversed from upwards to
downwards when about 65-70% of the moisture to be removed from the batch has been
extracted. Air reversal is accomplished by the closing and opening of two air valves. The
average downward air temperature is 35-45°C at a relative humidity of 15-30%, depending on
the ambient conditions. The total drying time is 50-100 hours.
Figure 12.8 depicts a typical ear-com dryer used in the hybrid seed com industry; it
consists of two drying bins which operate in tandem. At large ear-com installations, the number
of tandem units may be as high as 15-30.
276 Seed Drying

~--~~~~ll~pp~er~lt~--~
','
.. ·:.i·;
~
tunuel
'.
:.:.:.... .
~
fill'
..... :.~:.i;.·.'.• ~.~t-----I. .':~.

.... .. ...
I e'

...... '.
drlor sced ...... 1--1""" '., .•.• ,..,...-
•..••,.......... : •. ,;,.
. '·"~'::···:'·:··.·"""'·IIr---.-1Y'eu.er
~~ •.f .• :,~!& ,."
seed
..... : • '::.. r ~.-.
•• , t e" ...... ~ "
,
lower ....
~~ .. :
.,"

4- •
~..-:;~: .•..•. ' ••
•.•~ .•. ,,:~.. lUl111el ,~.;~:.~

L....t >~ . . .......A --~.~

Figure 12.8. Modified bin batch-dryer (Cabrera 1999).

Management of such a system is a challenge, Le., the logistics of filling/emptying the bins,
of the reversal time of the air, of the choice of the air temperature, etc.

Questions

1. Can all seeds be dried and stored under similar conditions? Explain.
2. Distinguish between the dry-bulb and the wet-bulb temperatures of air used for the drying
of seeds.
3. Why is the equilibrium moisture content of a seed of importance in the drying of the seed?
What is the difference between a column batch-dryer and a bin layer-dryer to be used for
seed?
4. Explain the design and operation of a modern bin batch-dryer for ear corn?
5. Why is aeration of seed during long-time storage necessary?

References

Brooker, D.B., F.W. Bakker-Arkema and C.W. Hall. 1992. Drying and Storage ofSeeds and
Oi/seeds. Van Nostrand Reinhold, New York, NY.
Cabrera, E. 1999. Personal communication. Pioneer Hi-Bred International, Inc. Johnston, IA.
Loewer, OJ., T.C. Bridges and R. A. Bucklin. 1994. On-Farm Drying and Storage Systems.
American Society of Agricultural Engineers, St. Joseph, MI.
Sauer, D,E. 1992. Storage of Cereal Grains and Their Products, American Association of
Cereal Chemists, St. Paul, MN.
Suhargo, S. 1993. Sun-Drying ofGrain. Unpublished Ph.D. Thesis, Michigan State University,
East Lansing, MI.
 13
Seed
E,nhancements

Seeds have evolved over time to respond to a variety of environments. As a result, these
adaptations generally result in satisfactory performance throughout a wide range of environ-
ments. Seed enhancement technology has a central objective to further improve seed
performance under very specific regimes and with certain planting equipment. Various
techniques have been employed to assure this supc~rior performance and most have found
commercial application. This chapter considers three of these seed enhancements: seed
hydration, biological seed treatments, and seed coatings. While not a direct manipulation of a
zygotic seed, another approach to improving seed performance that will be considered in this
chapter has been the propagation of somatic embryos that are then coated and marketed as
synthetic seeds.
Seed hydration is a process whereby seeds are hydrated using various protocols and then
redried to permit routine handling. This process results in increased germination rate, more
uniform emergence, germination under a broader range of environments, and improved
seedling vigor and growth. Biological seed treatments are a new approach to adding
biological organisms to seeds that effectively control soil and seed pathogens. This technique
demonstrates an industry sensitivity to the increasing use of synthetic pesticides which are
avoided by biological seed treatments. While seed hydration and biological seed treatments
improve the physiological performance of the set~d, seed coatings physically add external
substances to seeds that further enhance their performance. These range from surrounding the
seed with a pellet to improve precision planting to coatings that contain products which
protect the seeds against an array of pests or even modifY the time that water is absorbed by
the seed. One of the prospects emerging from recent innovations in biotechnology and plant
tissue culture has been the potential to produce synthetic seeds. These are "seeds" that are
essentially embryos produced without the benefit of sexual fertilization. Such seeds are
genetically uniform and offer the promise of greater economic yields and superior crop
products. All of these seed enhancement technologies are constantly being improved and offer
great hope for enhancing seed performance in the future.

L. O. Copeland et al., Principles of Seed Science and Technology

© Kluwer Academic Publishers 2001
278 Seed Enhancements
SEED HYDRATION

The objective of seed hydration technology is to increase the percentage and rate of
germination, expand the range of temperatures over which the seed will germinate, and
increase the uniformity of stand establishment. To accomplish these objectives, seeds must be
hydrated in some way at a moisture level sufficient to initiate the early events of germination
(Phase II of imbibition) but not sufficient to permit radicle protrusion (Phase Ill) (Akers and
Holley 1986). Three general approaches to hydration have been developed: prehydration,
priming, and solid matrix priming (McDonald 2000).

Prebydration

Soaking seeds in water prior to planting enhances germination and seedling growth by
controIling the imbibition conditions and reducing the vagaries of adverse weather and soil
conditions (Bradford 1986). While the soaking process can involve water, seeds are often
pregerminated in gels at 20°C and then planted; a process known as fluid drilling (Gray
1981). The gel both protects the germinated seed and maintains seed moisture. Examples of
gels used in fluid drilling include hydroxyethyl cellulose, magnesium silicate, and
polyacrylamide (Taylor and Harman 1990). In some cases, the gel may contain activated
carbon to detoxify soil herbicides (Taylor and Warlrolic 1987). In other cases, it may contain
added nutrients and pesticides to further improve seedling performance (Finch-Savage 1984).
Prehydration can also occur followed by redrying if the seeds have not visibly germinated
(Phase III) to facilitate subsequent handling, storage, and planting using traditional
agricultural equipment. In such cases, the seeds still germinate under a broader range of
temperatures than seeds which have not been prehydrated (Gelmond 1965; Guedes and
Cantliffe 1980; Gerber and Caplan 1989). Because the amount of water absorbed by the seed
is precisely controlled during prehydration to ensure that germination does not occur, the
physiological mechanism that results in greater seedling performance is considered the same
as that for priming or solid matrix priming (Karssen and Weges 1987).

Priming

The terms priming (Heydecker and Coolbear 1977) and osmoconditioning (Khan et al.
1978) have been used to describe the soaking of seeds in aerated low water potential
osmotica. Examples of such compounds include polyethylene glycol (PEG), KN0 3, K3 P04 ,
KH2P04, MgS0 4, NaCI, glycerol, and mannitol. The benefit of such salts is to supply the
seed with nitrogen and other nutrients essential for protein synthesis during germination.
Their disadvantage is their occasional toxicity to the germinating seedling. Even so, given the
correct concentration and time, primed carrot and tomato seeds perform better than unprimed
seeds when salts are used as the osmoticum (Haigh and Barlow 1987). Today, the most
preferred osmoticum is PEG. It is a high molecular weight (from 6,000 to 8,000 daltons) inert
compound whose large molecular size precludes it from entering the seed and creating toxic
side effects associated with the use of salts (Michel and Kaufmann 1973). A major
disadvantage of PEG is that oxygen solubility is inversely related to its concentration (Mexal
et al. 1975). As a result, when PEG is used as the osmoticum, the solution is often aerated to
ensure an adequate supply of oxygen to the seed (Akers 1990). In addition, it is difficult to
commercially treat large quantities of seeds using this technique. Generally, seeds are soaked
Seed Enhancements 279
at 15°C (Bradford 1986) in osmotica that possess a water potential of -0.8 to -1.6 MPa
(Khan 1992) for several hours (Guedes and CantIiffe 1980) to several weeks (Khan et al.
1980/1981) for optimum performance.
Priming has been successful for such crops as tomato, carrot, onion (Haigh and Barlow
1987; Alvarado et al. 1987; Dahal et al. 1990), pepper (Bradford et al. 1990), celery
(Brocklehurst and Dearman 1983), parsley (Pill 1986), wheat, barley, sorghum (Bods worth
and Bewley 1981), and ryegrass (Danneberger et al. 1992). In other cases, priming has been
able to overcome thermodormancy in crops such as lettuce by expanding the range of
temperatures at which the seed will germinate (C'antliffe et al. 1984; Valdes and Bradford
1987). These studies show that priming is consistently successful with small-seeded crops.
The technique, however, has been less successful with large-seeded crops such as soybean
(Helsel et al. 1986) and sweet corn (Bennett and Waters 1987). Armstrong and McDonald
(1992) showed that priming of soybean seeds without an intervening air-dry treatment
resulted in increased plumule and radicle length and weight. However, when these seeds were
air-dried, seed performance was decreased due to excess leakage of electrolytes from cracked
cotyledons.
After priming, the seeds are dried back to enable normal handling, storage, and planting.
The drying treatment slightly depresses the germination advantages gained during soaking
(Brocklehurst et al. 1984). In addition, use of rapid drying rates or excessive temperatures
can cause seed injury, therefore, priming must be conducted under carefully controlled
conditions. Primed seeds can be stored successfully for short periods without losing the
benefits gained from the treatment (Thanos et al. 1989). However, long storage periods cause
faster loss of vigor and viability compared to nontreated seeds (Alvarado and Bradford 1988;
Argerich et al. 1989).

Solid Matrix Primning

Another approach to controlled seed hydration is the use of solid carriers with low
metric potentials (Kubik et al. 1988; Taylor et al. 1988); a process called solid matrix
priming or matriconditioning. Some of the ideal (;haracteristics of the carrier are: (1) a low
matric potential, (2) negligible water solubility, (.3) high water holding capacity, (4) a high
surface area, (5) nontoxicity to the seed, and (6) the ability to adhere to the seed surface
(Khan 1992). Natural substances with these characteristics are vermiculite, peat moss, and
sand. Commercially available substances include Celite and Micro-Cel, which are
diatomaceous silica products, and Zonolite, which is a vermiculite (Khan 1992). Other
substances include a Leonardite shale and bituminous soft coal that also have an appreciable
osmotic effect (Taylor et al. 1988) and a calcined day (Kubik et al. 1988).
To accomplish solid matrix priming, seeds ar,e generally placed into the carrier at matric
potentials ranging from -0.4 to -1.5 MPa at 15°C for 7 to 14 days and brought to moisture
equilibrium. After that period, the solid material is sieved away from the seeds. The technique
has proven successful in enhancing seed performance of a number of crops (Table 11.1). The
technique has also been applied to large-seeded crops such as sweet com (Bennett and Waters
1987; Harman et al. 1989).
280 Seed Enhancements
Physiological Mechanisms Responsible for Priming

Seed priming reduces the imbibitional damage associated with planting in cold soils
(Bennett and Waters 1987). It also results in less secondary dormancy caused by planting
seeds such as lettuce in excessively warm soils (Valdes and Bradford 1987). Seeds which
have been primed also leak less metabolites (Styer and Cantliffe 1983). Morphological

Table 13.1. The Effect of Matriconditioning in Micro~Cel E on the Perfonnance of

Vegetable Seeds at 20°/1 O°C (Khan 1992).

Total
Seed cv. Treatmene emergence (%) Tso(d)2 Top fresh wt. 3
Red beet Matriconditioned 155a4 3.8c 1.29a (13)
Matrieonditioned +
Cardinal dried 156a 3.9c 1.25a (13)
-1.2 MPa PEG 8000 140b 5.5b 1.03b (13)
Untreated 131b 7.5a 0.81e (13)
Sugar beet Matriconditioned 88ab 2.3c 1.46a (13)
Matriconditioned +
E-4 dried 95a 3.4b 1.38a (13)
-1.2 MPa PEG 8000 88ab 3.6b 0.94c (13)
Untreated 82b 4.9a 0.72d (13)
Onion Matriconditioned 98a 3.9c 0.62a (15)
Texas early Matriconditioned +
grano dried 97a 4.0c 0.61a (15)
-1.2 MPa PEG 8000 92b 6.8b 0.48b (15)
Untreated 93b 7.9a 0.36c (15)
Tomato Matriconditioned 95a 4.3d 1.32a (15)
Matriconditioned +
FM jackpot dried 94a 5.3c 1.12b (15)
-1.2 MPa PEG 8000 88a 8.2b 0.80c (15)
Untreated 89a l1.8a 0.69d (15)
Pepper Matriconditioned 96a 7.4b 2.26a (21)
Rino Untreated 82b 14.1a 1.18b (21)
Carrot Matriconditioned 88a 5.ob 0.58a (18)
Matriconditioned +
Nantes dried 74b 8.5a 0.42b (18)
-1.2 MPa PEG 8000 78b 8.4a 0.36b (18)
Untreated 89a 9.3a 0.31b (18)
Celery Matriconditioned 92a 6.ge 0.19a (17)
Matrieonditioned +
FM 1218 dried 68c 9.4b o.lob (17)
-1.2 MPa PEG 8000 86a 9.3b 0.06be (17)
Untreated 78b 13.8a O.03c (17)
IMatriconditioning was conducted at 15°C in light in a mixture of seed:carrier:water as shown in Table
4.4. After matriconditioning seeds were planted with or without air-drying in Cornell Peat-Lite mix.
Seeds conditioned in PEG 8000 were washed and wipe-dried before planting. Emergence recorded at 12-
hour day, 20 o /10°C, night-temperature regime.
'Time to 50% emergence.
'Fresh weight of 15 tops. Data in parentheses are days after planting.
'Means in columns for each seed type separated by DMRT at 5% level.
Seed Enhancements 281
changes have been observed in seeds following priming. For example, a portion of the lettuce
seed endosperm is hydrolyzed during priming that permits faster embryo growth (Dahal et al.
1990). Increases in cell wall elasticity have also been noted (Karssen et al. 1989). It is also
believed that hydration of the seed to Phase II during priming permits early DNA replication
(Bray et al. 1989), increased RNA and protein synthesis (Fu et al. 1988; Ibrahim et al. 1983),
and more ATP availability (Mazor et al. 1984). [n addition, studies have suggested that an
increase in seed vigor may also occur following priming. These have shown that repair of
deteriorated seed parts occurs during the hydration phase of the process (Karssen et al. 1989;
Saha et al. 1990). Importantly, these changes are irreversible and are tolerant of the
subsequent desiccation that follows priming.

Factors Affecting Seed Priming

The factors which affect seed priming include (1) ambient conditions during hydration
(temperature, light), (2) type(s) of osmotica, (3) oxygen availability, (4) duration of the
treatment, (5) control of microbial contamination, and (6) drying.
Generally, lower temperatures require a decreased PEG concentration to obtain the
desired water potential as at a higher temperature, which leads to increased oxygen
concentration. Thus, priming is usually more successful when conducted at a lower
temperature. Light may be necessary in some priming treatments if the species requires light
for germination. The type of osmoticum employed also affects the degree of priming success.
For example, the use of a solid matrix such as vermiculite or a finely ground shale allows
more air to get to the seed than when an osmoticum is used. This is because the solid matrix
carrier is not completely saturated to achieve the desired water potentials and the seed is
therefore in an aerobic environment. Compounds used in priming also may change water
potentials since the seeds are actively absorbing the water from the solution which lowers the
osmotic water potential even further. This problem is of greater concern when
osmoconditioning larger seeds or where there is a high seed-to-solution ratio. Evaporation of
the water from the osmoticum should also be considered, particularly when germination
temperatures are high. Controlling microbial contamination during priming is also an
important quality control step. In the case of solid matrix priming, seeds are often surface
sterilized or a slurry fungicide treatment added. When priming, fungicides can be added
directly to the osmoticum to prevent microbial growth. Finally, the method of seed drying can
influence the degree of priming. Drying can be performed by ambient air, forced air, or
vacuum drying. Ambient air drying occurs by placing the primed seed on a flat uniform
surface and allowing it to dry slowly. This provides a uniform drying rate which, in some
instances, may be considered too slow. In addition, this approach requires considerable space.
Forced air drying is another option which reduces the requirement for space and is more
rapid. However, careful attention must be given to ensure that mechanical seed damage does
not occur as the seed dries (Armstrong and McDonald 1992). Vacuum drying provides the
most uniform drying environment for primed seed but its cost makes this option less
attractive.
282 Seed Enhancements
BIOLOGICAL SEED TREATMENTS

Biological seed treatments are those that use fungi or bacteria to control soil and seed
pathogens instead of a synthetic chemical seed treatment. These are gaining increasing
popularity because of safety concerns for humans and the environment as well as
phytotoxicity problems associated with excess use of pesticides. In addition, biological seed
treatments offer the potential for protecting the plant throughout its entire life cycle rather
than just during the seed/seedling stage.
In contrast to most synthetic pesticides which control a wide range of microorganisms,
biological seed treatments typically have a narrow range of specificity and often target just
one specific pest. As a result, they disrupt ecosystems to a smaller degree than broad-
spectrum pesticides (Cook and Baker 1983). This trend, however, has been reversed in recent
years. Biologicals such as Trichoderma can control a number of pathogens such as Pythium,
Rhizoctonia, and Fusarium species. Some biological seed treatments are able to colonize
plant roots and contribute to long-term pest control effectiveness (Harman et al. 1989) as
long as the soil pathogen does not affect the population of the biologica I seed treatment on the
roots (Mazzola and Cook 1991). Biological seed treatments have been reported to induce
growth increases that are persistent and ultimately lead to increased yield enhancements as a
consequence of long-term disease control (Ahmad and Baker 1987; Schippers et a1. 1987).
Biological seed treatments markedly enhance plant performance. Fungi such as
Trichoderma haraianum (Smith and Wehner 1987), Pythium oligandrum (Martin and
Hancock 1987), and Chaetomium globosum (Walther and Gingrat 1988) control damping off
and other diseases. Bacteria such as Rhizobia species have long been applied to seed coatings
to enhance root nodulation and nitrogen fixation. These beneficial bacteria (Jawson et al.
1989), and others such as Bacillus subtilis (Tschen 1987) and pseudomonads (Digat 1989)
are being examined for their incorporation into seeds using seed enhancement technologies.
Not all biological seed treatments, however, have proven successful under a wide range of
soil conditions. Certain Trichoderma species are ineffective in lower-iron soils, presumably
due to the more effective competition for iron from pseudomonads (Hubbard et al. 1983). In
other cases, the biological seed treatments are not persistent in the soil, seed, or the plant
under natural conditions. For example, a genetically engineered biocontrol, Pseudomonas
fluorescens was added to com seeds that were planted in nutrient solution and in the field.
The biological control was found 43 days after planting in the seed remnant and plant roots of
nutrient-grown plants, but decreased over time for those planted in soil (Fisher et al. 1993).
The application of biocontrol agents to seeds is influenced by the efficacy of the
biocontrol, the number of propagules added to the seed, the application of the biocontrol, and
control of other microbes in the application process. Biological seed treatments that are
effective in protecting a germinating seed do not require as high a population on the seed as
one which is less effective. The number of propagules per seed must be determined to ensure
adequate control of the pest and this number often must be adjusted since repeated
subculturing of biocontrol agents results in diminishing effectiveness (Callan et a1. 1990).
Other amendments that enhance biological seed treatments include food bases for the
microorganisms, pH of the seed coating, and the rapidity with which the microorganism
establishes itself.
Seed Enhancements 283
The handling and application of biological sec~d treatments, in contrast to traditional seed
treating equipment, may require sterile conditions to prevent contamination of pathogenic
organisms with the beneficial microbes. Equipment which has been used to apply several
biological control agents should be sterilized to avoid contaminating the desired microbe with
less efficacious ones. If conventional seed treatment equipment is used in the application of
biocontrol microbes, it should be thoroughly cleaned since remaining fungicide residues may
be harmful to the biocontrol agent. Conventional seed treatment equipment may also create
physical damage to the biological organism during application to the seed that make it less
active. The application of biological seed treatffil~nts to seeds to achieve a uniform coating
may present a problem of seed agglutination (clumping) during drying. Capper and Higgins
(1993) developed microgranule formulations of Pseudomonasfluorescens that were applied
in furrow at planting instead of direct application to the seed. This same concept was applied
to a fungal biological seed treatment, Trichoderma haraianum, where granular formulations
were made from hyphal segments, chlamydospores, and conidia mixed with sodium alginate
and polyethylene glycol (Dandurand and Knudsen 1993). Finally, it is important to surface
sterilize seeds with sodium hypochlorite and a few drops of an adjuvant to ensure complete
seed coat coverage prior to application of the biological seed treatment. This rids the seed
coat of any other pathogens that may compete with the biological control agent.

SEED COATINGS

One of the most useful areas of seed enhancements is seed coatings. Seed placement and
performance can be greatly enhanced by altering the shape of the seed or placing chemicals
on the seed coat which regulate and improve germination. Two types of seed coatings are in
commercial use: seed pelleting and seed coating (Figure 13.1).

Film Coating
Seed Pellet - Thin film t6 delay germination;
• Shape of seed is obscured to a speclfic
Size; free flow!ng and is lIsed in precision - Multiple layers of micronutrients,
planting, fungicides, and insecticides

f.IIiH~- Seed

Seed Coat
~ Does not obscure shape of seed;
contains micronutrients, fungicides.
and insecticides.

Figure 13.1. Diagrammatic illustration of various types of seed coatings.

284 Seed Enhancements
Seed Pelleting

A seed pellet is applied to a seed to improve its plantability and performance. Many
seeds, particularly vegetable seeds, are not uniformly round, which hinders precision planting
for optimum crop yields. In other cases, seeds are so small and light that their accurate
placement inion the soil is uncertain (Smith and Miller 1987). To facilitate the free flow of
these seeds in planters, many seed companies provide seeds with coatings of materials that
change the shape and size of the seed so that it becomes heavier and rounder. A seed pellet is
characterized by its abil ity to totally obscure the shape of the encased seed.
Seeds are introduced into a coating drum or pan that resembles a cement mixer. An
amalgam of fillers (clays, limestone, calcium carbonate, talc, vermiculite) and cementing
additives (gum arabic, gelatin, methyIcellulose, polyvinyl alcohol, polyoxylethylene glycol-
based waxes) are used to form the pellet and other compounds such as innoculants,
fungicides, etc. may be added to enhance seed performance (Taylor and Harman 1990). A
unique application of pellets is with the addition of calcium oxide and peroxides to the
pelleting materials. These are believed to release oxygen to the seed under flooded field
conditions thus minimizing anaerobic damage (Ollerenshaw 1985; Langan et al. 1986). An
example of pelleting formulations for clover seeds is provided in Table 13.2. As the drum
rotates, the seeds are first sprayed with water followed by the addition of the filler with
binder. The wet seed attracts and becomes coated with the dry filler and the pel1et gradual1y
increases in size with each turn of the coating drum. Longer rotation times with greater
amounts of filler lead to greater pellet size and roundness. At the end of the pelleting process,
a binder is added to harden the outer layer of the pellet. This also reduces the amount of dust
produced during handling, shipping, and planting. After pelleting is complete, the pelleted
seeds are dried and handled in the same way as unpelleted seeds.

Table 13.2. Composition of Pellets Containing Clover Seed Used in This Investigation.

Formulation
Component 1 2 3 4 5 6 7 8 9
Kaolin clay 59.0 57.5 57.5 57.5 75.0 51.0 58.5 52.0 57.5
Seed 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0
Inoculum 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0
PYA 205S 1.0 0.5 1.0
PYA 5238 1.0 1.0
PYA 5408 1.0
Sodium silicate 0.8
Calcium sulfate 8.0
DAp 1 15.0 SOpz
Water 38.0 38.5 38.0 38.0 21.2 38.0 38.0 39.0 38.0
'DAP: diammonium phosphate.
'SOP: sprayed dried pellet with saturated aqueous solution of OAP (100 ml solution/200 g pellets).
From Smith and Miller (1987).
Seed Enhancements 285
Certain technical problems must be monitored during the pelleting process. The pelleting
material must be compatible with the seed so that seed quality is maintained and germination
is not hindered. For example, pelleting materials are "wet" during pelleting so that inadvertent
seed hydration occurs which leads to increased respiration and possibly reduced seed quality.
In other instances, if the pelleting material is too hard after drying, it may be difficult for
radicle emergence through the pelleted material. While pellet strength is an important asset
which ensures that the pellet remains intact during routine handling, if the pellet is too strong,
germinability will be reduced. Successful pelleting also requires that a pellet form around an
individual seed. If this does not occur, precision planting success is compromised. While this
objective is often achieved, this is not always the case. For instance, some pellets may form
around multiple seeds. In other cases, particularly when the seed lot is not thoroughly
cleaned, pellets may form around inert matter such as sand or undesired plant material. The
size of the pellet can also be an issue to seed purchasers. Therefore, pellets are routinely sized
during and after the pelleting process. Seed analysts also have a greater responsibility when
conducting a purity test with pelleted seeds. They must ensure that each pellet contains a seed
and, furthermore, they must also identify the puri~y of the pelleted seed lot which will contain
a high percentage of inert matter contributed by the filler present in the pellet. Table 13.3
illustrates some of the variability in seed perfonnance and pellet attributes following seed
pclleting using differing formulations.
Despite these technical problems, pelleting of seed is recognized as an important
addition to the precision planting in many vegetables such as onion, lettuce, carrot, and
various flowers. The addition of other beneficial chemicals such as plant hormones,
micronutrients, microbes, and fungicides to the pellet further improves seed performance in
the field.

Table 13.3. Percentage Germination ofLadino Clover Seed Pellets at 21 Days After Planting,
Pellet Strength, and Pellet Destructability. Pellets Were Planted Immediately After Drying
(Planting I) and After Six Months Storage at Room Temperature (Planting 2).

Seedlings
Germination pellet- 1
Planting Planting Pellet strength
Longitu- Diamet- Pellet de-
Formulation 1 2 1 2 dinal rieal struetabilit y l
--(%)-- -(no.)- (g) -(%)--
1 85 86 1.6 2.3 270 293 3.3
2 82 89 2.0 2.5 3573 2363 0.8
3 89 91 1.9 2.6 6882 7061 0.4
4 82 80 1.9 2.1 6772 6200 0.4
5 74 2 62 2.0 1.8 795 776 1.2
6 58 52 1.7 1.5 362 442 1.2
7 82 89 1.8 2.2 2520 2974 1.0
8 1 1 0.1 0.1 1846 1515 1.8
9 48 16 1.4 0.7 4879 4988 0.8
LSD 10 10 0.3 0.5 751 825 0.8
'Pellet destructability was determined by weight lost when tumbled for 1 hour.
'Means for plantings 1 and 2 are significantly different (P = .05) for each formulation as determined by
student's T test.
From Smith and Miller (1987).
286 Seed Enhancements
Seed Coating

Seed coating is one of the most economical approaches to improving seed performance.
A seed coating is a substance that is applied to the seed but does not obscure its shape
(Figure 13.1). Often, the purpose of the coating is to apply substances such as fungicides,
insecticides, safeners, micronutrients, and other compounds directly to the seed. This enables
the seed company to precisely tailor the seed to avoid specific stresses anticipated in certain
planting environments. The ideal traits of a seed coating polymer are that it should (Rushing
1988) (1) be a water-based polymer, (2) have a low viscosity range, (3) have a high
concentration of solids, (4) have an adjustable hydrophiliclhydrophobic balance, and (5) form
a hard film upon drying. These traits should lead to excellent plantability, contain no "dust
off" of additives, and provide for excellent germination under all environmental conditions.
The application of seed coatings is very similar to slurry seed treatments and similar
equipment is used. One of the major benefits of such seed enhancements is that they are
placed directly on the seed and in the immediate vicinity of the germinating seedling. This
means that less chemical is required compared to broadcast or furrow applications with far
less cost, while avoiding environmental damage from excess pesticide use.
Another approach to seed coatings is film coating of seeds. This is the application of
additives dissolved in a dyed solution of a "sticky" polymer. The addition of the coating is
very different from pelleting since it only represents an increase of 1-10% of the seed weight
and the shape of the seed is still retained (Figure 13.1). The seeds are dipped or sprayed with
the dissolved polymer and then immediately dried. An interesting advantage of this approach
is that the minimal increase in seed weight allows the formulation to be changed several times
during the spraying and drying process so that the seeds can contain a multilayer film coat.
Other advantages of film coatings are the absence of "dusting off" problems and improvement
of seed flow in planting equipment compared to other coating processes.
Many of the seed coatings described to this point have emphasized the use of chemical
or biological controls to improve seed performance. A new area of seed coating research has
emphasized the application of water-impermeable plastic film coatings to delay germination
until a specified time. This approach permits planting of the seed in the fall and degradation
of the seed coating during the winter, with resultant germination in the spring. This avoids
problems often encountered in wet fields in the spring which consequently delay planting and
reduce performance. Another application of water-impermeable plastic seed coatings is the
delay in germination of incompatible male or female lines in hybrid seed production so that
synchronous flowering or nicking is achieved. While the potential for such seed coatings is
exciting, the mechanisms which control seed coat degradation are still difficult to precisely
predict. Further research in these areas is needed.

Integration of Seed Priming and Other Seed Enhancements

Seed priming treatments are often integrated with other seed treatments to enhance
performance. For example, pregerminating primed seeds further reduces emergence time and
increases shoot weight (Pill 1986). Seed coating and pelleting technologies have been
developed to improve plantability of flat seeds and also to permit the addition of bioactive
chemicals, nutrients, and beneficial microbes (Raimer 1988). Pelleting of primed seeds has
been shown to produce greater performance than either the pelleting or priming treatment
Seed Enhancements 287
alone (Valdes et al. 1985; Bennett 1988). Another integrative approach is to include growth
regulators and/or pesticides in the osmoticum or solid matrix carrier soak water. The addition
of GA3 to the PEG solution has reduced the time to germination, improved the rate of
germination, and prevented the induction of secondary dormancy in lettuce (Khan 1992).
Fungicides (Giammichele and Pill 1984; Osburn and Schroth 1989) and plant nutrients
(Finch-Savage and Cox 1982) also have been added to seeds during priming.

SYNTHETIC SEEDS

In some cases, the sexual reproduction of seeds is undesirable because it assures that
seeds are not alike genetically following meiotic recombination. In fact, many crops yield best
when their seed is produced by hybridization of male and female lines. This hybrid production
is expensive, particularly when conducted manually, and yields of hybrid seed are often low
and hybridization success not always assured. In addition, many crops are vegetatively
propagated when planting of seed might be preferred. For these reasons, the clonal or
vegetative production of synthetic seeds has been explored and is a rapidly expanding area of
activity. A review of progress on this subject has been presented by Redenbaugh (1993).
Synthetic seeds may have value for a number of traditional crops (Table 13.4). For
example, synthetic cultivars of alfalfa and orchardgrass are commonly used in which seeds
are nonuniform and each plant is a distinct genotype. Synthetic seeds would allow the
incorporation of specific new genes into single outstanding hybrids that could then be
produced asexually by somatic embryos. Synthetic seed production might also benefit the
planting efficiency of crops that are vegetatively propagated such as fruit and nut trees
because of self-incompatibility and long breeding c:ycles. Similarly, forest conifers such as
pine and spruce trees, which are currently planted only by seed, would benefit from synthetic
seed production because the long conifer life cyclt: before the trees are capable of bearing
seeds would be avoided. In certain self-pollinated crops such as cotton and soybean, the
production of hybrid seeds has been hindered because of the closed flowers at the time of
pollination and the requirement for hand pollination. Somatic embryogenesis would
circumvent this labor-intensive process. Other crops, such as tomato and seedless
watermelon, demand a high value for hybrid seeds, which makes the production of synthetic
seeds for these particular markets more competitive (Redenbaugh et a1. 1991).

Principle

Synthetic seeds rely on the production of somatic embryos instead of zygotic embryos
formed from sexual fertilization of male and female parents (Figure 13.2). Somatic embryos
are produced asexually from vegetative tissue that has been given specific nutrient and
environmental regimes causing the undifferentiated vegetative cells to form into the parts of
an embryo. As a result, somatic embryos are clones that possess identical genotypes. Both
288 Seed Enhancements
Table 13.4. Potential Applications of Synthetic Seed Technology (SST)
for Selected Crop Species.

Relative
Somatic Relative Need
Embryo Seed for
Crop Quality 8 Cost b Applicationc SST d
Alfalfa h 1 s m
Corn p m i m
Cotton p m h m
Grape h na s,g m
Loblolly pine p h c h
Norway spruce h c h
Orchardgrass h s m
Soybean p m h m
Hybrid Tomato n v d h
Seedless Watermelon n v d h
"Relative somatic embryo quality: h-highly developed embryos; p-poorly developed embryos; n-so-
matic embryos not obtained.
bRelative cost of seed: v-seed cost limits planting; h-seed is costly; m-moderate; I-relatively inexpen-
sive; na-seed is not used.
'Application for synthetic seed: c-circumvent long breeding cycles; d-decrease hybrid seed cost; g-
germplasm conservation; h-mass production of hybrids; i-eliminate need for inbreds; s-circumvent
self-incompatibility.
dRelative need: h-highly useful if implemented; m-existing methods are effective but implementation
should yield improvements.

somatic and zygotic embryos show typical embryonic development characteristic of monocots
(Gray and Conger 1985) and dicots (Becwar et at. 1989). However, they differ because
multiple somatic embryos are produced from a single callus while a single zygotic embryo is
produced from each fertilization.

Challenges

Multiple embryos are often found on a single callus, in which multiple stages of embryo
development are also observed. This causes the nonuniform embryos to be subjected to
changing nutrient conditions since the nutrients are depleted by the developing tissues and
then replenished. Consequently, many somatic embryos have organs developing at differing
rates which contribute to asynchronous embryo development (Conger et al. 1989). In some
cases, this leads to precocious germination, while in others the prevailing nutrient
environment may be conducive to shoot or root development, but not both. In still other cases,
somatic embryos often develop extra cotyledons or poorly developed apical meristems
(Ammirato 1987). This asynchronous embryo development makes harvest of uniformly
mature somatic embryos difficult in synthetic seed production. Attempts to better synchronize
somatic embryo development have included physical separation of proembryonal cultures to
assure uniform caJIus size and physiological synchronization by adding abscisic acid to the
culture medium. Abscisic acid appears to cause cell water (turgor) content to decrease,
thereby slowing embryo growth which inhibits germination of embryos that would tend to
germinate precociously (Gray 1989).
Seed Enhancements 289

Get Suspension

Fluid
Drilling

4
Bioreactor I

Bioreactor II ~~~1J Transplanting

Figure 13.2. Schemes for vegetative propagation via somatic embryogenesis were published as one
of the frontpieces for Volume 2 of the Handbook of Plant Cell Culture, and includes the growth of
embryos for transplants, fluid drilling, encapsulation" and seed tape technology. It is not clear if
these projected systems were researched (From Sharp, W R., Evans, A., Ammirato, P. v., and
Yamada, Y., Handbook of Plant Cell Culture. Vol. 2, Crop Species, Macmillian, New York, 1984.
With permission).
290 Seed Enhancements

Figure 13.3. Synthetic celery seeds coated in a calcium alginate gel. Note small somatic embryos
inside gel coating. Photograph courtesy of K. Redenbaugh.

Somatic embryos also do not typically exhibit a quiet resting or dormancy (quiescence)
phase characteristic of orthodox seeds (Gray 1987). Usually, somatic embryos continue to
grow into seedlings or they revert back into disorganized callus tissue. This inability to
produce a resting phase where all embryos are at the same arrested physiological and
morphological state also is a challenge to synthetic seed development. Without this arrested
growth stage, synthetic seeds cannot be successfully stored or treated using traditional seed
technology practices. Attempts to introduce dormancy into somatic embryos are being
explored using orthodox seeds as a developmental model. Since orthodox seeds express
dormancy after the seeds begin to dehydrate on the parent plant, studies have been initiated to
determine whether dry down of somatic embryos might induce this important trait and there is
evidence of success (McKersie et al. 1988; Carman 1988; Seneratna et al. 1989).

Encapsulations

Assuming that satisfactory techniques are developed for inducing dormancy into somatic
embryos, the dry, delicate embryos would benefit from a seed coating to protect against
injury incurred from traditional seed handling and planting. The protective encapsulations can
be designed to provide physical protection as well as essential nutrients, antibiotics, and
fungicides to assist the embryo during germination. Two approaches, hydrated and dry
encapsulation, have been studied. The hydrated encapsulation is most effective for nonquies-
Seed Enhancements 291
cent somatic embryos that can be planted directly without storage (Figure 13.3). Types of
hydrated encapsulations are composed primarily of calcium alginate (Redenbaugh et al.
1987) although the success of seedling establishment in the field using this approach has been
limited (Fujii et al. 1987). The preferred approach is the dry, hardened synthetic seed coating
surrounding somatic embryos which would permit conventional seed storage and handling.
To date, only a water-soluble plastic resin surrounding carrot embryos has been examined
and this did not result in satisfactory seedling establishment (Kitto and Janick 1985).

SUMMARY

The concept of direct genetic manipulation of the embryo through somatic em-
bryogenesis offers immense potential for the production of superior performing seeds of a
number of crops. However, numerous technical hurdles associated with dormancy imposition
and satisfactory encapsulation techniques to permit normal seed handling need to be better
developed. Overriding these technical concerns an~ the additional costs incurred for synthetic
seed production. It has been estimated that the cost of one synthetic alfalfa seed would be 3.3
cents (Redenbaugh et al. 1987), which is far in excess of conventional seed costs. Until the
cost of synthetic seeds becomes competitive with seeds that are naturally produced, this
promising technology will require further research to refine and automate synthetic seed
production systems.

Questions

1. Identify the central objective of seed enhancement technology.

2. What are the practical consequences of seed hydration? What seed moisture level is
necessary to obtain the benefits of seed hydration?
3. Name three general approaches to see:d hydration and describe how each is
accomp lished commercially.
4. Describe fluid drilling.
5. What is polyethylene glycol and how is it used as a seed enhancement technology?
What beneficial effects does it have on seed performance?
6. Outline six ideal characteristics of a solid matrix carrier and describe why each is
important.
7. Name six factors that affect seed priming success and explain why they are important.
8. What are biological seed treatments, why is the seed industry interested in continuing
their development, and how are they effective?
9. Distinguish between seed coatings, seed pelle1tings, and seed film coatings.
10. What are some of the commercial advantag(~s of synthetic seed production? How are
synthetic seeds produced? Is this seed enhancement technology practical from a seed
industry perspective?
292 Seed Enhancements
General References

Ahmad, 1. S., and R. Baker. 1987. Competitive saprophytic ability and cellulolytic activity of
rhizosphere-competent mutants of Trichoderma haraianum. Canadian Journal of
Microbiology 34:229-234.
Akers, S. W. 1990. Seed response to priming in aerated solutions. Search 19:8-17.
Akers, S. W., and K. E. Holley. 1986. SPS: A system for priming seeds using aerated
polyethylene glycol or salt solutions. Horricultural Science 21 :529-531.
Alvarado, A. D., and K. 1. Bradford. 1988. Priming and storage of tomato (Lycopersicon
lycopersicum) seeds. I. Effects of storage temperature on germination rate and viability. Seed
Science and Technology 16:601-612.
Alvarado, A. D., K. 1. Bradford, and J. D. Hewitt. 1987. Osmotic priming of tomato seeds:
Effects on germination, field emergence, seedling growth and fruit yield. Journal ofAmerican
Society Horticultural Science 112:427-432.
Arnmirato, P. V. 1987. Organizational events during somatic embryogenesis. In: Plant Tissue
and Cell Culture. New York: Alan Liss.
Argerich, C. A., K. J. Bradford, and A. M. Tarquis. 1989. The effects of priming and aging
on resistance to deterioration of tomato seeds. Journal of Experimental Botany 40:593-598.
Armstrong, H., and M. B. McDonald. 1992. Effects of osmoconditioning on water uptake
and electrical conductivity in soybean seeds. Seed Science and Technology 20:391-400.
Becwar, M. R., T. L. Noland, and 1. L. Wycoff. 1989. Maturation, germination, and
conversion of Norway spruce (Picea abies L.) somatic embryos to plants. In Vitro Cellular
and Developmental Biology 25:575-580.
Bennett, M. A. 1988. Evaluation of seed coating and priming treatments for stand
establishment of processing tomatoes. Proceedings of International Conference on Stand
Establishment Horticultural Crops. American Society of Horticultural Sciences, Lancaster,
PA.
Bennett, M. A., and L. Waters. 1987. Seed hydration treatments for improved sweet corn
germination and stand establishment. Journal ofAmerican Society of Horticultural Sciences
112:45-49.
Bodsworth, S., and J. D. Bewley. 1981. Osmotic priming of seeds of crop species with
polyethylene glycol as a means of enhancing early and synchronous germination at cool
temperatures. Canadian Journal of Botany 59:672-676.
Bradford, K. J. 1986. Manipulation of seed water relations via osmotic priming to improve
germination under stress conditions. Horticultural Science 21: 11 05-1112.
Bradford, K. 1., 1. J. Steiner, and S. E. Trawatha. 1990. Seed priming influence on
germination and emergence of pepper seed lots. Crop Science 30:718-721.
Bray, C. M., P. A. Davison, M. Ashraf, and R. M. Taylor. 1989. Biochemical events during
osmopriming of leek seeds. Annals ofApplied Biology 102:185-193.
Brocklehurst, P. A., and J. Dearman. 1983. Interactions between seed priming treatments and
nine seed lots of carrot, celery, and onion. II. Seedling emergence and plant growth. Annals of
Applied Biology 102:585-593.
Brocklehurst, P. A., J. Dearman, and R. K. L. Drew. 1984. Effects of osmotic priming on
seed germination and seedling growth in leek. Scientia Horticulturae 24:201-210.
Seed Enhancements 293
Callan, N. W., D. E. Mathre, and J. B. Miller. 1990. Biopriming seed treatment for
biological control of Pythium ultimum premergence damping-off in sh2 sweet corn. Plant
Disease Reporter 74:368-372.
Cantliffe, D. J., J. M. Fischer, and T. A. Nell. 1984. Mechanism of seed-priming in
circumventing thermodormancy in lettuce. Plant Physiology 75:290-294.
Capper, A. L., and K. P. Higgins. 1993. Application of Pseudomonas jluorescens isolates to
wheat as potential biological control agents against take-all. Plant Pathology 42:560-567.
Carman, J. G. 1988. Improved somatic embryogenesis in wheat by partial simulation of the
in ovulo oxygen, growth regulator and desiccation c~nvironments. Planta 175:417-422.
Conger, B. Y., J. C. Hovanesian, R. N. Trigiano., and D. J. Gray. 1989. Somatic embryo
ontogeny in suspension cultures of orchardgrass. Crop Science 29:448-452.
Cook, R. J., and K. F. Baker. 1983. The Nature and Practice of Biological Control of Plant
Pathogens. St. Paul, Minn.: American Phytopathological Society.
Dahal, P., K. J. Bradford, and R. A. Jones. 1990. Effects of priming and endosperm integrity
on seed germination rates of tomato genotypes. II. Germination at reduced water potential.
Journal of Experimental Botany 41 : 1441-1453.
Dandurand, L. M., and G. R. Knudsen. 1993. Influence of Pseudomonas jluorescens on
hyphal growth and biocontrol activity of Trichoderma haraianum in the spermosphere and
rhizosphere of pea. Phytopathology 83:265-270.
Danneberger, T. K., M. B. McDonald, C. A. Geron, and P. Kumari. 1992. Rate of
germination and seedling growth of perennial ryegrass seed following osmoconditioning.
Horticultural Science 27:28-30.
Digat, B. 1989. Strategies for seed bacterization. Acta Horticulturae 253:121-130.
Finch-Savage, W. E. 1984. A comparison of seedling emergence from dry-sown and fluid-
drilled carrot seeds. Journal of Horticultural Science 59:403-410.
Finch-Savage, W. E., and C. J. Cox. 1982. Effect of adding plant nutrients to the gel used for
fluid drilling early carrots. Journal ofAgricultural Science 99:295-303.
Fisher, P. J., S. A. Broad, C. D. Clegg, and H. M. Lappin Scott. 1993. Retention and spread
of a genetically engineered pseudomonad in seeds and plants of Zea mays L.-A preliminary
study. New Phytology 124:101-106.
Fu, J. R., S. H. Lu, R. Z. Chen, B. Z. Zhang, Z. S. Liu, Z. S. Li, and D. Y. Cai. 1988.
Osmoconditioning of peanut (Arachis hypogaea L.) seeds with PEG to improve vigour and
some biochemical activities. Seed Science and Technology 16: 197-212.
Fujii, J. A., D. T. Slade, K. Redenbaugh, and K. A. Walker. t 987. Artificial seeds for plant
propagation. Trends in Biotechnology 5:335-340.
Gelmond, H. 1965. Pretreatment of leek seeds as a means of overcoming superoptimal
temperatures of germination. Proceedings of International Seed Testing Association 30:737-
742.
Gerber, J. M., and L. A. Caplan. 1989. Priming sh2 sweet corn seed for improved emergence.
Horticultural Science 24:854.
Gray, D. 1981. Fluid drilling of vegetable seeds. Horticulture Review 3:1-27.
Giammichele, L. A., and W. G. Pill. 1984. Protection of fluid-drilled tomato seedlings against
damping-off by fungicide incorporation in a gel carrier. Horticultural Science 19:877-879.
Gray, D. J. 1987. Effects of dehydration and other environmental factors on dormancy in
grape somatic embryos. Horticultural Science 23 :786-791.
294 Seed Enhancements
Gray, D. J. 1989. Synthetic seed for clonal production of crop plants. In: Recent Advances in
the Development and Germination of Seeds, ed. R. B. Taylorson, pp. 29-45. New York:
Plenum Press.
Gray, D. J., and B. V. Conger. 1985. Somatic embryo ontogeny in tissue cultures of
orchardgrass. In: Tissue Culture in Forestry and Agriculture, ed. R. R. Henke, K. W.
Hughes, M. J. Constantin, and A Hollaender. New York: Plenum.
Guedes, A C., and D. J. Cantliffe.1980. Germination of lettuce (Lactuca sativa) at high
temperature after seed priming. Journal of American Society of Horticultural Science
105:777-781.
Haigh, A M., and E. W. R. Barlow. 1987. Germination and priming of tomato, carrot,
onion, and sorghum seeds in a range of osmotica. Journal of American Society of
Horticultural Science 112:202-208.
Halmer, P. 1988. Technical and Commercial Aspects of Seed Pelleting and Film Coating,
pp. 191-204. Thornton Heath, U.K.: British Crop Protection Council.
Harman, G. E., A G. Taylor, and T. E. Stasz. 1989. Combining effective strains of
Trichoderma hareianum and solid matrix priming to improve biological seed treatments.
Plant Disease Reporter 73 :631-63 7.
Helsel, D. G., D. R. Helsel. and H. C. Minor. 1986. Field studies on osmoconditioning
soybeans, Glycine max. Field Crop Research 14:291-298.
Heydecker, W., and P. Coolbear. 1977. Seed treatments for improved performance-survey,
an attempted prognosis. Seed Science and Technology 5:353-425.
Hubbard, J. P., G. E. Harman, and Y. Hadar. 1983. Effect of soilborne Pseudomonas spp.
on the biological control agent, Trichoderma hamatum on pea seeds. Phytopathology
73:655-659.
Ibrahim, A E., E. H. Roberts, and A H. Murdoch.1983. Viability of lettuce seeds. n.
Survival and oxygen uptake in osmotically controlled storage. Journal of Experimental
Botany 34:631-640.
Jawson, M. D., A 1. Franzluebbers, and R. K. Berg. 1989. Bradyrhizobium japonicum
survival in and soybean inoculation with fluid gels. Applied Environmental Microbiology
55:617-622.
Karssen, C. M., A. Haigh, P. van der Toorn, and R. Weges. 1989. Physiological mechanisms
involved in seed priming. In: Recent Advances in the Development and Germination of
Seeds, pp. 269-280. New York: Plenum Press.
Karssen, C. M., and R. Weges. 1987. Osmoconditioning of lettuce seeds and induction of
secondary dormancy. Acta Honticulturae 215: 165-171.
Khan, A. A. 1992. Preplant physiological seed conditioning. Horticulture Review 13: 131-
181.
Khan, A A., N. H. Peck, and C. Sammimy. 198011981. Seed osmoconditioning:
Physiological and biochemical changes. Israel Journal of Botany 29: 133-144.
Khan, A A, K. L. Tao, J. S. Knypl, B. Borkowska, and L. E. Powell. 1978. Osmotic
conditioning of seeds: Physiological and biochemical changes. Acta Horticulturae 83:267-
278.
Seed Enhancements 295
Kitto, S. L., and J. Janick. 1985. Hardening treatments increase survival of synthetically
coated asexual embryos of carrot. Journal of American Society of Horticultural Science
110:282-287.
Kubik, K. K., J. A. Eastin, J. D. Eastin, and K. M. Eskridge. 1988. Solid matrix priming of
tomato and pepper. Proceedings of International Conference Stand Establishment of
Horticultural Crops. American Society of Horticultural Sciences, Lancaster, PA, pp. 86-96.
Langan, T. D., 1. W. Pendleton, and E. S. Oplinger. 1986. Peroxide coated seed emergence in
water-saturated soil. Agronomy Journal 78:769-772.
Martin, F. N., and 1. G. Hancock. 1987. The use of Pythium oligandrum for biological
control of preemergence damping-off caused by Pythium ultimum. Phytopathology 77: 1013-
1020.
Mazor, L., M. Perl, and M. Negbi. 1984. Changes in some ATP-dependent activities in seeds
during treatment with polyethylene glycol and during the redrying process. Journal ofExperi-
mental Botany 35:1119-1127.
Mazzola, M., and R. J. Cook. 1991. Effects of fungal root pathogens on the population
dynamics of biocontrol strains of fluorescent pseudomonads in the wheat rhizosphere.
Applied Environmental Microbiology 57:2171-2178.
McDonald, M. B. 2000. Seed priming. In Seed Technology and its Biological Basis (eds. M.
Black and 1. D. Bewley). Pp. 287-325. CRC Press, Boca Raton, FL.
McKersie, B. D., S. R. Bowley, T. Senaratna, D. C. W. Brown, and 1. D. Bewley. 1988.
Application of artificial seed technology in the production of alfalfa (Medicago sativa L.). In
Vitro Cellular and Developmental Biology 24:71-76.
Mexal, J., J. T. Fisher, J. Osteryoung, and C. P. Reid. 1975. Oxygen availability in
polyethylene glycol solutions and its implications in plant-water relations. Plant Physiology
55:20-24.
Michel, B. E., and M. R. Kaufmann. 1973. The osmotic potential of polyethylene glycol
6000. Plant Physiology 51:914-916.
Ollerenshaw,1. H. 1985. Influence of waterlogging on the emergence and growth of Lolium
perenne L. shoots from seed coated with calcium p,eroxide. Plant and Soil 85: 131-141.
Osburn, R. M., and M. N. Schroth. 1989. Effi~ct of osmopriming sugar beet seed on
germination rate and incidence of Pythium ultimum damping-off. Plant Disease Reporter
73:21-24.
Pill, W. G. 1986. Parsley emergence and seedling growth from raw, osmoconditioned and
pregerminated seeds. Horticultural Science 21: 1134-1136.
Redenbaugh, K., ed. 1993. Synseeds: Synthetic Seeds to Crop Improvement. Boca Raton,
Fla.: CRC Press.
Redenbaugh, K., J. A. Fujii, and D. Slade. 1991. Synthetic seed technology. In: Cell Culture
and Somatic Cell Genetics of Plants. Vol. 8, t~d. l. K. Vasil, pp. 35-74. New York:
Academic Press.
Redenbaugh, K., D. Salde, P. Viss, and 1. A. Fujii . 1987. Encapsulation of somatic embryos
in synthetic seed coats. In: Proceedings of the Symposium on Synthetic Seed Technology for
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296 Seed Enhancements
Rushing, K. W. 1988. Additives to enhance seed quality and field performance. Proceedings
of Mississippi State University. Mississippi State, MS State Seed Short Course Vol. 30.
Saha, R., A. K. MandaI, and R. N. Basu. 1990. Physiology of seed invigoration treatments in
soybean (Glycine max L.). Seed Science and Technology 18:269~276.
Schippers, B., B. Lutenberg, and. P. J. Weisbeek. 1987. Plant growth control by fluorescent
pseudomonads. In: Innovative Approaches to Plant Disease Control, ed. 1. Chet, pp. 19~39.
New York: John Wiley & Sons.
Seneratna, T., B. D. McKersie, and D. C. W. Brown. 1989. Artificial seeds: Desiccated
somatic embryos. In Vitro Cellular and Developmental Biology 25:39-45.
Sharp, W. R., A. Evans, P. V. Ammirato, and Y. Yamada. 1984. Handbook of Plant Cell
Culture, Vol. 2, Crop Species. New York: Macmillan, NY.
Smith, A. E., and R. Miller. 1987. Seed pellets for improved seed distribution of small seeded
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Smith, E. M., and F. C. Wellner. 1987. Biological and chemical measures integrated with
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Taylor, A. G., D. E. Klein, and T. H. Whitlow. 1988. SMP: Solid matrix priming of seeds.
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Thanos, C. A., K. Georghiou, and H. C. Pasam. 1989. Osmoconditioning and aging of
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the Mycological Society ofJapan 28:483-494.
Valdes, V. M., and K. J. Bradford. ] 987. Effects of seed coating and osmotic priming on the
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Microbiology 34:631-637.
 14
Seed
Certification
Seed certification is a program to maintain and make available to the public high quality
seeds and propagating materials of genetically distinct crop varieties. Under this program,
certified seed is produced by outstanding farmers and seed producers using pedigreed planting
stock, carefu I quality control, field inspections during the growing season, and seed inspections
following harvest. Certification is an officially recognized method for maintaining varietal
identity of seed on the open market. Consequently, it has become especially important for field
crops (except for domestic hybrid corn) because most varieties offield crops traditionally have
been publicly released and their seed sold on the open market. It is oflesser importance for other
kinds of crops, whose varieties are most often privately released and seed production controlled
by private companies.
Certification is also widely used (including hybrid corn) for seed destined for international
sales. In the United States alone, over 100,000 tons of seed are produced annually for export
under the OECD certification program.

HISTORY

Seed certification in the United States and Canada dates back to the early 1900s when the
first new varieties appeared from state land grant colleges and government experiment stations.
Prior to this, most field crops originated from plant materials introduced from other countries.
When new varieties became available, they were distributed to farmers on a haphazard,
inefficient, and often inequitable basis. Frequently, such varieties were contaminated, lost, or
of poor physical quality.
During the period from ) 900 to 1920, organizations were set up in various states through
which seeds of new "college-bred" varieties were distributed to farmers. These organizations
were often outgrowths of state experiment associations and soon became known as crop

L. O. Copeland et al., Principles of Seed Science and Technology

© Kluwer Academic Publishers 2001
298 Seed Certification
improvement associations or seed certification agencies. These agencies were frequently
administered by experiment station or extension service staff of the land-grant institutions where
they were located.
During the 20s, 30s and 40s, under the guidance and influence of the universities, seed
certification became an established institution for increasing and making available to the public
high-quality seed of improved varieties--varieties that were almost without exception products
of the university or goverrunent agency breeding programs.

CERTIFICATION TODAY
Seed certification in the United States is the responsibility of each individual state; within
each there is an agency designated to certify seed. Regardless of the agency responsible, the
basic authority for certification is derived from the seed law of the individual state. Several
states have certification programs administered by state departments of agriculture. In a few
states, certification is administered by the Cooperative Extension Service. Most states have
certification programs administered by grower-controlled crop improvement associations,
although university personnel frequently act as their secretary-managers. In other states, a
secretary-manager is hired by the board of directors of the crop improvement associations.
Regardless of how the project is organized, certification programs in the United States and
Canada are generally non-profit programs, but must generate funds to cover salaries, overhead,
and operating expenses. Ordinarily, even these state agencies maintain close association with
university personnel who may serve on their boards of directors.
In Canada, the seed certification program is administered by the Canadian Seed Growers
Association, representing pedigreed (certified) seed growers from all Canadian provinces.

THE ASSOCIATION OF OFFICIAL SEED CERTIFYING AGENCIES

The Association of Official Seed CertifYing Agencies (AOSCA) is an organization of
certification agencies in the United States, Canada, and New Zealand. However, official
agencies from several other countries have indicated an interest in joining. Its purposes are: (1)
to establish minimum standards for genetic purity and recommend minimum standards for the
classes of certified seed, (2) to standardize seed certification regulations and procedures, (3) to
encourage cooperation with all individuals, agencies, groups, and organizations to accomplish
these purposes, and (4) to assist its member agencies in seed promotion, production, and
distribution.
The history of the AOSCA dates back to 1919, when representatives from Michigan,
Minnesota, North Dakota, South Dakota, Wisconsin, and the Canadian Seed Growers
Association met in St. Paul, Minnesota, to explore the possibilities of developing some type of
organization that would be helpful in solving mutual problems. After considerable discussion,
a Seed Improvement Federation was proposed, but the group felt that more states should be
represented. Another meeting was set for December 1919, at the International Grain and Hay
Show in Chicago. At that meeting, 13 states and Canada were represented, and the International
Crop Improvement Association (ICIA) was formed. The objectives of the ICIA were to promote
the agricultural interests of the various states as well as the provinces of Canada, emphasizing
especially the improvement offield crops in general and seed improvement in particular. These
objectives were to be attained by:
Seed Certification 299
I. Encouraging the breeding and improvement of field crops and seeds.
2. Husbanding, propagating, and disseminating elite, registered, certified, and improved
seeds.
3. Creating a more active interest in better seeds through circulars, reports, and other
publicity as well as by encouraging local, statle, national and international shows.
4. Assisting in the standardization of seed improvement and certification work being done by
member agencies.

From the beginning, the ICIA had a major influence on certification throughout the United
States and Canada. It has been instrumental in enunciating the fundamental concepts of
certification, establishing field and laboratory inspection standards, and encouraging uniformity
in certification procedures among its member agencies. Membership in the organization was
voluntary, as were its standards and policies; however, almost all certification agencies in both
the United States and Canada were members of the ICIA and were greatly influenced by it. In
1968, the name of the ICIA was changed to tht~ Association of Official Seed CertifYing
Agencies.

THE GENERATION SCHEME OF CERTIFICATION

Inherent in the certification concept is a generation system whereby the pedigree of superior
crop varieties is maintained through subsequent seed production. A four generation scheme
(Figure 14.1) has been devised to do this, and seed of each generation is identified by a special
color labeling tag.

1. Breeder seed is produced under the direct supervision or authorization by the plant
breeder and represents the true pedigree of the variety.
2. Foundation seed is the first generation seed fi'om breeder seed and is ordinarily produced
under contract by a foundation seed organization as authorized by the plant breeder.
Foundation seed is also labeled with white certification tags.
3. Registered seed is the seed from foundation seed and is intended for the purpose of
increasing seed another generation before the production of certified seed. Registered seed
is not intended to be a commercial class of seed. It is designated by purple seed tags. In two
states (Michigan and Wisconsin), all certified seed is the progeny offoundation seed, and
no registered class is used. Most states still maintain the registered generation. However,
this class is eliminated in many cross-pollinated crops, particularly with species where seed
is produced outside the area of adaptation.
4. Certified seed is produced from foundation or registered seed and represents the final
product of the certification program. It is labeled with the familiar blue tag, which has
become associated with the public image of clertification.

Although the four-generation scheme for seed certification has been an integral part of
certification since the very beginning, it has not always been applied in the strictest sense. It was
not until the mid-I 960s that the four-generation concept was adopted in practice by all certifYing
agencies. However, many variety developers have elected to restrict some or all of their varieties
to a three-class (Breeder, Foundation, Certified) system in order to better maintain varietal
purity and assessment of a royalty or research fee.
300 Seed Certification

VAJUETALDEVELOPMENT

(Plant breeding programs of

public and private agencies)

/ Comments
I. Available only in small quantities
Breeder Seed 2. Under control ofthe plant breeder
3. Labeled with a white tag
4. Planted to produce foundation seed

l. Available in limited quantities

2. Under control offoundation seed stocks
organization (public or private)
3. Labeled with a white tag
4. Planted to produce registered

!
(or certified) seed

I. Usually a noncommercial seed class,

available as planting stock for certified
seed producers
2. Under control of registered seed producers
3. Labeled with a purple tag
4. Planted to produce certified seed

I. Available in large quantities

2. Under control of certified seed producers
3. Labeled with a blue tag (often called "blue
Certified Seed
tag" seed)
4. Sold to commercial farmers for general crop
production

Figure 14.1. Diagrammatic scheme ~rthe overall limited generation seed certification program/rom
the development of a new variety to its availability to commercial farmers.

The Canadian generation system is the same as that in the United States except that there
is a select class between the breeder and foundation classes for wheat, oats, barley, rye, flax,
triticale, buckwheat, field peas, lentils, field beans, faba beans, and soybeans. Breeder seed of
these crops is allocated to members of the Canadian Seed Growers Association (CSGA) who
have become established as select seed growers after serving a three-year probationary plot-
production program. For select status, growers may grow no more than 2.5 acres of one variety
of select seed or no more than 5 acres of all crop varieties. The plots must have no more than
1 off-type plant per 20,000 crop plants. Seed from each plot is post-controlled for varietal purity
by Agriculture Canada. Select seed may produce select seed for five multiplications before the
grower is required to obtain new breeder seed. Select seed is used to produce foundation seed
crops which are also post-controlled for varietal purity. A Breeder Seed Crop Certificate is
Seed Certification 301
issued by the CSGA for the initial increase of breeder seed and each increase made thereafter
by the originating plant breeder. Crops of all classes, including breeder seed, are field inspected
for varietal purity by Agriculture Canada for the CSGA.

FOUNDATION SEED PRODUCTION

Foundation seed can be thought of as the "vital link" between breeder seed produced under
the control of the plant breeder and certified seed produced by the certified seed grower. It is the
seed stock from which Registered and Certified seed are produced. It is produced by a
foundation seed organization, which may be a private association of seed growers, a special
project within a university experiment station, or an independent private business.
Foundation seed agencies receive breeder seed of new crop varieties as they are released
and increase them to foundation seed. After the initial release of breeder seed, it must be
maintained and made available every year. This is done in one of several ways. First, a small
portion ofthe foundation seed field may be designated as eligible for breeder seed production.
This area is carefully inspected, rogued of off-types, and classified as breeder seed in
cooperation with the experiment station or releasing agency. Thus, foundation seed of annual
crops can be maintained on a permanent basis. An alternate, but less frequently used, method
is for the releasing institution to grow small lots of breeder seed each year for annual release to
the foundation seed organization. For perennial crops, maintenance of breeder seed is much
simpler, since small production blocks of each variety can be maintained to provide breeder seed
for use foundation seed regeneration.
If the foundation seed organization does not have adequate production and processing
facilities of its own, it may arrange for contract production, which is then made availabl.e to
registered or certified seed growers after the seed has been cleaned, conditioned, and bagged to
the proper standards.
Several factors should be considered when producing foundation seed. First, only the best
seed growers with the right combination of experience, land, facilities, and ability are accepted
as foundation seed growers. Second, the supply of foundation seed should not exceed the
demand. To anticipate demand and provide for foundation seed production calls for advance
planning. If excess foundation seed is produced, it must be carried over at extra expense,
downgraded to certified seed, sold as commercial grain, or destroyed at a considerable loss.
Third, the foundation seed should be available to all certified seed growers on an equitable basis
at a reasonable price. When the supply of foundation seed is limited, consideration is given to
the growers' production history, their facilities, and their ability to produce high-quality seed.
Although most foundation seed programs operate on a nonprofit basis, they must be self-
sustaining. Costs such as labor, overhead, buildings, and conditioning facilities must be covered
from the saleoffoundation seed-requiring operation on sound business principles. Responsibility
for this rests with the manager, who is employed to handle day-to-day activities. Often a board
of directors is responsible for establishing overall policy and to ensure that the aims and function
of the organization are met.
Foundation seed organizations usually have a formal agreement with the state agricul-
tural experiment station which defines the terms of breeder seed release of new crop
302 Seed Certification

varieties. Exceptions may occur when foundation seed organizations are private businesses not
associated with a university experiment station, or when the foundation seed project is
administered by an experiment station staff member.

HOW VARIETIES BECOME ELIGIBLE FOR CERTIFICATION

Varietal Release

Most agronomic varieties have traditionally been released from university experiment
stations; however, private seed companies also release new crop varieties. Privately released
varieties have long been available for vegetable crops, corn, sorghum, and cotton, and are now
common for other field crops.
Regardless ofthe releasing agency, a procedure must be available for evaluating potential
varieties and recommending their release. Most of our experience comes from release by
university experiment stations. When plant breeders have a candidate for release, they submit
to the appropriate review board a description of their variety, its identifYing characteristics, and
performance data. Release procedures may consist of one or two steps; however, it is quite
common to submit it first for consideration by a commodity committee comprised of persons
closely concerned and familiar with the crop. In experiment stations, this usually involves the
plant breeder, a plant pathologist, an entomologist, extension agronomists, and other trained
personnel. After favorable action, it often goes to a larger, more formal committee composed
of experiment station personnel from disciplines who are responsible for release of new varieties
of all crops. The first group provides closer knowledge and involvement with specific crop
areas, while the second committee observes uniformity of release procedures and provides
perhaps a more objective evaluation of candidates for release. The second committee also
advises on specific release procedures and other seed increase matters.

Definition of a Variety

To be eligible for certification a variety must be properly released, named, and described.
In the past, the term variety has been simply defined as "a specific close subdivision of a kind
with definite distinguishing genetic characteristics that can be maintained or inherited when the
plants are reproduced over a period of years." Because of the different kinds of crops and germ-
plasm available for certification, it has often been unclear whether many candidates for
certification actually qualified as varieties. To help clarifY this situation, an ad hoc committee,
representing the United States Department of Agriculture, the Association of Official Seed
CertifYing Agencies, the American Society of Agronomy, and the American Seed Trade
Association, has developed a comprehensive consensus defmition of different kinds of varieties.
These definitions are published and made available to all concerned organizations.

Varietal Review Boards

Individual certification agencies are aided in determining the eligibility of varieties for
certification by national variety review boards which have been established by AOSCA.
Seed Certification 303

Four review boards have been established representing alfalfa, grasses, soybeans, and small
grains. Each board is composed of six members representing the: (1) American Seed Trade
Association, (2) Association of Official Seed CertifYing Agencies, (3) Crop Science Society of
America, (4) National Council of Commercial Plant Breeders, (5) United States Department of
Agriculture, and (6) Agricultural Research Servic~~, USDA.
The functions of the boards are to review and evaluate information presented by breeders
of the respective crops in industry and in public agencies who request certification of new
varieties and to advise the AOSCA on their acceptability as bona fide varieties that have been
properly released and described. The goal of this arrangement is the acceptance by any member
agency for certification of all varieties given favorable action by these boards. However, in
actual practice, many individual agencies still require varieties to meet adaptability and
performance standards established for their particular state.

CERTIFICATION ItROCEDURE

Planting Stock

The proper planting stock is essential for certified seed production. It provides a pedigree,
which is central to the certification concept. Certifiied seed is usually produced from registered
seed, although it may be produced from foundation or breeder seed. Likewise, other classes of
seed may be produced from earlier seed generations.

Application

An application for certification must be submitted to the appropriate state or country

certifying agency requesting certification as foundation, registered, or certified seed. The
application must be accompanied by at least one official tag substantiating the class of seed
planted: breeder, foundation, or registered. Each c:ertification agency has its own application
procedures that must be followed.

Field Inspections

Inspections are performed on all fields for whieh applications are received. These are timed
so that varietal off-types and other crop and weed contamination are most easily detected. In
some crops (e.g., clover and alfalfa), a seedling inspection may be required to check for
volunteer crop plants. These are normally made a few weeks following seeding. Small grain
inspections are normally made after the chaff color has changed, and during the hard dough
stage when different chaff color of off-types is most easily distinguished. However, oats are
often inspected while the plants are green and the seed is in the soft dough stage. Grass and
legume seed fields are usually inspected during poHination, when off-types and weeds are most
easily seen and when isolation from adjacent cross-fertile fields is apparent. Table 14.1 shows
the AOSCA genetic standards for several crops. Note that in this table, the cross-pollinated
crops must be separated (isolated) specified minimum distances from other fields with which
they could potentially outcross. No isolation is required between self-pollinating crops, except
for short distances (e.g., 5-10 m) to help prevent mechanical mixing.
304 Seed Certification

Harvesting

A crop is harvested for certified seed in the same way as for other purposes except that
more care is given to moisture content, purity, and prevention of mechanical damage. Seed
harvested with excessive moisture will not maintain its quality during storage, while excessively
dry seed is more susceptible to mechanical injury. Particular attention is directed to thorough
cleaning of combine and handling equipment before starting to harvest a field in order to avoid
both genetic and mechanical contamination.

Conditioning

All seed must be cleaned thoroughly to remove other crop and weed seed, chaff, straw, and
other inert matter before it will meet the purity standards for certification in most states. The
amount and kind of cleaning depends on the kind of seed and its composition. It is absolutely
essential that the conditioning facilities be thoroughly cleaned between seed lots. Although this
is a difficult and exacting task, its importance can hardly be overemphasized. Utmost care
should be given to avoid mechanical damage during conditioning. Seeds of certain species are
especially fragile and must be handled with extreme care. Seed must be elevated at least once
to reach the top of the conditioning flow; however, if possible, it should be elevated only once,
and gravity should be used to move it through the different steps in cleaning and bagging. The
use of augers to move seed should be avoided whenever possible, because they tend to damage
the seed, especially that of large-seeded legumes.

Sampling

The sample that is tested for determining seed quality and acceptance for certification is
taken after the last conditioning operation. It may be taken by an automatic (mechanical)
sampling device, but is more commonly taken from the bagged or bulk seed by hand sampling
methods by an official of the certifying agency, however, some agencies permit producers to
draw their own samples. The sample should be taken at a time and by a procedure that will yield
a sample that will represent accurately the seed to be marketed. If the seed is treated with a
fungicide, the submitted sample should be drawn from the treated seed.

Seed Inspection

A sample of the conditioned seed to be certified is tested for purity, germination and
noxious weed seed content. Sometimes phytosanitary tests are also required by the certification
agency for disease assessment. Whether the analysis is performed by the certification agency
or by an official state seed laboratory, it is used to determine whether the lot qualifies for
certification. Table 14.2 shows the suggested nongenetic AOSCA seed quality standards for
alfalfa. Unlike genetic standards, they are only suggested standards, and are not required for
certification, except when required by the state certification agency.
 ~
(';:
~
(""'J
(';:
Table 14.1. AOSCA Minimum Genetic Standard&(Used as an Example). .......
'S
r::
Foundation Registered Certified t:l
g.
Crop Kind Landt Isolation2 Field 3 S~d. Landt Isolation 2 Field 3 Seed. Land 1 Isolation 2 Field 3 Seed· :s
Alfalfa 4 600 1000 0.1 3 300 400 0.25 1 165 100 1.0
Barley 1 0 3000 0.05 1 0 2000 0.1 1 0 1000 0.2
Hybrid 1 660 3000 0.05 1 660 2000 0.1 1 330 1000 0.2
Bird's-foot
Trefoil 5 600 1000 0.1 3 300 400 0.25 2 165 100 1.0
Clover
(all kinds) 5 600 1000 0.1 3 300 400 0.25 2 165 100 1.0
Corn
Inbred lines 0 660 1000 0.1
Foundation
Single Cross 0 660 1000 0.1
Hybrid 0 660 0.5
Open-
Pollinated 0 660 200 0.5
Sweet 0 660 0.5
Cotton 0 0 0 0 0 0 35000 0.01 0 0 7000 0.1
Cowpeas 1 10 1000 0.1 1 10 500 0.2 1 10 200 0.5
Crambe 1 660 2000 0.05 1 660 1000 0.1 1 660 500 0.25
Crown vetch 5 600 1000 0.1 3 300 400 0.25 2 165 100 1.0
Field & Garden
beans 1 0 2000 0.05 1 0 1000 0.10 1 0 500 0.20
lNumber ef years that must elapse between destruction of a stand of a variety and establishment of a stand of a specific class of a variety of the
same crop kind.
2Distance in feet from any contaminating sources.
"Minimum number of plants or heads in which 1 plant or head of another variety or off-type is permitted.
'Maximum percentage of seed of other varieties or off-types permitted.
Data from Certification Handbook, AOSCA. p. 9.

w
0
Vl
306 Seed Certification

Table 14.2 Suggested AOSCA Nongenetic Seed Standards for Alfalfa.

Standards for each class
Factor Foundation Registered Certified
Pure Seed (minimum) ......................... . 99.00% 99.00% 99.00%
Inert Matter (maximum) ....................... . 1.00% 1.00% 1.00%
Weed Seeds (maximum) ....................... . 0.10% 0.20% 0.50%
"Objectionable or Noxious Weed
Seeds (maximum) ......................... . None None None
Total Other Crop Seeds (maximum) ............. . 0.20% 0.35% 1.00%
Other Varieties (maximum) .................. . 0.10% 0.25% 1.00%
tOther Kinds (maximum) .................... . 0.10% 0.10% 0.50%
Germination & Hard Seed (minimum) ........... . 80.00% 80.00% 80.00%
'Objectionable or noxious weed seeds shall include the following: bindweed (Convolvulus arvensis), Canada
thistle (Cirsium arvense), dodder (Cuscuta spp.), dogbane (Apocynum cannabinum), Johnson grass (Sorghum
halepense), leafy spurge (Euphorbia esula), perennial sow thistle (Sonchus arvensis), quackgrass (Agropyron
repens), Russian knapweed (Centaurea repens), and white top (Lepidium draba, L. repens, Hymenophysa,
pubescens ).
tSweet clover seed shall not exceed 9 per lb for foundation seed; 90 per lb for registered seed; and 180 per lb
for certified seed.
Data from Certification Handbook, AOSCA, p. 20.

Seed Tagging

A few certification agencies have a one-tag system in which the analysis information
(purity, germination, etc.) is printed on the certification tag. However, most agencies have
adopted a two-tag system, in which the analysis tag and certification tag are different. Some
agencies maintain laboratories that determine only if seed meets certification requirements;
labeling information is obtained from tests performed in an official state or commercial
laboratory. With the two-tag system, the seed quality information can be changed or updated
without removing the official blue certification tag.
Regardless of the system used, the tags should be attached in a way that will reveal
evidence of any opening, reclosing, or other tampering with the contents of the container. It
should be impossible to open the container without breaking or defacing the tag. This is easily
accomplished by sewing the tag into the seam or by attaching the tag to the stitching with a
metal seal. Metal seals were very common in the earlier days of certification but are seldom used
today.

Marketing

The marketing of certified seed is the responsibility of the producer. It requires both
promotion and a reputation for delivering quality, and most experienced growers have
established regular customers through which most of their seed is marketed. Many certification
agencies have advertising programs to help promote the image of certified seed and to carry out
educational programs in seed improvement.
Many growers avoid marketing problems by growing seed on contract for elevators, seed
dealers, or other larger seed growers. The contractor may submit the application, pay all
certification charges, and even condition, bag, and tag the seed. Normally, the grower
Seed Certification 307

only plants the seed, harvests the crop, and delivers it to the contractor, although various other
arrangements may be used. Contract seed production is becoming more important as smaller
growers find it increasingly difficult to compete with larger seed dealers. It is common in the
western grass and legume seed production area, which is outside the major area of utilization
and requires extensive transportation and marketing arrangements. Contract seed production
offers the security of a fair and stable price for the individual seed grower and provides the seed
dealer the security of having adequate supplies of seed at competitive prices.
Certitied seed may be marketed cooperatively with that of other growers. Successful
marketing cooperatives usually have either a unique product, a common geographical location,
or some other factor that promotes the common interest of the members and allows them to
compete with other seed providers. Furthermore., by group action they can handle larger
volumes, promote their product, offer it at a stable price, and generally be more competitive in
the seed market.
Crop improvement associations in North America commonly assess additional fees to
provide revenues to support programs to help promote certified seed. Such programs generate
funds to promote certified seed in the broadcast and print media as well as other kinds of
promotional methods. Such promotion may focus on general advantages of certified seed or be
used to promote particular certified varieties. Some are done in cooperation with individual
certified seed growers or groups of growers.

Benefits of Certification

Benefits for the farmer. There are several benefits for the average farmer in planting
certified seed. First, it provides access to seed of excellent varieties with good assurance of high
genetic purity. Thus, it helps avoid unnecessary losses in yield from planting seed of unknown
or contaminated varieties. Such off-types are likely to yield plants of different maturity,
susceptibility to diseases and insects, or be less productive. Similarly, certified seed which is
high in mechanical purity provides assurance to the user against the introduction of weeds,
diseases or other crop seeds. Contamination by undesirable plants of any kind can reduce
productivity and lower crop quality.
Benefits for Certified Seed Producers. Historically, only outstanding farmers in each
state have produced certified seed on a sustained basis. There are several reasons for this in
addition to the increased income potential from seed production. They tend to be generally more
will ing to accept greater effort and timely management required for success. They also recognize
the inherent advantage of having early access to new varieties. Finally, they demand the highest
quality seed possible for their farming operation and take pride in meeting these demands.

INTERAGENCY CERTIFICATION

Interagency certification is the participation of two or more certifying agencies in

performing the services required to complete certification on an individual lot of seed. It may
be useful when a demand for seed exists in a state other than that where the seed is produced.
The plan was first proposed in 1943 to facilitate the interstate movement of hybrid corn
seed, where the production and field inspection would be done in one state, and the
308 Seed Certification

seed inspection and final certification completed in another. Under this scheme, seed growers
in Michigan have historically produced several thousand acres of certified oat seed annually,
which have been field inspected and then shipped into New York and Ohio for fmal certification.

OECD-AN INTERNATIONAL CERTIFICATION PROGRAM

The Organization for Economic Cooperation and Development (OECD) provides a scheme
for the varietal certification of seed moving in the international market and is the nearest existing
program to a completely international seed certification organization. Instituted in 1960, the
OECD is an outgrowth of the Organization for European Economic Cooperation (OEEC),
which included membership from several European countries, Japan, and North America. The
entire program involves trade agreements, economic expansion, financial stability, and overall
economic well-being of the member countries. This program is funded in the United States and
Canada by an additional assessment on each unit of seed certified.
Certification under the OEeD scheme is on a basis of varietal purity only, and standards
have been established to help maintain varietal purity. Requirements include: (1) authentication
of the proper planting stock, (2) documentation of previous cropping history, (3) minimum
isolation between adjacent seed fields, (4) the number of harvest years during which a field can
produce certified seed, and (5) field inspection criteria. As long as the varietal purity is intact,
no fields or seed lots are rejected because of nongenetic seed quality factors such as germination
and pure seed content.
The OEeD has an agreement with the U.S. Agricultural Research Service (ARS) to
provide for the program in the United States. State agencies may negotiate a memorandum of
agreement for certifying seed according to OEeD standards and attaching official OEeD
certification tags. Ordinarily, this procedure is followed when the seed is to be shipped to
another OEeD member country. Under this program, thousands of acres of grass and legume
seed crops (and hybrid corn) are certified each year in the United States and are sold (usually
under contract) by U.S. seed companies to companies in other countries. This is possible
because the OEeD tag assures that the seed has met internationally recognized standards for
genetic purity. The program has greatly aided international trade in seed. Table 14.3 shows the
tonnage of total OECD certified seed produced in 1991192 and the relative production in
different countries. The United States is by far the largest participant in this program and its
production is about equally divided between domestic and foreign cultivars.
The OEeD certification standards arc printed in OEeD Publication Documentation in
Food and Agriculture.
CHANGING CONCEPTS AND SERVICES

From its origin in the early 1900s up to the late 1950s, seed certification was built around
three primary concepts: superior varieties, genetic purity, and high seed quality standards. These
concepts were seldom criticized or questioned, and over the years, they became almost
synonymous with certification. However, in the late 1950s, are-evaluation ofthis philosophy
by the certification agencies greatly changed both the concept and services of certification.
Seed Certification 309

Table 14.3. Weight (Tons) of Seed Certified Under OECD Seed Schemes (1991/02).

Homel
Foreign Herbage Total All Country
Country Cultivars and Oil Cereal Maize Beet Schemes Totals Percent
Argentina H 334 1,500 1,834
F 88 3,627 3,715 5,549 2.0
Australia H 5,932 2,188 8,120
F 837 837 8,957 3.0
Austria H 37 104 21 162
F 653 71 6,134 244 7,102 7,264 2.0
Belgium H 291 132 423
F 364 10 1,191 7 1,572 1,995 1.0
Canada H 14,312 51 14,363
F 16,284 2,784 19,068 33,431 11.0
Czech Republic H 7,790 1,872 9,662
F 9,662 3.0
Denmark H 81 150 231
F 258 275 0.7 534 765 0.2
France H 2,595 297 2,892
F 2,892 1.0
Germany H 1,818 503 156 256 2,733
F 158 25 73 50 306 3,039 1.0
Hungary H 1,666 3,074 2,457 7,197
F 27,161 582 35,502 205 63,450 70,647 23.0
Ireland H 50 50
F 21 21 71 0.2
Israel H 2,719 2,719
F 2,719 1.0
Italy H 27 0.0 35 50 112
F 339 101 440 552 0.2
Japan H 11 11
F 11 0.004
Netherlands H 1,344 67 1,411
F 73 15 88 1,499 0.5
Norway H 0.9 1.1 2
F 2 0.006
Poland H 9,334 1,788 10 11,132
F 3,076 3,076 14,208 5.0
Romania H 261 261
F 863 12:,143 13,006 13,267 4.0
South Africa H 25
F 25 25 0.008
Spain H 0.9 0.9
F 1,040 54 1,938 332 3,364 3,365 1.0
Sweden H 338 84 422
F 9 0.8 10 432 0.1
Tunisia H 16,099 16,099
F 16,099 5.0
Turkey H
F 442 1.,902 25 2,369 2,269 1.0
United States H 47,371 14,491 38,286 100,148
F 8,817 .~,151 10,968 111,116 36.0
Total Domestic 179,983
Total Foreign 129,926 309,909 100.0
Subterranean Clover Seed Scheme
Australia 4,083
310 Seed Certification

Performance and Recommendation Criteria

Traditionally, crop varieties became eligible for certification only after they were released
and recommended to growers within the jurisdiction of a given certification agency. Such a
requirement evolved quite naturally, since only superior varieties were eligible for release by
state experiment stations. As plant breeding programs grew and new varieties were developed,
older varieties were removed from the recommended lists and new ones added. This policy was
strengthened by strong crops extension programs that complemented the varietal release
programs. It became customary for state universities to publish annual lists of variety
recommendations, and the appearance of a variety on these lists normally meant eligibility for
certification.
In the late 1950s, recommendation and performance standards came under question as
valid criteria for certification. Two factors were primarily responsible for this: (I) the volume
of seed produced away from the area of consumption, and (2) the appearance of private field
crop varieties. Today, evidence of varietal performance (merit) is usually not a factor in
certification. Most agencies will certifY any variety that has been properly identified and
described by any public or private agency and has met the criteria described on page 302.

Varietal Purity Only (VPO) Certification

One of the more controversial issues to confront certification in the United States has been
the concept of varietal purity only certification. Under this concept, seed is certified if the field
and seed inspections show the crop to meet minimum standards of varietal purity; seed lots are
rejected only for excessive contamination by off-types, inadequate field isolation, or other
genetic purity factors. The occurrence of weeds or other crops, disease infestation, or even low
germination does not constitute cause for rejection of any field or seed lot. Under the practice
of VPO certification, consumers are assured of variety purity by the certification agency and
choose seed lots that meet their own seed quality (purity, germination, etc.) criteria on the basis
of information on the seed tag. Several state certification agencies have adopted VPO
certification, though most still require certified seed to meet minimum seed quality standards.
Certification of Blends

Varietal blends of seed are eligible for certification by several certification agencies. Where
this is permitted, all components of the blend must represent certified seed and must be blended
in specific, predetermined, and commercially acceptable proportions. The components of the
blend are confidential between the producer and the certification agency. Some agencies are
reluctant to certifY blends on the basis that they do not represent a pure variety.

Sod Certification

Some certification agencies have been certifYing turf sod for many years. Certification
is usually performed on the basis of elite varietal and mechanical purity of seedstocks,
and verification of vegetative varietal composition, freedom from diseases and insects, and
Seed Certification 311

absence of weeds and other crop plants. [n some states, sod certification is principally for
phytosanitary condition.

Tree Seed Certification

Progress in the breeding and improvement of trees has caused an interest in the seed
certification of improved tree varieties. Seed orchards of new tree varieties have been established
from which certified seed and seedlings are harvestc~d and sold to help establish improved tree
stands. Certification agencies have responded to forestry industry requests for help in developing
procedures by which customers can be assured of varietal purity and high seed quality. The
AOSCA standards include trees, shrubs and native plant species.
Some agencies also certifY the source or origin of forest tree seeds and seedlings. This type
of certification is performed in the absence of, or as a supplement to, seed from established seed
orchards. Certification of the origin of seed is important when it is desirable to obtain seed from
locations of climate, elevation, and exposure similar to the sites where the seed is intended for
planting. This assures that the resulting plants will be ecologically suited for the planting site
and also ensures their survival and performance.

Phytosanitary Certification

Phytosanitary certification does not qualifY as seed certification in the usual meaning of
the term. It certifies only that the seed and the fielld from which it came is free of specified
diseases. Phytosanitary certification is normally performed by pathologists from the official
government agency, and a tag is attached to specifY what has been done. Usually, a
phytosanitary certificate is issued for the seed. This enables seed suppliers to provide the
certificate to their customers and to officials of the state or country into which the seed is
shipped. When seed is shipped internationally, phytosanitary certification is usually required by
the receiving country. Most phytosanitary certificates expire within 14 days after shipment of
the seed or plant materials.

The United States (USA) Certification Concept

A diversity of certification programs in the United States and different standards and
procedures remain in spite of efforts toward standardization. This lack of uniformity has
frequently caused problems for certified seed produced in the United States in spite of progress
made under AOSCA certification standards which sets minimum genetic standards for all seed
produced by member agencies. This has caused interest in the formation of a "USA certification"
program in the United States. The goal of such an organization would be to certifY seed by
standardized procedures that would make the seed acceptable in international in commerce and
also appeal to the private seed industry in the United States who normally does not support
certification or who would like to see more uniformity in certification procedures among U.S.
certification agencies. Although at this point (2001), the USA seed certification concept is still
in its infancy, it appears to have good potential for the future.
312 Seed Certification
Ancillary Programs

Because of financial difficulties, many certification agencies have had to consider other
ways to maintain profitability in addition to providing traditional certification services.
Consequently, they have explored the possibility of providing ancillary programs to help utilize
their talents or services and to help make ends meet. Two of these programs are discussed
below.
Identity Preserved Programs. The identitlcation and maintenance of genetic purity has
been the strength and focus of seed certification programs since their inception. This has enab led
improved varieties to be made available to farmers both quickly and efficiently, and has
contributed enormously to crop production in the United States and around the world. In recent
years, it has become evident that this same expertise could also be extended beyond the farmer
level to the consumer and end-user of agricultural grain products. For example, millers of
certain types of soybeans prefer high oil varieties and are willing to pay a premium for soybeans
that are documented to be of the preferred type. This provides a way to avoid the necessity of
the mixing of different varieties or quality levels of grains, oil seeds, or other farm products
during storage or marketing. The further development of this concept could provide a valuable
service to agriculture and food-related industries which require high levels of product quality
and uniformity.
Identity preserved programs for grain products have been established by seed certification
agencies in several states. For the most part, these programs utilize the same procedures and
practices of conventional certification, including careful record-keeping, field (in some cases),
and post-harvest inspections for genetic purity and other aspects of quality, including
uniformity.
Quality Assurance. Many certification agencies offer quality assurance programs to the
seed industry. These provide field inspection and evaluation services when there is no interest
in completing the certification process. This kind of service is used by seed companies that
desire the expertise of the certification agency in providing field or laboratory inspections and
advice on assuring varietal purity and overall quality. This may include verification of GMO
(genetically modified organism) or non GMO status. [n many ways, it is not unlike a pest
scouting or crop consulting service by an agency with a particular expertise in quality seed
programs. In such cases, the seed may be labeled with a specially developed quality assurance
tag which indicates the kind of services performed.

THE FUTURE OF SEED CERTIFICATION

Past Contributions

There is little doubt about the contribution of seed certification to the development ofNorth
American agriculture. [t has provided a rapid and highly efficient way for seed increase and
distribution of superior varieties developed and released by state experiment stations. This has
had great impact on both the seed industry and North American agriculture, and has provided
a model for similar development in countries around the world. However, many changes have
recently occurred in the seed industry that give cause for concern about the changing form of
certification and perhaps its very survival.
Seed Certification 313

The Changing Seed Industry

The nature of the North American seed industry has developed and changed dramatically
in the last 100 years. Today, it is well supplied with a wide range of varieties of all major crops.
For the most part, it has sophisticated production, quality control, and marketing programs.
Furthermore, those in the industry are well aware that farmers have become more discriminating
in their needs and demands. This has made the seed industry highly competitive, efficient, and
responsible to seed customers.
Today's seed industry is showing greater interest and ability to provide its own quality
assurance programs, with decreasing dependence on third-party umpire services such as
certification to evaluate the quality of their product. The present interest in the International
Standards Organization (lSO-9000) program is an indication that the seed industry feels that
it can provide quality assurance programs for itself. Such programs have rigid guidelines and
requirements for quality assurance regarding facilities, qualification and training of inspectors
and precision and instrumentation, as well as continuous monitoring to assure that standards are
met and quality is maintained. While large seed companies are better able to afford such self-
policing quality assurance programs without the need for third-party certification, smaller
companies are more likely to use the quality control and referee services of crop improvement
agencies.
The passage ofthe Plant Variety Protection Act (PVPA) in 1970 and the varietal explosion
that followed has had a profound effect on seed certification in the United States. Seed buyers
today have a great many choices of both private and public varieties compared to the relatively
few, mainly public varieties of a half century ago. Varietal protection has allowed commercial
companies to invest in genetic improvement programs and protect their varieties against
infringement by other parties. Thus, many seed companies have invested heavily in variety
development and have strong production and sales programs involving hundreds of varieties
across most field crops.
The professionalism of the modern seed industry, along with its arsenal of high quality,
productive varieties has allowed the private seed industry to thrive and prosper in competition
with certified seed producers, who for the most part produce only public varieties. Although the
private seed industry may also produce seed of publi,; varieties, private varieties tend to be more
profitable because of the marketing advantage provided by exclusivity and promotional
programs. This off-sets many of the traditional advantages that certification has in the past
offered for public varieties. Consequently, there may be little incentive for commercial
companies to use the certification process. This is especially true for large companies with well-
established research and quality control programs. Furthermore, private companies promote and
market their seed aggressively and use their resources and reputation to stand behind their
products. The use of brands and special marketing ]programs is also an important part of their
programs. These factors have led to the perception that private varieties or brands are superior
to public varieties and have helped to erode into the market that certified seed has had in the
past. Perhaps the crop that reflects this trend more than any other is soybean where seed of
private varieties command as much as 30% hight:r price than seed of comparable certified
varieties.
314 Seed Certification
Competition and Survival

Certified seed producers have competed with the private seed industry in various ways.
One is by aggressive marketing programs that promote the advantages that certified seed has
always enjoyed, i.e., excellent varieties, third person quality verification, and cooperative
promotional efforts. Two examples of this are Public Varieties ofIndiana (PVI) and Public
Varieties of Minnesota (PVM). These programs, composed of certified seed producers, assess
themselves additional fees that are used to collectively promote certified public varieties. Several
other agencies have similar programs.
Certification can exist only when it provides a real or perceived quality advantage or a
service to the seed industry. In the past, its major advantage has been that it represented access
to superior germplasm as new public varieties were released from public research institutions
and made available through certification. Certification represents an unbiased and official third-
party assessment of a high level of genetic and mechanical quality. It is ideally suited for seed
of public varieties which is produced and made available by many different seed producers in
competition with one another and with uncertified commercial or bin-run seed of the same or
different varieties. It has been less successful for seed of private varieties.
Certification is very important for almost all kinds of field seeds moving in international
commerce, since most countries require seed to be certified in order to be imported.
Consequently, even hybrid com seed moving in international commerce is certified. Otherwise,
little, if any, hybrid com seed sold domestically is certified, since it offers no particular
advantage over uncertified hybrid com seed. Forage and turf seed certified under the OECD
program also move easily in international commerce, although, unlike hybrid com, this seed is
also certified for domestic use.

New Dimensions; New Horizons

As the 21 st Century begins, the future of the role of certified seed in a modem,
sophisticated seed industry appears somewhat uncertain. Although certification continues for
publicly released field crop varieties, the increasing prominence of private varieties and the
ability of the private seed industry to provide many of the benefits of certification has led to an
uncertain future for certification programs. As a result, many U.S. certification agencies are
broadening their role by offering other services to help provide a strong fmancial base in order
to secure their future. These include (l) quality assurance programs, including their GMO or
non-GMO status (2) identity preserved programs, (3) pest scouting services, and (4) seed
sampling and seed testing services. Though none of the programs are necessarily identical to
past certification roles, all can make valuable contributions to the seed industry and to
agriculture, especially for smaller seed companies.
While it is evident that certification will continue to evolve and adapt to a changing seed
industry, it also seems clear that it can continue to have an important role in the seed industry.
First, it should be a vehicle in the seed increase and availability of public varieties. Second, it
should continue its role in the collection of research assessments or royalties on public varieties
and thus continue to fund variety development by public institutions. This partnership with
public research institutions should be mutually beneficial while providing a valuable service to
the seed industry.
Seed Certification 315
Questions

1. What basic authority defines the responsible seed certification agency in any given state?
Can you name three types of agencies that are responsible for certification in various
states? Why do these differ? What agency certifies seed in your state?
2. What is meant by OECD certification?
3. What are the advantages and disadvantages of certification on the basis of varietal purity
only? Do you approve of CVPO? Why or why not?
4. What is the importance of certification for vegetable and hybrid corn seed?
5. Do you believe certification will eventually outlive its usefulness? Why or why not?
6. If you were a farmer, would you buy certified seed? Why or why not?

General References

Armstrong, J. 1994. The Role ofCertified Seed in the Twenty-First Century. Presented at the
1994 Annual Meeting of the Association of Official Seed Certifying Agencies, Fort Mitchell,
Kentucky.
Clapp, A. L. 1970. The Kansas Seed Grower: A History of Seed Certification in Kansas.
Manhattan, Kansas: The Kansas Crop Improvement Association.
Cowan, J. R. 1972. Seed Certification. In Seed Biology, Vol. 3. ed. T. T. Kozlowski, pp. 371-
398. New York: Academic Press.
Douglas, 1. E., ed. 1980. Succes~ful Seed Programs: A Planning and Management Guide.
Boulder, Colo.: Westview Press. 302 pp.
Hackleman, J. C. History of the International Crop Improvement Association, 1919-1961.
International Crop Improvement Association.
McDonald, M. B. and W. D. Pardee (eds.). 1985. The Role ofSeed Certification in the Seed
Industry. CSSA Spec. Publ. No.1 0, Madison, WI.
Thompson,1. R. 1979. An Introduction to Seed Technology, Toronto: John Wiley and Sons.
252 pp.
U. S. Department ofAgriculture. 1961. Seeds: The Yearbook ofAgriculture. Washington, D.C.:
U.S. Department of Agriculture.
Wheeler, W. A., and D. D. Hill. 1957. Grassland Seeds. New York: D. Van Nostrand
Company, 734 pp.
 15
Seed
Testing

Seed testing is the art and science of evaluating seed quality for agricultural purposes.
Although initially developed for evaluating the planting quality of field and vegetable seeds, it
is also valuable for determining the quality of lawn, flower, and tree seeds.
Even though humanity's use of seeds dates back to prehistoric times, the art and science
of seed testing have only developed in the last century or so. Until about 300 years ago, our
knowledge concerning seed morphology and physiology was limited. As a result, it was possible
for unethical vendors to market seed of such crops as alfalfa mixed with sweet clover or other
contaminants. Such practices became so widespread and of such serious concern that laws were
passed and seed testing procedures established. Berne, Switzerland was the first city to enact
seed legislation prohibiting the sale of adulterated clover seed in 1816. The first seed testing
laboratory was established in Saxony, Germany, in 1869, and the first one in America was set
up at the Connecticut Agricultural Experiment Station in 1876. Today, official seed testing
laboratories are found in almost aliSO states and in nearly every country in the world, and many
privately operated commercial laboratories are located throughout North America and Western
Europe.
Other important milestones in seed testing include the publication of the first "Rules and
Apparatus for Seed Testing" by the United States Department of Agriculture in 1897 and the
first drawings of seeds in the publication "The Viability and Germination of Seeds" by F. A.
Hillman in 1902. In 1915, E. G. Boerner developed the first divider for separating grain, seed
and other material; in 1916, H. D. Hughes developed the first seed counter for preparing
germination tests; and in 1917, G. N. Collins developed the fIrst seed blower.
The expression seed quality is used loosely to reflect the overall value of seed for its
intended purpose. Seed quality is usually a composite of several factors, all of which contribute
to the desirability, or planting value, of the seed. The key question is "Why do we test seeds?"
There are several reasons. First, and most obvious, is that the dry seed's potential to establish
a seedling cannot be determined until the seed has been germinated. However, we also test seeds
to determine the genetic (varietal) and mechanical (weed/other crop) components ofthe seed lot.

L. O. Copeland et al., Principles of Seed Science and Technology

© Kluwer Academic Publishers 2001
Seed Testing 317
Seed testing results provide important information to both the seed producer and
purchaser. The seed producer wants to ensure that only a quality product is marketed so that
consumers will return for their future seed needs. Seed purchasers need assurance that
associated expenses such as field tillage, pesticide applications, and other costs are not lost due
to stand failure as a result of planting poor quality seeds. Finally, both buyers and sellers
recognize that seed laws have been enacted to aid in the orderly marketing of seed based on the
principle oftruth-in-Iabeling. In some cases, differences in reported values on the seed label exist
and are litigated in court. The seed testing information and how it was acquired form the
foundation of these legal cases and emphasize the need for proper conduct and interpretation of
seed test results.
Because of the universal importance and value of seed, many organizations rely on the
results of routine seed testing. These organizations vary from local or state agencies to national
and international agencies. As a result, it is important that test procedures be standardized and
that results be widely reproducible. This means that the tests must be conducted under the same
conditions with uniform interpretations. The process toward standardization in the United States
began in 1905 when the Annual Appropriations Act was passed which gave the U.S.
Department of Agriculture authority to purchase s~:eds on the open market, to test and publish
the results, and to identify individuals or organizations found to market mislabeled seed. The
standardized testing of seeds required that a compendium of test procedures be developed. The
Rules for Testing Seeds of the Association ofOffic:ial Seed Analysts were developed to meet
this objective. The following considerations discuss some ofthe specifications put forth in those
rules. Individuals interested in a greater knowledge of the procedures for conducting a seed test
should consult the AOSA Seed Analyst Training Manual (McDonald et al. I 992).

OBTAINING THE SAMPLE

No seed analysis, regardless of how carefully or accurately accomplished, is any better

than the sample on which it is performed. The importance of a representative sample cannot be
overemphasized; that is, the sample must truthfully represent the quality of the seed lot from
which it is drawn. It is generally assumed that a seed lot is homogeneous. If this were the case,
it would be satisfactory to extract a portion (sample) from any portion of the seed lot and
presume that it represents the bulk of the seed. However, this seldom occurs. Seed lots are
almost never completely homogeneous for at least four reasons. First, heavy and light seeds
tend to segregate within the bulk or bag due to gravity, with heavier seeds being found
predominantly at the bottom of the container. St~cond, harvesting of the crop in the field
combines seed from differing locations, thus altering the composition of the seed as a result of
variations in maturity, lodging, disease, or the occurrence of weeds. Third, failure to adequately
blend two or more lots from differing locations at the time of bagging can result in seed lot
heterogeneity. Fourth, lack of uniformity in harvesting, storage, and conditioning results in seed
lot heterogeneity.
As a result of this heterogeneity in most seed lots, a lot must be sampled and the sample
must be representative ofthe lot. Sampling is usually done in two steps. First, the sample to be
submitted to a seed laboratory is drawn from the bulk seed lot and sent to the laboratory for
analysis. This is known as the submitted sample. Second, when it reaches the laboratory, it must
be divided further to a size that can be analyzed . This latter sample is used for the actual
analysis and is called the working sample.
318 Seed Testing

The Submitted Sample

The sample may be drawn at any time during seed conditioning or after the seed is offered
for sale. If drawn during conditioning, it may be taken by hand or small container at frequent
intervals during the conditioning operation or automatically drawn at specified intervals by a
mechanical sampler. Either technique, if done properly, will give a representative sample. Seed
is usually sampled for testing while still in storage or as it is offered for sale (Figure 15.1).
Because ofthe variety of ways in which seeds are stored and offered for sale, they may be found
in various types of containers, from small vegetable and flower seed packets, to boxes and cans
of grass seed, to large bulk lots of cereal grain seed. Regardless of the container, the seed lot
must be properly sampled so the sample is representative. Rules and procedures for sampling
under various conditions have been established by the Association of Official Seed Analysts and
the International Seed Testing Association. These rules provide for sampling by mechanical
samplers, by use of standard sampling probes or triers (Figure 15.2), by hand, or by taking the
entire container as the submitted sample.

The Sampling Process

Bulk Seed. A trier, or probe, is recommended for sampling bulk seed, although hand
sampling may also be performed if handsful are carefully and randomly taken from
well-distributed points throughout the bulk. Hand sampling is limited by the difficulty of
reaching all portions of large bulk lots, whereas large probes up to 72 in. in length can be used
to sample hard-to-reach locations within the seed lot.

Seed in Bags

When a seed lot consists of six bags or less, each bag should be sampled from
well-distributed points throughout the bags. When lots consist of more than six bags, samples
should be taken from five bags plus 10% of the remaining bags. Regardless of the lot size,
however, it is not necessary to sample from more than 30 bags. Here are some examples:

No. of bags in lot 5 7 10 23 50 100 200 300 400

No. of bags to sample 5 6 6 7 10 15 25 30 30

Seed in Small Containers. Seed in small containers should be sampled by taking at

random an entire unopened container from the supply in order to obtain the minimum amount
required for the working sample.

Subdividing the Sample

The sample drawn by any of the various techniques may be too large for the submitted
sample and should be divided further before submitting it to the laboratory. Further subdivision
should be done by a mechanical halving device, such as the Boerner or Gamet dividers. Absolute
care should be taken at this point to guard against introducing bias into the sample to be
 Seed Testing
319

Opens
periodically
to catch -~_~~~
sample.

Automatic
Sampler
To Sample Room -'~t+---.L.
c

Figure /5.1. Seed sampling techniques: (A) bag sampling; (B) bulk sampling; and (C) mechanical
sampling (A, courtesy of Bob Neumann).
320 Seed Testing

I
I
~
U II
I I iJ
Figure 15.2. Examples of sampling probes. On the far right is a "thiefprobe. "

submitted. During the subdividing process, there may be a tendency to unconsciously remove
stones, stems, damaged seeds, or even noxious weed seeds. Such deviations from the correct
sampling procedures can make all subsequent testing results meaningless.

Mailing the Sample

After the properly sized sample is obtained, it should be carefully labeled and placed in a
container suitable for mailing to the seed laboratory. Cloth, plastic, or paper bags are
acceptable; however, these should be placed inside a sturdy cardboard box that can be labeled
and can withstand the rigors of mailing. Each sample should be labeled as follows: (1) name and
address of owner, (2) crop kind and variety, (3) tests requested, and (4) lot number and number
and weight of containers (bags) in the lot.
Seed Testing 321
SUBSAMPLING

When the submitted sample arrives at the seed laboratory, it is entered in the official
logbook, assigned a number, and the accompanying information is recorded. The sample then
goes to the subsampling area of the laboratory where it is divided into working samples for the
various tests that will be performed. Here, the working samples for the purity examination,
noxious weed examination, germination, and other special tests are obtained. The remaining
portion of the sample is retained as an official sample in case future tests are desired. The weight
of the working sample for purity analysis is determined by the weight of seed required to
comprise a minimum of 2500 seeds and will vary greatly from small- to large-seeded species.

The Importance of Subsampling

Dividing procedures must be absolutely precise and unbiased if the test results are to be
meaningful. The working sample must accurately represent the sample submitted to the
laboratory, which in turn represents the seed lot only if sampling procedures were properly
followed. In contrast to sampling, sample dividing procedures are generally dependable, because
this operation is performed in the laboratory by professional, trained personnel, while sampling
from bulk seed lots is often done by persons who may not realize the importance of a
representative sample.

Subsampling Techniques

The Rules for Testing Seeds state only that the working sample shall be taken from the
submitted sample in such a manner that it will be representative. The actual procedure may be
either by manual or mechanical methods. Several hand methods are used. One very simple
method, often called the pie method, consists of spreading the sample on a clean, flat surface,
and dividing it into sections as if cutting a pie. Any of the sections, if randomly selected, may
be used alone or in combination with other sections as the working sample. Another hand
technique, called the random cup method, consists of placing a number of uniformly sized
thimbles or cups on a clean, flat surface and slowly pouring the sample so the seed is distributed
evenly over the flat surface filling the cups as the sc(!d is distributed. The working sample may
then be obtained by randomly selecting several of the cups until sufficient seed is obtained.
Mechanical halving devices (Figure 15.3) are most often used for subdividing and are
dependable for providing a representative sample. These are devices that divide the sample into
two equal portions, both in size and content. The working sample is obtained by dividing the
submitted sample one to several times until the prop~~r weight of the working sample is obtained.
Any of the divided portions may be combined and redivided to yield the proper-siZed working
sample.
Three mechanical dividers are commonly used for subsampling. Any ofthese dividers will
yield a representative sample; however, each has certain advantages over the others. The
Boerner divider is probably the most common; however, some chaffy grasses and other
non-free-flowing seeds will not flow through it. A Boerner divider consists ofa hopper, inverted
cone, and a series of baffles directing the seed into two spouts. The baffles form alternate
channels and spaces of equal width. They are arranged in a circle at their summit and are
directed inward and downward, the channels leading to one spout and the spaces to an opposite
322 Seed Testing

spout. A valve or gate at the base of the hopper retains the seed. When the valve is opened, the
seed falls through the spouts into the seed pans. The Gamet Precision divider requires electrical
power to operate and is more suitable for seeds that do not flow through the Boerner divider.
The Gamet Precision divider makes use of centrifugal force to mix and scatter the seeds over
the dividing surface. In this divider, the seed flows downward through a hopper onto a shallow
rubber cup. Upon rotation of the rubber cup by an electric motor, the seeds are thrown out by
centrifugal force and fall downward. The circle or area where the seed falls is equally divided
into two parts by a stationary baffle so that one-half the seeds fall into one spout and one-half
into the other. For non-free-flowing chaftY grass seeds, such as gramma grass and needle grass,
the Hay-Bates or a similar divider should be used. For seeds of nondelinted cotton and certain
other species, hand-dividing methods of subsampling may be necessary; however, extreme
caution must be taken to obtain a representative sample.

PURITY TESTING

When individuals purchase seed, one of the primary decisions that they make is what kind
of seed is needed. Their purchase is made with the understanding that the species and variety are
the principal constituents of the seed lot. Yet, we know that harvesting and cleaning of seed are
not exact sciences. Other types of seed and materials are almost always present. The type and

Figure 15.3. Subsampling dividers: on the left is a Gamet Precision divider and on the right is a
Boerner divider (Courtesy of Bob Neumann).
Seed Testing 323
level of contamination of these other components can significantly influence the value of the
seed. The purity test, therefore, is designed to identify what these contaminants are and how
much of them are present.
Seed purity denotes the composition of a particular seed lot. It is based on a physical
determination of the components present and includes percentages by weight of: (1) pure seed,
(2) other crop seed, (3) weed seed, and (4) inert matter. Pure seed is the portion of the working
sample represented by the crop species for which the lot is being tested; in actual practice, it
includes the percentage of each crop species present in levels of 5% or more. Other crop seed
is the percentage of crop seeds, other than the species being tested, present in concentrations of
less than 5%. Weed seed indicates the percentage of seeds present from plants considered as
weeds. Sometimes this designation may be arbitrary, since a plant may be considered a crop in
one state or country but a weed elsewhere. For any particular region, however, the analyst uses
well-accepted guidelines for classifying as crops or weeds. Inert matter denotes the portion of
the sample that is not seed. It usually consists of chaff, stems, and small stones, but may include
pieces of broken, damaged, or immature crop or weed seeds that do not qualifY as entire seeds.
The criteria for this distinction are explicit and defined in the Rules for Testing Seeds (AOSA
2000).
The size of the sample on which the purity examination is performed is given in the Rules
for Testing Seeds. The sample size (weight) is determined by the size of seed being tested;
approximately 2500 seeds are considered acceptable to yield a statistically acceptable test
result. The size of the working sample specified in the Rules is larger for large-seeded crops than
for small-seeded crops; however, the actual number of seeds in the test is not greatly different.

Philosophy of Purity Testing

The philosophy of purity testing is to avoid judging whether seeds are capable of
germinating when performing the test. Consequently, shriveled, immature, frosted, or otherwise
damaged seeds are considered as pure seed. This may be called the quick method, in contrast
to the strong method used earlier by European seed analysts, who classified crop seed as pure
seed only if it appeared to be capable of germination. The quick method left the determination
of germination capability to the germination analyst, but is no longer used.

Procedures for Purity Separations

The purity test is perhaps the most complex and exacting of all tests for seed quality. A
seed analyst must have a comprehensive knowledge of seed structure and function and must be
able to identify a wide array of differing species. For this reason, it is not uncommon to find
that many seed analysts have their own working seed herbaria to assist in the
identification of unknown samples. Seed herbaria are useful because published descriptions of
seeds are rarely as comprehensive as those for plants and viewing actual specimens can be very
helpful. An average seed herbarium contains 2,000-3,000 specimens, but varies in size
depending on the range of seed materials typically ,encountered in the seed testing laboratory.
The herbarium can be arranged alphabetically or by phylogeny. Phylogenetic arrangements are
by plant families and then according to species within the family. This approach assures that
specimens which are closely related are placed together. Its disadvantage is that many seed
analysts are not familiar with taxonomic relationships which vary according to the authority
324 Seed Testing

Figure 15.4. A purity testing station (Courtesy of Bob Neumann).

used to classifY them. As a result, some analysts simply file specimens alphabetically according
to family and then species. This approach offers the advantage of rapid retrieval of specimens
but does not afford the opportunity for direct comparisons with related groups.
Purity separations are usually made by hand, although mechanical aids may be used to
speed up the analysis or make it less tiring for the analyst. Purity analysis equipment usuaIly
includes a work board, adequate light, forceps, a hand lens, and a stereoscopic microscope for
identifYing small seeds.
Seed Testing 325
Various mechanical devices are frequently used to aid in the purity analysis. The use of
seed blowers (Figure 15.5) has contributed to speeding up and improving uniformity of purity
analyses, especially for grass seeds, for use in separating empty florets, bits of stems and leaves,
chaff, and other inert material. The Rules for Testing Seeds provide uniform blowing procedures
for the purity determinations of small-seeded grass species in lieu of the more time-consuming
hand separation procedures. The blowing procedun~ not only speeds up the test but reduces
variability among seed laboratory results.
Another difficult area for purity testing is caused by the increased use of coated and
pelleted seeds (see Chapter 13 on Seed Enhancements). The Rules for Testing Seeds specify
coated seeds as a seed unit covered with any substan(:e which changes the size, shape or weight
of the original seed (seeds coated with ingredients such as, but not limited to, rhizobia, dyes and
pesticides are excluded). This process alters the shape of flat seeds that are difficult to
mechanically plant, for example, and makes them round by adding clay fillers so that the
pelleted seed can easily roll into planters. While seed coatings are extremely valuable from a
practical perspective, they make the purity analysis more difficult. The analyst must first
separate the coating material (often 90% of the dry weight of the seed sample) to determine what
is actually present in the clay pellet. This separated portion is weighed as other components of
a purity test and recorded as "coating material." To aid in purity testing, seed scientists at

Figure 15.5. Seed blowers: (left) the General seed blower, (middle) the Ottawa blower, and (right) the
South Dakota blower (Courtesy of Bob Neumann).
326 Seed Testing
Oregon State University have developed a microscopic inspection station, as well as a
vibrator-separator to help purity analysts in making their separations. This type of equipment
is used in several North American seed laboratories.

NOXIOUS WEED SEED EXAMINATIONS

Each state has established an official list of noxious weed seeds. In general, the plants
from these seeds are particularly troublesome and objectionable. Such lists are a part of the
state seed law (or regulation) and are usually defined in two categories~ primary (or prohibited)
and secondary (or restricted) noxious weed seeds. Sale of seed lots containing primary (or
prohibited) noxious weed seeds is prohibited), while the sale of lots containing secondary (or
restricted) noxious weed seeds is permitted, but their number per pound (of crop seed) is limited.
Since each state has its own seed law, the weed seeds listed as noxious are not necessarily the
same from state to state.
The noxious weed seed examination is an attempt to provide special information about
noxious weed seed content of seed lots. Because of the limited occurrence of noxious weed
seeds, a large working sample is used for the noxious weed seed examination-often as much as
500 grams for large-seeded types down to 2.5 grams for small-seeded types. The occurrence
of noxious weed seeds in the United States is reported in number of seeds per pound or ounce.
Since state seed laws and noxious weed seed lists vary among states, the results of this
examination may be reported for only one state, or it may be reported for two or more states.
Crop seeds transported in interstate commerce should have an "all-state" noxious weed seed
examination.

GERMINATION TESTING

Probably the single most convincing and acceptable index of seed quality is the ability to
germinate. Seeds are tested for germination because a seed lot is composed of a population of
individual seed units, each possessing its own distinct capability to grow and produce a mature
plant. A seed germination test is an analytical procedure to evaluate seed viability and
germination under standardized (favorable) conditions. It enables a seed vendor to determine and
compare the quality of a seed lot before it is marketed to the consumer. Furthermore, the percent
germination can be used to determine the planting value of a seed lot, its storage potential, and
labeling information required to provide for standardized marketing of seed lots. Thus,
germination testing is perhaps the single most important function of a seed testing laboratory.
Since the process of seed germination is covered in Chapter 5, this discussion will cover only
the laboratory techniques used for performing the analysis.

Procedures for Germination Testing

The germination test is ordinarily performed on the pure seed of the crop kinds that
constitute 5% (or more) of the sample after all inert matter and other crop and weed seeds are
removed. Each pure seed kind is germinated and reported separately. A minimum sample of 400
seeds is recommended for a statistically dependable germination test. These are usually planted
in four replicates of 100 seeds each, although various other arrangements are sometimes used
(Figure 15.6). Each replicate is evaluated separately, but the official germination report is an
Seed Testing 327

Figure 15.6. Using a vacuum headfor preparing 1OO-seed replicates for germination testing
(Courtesy of Bob Neumann).
328 Seed Testing
average of all replicates. The exact procedures and regimes under which different kinds of seeds
are germinated have been developed throughout more than 100 years of experience in
germination testing and have been augmented during the last 40 years by a systematic program
of referee testing involving interchange of samples and results among laboratories. The testing
instructions given in the Rules for Testing Seeds include the germination media (substrata), the
temperature required, the duration of the test period, and additional suggestions for optimal
resu Its (F igure 15.7).
The time required for germination tests varies among species. Some seeds require less than
seven days, while others may require a month or longer. The seeds of some trees and woody
shrubs are notorious for their long germination requirements. The Rules for Testing Seeds also
specify germination requirements for tree, vegetable, woody shrub, and flower species.
However, knowledge of germination requirements, especially for wild and exotic species, is not
complete.

Evaluation of Germination

At the end of the prescribed germination period, the tests are evaluated; however, it is
sometimes desirable to make preliminary evaluations, called first counts. Seedlings that have
germinated and are normal are counted and removed from the substrate at the time of the first

Figure 15.7. A light-equipped seed germinator for testing seeds that require light for best
germination (Courtesy of Bob Neumann).
Seed Testing 329
count. This procedure helps subsequent counts, because early-germinating seedlings often tend
to grow profusely, causing difficulty in evaluating later-germinating seedlings. Seeds that remain
ungerminated at the end of the prescribed period are considered dead or dormant (refer to the
discussion on dormant seed in Chapter 7).
The 'Normal Seedling.' The seed analyst has a somewhat different concept of seed
germination than the layperson, to whom germination implies the rupture of the seed coat and
the emergence of the root and shoot apices. The Rules of the AOSA, which most seed analysts
in North America follow, prescribe the following defmition of germination, embodying the
normal seedling concept: "the emergence and development from the seed embryo of those
essential structures which, for the kind of seed in question, are indicative of the ability to
produce a normal plant under favorable conditions." Not only does this concept include the
layperson's definition of germination, it also reflects the agronomic value of the seed (i.e.,
capacity to produce normal plants under favorablt: conditions).
Abnormal Seedlings. Any seedling that is not classified as a normal seedling is
considered abnormal. The germination analyst may classify seedlings as abnormal for various
reasons; for example, the absence of certain essential structures (such as radicle, epicotyl,
twisted, or otherwise abnormal shape, to greatly reduced growth or seedling vigor. The ability
of a seed analyst to discriminate between and classify normal and abnormal seedlings is one of
the most subjective aspects of seed testing. Therefore, constant education and training are
required to assure uniformity in interpretations. To help provide uniformity, the AOSA
developed a Seedling Evaluation Handbook in 1992 which is now recognized as a formal
component of the Rules for Testing Seeds. The Handbook provides line drawings depicting
differences between normal and abnormal seedlings to help analysts in discriminating among
questionab Ie seedlings (Figure 15.8).
Firm, Ungerminated (Dormant) Seeds. Seeds other than hard seeds that remain firm
(nondecayed) and ungerminated at the end of the prescribed germination period are called firm,
ungerminated seeds. This is a type of dormancy commonly found in certain grasses, and should
be treated appropriately to stimulate germination.
Hard Seeds. Hard seeds are those that do not imbibe water and therefore remain hard at
the end of the prescribed germination period. Hard-seededness is a type of dormancy that
prevents germination of viable seeds because they cannot absorb water through their
impermeable seed coat. The percentage of hard seeds is reported as part ofthe total percentage
germination.

Laboratory Methods of Breaking Dormancy

Any time a seed fails to germinate in the time: specified in the Rules for Testing Seeds, the
analyst must determine whether the seed was ungelminable due to lack of viability or dormancy.
If the seed does not appear diseased, it is probably dormant. Once it is recognized that
dormancy exists, the challenge to the seed analyst is to determine approaches that can break the
dormancy. Since seeds have evolved many unique ways to maintain dormancy, the analyst must
employ various approaches to break the dormancy-imposing mechanism(s). In some cases, a
single treatment may be effective. In others, a combination of techniques may be necessary. The
Rules for Testing Seeds specify appropriate dormancy-breaking techniques for species where
dormancy commonly occurs. These usually are either by prechilling or the use of KN0 3 •
330 Seed Testing

,.---___ Tenninal bud

Epicotyl
~._. .,,- Primary leaf

_____ Cotyledons

- -_ _ _ Hypocotyl

11.1 a Sand test 11.1 b Towel test

Secondary root

8----11-- Primary root

Figure 15.8. Soybean seven-day seedlings, sand test and towel test. (From AOSA Seedling Evaluation
Handbook, 1992. Contribution No. 35 to the Handbook on Seed Testing. Association of Official Seed
Analysts. 101 pp.).

Prechilling. Viable seeds other than hard seeds can often be stimulated to germinate by
a cold treatment of the water-imbibed seeds, commonly called prechilling, or stratification. This
is accomplished by placing the seed on or in moist substrata at relatively low temperatures
(about 5°C) for a specified time-usually about five days; longer durations may be necessary for
the seeds of some species (e.g., woody species). The experienced seed analyst recognizes those
species in which dormancy is likely to occur and routinely prechills them as a standard part of
the laboratory procedure.
Seed Testing 331
Potassium Nitrate (KN03). Seed germination in many species, such as turf grasses, can
be stimulated by using a dilute solution (0.1 % to 1.0%) of potassium nitrate as moisture for the
germination test. Like prechilling, the use of KN0 3., is a valuable aid in germination ofthose
species benefited by it and has become a routine procedure in the germination testing of many
species.

SPECIAL TESTS FOR SEED QUALITY

Although purity, germination, and noxious weed evaluations are routinely performed on
almost every seed sample submitted to the laboratory, many additional tests also reflect seed
quality. Such special tests are usually performed only when requested; however, they may be
done routinely for certain species or for law enforcement or certification samples. These special
tests have been developed as byproducts of routine testing procedures in the seed technicians'
attempts to learn more about the quality of seed lots. Today, most modem, well-equipped seed
laboratories have the capability of conducting such tests.

Genetic Purity Testing

Changes are rapidly occurring in agriculture, many of these at the level of the seed
industry. The ability to develop new varieties that differ in all but a single or several genes
places an even greater burden on genetic purity testing. It seems certain that seed products
developed from molecular biology will become increasingly common because they benefit
numerous people. For example, farmers will obtain higher crop yields from improved insect,
weed and disease control. Because these controls are obtained without chemical use, less
concern will exist about environmental pollution. Farmers will also benefit from lower input
costs for pest/weed control and will likely obtain premiums for seeds with selective output traits.
Seed companies also will benefit from increased seed premiums that will enhance seed margins.
Those companies that are the research and development leaders wi1llikely enjoy a market share
advantage from being the first to offer these new products. Finally, gene providers will obtain
additional income from per acre technology fees and., in some cases, increased herbicide market
share for companies selling seeds of herbicide tolerant varieties.
The continued development of new and improved varieties is the cornerstone of increases
in crop yield and agricultural productivity. By definition, a variety of a cultivated crop differs
from other varieties of the same species in one or more specific characteristic(s). Such
characteristics as maturity, lodging resistance, disease resistance, plant height, and market
quality make varieties distinct from one another. More recently, advances in molecular biology
have led to the release of new varieties that may differ in as little as one gene for a specific trait
such as herbicide tolerance or insect resistance. Farmers and growers are vitally interested in
the selection of the variety best suited to their particular field/greenhouse conditions because
they recognize that this single decision can have a marked effect on their yields and profit.
Genetic purity testing is so important that it has been the subject of a recent book (Wrigley
1995), several reviews (McDonald 1995; 1998; Smith and Register 1998; Cooke 1995; 1998)
and specific genetic purity testing protocols hav(~ been outlined in the Seed Technologist
Training Manual (Society of Commercial Seed Tec:hnologists, 2001) and the Cultivar Purity
Testing Handbook (AOSA 1991).
When new varieties are developed by plant bre:eders, a limited amount of seed is increased
to quantities sufficient to supply larger grower needs. As this seed is increased, it must be
332 Seed Testing
monitored to ensure that the genetic purity of the breeder seed is not compromised. Two
principal concerns exist in maintaining genetic purity. First, the genetic composition of the
variety initially developed by the breeder must be the same as that marketed to the grower after
several regenerations of seed increase. Second, for hybrid seed crops, the success of
hybridization must be ensured by minimizing the percentage of selfing and outcrossing.
In the early days of seed testing, varietal tests, when conducted, were relatively simple for
two basic reasons: (1) there were fewer varieties, and (2) there were usually greater differences
among varieties. Because of the success of modern plant breeding, the resulting variety
explosion, and the appearance of many closely related varieties, seed analysts have been obliged
to find newer, more sophisticated ways of distinguishing among varieties in the seed laboratory.
Within varieties, genetic purity testing is important so that (Smith and Register, 1998): (I)
intellectual property protection through Plant Variety Protection or Utility Patents can be
obtained and then subsequently maintained, (2) varieties can be created with uniform appearance
and agronomic performance that meet the demands of farmers, processors and consumers, (3)
varieties with stable genetic identities can be created so that plant performance can be as
predictable as possible given unpredictable environmental fluctuations, and (4) breeders can
more completely and precisely characterize and measure genetic diversity so that genetic
resources can be thoroughly evaluated in terms of plant performance and effectively utilized for
the creation of improved varieties.
Recognizing the importance of these new markets and new agricultural products, seed
technology will necessarily be at the forefront of ensuring the genetic purity of these seed
products. Moreover, the increasing value of seeds in the future portends that high quality seeds
will be paramount to avoid litigation concerning inaccurate identification of varieties. So, seed
technologists are challenged to develop an array of more sophisticated genetic purity tests. In
some cases, this may be a rather simple process such as imbibing seeds/seedlings in an herbicide
to determine their tolerance to the compound. In most cases, however, when only a single gene
is modified, more powerful genetic purity tests may be required. Some of these may include the
use of immunoassays to detect the proteins produced by the inserted genes. Other approaches
may employ the use of electrophoresis on starch or polyacrylamide gels to separate an array of
specific proteins/enzymes. Even more sophisticated approaches include newer DNA-based
technologies that use the polymerase chain reaction to allow even more discrimination and faster
identification of varieties. At the moment, this area is rapidly changing and it is difficult to
anticipate which of these tests will provide the seed technologist the greatest benefit in genetic
purity testing.

Criteria for a Genetic Purity Test

The ideal genetic purity test must meet four criteria (Payne 1986). First, results must be
easy to reproduce, not only within a laboratory, but also among different laboratories. Second,
it should be technically uncomplicated so seed technologists can be successfully trained to
conduct the test in a minimal amount of time. Third, it should require only a short time to
complete. Finally, it should be inexpensive to conduct.
The basic objective of a genetic purity test is to test for the occurrence of traits that help
identify a particular variety when grown in different environmental conditions and generations.
Thus, it is assumed that these characteristics are environmentally stable and will not change
from one generation to the next. The following approaches represent some of the most
promising genetic purity tests currently conducted by the seed industry.
Seed Testing 333
Types of Genetic Purity Tests

Field Testing. Traditionally, breeders, seed companies and certification agencies

determine genetic purity using physical traits expressed by the seedling and/or mature plant.
However, the success of field tests is limited because environmental stress often masks or
alters specific seedling and plant anatomical/morphological features. For field testing to be
successful, genetic purity tests must be grown (1) in areas where the crop is well adapted, (2)
under the best possible cultural practices, and (3) during the proper growing season.
Otherwise, the development ofthe crop may be altered to such an extent that accurate genetic
purity results become uncertain. Field testing is also expensive: requiring equipment, planting
and harvesting personnel, land use, and training of competent technicians for detecting
specific plant traits. Most important, the number of morphological traits available today is
limited and many supplemental laboratory tests havl~ been developed and, in some cases, have
completely replaced greenhouse and field testing.
Morphological. Distinguishing morphological features has been a major component of
genetic purity testing in the laboratory. Morphological studies of the seed, seedling, and
mature crop are used.
Seed. This is the simplest type of genetic puri~y test and probably the earliest used. Such
characteristics as seed size, hilum color (Figure 15.9), seed shape, shape of rachilla, lemma
and palea characteristics and presence or absence of awns are often evaluated. Although still
useful, visual observation of the seed is usually not reliable for positive genetic purity testing
and should be used only in conjunction with other tests. For example, Chippewa 64 soybean
can easily be distinguished from soybean varieties that do not have a black hilum. However,
more sophisticated techniques must be used to distinguish it from other varieties that also have
black hila. Seed size is also a useful index of variety; however, it is so variable and so
environmentally influenced that it must be used only with extreme caution.
Seedling. Many useful genetic purity tests can be performed on seedlings. Such
characteristics as hypocotyl color (Figure 15.10), leaf coloration pattern, vernation (folded

Figure 15.9. Five different hilum colors of soybean seeds. Differences can be observed, even in this
black and white illustration, hut are much more distinct when shown in color.
334 Seed Testing

Figure 15.10. Soybean hypocotyl pigmentation patterns: Dark purple, intermediate purple, bronze,
and green (left to right;from AOSA Cultivar Purity Testing Handbook, 1991).

or rolled) pattern of the leaf, length of internodes (dwarf vs. normal), pubescence and leaf shape
are often examined. Such tests are useful because they may yield more information than do
observations ofthe ungerminated seed and do not require as much time as field grow-out tests.
Crop. Traditionally, distinctions in flower color, stem pubescence color, leaf shape,
photoperiodic responses, disease resistance, maturity date, and growth habit are genetic purity
traits examined in the greenhouse or field. Greenhouse tests are usually performed in
conjunction with seed and seedling tests to substantiate decisions made earlier. Growing plants
in the greenhouse, however, involves many of the undesirable characteristics of field testing,
e.g., space, time and expense.
Other types of genetic purity tests conducted in the laboratory include ultraviolet light tests,
chemical assays, chromosome counts, chromatographic methods, herbicide tolerance, ELISA,
electrophoresis of proteins/enzymes, and polymerase chain reaction technologies.
Ultraviolet Light Tests. Response under ultraviolet light has been used for both seed and
seedling variety tests with varying success. The lemma, palea, and glumes of certain oat
varieties contain substances that fluoresce when exposed to ultraviolet light. The ultraviolet
light test, however, has limited usefulness, because many oat varieties show the same response
- either fluorescence or nonfluorescence; therefore, the test is useful only when two varieties
with opposite responses are being compared.
Seed Testing 335

For many years, the fluorescence test of ryegrass has been used with great success to help
distinguish between two species - annual or Italian (Lotium multiflorum) and perennial or
English (Lolium perenne) ryegrass. The past usefulness of this test was due to the fact that
seedling roots of all known annual ryegrass varieties exhibited a positive response, while
seedling roots of perennial varieties were nonfluorescent. It may be otherwise impossible to
distinguish between annual and perennial ryegrass seeds, although the presence of an awn
usually indicates annual ryegrass. However, this characteristic is not dependable, since the awn
is quite fragile and may be detached by handling. The accuracy of the fluorescence test has been
good enough in the past to use as a measurement of the percentage of the two species when
found together in mixtures. Today, however, it is believed that crossing between annual and
perennial species in the field has made the fluorescence test of seedlings less meaningful and a
search for new tests to distinguish between annual and perennial ryegrass is underway.
Unlike the fluorescence test of oats, which dt::pends on the fluorescence of the seed coat
under ultraviolet light, the test for ryegrass is conducted on five- to ten-day-old seedlings grown
on white filter paper. The fluorescent substance has been isolated and designated as annuoline.
[t appears as an exudate from the ryegrass roots that are in contact with the paper medium.
Chemical Assays. The ideal genetic purity test would utilize the exposure of a seed to
some chemical that would clearly reveal its varietal identity in comparison with related varieties.
Unfortunately, such a test does not exist. Several easily performed chemical assays have been
developed for use in genetic purity testing. F or example, the phenol test (Elekes 1980) is
dependent upon a flavenoid reaction in the seed pericarp and has been used successfully for
distinguishing wheat and bluegrass varieties. The test is performed by placing the seeds on a
paper medium moistened with approximately I % carbolic acid (phenol solution) for about four
hours. Tests are evaluated according to the darkness of staining that occurs; the seeds of some
bluegrasses stain in the embryo area and can be distinguished from nonstaining varieties, while
in wheat the entire pericarp is observed for the degree of staining. Wheat varieties can be
categorized according to whether they stain very dark, medium dark, very light, or remain
unstained (Figure 15.11).
Another very useful chemical test for distinguishing soybean varieties is the peroxidase test
that separates varieties into two groups based on the presence or absence (Figure 15.12) of the
enzyme in the seed coat (Buttery and Buzzell 1968). The test is conducted by removing the seed
coat from the seed and placing it into a test tube to which is added 10 drops of a 0.5% guaiacol
solution for 10 minutes. After that period, one drop of a 0.1 % H20 2 is added to the test. A
negative test will be indicated by a colorless solution.
Other useful chemical tests include the hydrochloric acid test for oats, sodium hydroxide
test for wheat, and potassium hydroxide test for red rice and sorghum. The procedures are
provided in the AOSA Cultivar Purity Testing Handbook (AOSA 1991).
Chromosome Counts. During the 1960s, many new tetraploid grass and legume varieties
were released in the United States. The double chromosome complement of these varieties
compared to their diploid counterparts has provided seed analysts with a built-in genetic purity
test by merely counting the number of chromosomes in seedling root tips (Figure 15.13; Will,
et al. 1967). Like many other genetic purity tests, chromosome counts cannot be used to
distinguish between different varieties with the same chromosome number. However, they are
useful in detecting contamination, especially diploiid contamination of tetraploid varieties. The
tests are a valuable aid in monitoring the genetic purity of certain varieties of certified tetraploid
grasses and legumes.
336 Seed Testing

Ivory Fawn

Figure J5. 1 J. The phenol test for wheat: Examples ofthe five different color categories (From A OSA
Cultivar Purity Testing Handbook /99/).
Seed Testing 337

Figure 15.12. The peroxidase test for soybean. Peroxidase positive (left two rows), peroxidase
negative (right two rows) (from AOSA Cultivar Purity Testing Handbook, 1991).

The use of chromosome count tests is more limited for higher polyploid species, such as
wheat and bluegrasses, because oftheir difficulty associated with counting the chromosomes and
the complexity of their polyploidy.
Chromatographic Methods. Both thin-layer and paper chromatography have been used
with varying success in genetic purity testing. Readily observable differences have been
reported in thin-layer chromatographic bands among both ryegrass and soybean varieties, and,
to a lesser extent, in oat varieties. Thin-layer chromatographic methods have also been used to
distinguish soft white from durum wheat and between closely related members of the genus
Trifolium. Paper chromatography has been used to aid in genetic purity testing of Brassica and
other species (Payne 1986).
In routine genetic purity testing, the usefulness of chromatographic techniques is limited,
primarily because it is difficult to use, a long time is required for the test, and also because
different varieties do not always have observable differences in chromatographic bands. This
latter problem can sometimes be overcome by refinement of the technique, for example, by use
of different absorbents (thin layer), developers, and ultraviolet light to help distinguish
differences on the chromatograph.
Herbicide Tolerance. The first herbicide tolerance test was a seed soak method for
Sulfonylurea tolerant soybean (STS) seeds (Sebastion and Chaleff 1987). However, following
the introduction of Roundup ReadyTM soybean seeds in the 1990s, an increased emphasis on
detection of those soybean seeds that possessed the trait was essential. For example, should a
farmer believe that Roundup Ready soybean seed had been planted, but were not, subsequent
338 Seed Testing

A B

Figure 15.13. Metaphase chromosomes ofryegrass: (AJ diploid ryegrass, with 14 chromosomes, and
(B) tetraploid ryegrass, with 28 chromosomes (From Will et al. 1967).

spraying of the nonselective herbicide would result in a complete crop failure. Seed
technologists quickly identified three approaches to successfully determine herbicide tolerance.
These included presoak, substrate imbibition and seedling spray tests.
Presoak. This method allows the seeds to soak in a solution of the herbicide for a
predetermined interval. The seeds are then planted and germinated under normal conditions and
susceptibility to the herbicide determined. The advantage of the presoak method is that there
is less contamination of facilities, equipment, and waste with the herbicide.
Substrate Imbibition. Grote (1992) developed the original substrate imbibition test for
testing imidazolinone tolerant corn and Gutormson (1999) published a 13-step method for
testing Roundup Ready corn. In this method, the germination medium/substrate is soaked with
the herbicide followed by placing the seeds on the moistened medium. This approach allows the
seeds to be exposed to the herbicide throughout the duration of the test. The herbicide
concentrations used are usually less than those used in the field since the non-trait
seeds/seedlings must emerge to express non-trait symptoms in their growth and anatomy. The
advantages of the substrate imbibition test are the automation of the method, ease of including
a check sample with each replicate, and less steps to plant. Important disadvantages include the
requirement for dedicated equipment to avoid herbicide toxicity to other seedlings and
environmental concerns regarding the disposal of media containing the herbicides.
Seedling Spray. Seedling spray methods involve growing seedlings in a laboratory or
greenhouse and spraying the normal seedlings with the herbicide solution. After several days,
the susceptibility or tolerance to the herbicide can be determined (Figure 15.14). Advantages
of this test are that it relates well to field conditions and seed quality is of a concern since
only emerged seedlings are sprayed. Disadvantages include the increased cost of a
laboratory/greenhouse test and the additional time required for seedling development.
Seed Testing 339

Figure 15.14. Seedling spray tests conducted in a greenhouse on Roundup Ready soybeans. Seedlings
in the front are non-trait and susceptible to the herbicide while seedlings in the back possess the
tolerance trait and are not susceptible to the herbicide (Courtesy of Illinois Crop Improvement
Association).

Enzyme Linked Immunosorbant Assay (ELISA). Antibodies are substances that

recognize the unique molecular structure of proteins or antigens and then bind to them. These
antibodies are very specific and bind on Iy to specific types of proteins. Thus, if a corn seed has
the 8t gene for insect resistance, it will produce a specific 8t protein that an appropriate
antibody binds to. However, since the bound antibody and antigen are too small to visualize,
an enzyme label is added to the mixture that generates a visible color change in a substrate if
the enzyme successfully combines with the antibody/antigen complex (Figure 15.15). The color
reaction for each seed can be read by eye in 96-well microtiter plates. Commercially available
ELISA kits and reagents are available for genetically modified crops such as 8t corn and
Roundup ReadyTM soybeans.
Electrophoresis. Electrophoresis of seed proteins/enzymes is a highly versatile approach
to genetic purity testing of seeds--so much so that this technology has been incorporated into the
Rules of the International Seed Testing Association and has been described in the Association
of Official Seed Analysts' Cultivar Purity Testing Handbook (1991). Most electrophoretic
systems employ either starch or polyacrylamide gels as the preferred media in which protein
separations based on molecular size and charge density are made. It produces a separation of
seed proteins or isoenzymes in the media by establishing an electric field and permitting the
proteins to arrange themselves according to their polar (positive or negative electrical charge)
nature; those with a more positive charge will align themselves near the positive pole. After the
protein pattern has been established, it can be photographed and compared with the patterns of
340 Seed Testing

1. 2. 3. 4.

+ +>-E ~>-E -<+)-E.

•
•+
0
a

-<.
~
*-£ a
a
~)-E.
•
• I
a
•

+•
m (.--f 0 He
•
--< Antibody Sample
(antigen)
)--E Enzyme
conjugate a Substrate .Catalysed
substrate

Figure 15.15. Diagram of a double antibody sandwich ELISA test (Courtesy of Ag Dia, Inc.).

known varieties. Another approach is the use of isoelectrically focused gels that separate
proteins based on their position within a pH gradient; the protein ultimately coming to
equilibrium at a pH where the molecule is no longer charged (isoelectric point or pl). All of
these systems possess advantages and disadvantages from a commercial perspective (Table
15.1). Principal among these is the ability to evaluate sufficient numbers of seeds at the lowest
cost. Starch gel electrophoresis systems have been described that allow adequate numbers of
seeds (usually 80) to be run at one time (Stuber et al. 1988). More importantly, starch gels can
be molded that permit up to five slices of the parent gel, each gel slice being evaluated for a
different enzyme staining pattern (Figure 15.16). This system enables the equivalent of five
electrophoretic evaluations of the same seed samples, thereby significantly reducing costs and
analytical time. Starch gel electrophoresis of maize seed proteins provides reproducible and
standardized results within and among seed testing laboratories and is commonly used in
quality control programs in the maize seed industry (McDonald 1990). Other approaches have
been identified to make electrophoresis more efficient and cost effective. One technique is to
miniaturize and computerize the electrophoresis process. This results in less time being
committed to electrophoresis, staining and destaining of gels. Such an approach has been
developed, evaluated and found commercial application (McDonald and Drake 1990). Another
strategy is to utilize non-denaturing isoelectric focusing gels that cannot be sliced and blot the
proteins from the parent gel onto a nitrocellulose transfer membrane. This blotting technique
allows multiple enzyme staining patterns from one electrophoretic run (Figure 15.17) and
provides greater resolution of banding patterns than obtained on a starch gel (McDonald 1991).
Seed Testing 341
Table 15.1. Advantages and disadvantages of polyacrylamide, starch, and IEF (isoelectric focusing)
electrophoresis for genetic purity testing of seeds (McDonald, 1995).

Polyacrylamide Starch IEF

Advantages Technical system Technical system Technical system

well defined well defined well defined

Excellent band Inexpensive Uses charge (pI) rather

resolution than charge density
and size of proteins

Gels can be sliced Gels can be blotted

Short running time (1.5 h)

Disadvantages Expensive Standardization of gels Expensive

Cannot slice gel Long running time (5-6 h) Cannot slice gel

Potentially toxic Poor band resolution

Figure 15. 16. A starch gel stained for malate dehydrogenase isozymes. The banding patterns appear
vertically on the gel as lines. Each line represents isozymes from a single maize seed (Courtesy of
Novartis Seeds, Inc.).
342 Seed Testing

Figure 15.17. Blotting of oat cultivar seed proteins assayed for total protein (left) on the parent
isoelectrically focused gel with blotted membranes demonstrating peroxidase (middle). and esterase
(right) enzyme activity (McDonald 1991).

Polymerase Chain Reaction. Still, these techniques fail to differentiate a number of

varieties in some crops. Greater sensitivity in genotypic identification is desired. In 1990, a
new genetic assay called random amplified polymorphic DNA (RAPD) based on the uses
a single arbitrarily chosen oligonucleotide primer that hybridizes to the genomic DNA template
at two different sites, one on each strand of the complementary DNA. Under appropriate
temperature alternations, a thermostable DNA polymerase is able to synthesize discrete DNA
products (usually 200 to 2,000 base pairs long) that can be resolved on an agarose gel
following electrophoresis. Each primer has the capability to consistently direct amplification
of several unique DNA fragments in the genome. Some amplified fragments or patterns of
fragments may be unique to a genotype (Figure 15.18; Jianhua et al. 1996) and hence useful in
genetic purity testing. It should be emphasized that this area of seed technology is rapidly
advancing and the development of more robust and standardizab Ie genetic purity tests based on
DNA technologies is in the immediate future.
Other Cytological Methods. Although other cytological methods have not been used
extensively in genetic purity testing, they offer considerable potential. Cytological testing
methods should become more valuable as new ways are found to introduce and direct
chromosomal aberrations for creating new plant varieties having new predetermined
characteristics (for example, disease resistance). In such instances, the presence of known
chromosomal aberrations, such as deficiencies, duplications, inversions, and trans locations,
might be used for positive varietal identification.
Seed Testing 343

Figure 15.18. RAPD polymorphisms with primer 244 for 18 soybean cultivars. Mis the base pair lane
(Jianhua et al. 1996).

Disease Resistance for Varietal Identifica1:ion. A pathogen inoculation test can be

easily used to distinguish between disease-resistant and disease-susceptible crop varieties that
otherwise appear similar. The test has been sUlccessfully used to distinguish between
phytophthera- resistant and susceptible soybean varieties (Figure 15.19) and also in com
hybrids with different responses to southern com leaf blight - for example~ Texas male sterile
varieties versus normal cytoplasm varieties.

Determining the Effectiveness of Seed Treatments

This test is used to determine the effectiveness of seed treatment with chemical
pesticides. Although state and federal laws require treated seed to be dyed a contrasting color
(see Chapter 18), the effectiveness ofthe coloration does not indicate the completeness and
effectiveness of the treatment. The common method of testing the effectiveness of fungicide
treatment is to plate treated seed on agar media and apply a covering of Gibberella,
Glomerella, or Cingulata spores over the entire media surface. If seeds are ineffectively
treated, the spores should germinate and grow around the seeds. A clear zone soon appears
around each effectively treated seed where spore germination is prevented (Figure 15.20).

Effectiveness of Inoculation of Legume Seed

Tests for effectiveness of legume seed inoculation can be performed by the grow-out of
seeds in the greenhouse or in growth chambers and their comparison to well-inoculated control
samples. This test has been routinely performed in ]Indiana, and the information obtained used
in the enforcement ofthat state's seed law pertaining to preinoculated seed (Figure 15.21).
344 Seed Testing

Figure 15. 19. A test for resistance to Phythphthora root rot in two soybean varieties. The one on the
left is susceptible (Courtesy ofA. F. Schmitthenner).

Seed Moisture Test

Evaluation of seed moisture content is an extremely important determination in seed testing.

Knowledge ofthe seed moisture content is useful because it provides information regarding the
potential for harvesting, cleaning, and planting injury(ies) as well as the likelihood for successful
long-term storage. A seed moisture test is conducted by first weighing the seeds to determine
their "wet" weight. Then, the seeds are placed into an oven set for 100°C for grass, legume, and
cereal seeds or 85°C for tree seeds for 24 hours except for large or thick-coated pine seeds and
oily seeds such as Brassica species in which 48 hours at 85°C is required. After the drying
period, the seeds are removed from the oven, placed in a desiccator for 15 minutes to cool, and
the "dry" weight of the seeds determined. Two methods are used to express seed moisture
content and are calculated in the following way:

Wet Weight

Weight before drying - Weight after drying x 100 = % Moisture content

Weight before drying (Wet weight basis)

Dry Weight

Weight before drying - Weight after drying x 100 % Moisture content

Weight after drying (Dry weight basis)
Seed Testing 345

Figure 15. 20. Inhibition zones around seed q{an agar plaring [(!St indicate ellcctive seed trealment.

Figure 15.21. Left: plants from an effective humus inoculum applied to red clover seed at time of
planting. Right: ineffective pre inoculated seed (Courtesy of L. C. Shenberger).
346 Seed Testing

In the seed trade, seed moisture is described on a fresh weight basis. The value will never
exceed 100%. Research scientists often determine this value on a dry weight basis and the value
can often exceed 100%. Thus, it is important that the analyst be a ware of the specific procedure
followed when percent seed moisture content is determined, since the calculations can lead to
divergent results and interpretations.

SEED TESTING TOLERANCES

The importance of having the working sample be representative of the entire seed lot can
hardly be overemphasized since a nonrepresentative sample would provide erroneous results
about the quality of the seed Jot. Moreover, each time the working sample is reduced,
determinations of the seed lot quality can tend to become less accurate. In addition, a seed
sample is composed of individual biological units with their own inherent quality and
performance characteristic, so it is not surprising that variability in test results between two
working samples obtained from the same submitted sample would be obtained. Thus, variation
in test results from the same seed lot is expected and normal. But, how much variation in test
results is acceptable or tolerable? That is the purpose of seed testing tolerance.
Tolerances are used to define statistically acceptable limits within which different test
results may be expected to vary. Tolerances have been established for the more common tests
performed on seeds. They usually provide for the variability expected from random sampling
error and some account for variability caused by interpretational errors or seed lot heterogeneity.
Most studies of variation have shown that actual variability among different test results often
exceeds that accounted for by existing tolerances. Because of this, seed law enforcement
agencies sometimes allow "administrative tolerances" when determining if seed is improperly
labeled. When properly applied, tolerances specify when results are "out of tolerance" or if a
retest is necessary. Tolerances are based on the fact that the values reported have a probability
of error of 5%. Tolerance is defined as the difference permitted between a labeled percentage
(the first analysis) and the test results obtained by a laboratory when checking the accuracy of
the labeled information. Tolerances for purity, germination, fluorescence, and noxious weed seed
examinations are included in the Rules for Testing Seeds. The following example illustrates the
use of germination tolerances. A state seed inspector picked up a Merion bluegrass seed sample
from a lawn and garden store which was labeled as germinating 95%; an official germination
test in the seed laboratory showed the sample to germinate 87%. To determine if the sample is
mislabeled, both tests are given equal chance of being correct; thus they are averaged (95+87
12) to give a weighted mean of91%. The tolerance for the weighted mean of91% is 6. The
difference between the labeled germination and the second test is 8; consequently, the sample
is out of tolerance and the seed lot is considered to be mislabeled.

SEED TESTING ORGANIZATIONS

The importance of seeds as an agricultural commodity is mirrored in the complexity of

international, national, and state organizations concerned with assessing its quality (Figure
15.22). Many of these organizations are interested in the orderly movement of seeds from one
country or state where the seeds are produced to the next where they are used. Of particular
interest to seed analysts are those organizations devoted exclusively to seed testing. These
include ISTA, AOSA, the Society of Commercial Seed Technologists (SCST), the Commercial
Seed Testing 347

(Laws) (Certification) (Testing) (Testing) (Tr3de)

(Laws) (Growers) (Testing)

School of Agriculture

SELECTED SEED ORGANIZATIONS

AASCO - American Association of Seed Control Officials (Uniform Laws)

AOSA· Association of Official Seed Analysts (Standardize Testing)
AOSCA - Association of Official Seed Certifying Agencies (Standardize Certification)
ASTA - American Seed Trade Association
EEC· European Economic Community (Marketing)
FlS - Federation of International Seedsmen
FSA - Federal Seed Act (Regulates Interstate Seed Movement)
ISTA - Interernational Seed Testing Association (Standardize Testing)
OECD - Organization for Economic Cooperation and Development (International Certification)
RUSSL - Recommended Uniform State Seed Law (Standardize State Seed Laws)
SCST - Society of Commercial Seed Technologists
SR&T - Seed Regulatory and Testing

Figure 15.22. A typicaljlowchart illustrating important seed organizations at international, national

and stale levels (Courte,\y of Oregon State University Seed Lab).

Seed Analysts Association of Canada (CSAAC), and the International Society of Seed
Technologists (ISST).

International Seed Testing Association (www.seedtest.org)

The International Seed Testing Association (ISTA) is the only worldwide organization of
national laboratories dedicated to seed testing on an international scope. Its goals include: (1)
348 Seed Testing

development ofrules for seed testing, (2) standardization oftesting techniques, (3) seed research,
and (4) cooperation with other international agencies for seed improvement.
The 1STA had its beginnings in the early 1900s, when seed technicians from several
European laboratories felt the need for more exchange of seed testing information and
communication among seed laboratories in different countries. During this period, the
international seed trade was becoming established, creating the need for standardization of seed
quality concepts across national borders.
This need was first put into action at the 1905 Botanical Congress in Vienna, during
which several people met informally to plan a European seed testing association. Plans were
made for a Seed Testing Congress in Hamburg, Germany, in 1906. Another Seed Testing
Congress was held in 1910. Due to conditions in Europe, another meeting was not held until
Professor K. Dorph Peterson, of Copenhagen, called a Third Testing Congress in Copenhagen
in 1921, where the European Seed Testing Association was formed. Under the auspices of this
group, the Fourth International Seed Testing Congress was held in Cambridge, England, in
1924. At this meeting, the name was officially changed to the International Seed Testing
Association. Since its beginning, the 1STA has had great growth and accomp lishments. It has
become truly worldwide in both scope and representation. Membership in ISTA now includes
117 laboratories from 53 countries. Some of its notable accomplishments are:

1. In promoting uniformity of seed testing results among laboratories, it has facilitated

movement of seed across international boundaries and helped farmers get the best possible
seed regardless of the country of origin.
2. It has arranged for seed scientists and technicians to meet and discuss their problems and
to find solutions for them. By drafting seed testing rules and by discussing their
interpretations, they have provided a sound basis for enactment of seed laws to protect the
farmer.
3. It has helped to achieve closer association between test results and field performance,
assisting farmers to recognize seed of high planting value.
4. It has organized training courses and workshops in Europe, Africa, Asia, and South
America to help promote seed testing in areas of rapidly emerging agriculture.
5. It has provided a focal point of seed knowledge.

The 1STA holds a Congress every three years at different locations throughout the world
to hear scientific and technical papers from its members and to provide forums and committee
meetings for the exchange of information and the finding of solutions to mutual problems. The
complete activities at each congress are published in its official journal-Seed Science and
Technology (prior to 1972-ISTA Proceedings).

Association of Official Seed Analysts (www.aosaseed.com)

The Association of Official Seed Analysts (AOSA) is an organization composed of

analysts from official state, federal, and university seed laboratories throughout the United
States and Canada. Its contribution in bringing seed testing to a respected and highly
sophisticated level in these two countries has been enormous. Perhaps its greatest contribution
has been the development of rules and procedures for seed testing, and the standardization of
their interpretation. It also has had great influence on seed legislation in every state as
Seed Testing 349

well as at the federal level. The Referee Committee of AOSA distributes problem seed samples
to different laboratories for testing. Such activity hellps attain standardization in procedures and
interpretation among different laboratories.
The AOSA was formally organized in Washington, D.C. in 1908, with 16 states
represented. Since its early days, the AOSA has held annual meetings almost every year. The
minutes of its annual meetings and the papers presented are published in Seed Technology
(formerly Journal of Seed Technology; AOSA Proceedings). It also publishes a newsletter in
conjunction with SCST three times a year, which includes articles on seed testing topics. The
Association has published many special publications, among which are a series of handbooks
on selected topics.

Society of Commercial Seed Technologists (www.seedtechnology.net)

The Society of Commercial Seed Technologists (SCST) is a society of seed analysts from
private or commercial seed laboratories throughout the United States and Canada. This includes
self-employed seed analysts who test seed on a custom-fee basis and analysts from seed
companies, who ordinarily are salaried or on a commission, and who test seed handled in the
company's business.
The SCST originated in the early 1920s, largdy as a liaison between the AOSA and the
American Seed Trade Association (ASTA) because of their mutual need for better acquaintance
and communication. The AOSA had regarded the ASTA suspiciously because they felt that
some seed producers had flagrantly violated seed labeling laws and occasionally used fraudulent
merchandising schemes. ASTA members regarded the AOSA as a well-meaning, but highly
technical and regu latory-minded organization that promoted complex and often conflicting seed
legislation. By 1922, some of the larger seed companies had their own seed testing laboratories
and analysts. At the combined AOSA and ASTA meeting in Chicago in 1922, 13 commercial
seed analysts met to form what was first caned the American Society of Commercial Analysts.
From the time SCST was first organized, there was good cooperation between the SCST and
AOSA. These two organizations held their annual meetings at a common place, presented
papers, exchanged ideas, and participated in referee testing together. The AOSA welcomed the
new organization because it created a new bond of communication with the ASTA on a more
technical and professional level. The respect for the SCST was strengthened by the high
standards it established for society membership. In 1947, membership standards were further
strengthened by the estab lishment of a comprehens ive examination for membership. A minimum
score of 80 was established for passing the test, which included: (1) seed identification, (2)
purity and germination techniques, (3) evaluation of normal and abnormal seedlings, (4)
knowledge of botany, (5) Canadian and United States federal seed laws, (6) official rules for
seed testing and tolerances.
In addition, a combination of minimum college credit in the biological sciences and
experience in seed analysis was established as a requirement. Analysts who pass all
requirements and are accepted by a two-thirds vote by SCS T members have the right to use the
Society Seal and Insignia. The official seal is proofthat the SCST member is a Registered Seed
Technologist, and this becomes part of the analyst's credentials. Thereafter, the seal
accompanies the results of any test performed under his or her supervision.
350 Seed Testing

Commercial Seed Analysts Association of Canada (www.seedanalysts.com)

In 1944, six commercial seed analysts, formerly with the Toronto Seed Laboratory of the
Canada Department of Agriculture, met in Toronto and formed the (www.cdnseed.org) Ontario
Commercial Seed Analysts Association. The purposes of this organization were (1) to keep
abreast of new methods of seed testing, and (2) to assist analysts in overcoming any problems
that might arise in their work. Analysts from other Canadian provinces quickly showed an
interest in this association, and at the second meeting in 1945, the name was changed to the
Commercial Seed Analysts Association of Canada (CSAAC). The association has since grown
to around 40 members, with representatives from Ontario, Alberta, Quebec, the United States,
and England.
The Association holds its annual meetings in Toronto, and proceedings ofthis meeting are
published in the Maple Leaf, the official publication of CSAAC.
Most members of the GSAAC are also members of the Society of Commercial Seed
Technologists, so close communication is maintained between these two organizations. Members
ofCSAAC also attend meetings of the Association of Official Seed Analysts.

International Society of Seed Technologists (ISST) (www.seedtest.org)

The International Society of Seed Technologists (ISST) was formed in 1997. Its purposes
are to maintain and encourage the highest proficiency and professional standards among its
members; to promote seed technology research, teaching and extension activities; to promote
improvements in seed testing rules and procedures; to promote the best interests of domestic and
international seed trade; and to encourage cooperation between regulatory and commercial
agencies. The organization of ISST is based on the establishment of world-wide chapters
composed of seed technologists from differing countries. While ISTA is composed of
government-supported regulatory seed testing laboratories, ISST's emphasis is in support of
seed technologists around the world.

Questions

1. Do you consider seed testing to be a science, a skill, or an art?

2. What four components are considered to be part of the purity separation?
3. Define a noxious weed. How is the incidence of noxious weed seeds recorded in the purity
test results? Why is a larger seed sample examined for noxious weed seeds than for the
purity test?
4 Explain the normal seedling concept in interpreting laboratory germination tests.
5. What is the difference, if any, between hard seed and firm ungerminated seeds?
6. List several ways of stimulating faster germination in the seed-testing laboratory.
7. Describe several ways of distinguishing among varieties in a seed testing laboratory.
Which do you consider to be the most practical in routine seed testing? The least
practical?
8. Do you feel there should be more emphasis on pathological testing in the United States
and Canada? Can more emphasis be justified in view of its costs and results? If so, which
kind of pathological tests should be used? Which are presently in use? (See Chapter 16).
Seed Testing 351

9. Do you think most seed samples submitted by seed growers for testing are properly
drawn? Do you believe a grower would ever purposely misrepresent a sample to make a
seed lot appear better? How many bags should be sampled from a seed lot containing 250
bags of seed?
10. What is the difference between sampling and subsampling?
11. What are the purposes of tolerances? Do you know how they are applied for the various
test results?
12. Name several seed testing organizations. What is a Registered Seed Technologist?
13. Define a variety and identify two principal reasons why genetic purity testing is
important.
14. List four advantages of performing genetic purity tests.
15. What are the criteria for a successful genetic purity test?
16. What are some disadvantages of field testing for genetic purity?
17. Describe three types of morphological tests for genetic purity.
18. Explain the advantages and disadvantages of presoak, substrate imbibition and seedling
spray herbicide tolerance tests.
19. Identify at least three different electrophoretic approaches that can be used in genetic
purity testing.
20. What are the specific advantages of polymerase chain reaction technologies for genetic
purity testing?

References

Association of Official Seed Analysts. 1991. Cultivar Purity Testing Handbook. (eds. M. B.
McDonald and R. Payne). Contribution No. 33 to the Handbook on Seed Testing. Association
of Official Seed Analysts. 78 pp.
Association of Official Seed Analysts. 2000. Rules for testing seeds. Lincoln NE: AOSA
publication.
Buttery, B. R., and R. 1. Buzzell. 1968. Peroxidase activity in seeds of soybean varieties.
Crop Science 8:722-725.
Cooke, R. J. 1995. Gel electrophoresis for the idientiiication of plant varieties. Journal of
Chromatography. 698:281-299.
Cooke, R. J. 1998. Modern methods for cultivar verification and the transgenic plant challenge.
Presentation, 1998 International Seed Testing Association Symposium, Pretoria, South Africa ..
Douglas, J. E., ed. 1980. Successful Seed Programs: A Planning and Management Guide.
Boulder, Colo.: Westview Press.
International Seed Testing Association. 1993. International Rules for Seed Testing. Seed
Science and Technology 21: 1-288.
Elekes, P. 1980. The nature and improvement ofthe phenol reaction of wheat seeds. 19th 1STA
Congress Preprint No. I3-S.V.
Grote, S. R. 1992. Media imbibition method for detecting ALS trait in corn. Personal
communication.
Gutormson, T. J. 1999. Bioassay procedure for determining the presence of the Roundup
ReadyThf gene in corn. Pp. 18-19. In: Testing methodologies for hybrid parentage and trait
determination (ed. F. Zaworski). Corn, Sorghum and Soybean Technology. 1999. A special
publication of Seed Trade News.
352 Seed Testing

Jianhua, Z., M. B. McDonald, and P. M. Sweeney. 1996. Soybean cultivar identification using
RAPD. Seed Science and Technology 24:589-592.
Justice, O. L. 1972. Essentials of seed testing. In: Seed Biology, vol. 3, pp. 301-370. New York:
Academic Press.
McDonald, M. 8., L. J. Elliot, and P. M. Sweeney. 1994. DNA extraction from dry seeds for
RAPD analyses in varietal identification studies. Seed Science and Technology 22: 171-176.
McDonald, M. B. 1990. Validation of starch gel electrophoresis for corn seed purity
determinations. Proceedings, 45'h Annual ASTA Corn and Sorghum Research Conference
45:43-53.
McDonald, M. B. 1991. Blotting of seed proteins from isoelectrically focused gels for cultivar
identification. Seed Science and Technology 19:33-40.
McDonald, M. B. 1995. Genetic purity: From protein electrophoresis to RAPDs.
Proceedings, 50,h Annual ASTA Corn and Sorghum Research Conference 50:256-276.
McDonald, M. 8. 1998. Seed quality assessment. Seed Science Research 8:265-275.
McDonald, M. 8. and D. M. Drake. 1990. An evaluation of a rapid and automated
electrophoresis system for varietal identification of seeds. Seed Science and Technology 18:89-
96.
McDonald, M. 8., R. Danielson, and T. Gutormson. 1992. Seed Analyst Training Manual.
Association of Official Seed Analysts, Lincoln, NE.
Payne, R. C. 1986. Variety testing by official AOSA seed laboratories. Journal of Seed
Technology 10:24-36.
Sebastion, S. A., and R. S. Chaleff. 1987. Soybean mutants with increased tolerance for
sulfonylurea herbicides. Crop Science 27:948-952.
Smith, J. S. C., and J. C. Register, III. 1998. Genetic purity and testing technologies for seed
quality: A company perspective. Seed Science Research 8:285-293.
Society of Commercial Seed Technologists. 2001. Seed Technologist Training Manual. In
press.
Stuber, C. W., J. F. Wendel, M. M. Goodman, and J. S. C. Smith. 1988. Techniques and
scoring procedure for starch gel electrophoresis ofenzymes from maize (Zea mays L.). N. C.
Agr. Res. Serv., N. C. State Univ. Tech. Bull. 286:1-87.
Thompson, J. R. 1979. An Introduction to Seed Technology. New York: John Wiley and Sons.
U. S. Department of Agriculture. 1961. Seeds,' The Yearbook ofAgriculture. Washington, D. C.:
U.S. Government Printing Office.

1. Andersen, A. M., and C. M. Leach. Testing seeds for seedborne organisms, pp. 453-457.
2. Carter, A. S. In testing, the sample is all-important, pp. 414-417.
3. Colbry, V. L., T. F. Swofford, and R. P. Moore. Tests for germination in the laboratory,
pp.433-443.
4. Davidson, W. A., and B. E. Clark. How we try to measure trueness to variety, pp. 448-
452.
5. Justice, O. L. The science of seed testing, pp. 406-413.
6. Justice, O. L., and E. C. Houseman. Tolerances in the testing of seeds, pp. 457-462.
7. Musil, A. F. Testing seeds for purity and origin, pp. 417-432.
8. Zeleny, L. Ways to test seeds for moisture, pp. 443-447.
Seed Testing 353

u.s. Department of Agriculture. Production and Marketing Administration in cooperation with

Plant Industry, Soils, and Agricultural Engineering. 1952. Testing Agricultural and Vegetable
Seeds. Washington, D.C.:U.S. Government Printing Office.
Welsh, 1., and M. McClelland. 1990. Fingerprinting genomes using PCR with arbitrary
primers. Nucleic Acids Research 18:7213-7218.
Wheeler, W. A., and D. D. Hill. 1957. Grassland Seeds. Princeton, N.J.:D. Van Nostrand
Company.
Will, M. E., W. E. Kronsted, and D. M. TeKrony. 1967. A technique using lindane and cold
treatment to facilitate somatic chromosome counts in Lolium species. Proceedings 0/the A OSA
57:118.
Williams,1. G. K., A. R. Kubelik, K. 1. Livak, J. A. Rafalski, and S. V. Tingey. 1990. DNA
polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids
Research 18:6531-6535.
Wrigley, C. W. 1995. Identification o/rood-Grain Varieties. Amer. Assoc. Cereal Chern.,
St. Paul, MN. 283pp.
 16
Seed Pathology
and
Pathological Testing

In many parts of the world, testing for seedborne diseases has long been considered an
integral part of the routine inspection for seed quality. However, in North America,
pathological testing has not been as important as purity and germination testing. This is
partially because of uncertainty about whether the analytical results are significant enough to
justify the expense of maintaining a pathological testing program. Additionally, large samples
may have to be screened for results to be meaningful. Some pathogens can cause severe
losses if as few as one seed in 10,000 to 50,000 seeds are infected. Problems in reliably
detecting such small levels of infection and relating them to field losses are formidable, and
these tests inevitably require larger space and labor investments than do most other types of
seed quality tests. Consequently, pathological testing of seed in North America has not
developed to the level needed and is too often not a part of the routine seed laboratory
analysis.
For pathological testing of seeds to be a reasonable requirement for sale of seeds,
several conditions must be met. First, it must be established that seedborne infestation causes
reduction in plant stands, leads to field diseases, or causes other problems. The mere
existence of pathogens in seeds indicates little about the likelihood of the pathogen causing
problems for the ensuing crop. Second, the level of acceptable infestation must be established.
For example, if 5% of the seeds in a given seed lot must be infected for significant economic
loss to occur, it makes little sense to reject lots in which only 0.1 % of the seeds are infected.
Third, if the disease can develop explosively, seed pathogen testing may have to be
accompanied by legal restrictions on the sale of untested or infested seed lots. Grower A may
plant clean, healthy seeds, but may still suffer substantial losses if a neighbor plants infested
seeds and an efficient vector is present (e.g., a tractor, an insect, or the farm personnel).
However, if a disease is not present in an area, seeds infested with the pathogen should not be

L. O. Copeland et al., Principles of Seed Science and Technology

© Kluwer Academic Publishers 2001
Seed Pathology and Pathological Testing 355
planted in that area, regardless of whether the pathogen can be demonstrated to infect plants
from seedborne inoculum. Many diseases have no doubt been spread in this fashion and are
still reaching new areas today. For example, pea seedborne mosaic virus reached the United
States this way, and snapdragon rust probably arrived in Australia via the same route.
Several developing forces promise to change the status of pathological testing in North
America. One such force is the emergence of pathological testing in species for which
seedborne diseases seriously endanger crop productivity. For example, seedborne bacterial
diseases occur in several important crops in which pathological test results can be related to
crop productivity and translated into profit and loss. Another factor causing increased
attention to seed health testing is the expanded international trade in seeds and development of
uniformly-applied detection methods. Almost all countries require phytosanitary certificates
for imported seed to ensure that seedborne pathogens are not introduced from another
country. Thus, domestic as well as international forces are contributing to the recognition of
seed health testing as an important part of the seed testing process.

THE SEED-A MICROCOSM OF MICROBES

The seed has been described by Sinclair (1979) as a microcosm of microbes, with the
potential for carrying a wide variety of fungi, bacteria, viruses, and (sometimes) nematodes,
many of which can cause diseases in seedlings or plants. Some live on the seed surface and do
not visibly affect the seeds' appearance. They may become harmful only when environmental
conditions favor their growth and reproduction. Since such conditions normally exist in
germination chambers, these may cause problems with seed rots and seedling blight and
contribute to variability in germination results. Other microorganisms live in nonliving outer
tissues of the seed such as bracts, pericarps, or seed coats and attack the germinating seedling
when conditions become favorable. Still others are borne inside the seed, either on or inside
the embryo or endosperm tissue. Although these do not usually kill the seed, they may delay
germination and result in weak seedlings. Others survive in the embryo and resulting seedling
to infect plants and crops when grown from such seed. Detailed considerations of seed
pathology are provided by Maude (1996), Hutchins and Reeves (1997) and Agarwal and
Sinclair (1997).
Seed microbes can be divided into fungi, bacteria, and viruses.

Fungi

Fungi have been present on the earth as long as any other living organism as evidenced
by over 100,000 distinct species. Yet, with all of this diversity, they have several common
characteristics that help us understand their important role as seed pathologens. First, they
have no chlorophyll like higher plants so they cannot make their own food. As a result, they
live on substances provided by other plants or plant parts such as seeds. When these
organisms used for energy are living, the fungi are parasitic. In other cases, the organisms or
tissues they invade are dead in which case the fungi are saprophytes that cause rots or decay.
Second, fungi produce rapidly growing mycelia which are microscopic branches that produce
digestive enzymes and acids. These substances digest the nutritive material in which the
mycelia are present and provide sustenance to the fungus for continued growth. Third, fungi
produce spores which are the mycological reproductive equivalent of seeds from plants.
356 Seed Pathology and Pathological Testing
These spores are produced in such astronomical numbers that fungal contamination can
increase exponentially. Fourth, fungi require water, a favorable temperature, food, and
oxygen for growth. Most fungi require a high water content. Ifwater can be squeezed out ofa
tissue, then fungal growth is likely. They also grow best at fairly high temperatures, usually
about 30°C for optimum growth. When temperatures are below 5-10°C or above 35°C,
fungal growth is inhibited although some exceptions exist. Fungi that attack seeds can be
divided into field and storage fungi based on their environmental requirements.
Field fungi invade seeds almost exclusively during development or after physiological
maturity and usually have completed their damage prior to harvest. Common and serious
examples of field fungi include Alternaria, Cladosporium, Fusarium, and Helmin-
thosporium. For field fungi to cause seed damage, they require seed moisture contents in
equilibrium with relative humidities greater than 90%. For cereal seeds high in starch such as
barley, com, oat, and wheat, this is usually at a seed moisture content of 20-25% or higher on
a wet weight basis. These moisture contents are far above those encountered when seeds are
stored. When the seeds dry down below 20% moisture and eventually go into storage, the
continued growth of field fungi is reduced.
In contrast to field fungi, storage fungi actively invade seeds and cause damage under
conditions that can be encountered during storage. Among the most prevalent and important
storage fungi are those from the genera Aspergillus and Penicillium. They invade seeds in
equilibrium with relative humidities between 65 and 90%. For starchy seeds, this is at
moisture contents of 13-20%; soybeans at moisture contents of 12-19%; and for oil seeds at
moisture contents of 5-12% (Table 16.1). Almost all storage fungi grow during conditions of
seed storage that are favorable for their growth. The one exception is Aspergillus flavus
which sometimes invades seeds of com, peanut, and cotton in the field. Under normal
conditions, the activity of storage fungi is not specific to seeds as a substrate. They will grow
on almost anything, including chair stuffings, mattresses, pillows, and other materials when
provided the appropriate environment.
Storage fungi are almost always classified as saprophytes (plants that live on dead
tissue). They do not produce mycelia that release exocellular digestive enzymes and acids to
digest invaded living tissue. So how do they create such widespread damage, including the
decay of living tissue? The answer seems to be that these saprophytes initially colonize only
those seed parts which are dead such as the seed coat. There they proliferate and through
their normal metabolism synthesize toxins that are secreted to the outside of the fungus. The
toxin kills the living tissue which provides additional substrate for digestion by the fungus.
Perhaps the best known example of this process is the toxin produced by Aspergillus flavus
called aflatoxin. This toxin not only is toxic to the seeds on which it grows but is also highly
toxic to animals that digest the infected seed or grain. Because of the seriousness of this
toxin, peanuts, com, and cotton intended for feed are routinely inspected by the Food and
Drug Administration for the presence of aflatoxin. Similar health problems exist with toxins
produced by Fusarium species, Aspergillus ochraceus, and Claviceps purpurea.

Bacteria

A large number of bacteria exist in nature but only about 200 species are known to
cause plant diseases. In general, very few of these cause disease in seeds because free water
(high seed moisture content) and moderate to warm temperatures are required for bacterial
growth and disease development. As a result, seeds typically are invaded by bacteria and used
Seed Pathology and Pathological Testing 357
Table 16.1. Equilibrium Moisture Contents of Common Grains, Seeds, and Feed Ingredients
at Relative Humidities at 65-90% and Fungi Likely to Be Encountered.
Starchy Cereal
Seeds, Defatted
Soybean and
Relative Cottonseed Meal, Sunflower,
Humidity Alfalfa Pellets, Safflower Seeds,
(%) Most Feeds Soybeans Peanuts, Copra Fungi
65-70 13.0-14.0 12.0-13.0 5.0-6.0 Aspergillus halophilicus
70-75 14.0-15.0 13.0-14.0 6.0-7.0 A. restrictus, A. glaucus,
Wallemia sebi
75-80 14.5-16.0 14.0-15.0 7.0-8.0 A. candidus, A. ochraceus,
plus the above
80-85 16.0-18.0 15.0-17.0 B.O-I0.0 A. jiavus, Penicillium, plus
the above
85-90 18.0-20.0 17.0-19.0 10.0-12.0 Penicillium, plus the above
From: Christensen and Sauer (1982).
'Percentage wet weight. The figures are approximations; in practice, variations up to ± 1.0% can be ex-
pected.

to assist in dispersal of the pathogen. They enter the seed through wounds created by insect
vectors or they are translocated by the infected mother plant into the developing seed. As the
seed matures and dries down, the bacteria become dormant. Then, during imbibition and
subsequent germination, the bacteria rapidly resume growth and multiply exponentially in the
seedling and plant tissue. They cause necrosis of cells, abnormal growth of galls and tumors,
breakdown of tissues (soft rots), and blockage of water-conducting vessels (wilts). Some of
the worst bacteria that are seedborne and cause plant disease come from the genera
Agrobacterium (crown gall), Bacillus (seed rot), Corynebacterium (wilt), Erwinia (soft rot),
Pseudomonas (blight), and Xanthomonas (black rotlblight).

Viruses

Viruses are extremely small organisms uSlllally composed of a nucleic acid strand
encased in a proteinaceous shell. By definition, a virus is considered a transmissible parasite
whose nucleic acid genome is less than 3 x 106 daltons and needs ribosomes and other
components of the host cell for multiplication. Like bacteria, most viruses do not cause direct
seed damage but use the seed for dispersal and subsequent plant infection. About 20% of the
known plant viruses are seed transmitted. Plants can become infected during pollination and
through wounds often caused by insects. The developing seed is then systemically infected by
the mother plant. It is sometimes difficult to identifY plant diseases caused by viruses because
they often mimic nutritional deficiencies or plant response to other environmental stress.
Typical conditions include chlorosis, stunting, wilting, mosaic patterns, and necrosis. The
bleeding hilum found in soybean seeds is an example of a seed borne virus caused by the
soybean mosaic virus.
358 Seed Pathology and Pathological Testing
CONTROL OF SEED BORNE DISEASES

Preharvest Control

Preharvest control of seedborne diseases may be accomplished by one of three different

methods: (I) selection of disease~free seed production areas, (2) cultural practices, and (3)
point of origin inspection. In the first case, seed is produced under environmental conditions
that restrict the occurrence of diseases. Thus, dry edible bean and snap bean seed produced in
dry, irrigated areas of Idaho or California are more likely to be free of bacterial blight
(Pseudomonas vulgaris) than seed produced in the Great Lakes region where the weather is
more humid and favorable for disease development.
Regardless of seed production area, cultural practices are crucial in the prevention and
control of seedbome diseases. These include the following:

I. Planting disease~free seed.

2. Treatment of seed with antibiotics.
3. Spraying seed fields with fungicides, bacteriocides, and other antibiotics to prevent
disease buildup.
4. Hand roguing of diseased plants.
5. Avoiding overhead irrigation which might otherwise create conditions favoring disease
buildup.
6. Use of resistant cultivars.
7. Crop rotation.
8. Isolation of seed fields from sources of potential infection.
9. Chemical or biological control of insect vectors.

The third preharvest control of seedborne diseases is inspection of seed fields so that
potential problems may be detected and eliminated prior to harvest. Diseased areas may be
destroyed or diverted from seed use, or the entire field can be diverted. While these
precautions may not completely prevent contamination by surfaceborne dusts, they do lower
the probability of seed infection.

Postharvest Control

Postharvest control of seed borne diseases should be considered only as a last resort,
since it is better to prevent the occurrence of seedborne diseases than to eradicate disease
infection (or infestation) that is already present. However, several methods may help upgrade
the phytosanitary quality of seed after harvest. These include (1) surface disinfectant by
chemical seed treatment, (2) separation of diseased seed and foreign material, (3) hot-water
treatment, and (4) organic solvent infusion of antibiotics.
Treating seed with antibiotics is usually effective only against surfaceborne pathogens,
but in some cases systemic antibiotics (e.g., carboxin) can penetrate into the seed and
eradicate internal infection. Sometimes penetration of antibiotics can be improved by organic
solvent infusion. Separation measures are effective for eliminating seeds or foreign material
in a seed lot that is disease infested. An example of this is ergot sclerotia in cereal seed, which
can be eliminated by cleaning.
Seed Pathology and Pathological Testing 359
Hot water treatment can be used to kill infection in the seed without destroying seed
viability. Prior to the development of systemic fungicides (e.g., carboxin), this was the only
effective method of controlling loose smut in wheat and barley seed.

FUNGI ASSOCIATEU WITH SEED

Fungi cause the largest number of plant diseases and occur more commonly in or on
seeds than bacteria or viruses. More than 8000 spt~cies of fungi have been identified as plant
pathogens. Fungi associated with seeds consist of both saprophytic and pathogenic fungi.
Saprophytic fungi are not specific to any particular host and may be found on seeds of
various plants, whereas pathogenic fungi are usually confined to a limited host range. Both
types may occur on the seed surface, in cracks, or inside the seed coat, but pathogenic fungi
may also occur within the seed itself. While saprophytic fungi may cause problems in the
seed testing laboratory by contaminating germination media, pathogenic fungi endanger crop
productivity and are of great economic importance to agriculture because: (1) infected seeds
may not germinate, (2) infected seeds provide inoculum for further spread of the disease, and
(3) seed infection prior to harvest may cause reduction in both crop yield and seed quality.
Fungi are composed of a threadlike vegetative body called mycelium. They reproduce by
means of spores that have a function similar to seeds in higher plants. Like seeds, fungal
spores vary greatly in size, shape, and color; however, they are much smaller than seeds and
microscopic in size. Some fungi do not produce sexual spores, but reproduce by means of
vegetative structures such as sclerotia, which are hardened, compacted masses of mycelium.
An example of a sclefC'tium is ergot, such as that formed by the fungus Claviceps purpurea.

METHODS OF DETECTION

Agar Testing

One of the oldest and most common pathogenic tests is the agar test for identification of
seedborne fungi (Figure 16.1). Agar is a carbohydrate medium prepared from certain species
of seaweed. It contains few nutrients for the grovvth of fungal pathogens; thus, it is usually
supplemented with extracts from plants such as pOltato or various fruits and vegetables, which
are in tum supplemented by sugars, salts, antibiotics, or other agents. The agar medium is
prepared by mixing agar powder with an appropriate quantity of water and nutrient additives.
This mixture is sterilized in an autoclave for 15 to 20 minutes and cooled to about 50°C, at
which time an antibiotic may be added. This mixture is carefully poured into petri dishes by
lifting the lid only enough to pour in the agar, thus avoiding contamination. It is then allowed
to cool and solidify for 20 minutes after which it is ready for use.
Seed to be tested should be surface-disinfected by pretreating for one minute in a 1%
available chlorine solution of sodium hypochlorite (NaOCI) prepared by diluting 20 parts of
laundry bleach (5.25% NaOCI) with 85 parts of water. For deep-seated infection, seeds may
be treated with stronger solutions (e.g., 5.25% available chlorine). This eliminates
contamination of the seed coat by saprophytes which tend to develop rapidly on the agar and
may inhibit or completely obscure the slower-growing pathogens. Usually about 10
(depending on size) seeds to be tested are surface-sterilized and individually placed on the
agar surface with a forceps. After placing each seed, the forceps tip is carefully disinfected by
dipping into a 70% alcohol solution and passing through a flame.
360 Seed Pathology and Pathological Testing
Figure J6.1. Agar plate test for seedborne fungi.

A. Mycosphaerella spp. On pea, agar plate test after 7 days: (1) Colonies of Mycosphaerella spp. at
II and 12 o'clock, large contaminant colony of Alternaria tennusis at 6 o'clock; (2 & 3) Close-up of
colony at 12 o'clock, note peppery appearance oftlask-shaped pycnidia (fruiting structures) formed
in agar surrounding the seed; (4) Close-up of colony at 12 o'clock showing pycnidia and pycnidial
ooze containing spores (Courtesy of Jim Sheppard, Agriculture Canada).

:2

3 4

B. Same as in A, btlt Ascochyta kmtis on lentil; note characteristic pycnidial ooze,

2
Seed Pathology and Pathological Testing 361
Sometimes bacterial colonies develop on the agar or blotter (see below) and inhibit
fungal growth, making identification difficult. This can be overcome by adding an antibiotic
such as streptomycin sulfate to the autoclaved agar medium (after it cools to SO° to 55°C) or
to the water used to moisten blotters in blotter tests.
After plating, the petri dishes are incubated at 20 to 2S0C for about five to eight days, at
which time seedborne pathogens (fungi) are identified on the basis of colony (vegetative) and
spore characteristics.
Recently, disposable plastic petri dishes have almost completely replaced the glass petri
dishes that had been used previously. These offer savings in labor, are convenient, and are
more energy-efficient than glass dishes, which n(~ed to be emptied, cleaned, and sterilized
before each use.

Blotter Tests

Blotter tests (see Figure 16.2) for pathogenic fungi are similar in technique to
germination tests in that seeds are placed on moisltened layers of blotter paper or filter paper
and incubated under conditions that promote fungal growth. The procedure consists of
saturating the blotters with sterile water, then allowing excess water to drain off briefly
before the seeds are planted manually with a forceps, a vacuum counter, or planting board.
Regardless of the placement method, the seeds should be evenly spaced to avoid contact with
each other.
Profuse seedling growth in blotter tests may tend to make interpretations difficult. This
difficulty can be overcome by adding enough 2,4-D sodium salt to provide a 0.2% moistening
solution. Seedling growth can also be inhibited by a freezing technique that allows the seeds
to imbibe for 24 hours (48 to 72 hours for corn) followed by a 24-hour exposure at -lS0C
before normal incubation. (Normal incubation usually consists of temperatures of20 to 25°C
with a 12-hour day-night NUV light cycle for five to seven days). Sporulation of many fungi
is stimulated by alternating cycles of blue light and darkness. Following this period, the test is
interpreted on the same basis as agar tests. However, killed seeds support luxuriant fungal

1 2 3

Figure 16.2. Blotter testfor Phoma lingam on Brassica seed (Rapeseed): 1, Arrangement of seed
on blotter; 2 & 3, Pycnidia growing on blotter after 10 days incubation. Note characteristic spore
exudate (Courtesy ofJim Sheppard, Agriculture Canada).
362 Seed Pathology and Pathological Testing

growth and avoid the necessity of examining seeds through a tangle of seedlings. Since
freezing makes agar mushy, seeds must be placed on a paper substrate for freezing and
afterwards transferred to blotters.

Virulence Tests

Virulence tests consist of making further isolations of suspected pathogens from blotter
or agar tests, culturing them on agar for identification (spores, vegetative growth), inoculating
the healthy plants (usually seedlings) for observation of pathogenecity symptoms, and
subsequently reisolating and culturing the pathogen on a suitable substrate. This procedure
follows Koch's postulates and is a method of positive identification of any pathogen.
Inoculations may be by injection or other methods (e.g., spraying or dusting following
mechanical abrasion).

Noncultural Tests

Several seedborne fungal pathogens can be detected by visual inspection of the seed
sample, or special noncuIturaI techniques. For example, ergot (Claviceps purpurea)
contamination may be detected by the presence of large dark sclerotia. These sclerotia, when
planted, produce fruiting bodies called apothecia. Apothecia further produce spores that
provide inoculum in crops planted with ergot-infested seed.
Visual examination can also be effective for detecting smut balls of bunt, or stinking
smut (Tilletiafoetida or caries) of wheat. These consist of the pericarp of the caryopsis, the
inside of which has been completely replaced by black smut spores. Normally, many of the
smut balls break open during threshing and spread spores to other seeds throughout the lot.
These adhere to the caryopsis and serve as inoculum for the next generation. Although such
spores may be quite visible on the seed surfaces, the following procedure provides a
systematic way of detecting and quantifYing stinking smut infestation.

1. Wash the spores from the seed surface by shaking the seed sample in a known amount
of water containing a small amount of detergent.
2. Centrifuge the washings from the sample and resuspend the resulting volume in a
small, measured quantity of fluid.
3. IdentifY and quantifY the spores under a high-powered microscope. Quantification of
the spores may be made by counting the number of spores present in a given volume or
over a given surface area of liquid.

One common noncultural test for seedbome fungal pathogens is for loose smut infection
of wheat or barley (Figure 16.3). Infection is present as hypha I strands that infect the embryo
internally. The presence of such infection and its incidence in a seed lot is determined by the
following method:

1. Soften the seeds by soaking in sodium hydroxide overnight.

2. Isolate the embryo by washing in a stream of warm water and separate by a fine wire
mesh sieve.
Seed Pathology and Pathological Testing 363

INFECTION

Smut Free Infected

Embryo Embryo

Figure 16.3. The loose smut test of wheat showing noninfected versus infected embryo
(Courtesy ofGustafwn Chemical Company).

3. Repeat washing with a solution of lactop heno I and water. The embryos will float while
the chaff and endosperm tend to sink to the bottom and can be drawn off.
4. Place the separated embryos into thick-bottomed glass dishes and clear further by
boiling in lactophenol for 10 to 20 minutes.
5. Arrange cleared embryos and examine under magnification for the presence of hypha.

IDENTIFICATION

Identitication of fungal pathogens requires considerable study and expertise in

mycology. Seed testing laboratories inspecting samples for fungal pathogens should have
trained mycologists on their staff. Official laboratories in some countries do have such
qualified staff and are well equipped for pathological testing. Unfortunately, few laboratories
in North America have qualified staff available for routine detection of fungal pathogens, and
are thus not equipped for identi-fying pathogens except for certain tests. However, tests for
examining barley embryos for loose smut infection are routinely performed in several state
and official laboratories.
If qualified staff are not available, technicians may be trained to make routine inspection
of samples and identify at least commonly occurring fungal pathogens. In any case, a
dissecting (i.e., stereoscopic) microscope capablc~ of 10 to 90 x magnification and high-
intensity lighting are essential, along with basic laboratory supplies and equipment for
preparation and incubation of agar and blotter tests.
The following descriptions were given by Kulik and Schoen (1977) for seven important
genera of fungi that commonly invade seed of major crops and are relatively simple to detect
and identify. Though separation of these genera into species can be made for a particular
364 Seed Pathology and Pathological Testing

sample, such identification is usually not necessary when they are associated with the primary
host crops listed below

l. Alternaria. Alternaria spp. are found in seeds of crucifers (Brassica spp.), carrot
(Daucus carota L. var. sativa DC), and flax (Linum usisatissimum L.), and many other
species. Colonies range in color from gray through greenish-olive brown to black.
Typically, an Alternaria spore is club-shaped and has transverse and longitudinal septa.
The common saprophyte A. alternata (Fr.) Keissler (syn. A. tenuis C. G. Nees) is
difficult to separate from the several pathogenic species on the basis of colony
characteristics. There is also some overlapping of spore features among the various
species. However, the more important pathogenic species generally produce long-beaked
spores that are solitary or in short chains and contrast with the longer chains of A.
alternata. An important exception is A. brassicicola (Schw.) Wiltshire, a frequently
isolated seedbome pathogen of crucifers. This fungus differs morphologically from A.
a/ternata in generally having spores that are smooth and with fewer transverse
septations.
2. Ascochyta (Perfect state: Mycosphaerella). Ascochyta spp. are often found associated
with seeds of various legumes, including Melitotus spp., Phaseo/us spp., Pisum spp.,
Trifolium spp., and Vicia spp. Seeds of the garden pea (Pisum sativum L.) may be
invaded by three species of Ascochyta: A. pin odella, Jones (syn. Phoma medicaginis
Malbr. & Roum. var. pinodella (Jones) Boerema) forms light to dark brown pycnidia
from which spores are exuded in the light buff to pinkish brown mass. The spores
average 3.5 x 8f.!. A. pinodes Jones [perfect state is Mycosphaerella pinodes (Berk. &
Blox.) Vestergr.] forms light to dark brown to black pycnidia with a light buff to pinkish
spore exudate. The spores average 5 x 14f.!. The pseudothecia formed by the perfect-
state are dark brown. To separate A. pinodes from A. pinodella it may be necessary to
compare the size of the spores under a compound microscope. A. pisi Lib. forms brown
pycnidia from which the spores are exuded in a carrot red mass.
3. Diaporthe. The most commonly encountered member of this genus is Diaporthe
phaseolorum (Cooke & Ellis) Sacc., which is frequently seedbome in soybean. Colonies
are wooly white, more compact and slower growing than those of Fusarium spp., which
also are often present in soybean seeds. Mature colonies usually contain embedded,
dark-colored perithecia or pycnidia (the latter produced by Phomopsis sojae Lehman,
the imperfect state of Diaporthe phaseolorum) that exude milky or yellow droplets of
spores. In contrast, these fruiting bodies may also occur scattered or more or less
regularly spaced on or just below the surface of the seed coat, with scant development of
mycelium or spore exudation. Although D. phaseolorum on soybean has been reported
to consist of two varieties, i.e., D. phaseolorum var. caulivora Athow & Caldwell, and
var. sojae (Lehman) Wehmeyer, it probably is not necessary in routine seed health
testing to differentiate between them.
4. Fusarium (Perfect states include Gibberella and Griphosphaeria). Fusarium spp. are
associated with seeds of many members of the Poaceae, including com, rice (Oryza
sativa L.), and wheat (Triticum aestivam L.); also with seeds of cotton (Gossypium
hirsutum L.), and various legumes such as the soybean and Phaseolus spp. Colonies
usually consist of fluffy white to pink or coral-colored mycelium, often reaching 25 mm
in diameter after one week of incubation. Spores are transparent hyaline, canoe- or
Seed Pathology and Pathological Testing 365

banana-shaped with pointed or rounded tips, usually subdivided into several cells with
thin walls (septa), sometimes in spindle-shaped (elliptical) translucent to orange clusters,
up to 0.5 mm in diameter. F. moniliforme Sheldon, a pathogen of com, is an atypical
species having spherical spores borne in long chains. Colonies of Epicoccum spp., found
mainly on small grains, and of Cephalosporium spp., on com, are similar in appearance
to Fusarium spp., but Epicoccum spp. are easily separated by their dark spore clusters.
Identification of Cephalosporium sp. is mOTl;: difficult because the spore clusters may
resemble those of Fusarium spp. although they are actually more globular. Individual
spores of Cephalosporium sp. are capsule-shaped or sometimes ovoid. Its occurrence is
relatively infrequent in most lots of seed com.
5. Helminthosporium (Synonyms for the imperf(~ct state include Bipolaris and Drechslera,
Cochliobolus, Ophiobolus, and Pyrenophora, which are perfect states of this common
imperfect genus). Helminthosporium spp. invade seeds of many of the Poaceae,
including cereals such as com, rice, and wheat, and a variety of other plants. Colonies
range in color from light to dark gray, but unless they are grown on certain agar media,
their positive identification to genus based solely on colony characteristics is nearly
impossible. This is due principally to the similar colony appearance of the ubiquitous
and omnipresent saprophyte, Alternaria alternata. In both Helminthosporium and
Alternaria, spores borne on short conidiophores often develop on the seed surface with
little or no production of mycelium. In Helminthosporium, spores are usually dark-
colored, cigar- or club-shaped, straight or slightly curved, and are subdivided into
several cells having relatively thick, transv(~rse septa. A. alternata has dark, club-
shaped, rough spores (sometimes ovoid) diviided into several cells, usually with both
transverse and longitudinal septa. The spores may occur separately, but usually form
various-sized chains with the tapered end of each spore (the beak) giving rise to the next
spore above it.
6. Phoma (Synonyms Phenodomus; perfect states include Leptosphaeria and Pleospora. ).
Phoma spp. are associated with seeds of crucifers, beets (Beta vulgaris L.), and flax.
Colonies growing from seeds of crucifers may be thin, or profuse, silvery white, and
after several days are penetrated from beneath by the relatively large (200f.l) flask-
shaped, dark-colored pycnidia that eventually exude pink to purple masses of spores.
The single pathogenic species on Brassica spp., P. lingam (Tode ex Fr.) Desm., can
sometimes be separated from the saprophyte P. herb arum Westend, based on the
smaller pycnidia and nearly colorless spore masses produced by the latter. An extremely
low incidence of P. lingam in a seed lot may lead to extensive plant loss in the field.
Phoma betoe Frank, found in seeds of sugar beet, produces black pycnidia that may be
almost spherical or flattened, up to 180f.l in diameter, with colorless spores. Seeds of
flax may be invaded by Phoma exigua Desm. which produces a colorless to gray myce-
lium, dark brown pycnidia, and colorless spores that may exude from the pycnidium in a
cream-colored ribbon.
7. Pyricularia. Pyricularia spp. are found in seeds of rice and finger millet [Eleusine
coracana (L.) Gaertn.] grown outside the United States. Colonies are very small,
inconspicuous, grayish or icy green, consisting of groups of conidiophores bearing pear-
shaped spores in terminal clusters. These clusters are usually restricted to the embryo
end of the seed. Under low magnification, they are similar in appearance to the common
seedborne saprophyte Cledosporium spp., but may be differentiated at higher
366 Seed Pathology and Pathological Testing

magnification (80-90x) due to the olive-green, dry appearance of this saprophyte.

Spores of Pyricularia spp. usually are pear-shaped and have two septa that divide them
into three unequal-sized cells while Cladosporium spp. have smaller single-celled spores
generally borne in short chains.

SEEDBORNE SAPROPHYTIC FUNGI

Saprophytic fungi are those that grow in dead tissue. Although they exist on all seed,
they do not cause plant diseases as do parasitic fungi. Spores of saprophytic fungi are almost
ubiquitous, occurring on seed as well as in the air. They are especially numerous on seed in
storage, and will germinate and grow profusely any time the storage environment exceeds
75% relative humidity and 15°C.
According to Kulik and Schoen (1977), seedborne or contaminating colonies of Mucor,
Rhizopus, Aspergillus, and Penicillium spp. are often encountered when seeds are tested on
agar or blotters for pathogenic fungi. Some Aspergillus spp. are pathogenic on stored seeds,
and their presence may be indicative of improper (i.e., relatively moist) storage conditions.
Mucor and Rhizopus are easily recognized by fast spreading cobwebby networks of aerial
mycelium that produce large spore clusters borne on heavy stalks called conidiophores.
Aspergillus and Penicillium produce slow-growing granular or velvety textured colonies.
Aspergillus colonies may be white or creamy at first, then change rapidly to a wide variety of
green or bluish-green hues; this is accompanied by a profuse production of highly pigmented,
powdery spores. Aspergillus niger, a common contaminant of cereal and cotton seeds,
produces black spores (Figure 16.4).
Another common seedborne saprophyte, Chaetomium spp., produces spore-bearing
bodies called perithecia which are ornamented by various types of bristles or hairs. Although
not as common as the genera cited above, Chaetomium might be mistaken for a pathogenic
species.

FUNGAL ENDOPHYTE

Another interesting fungus that infects plants of interest to seed scientists is the fungal
endophyte. Some cool-season grasses are infected by Acremonium coenophiallum, which
causes forage grasses to be toxic to grazing animals but confers insect resistance to grasses.
Tall fescue endophyte is the best known of these and is disseminated only by the seed. The
fungal hyphae grow into the developing seed and are primarily located between the scutellum
of the embryo and the aleurone layer of the mature seed (Bacon 1983). Biochemically, the
fungus does not appear to alter the amino acid composition of the developing and mature seed
(Bolesky et al. 1985). Developmentally, the fungus provides an advantage to the infected
plants and seeds. Seeds from infected plants of Lotium perenne and Festuca arundinacea
have higher rates of germination and produce more biomass and tillers than seeds from
noninfected plants (Clay 1987; Rice et al. 1990). The fungal endophyte has been shown to
enhance the production of indoleacetic acid in plant materials (DeBatista et al. 1990).
Recently, because of the developmental advantages provided by the fungus, endophyte-
enhanced seeds are being marketed for turf purposes. On the other hand, tall fescue infested
with the fungal endophyte, can cause feeding animals to become lame, a condition known as
"fescue foot."
Seed Pathology and Pathological Testing 367

Figure 16.4. Species of Aspergillus grown ji'om maize, rice, wheat and barley {Courtesy of C. M.
Christensen}.

Testing for Fungal Endophyte

Endophyte-free pastures may be established by planting endophyte-free seed. As a

result, seed testing procedures have been developed to detect the presence of the fungal
endophyte. These include taking the seed and allowing it to be digested in a 5% NaOH
solution for 12 hours. After digestion, the glumes are removed from the seed, which is placed
on a microscope slide, macerated, and stained with a one part 1% aniline blue solution in two
parts 85% lactic acid. The aniline blue solution allows resolution of the fungal hyphae in the
seed tissue under microscopic examination.

BACTERIAL PATHOGENS

Testing for seedborne bacterial pathogens is conducted in perhaps as many as 20 North

American laboratories on a routine basis. While most procedures involve tests for detecting
bacterial blight pathogens of field and garden beans, tests also exist for other crops,
particularly for seeds of vegetables such as tomato and cabbage. Most of the tests used for
the detection of seed borne bacterial pathogens can be considered modifications of one of four
368 Seed Pathology and Pathological Testing

basic methods: (I) detection of infected seed by means of external characteristics visible on
the seed coat, (2) diagnosis of plant symptoms resulting from inoculation (or injection) by
material from suspect seed, (3) isolation of bacterial pathogen to identifY the organism itself,
and (4) a combination of the above methods.

Common Tests for Seed borne Bacterial Pathogens

Serological Techniques. Serological techniques are based on the reaction between

antigens and antibodies. These techniques can be used for positive identification of both
bacterial and virus organisms. Antisera containing the specific antibodies is prepared by
injecting an antigen (the bacteria or virus) into the bloodstream of live animals (usually
rabbits). The blood immediately fights off the effect of the antigen by building up antibodies
in the bloodstream (which can be recovered by bleeding the animal). The antiserum is used to
test for the presence of the same bacteria in the homogenized seed by a precipitation test in
agar. When antibodies and antigens are present, a precipitate occurs; if no precipitate
appears, the causal agent is not present.
Different antisera are needed for serological testing for different pathogens. The serum
may be prepared and kept for further use by freezing.
Although serological tests are used for detecting seedborne bacterial pathogens in
several crops (e.g., bacterial blight of Phaseo/us vulgaris), several problems exist. First,
bacteria possess many antigenic materials, some of which are similar to those contained in
other, nonpathogenic bacteria. This can cause confusion in identifYing the precipitate
designating the target pathogen. Second, the test is too detailed and time-consuming to be
practical on an individual seed basis; thus, a homogenate of a seed sample is usually used.
For testing bulk samples, techniques such as ELISA (enzyme-linked imunosorbent
assay-page 372) offer promise of making serological tests quicker and more accurate,
providing the problems of confusion over other antigenic materials cited above can be
resolved.
Bacteriophage Technique. The bacteriophage technique is similar to the serological
test in that it uses a specific reaction between two organisms to positively identifY disease
organisms. In this case, bacterial organisms can be identified by using special phage viruses
which are added to homogenized seed material on the agar. If the bacterial organism is
present, a clear plaque area appears on the agar due to attack on the bacteria by the virus.
The bacteriophage technique works well if contaminating bacteria are not present.
Unfortunately, they usually are and will obscure the plaque area.
Plant Injection Tests. Plant injection tests can be useful for identifying certain
seedborne organisms, particularly bacteria (Figure 16.5) and viruses. The test is simple and
easy to perform. Seeds to be tested are soaked in sterile water for a few hours, after which the
leachate is injected into a young, healthy seedling. Sometimes individual seeds are
homogenized in a liquid solution for injection into the test seedling. The injection may be
placed in the vascular system (phloem) of the plant, which transports it throughout the plant.
The entire plant is closely observed for development of disease symptoms.
Another method often used, especially for virus detection, is to rub a mixture of a
homogenized seed (or a seed sample) and metal filings on the leaf surface. The metal filings
puncture the plant tissue providing entry for any disease inoculum present, resulting in
disease symptoms.
Seed Pathology and Pathological Testing 369
Young healthy plants of the species being tested are required for plant injection tests.
Usually these are grown in a greenhouse under phytosanitary conditions and should be two to
four weeks old when inoculated. About two weeks should be allowed for symptom
development.

SEED BORNE VIRUSES

According to Carroll (1979) about 200 plant viruses are known to cause diseases of
plants, of which 100 are fairly well known. An additional 500, though nonpathogenic, are
thought to be transmissible plant viruses. Of all these, only about 80 different viruses or
viruslike organisms are considered to be seed-transmitted. A few of these, such as tobacco
mosaic virus (TMV) on tomato, are carried on the surface of the seed or inside the seed
outside the embryo. However, most, such as bean common mosaic virus (BCMV) and barley
stripe mosaic virus, are carried inside the embryo.
Next to tests for seedborne bacterial pathogens, those for seedborne viruses represent the
most common tests for seedborne pathogens in North America. Two kinds of tests are
commonly used-biological and serological.

Biological Tests

Biological tests include both grow-out tests and direct seed tests. Grow-out tests usually
begin with a visual examination to detect abnormal-appearing seed. These, plus nonnal-
appearing seed, are then planted in an appropriate sterile medium and placed in a favorable
light and temperature environment for seed germination and seedling growth. Test seedlings
are observed carefully for symptom development. This test is simple, inexpensive, and
suitable for testing individual seeds. Biological tests may also be performed by grinding up
seeds or seedlings from suspected seed lots and mechanically inoculating a sample of the
homogenate into sensitive, healthy test seedlings and observing them for symptom
development.
Direct seed tests are those in which both normal- and abnormal-appearing seeds are
soaked in an aqueous medium and ground into a slurry that is inoculated onto indicator or test
plants. Seedborne viruses are indicated by the development of characteristic symptoms on the
test seedlings. Single seeds or a composite sample from a seed lot may be tested. According to
Carroll (1979), this test is suitable for detecting a seedborne mosaic virus in pea seed, using
purple pigweed (Chenopodium amaranticolor) as a test plant. Similar tests are used to
screen lettuce seeds for the presence of lettuce mosaic virus; Chenopodium quinosa is the
indicator plant in this case. It is illegal to plant seeds found to be infected by this or other
assays in parts of California or Florida. All seeds planted in these areas must be tested.

Serological Tests

Serological tests for seedborne viruses are similar in principle to those described for
detecting bacterial pathogens. They are based on the physiochemical reaction between the
virus in the seed or homogenate from the sample to be tested and blood sera prepared from
laboratory animals (usually rabbits). Five serological techniques were described by Carroll
(1979); these differed slightly, depending on the method used for meeting of pathogens from
virus-infected seed and antigens from the blood serum. These are described below:
370 Seed Pathology and Pathological Testing

A."·~Sterilizing beans. C.-.. 'pouring off solution to be injected

into test plants.

B.-Soaking beans.

D.-Filling syringe.
Figure 16.5. The seedling irljection procedure used in Michigan for detection of seedborne
bacterial blight in dry edible bean seed (Courtesy C?f William Young, Michigan Department oj
Agriculture).
Seed Pathology and Pathological Testing 371

E.-Injecting bean test plants. F.-Point of injection and lesion.

Figure 16.5. (Continued)

Double Diffusion Test. The double diffusion technique has been used more extensively
than any other serological method. It can assay a single seed or part of a seed. Usually the
seed to be tested is soaked in tap water. Then the whole seed or its parts (e.g., embryo) is
triturated and the resulting triturate is transferred to a well cut into the diffusion medium
(commonly, agar gel). Thereafter, an antiserum specific for the suspect virus is placed in a
separate well. In time, the virus particles (antigen) and the antibody molecules diffuse toward
one another. Since this diffusion is in two different directions, it is called double diffusion.
When the two seroreactants reach a point in the gel at which the relative concentration of
each is serologically equivalent, antigens and antibody molecules complex, precipitate, and
become immobilized. The precipitate appears as a white band at some point between the two
wells.
To detect viruses that have an elongated structure by the double diffusion technique,
seed preparations must be treated with some kind of agent that breaks up or degrades the
virus particles to enhance the diffusion of the virus particles in agar.
The double diffusion method is generally quite specific, reasonably sensitive, and can
detect virus concentrations of 10 to 25 Jlg/ml. This test incorporates sodium dodecyl sulfate
as a virus-degrading agent, and is used routindy at the Montana State Seed Testing
Laboratory for serodiagnosis of barley stripe mosaic in barley embryos (Figure 16.6). Two
hundred embryos from each seed lot are tested using filter-paper disks as seroreactant depots.
Radial Diffusion Test. The single or radial diffusion method is similar to the double
diffusion technique except that only the virus antigen diffuses from a well out into the agar
diffusion medium. The serum containing antibodies to a specific virus has been included in
the agar in this test. Therefore, the procedure is to charge the wells with the seed or seedling
preparations being tested. Ifvirus molecules are present, they diffuse radially from the wells,
and when they have reached a certain distance from the well surface, they complex with the
antibody molecules and precipitate in this region, forming a ring or halo around the charged
well.
372 Seed Pathology and Pathological Testing
Normally, a subunit of antiserum (prepared by injecting rabbits with a degraded virus
preparation) is used with the radial diffusion method, and therefore only degraded viruses can
be detected. This method is sensitive, rapid, and can detect as little as 1giml of degraded
virus. In a matter of minutes or a few hours, results can be obtained with preformed agar
diffusion plates. This method has been used for the detection of seedborne barley stripe
mosaic VIrUS.
Latex Flocculation Test. Latex flocculation is another serological method in which 100
seeds are ground in a Wiley mill. Then about 0.1 g ofthe ground seed material is placed in 2
ml of buffer and further ground with mortar and pestle. About 20 111 of this seed extract are
then placed in a 100-ml pipette to which 10111 of tagged latex had been previously added. The
tagged latex consists of a suspension of polystyrene latex spheres (about 0.81 11m in
diameter). The latex spheres are sensitized, or covered with antibody molecules specific for a
given virus. The pipette is oscillated for about 15 minutes and observed under a dissecting
microscope. When virus is present in the sample, the latex suspension becomes flocculated or
aggregated (See Figure 16.7).
The latex method is rapid, specific, and sensitive. Results can be obtained within 15
minutes to 1 hour. The method can detect one infected seed per 100 seeds or 1 I1gimlofvirus.
Enzyme-linked Immunosorbent Assay (ELISA). In this procedure, antisera specific
for a given virus is used to coat the polystyrene plate, and the antibody molecules become
absorbed. Next, excess antibody molecules are washed with the plate, the seed sample is
added, and the excess sample is washed away. This is followed by adding an enzyme-labeled
specific antibody and the enzyme alkaline phosphatase is conjugated to the antibody
molecules specific for the virus for which the examination is made. The excess labeled
antibody molecules are washed away, and finally, an enzyme substrate is added. This enzyme
hydrolyzes the substrate. The amount of substrate hydrolyzed is determined by measuring the
extinction of 405 nm wavelength spectrophotometrically or by visual observation. For this
assay, dry-milled or water-soaked seed is extracted with buffer and the seed preparations are
then ground further with another buffer and a surfactant. Afterwards, the seed extracts are
applied to polystyrene plates following the standard ELISA procedure. Virus-infected seed
preparations should produce a yellow color.
High sensitivity is the main attribute of the ELISA method. Some workers claim to
detect 0.1 giml of virus. The method has been used experimentally to detect tobacco ring spot
and soybean mosaic viruses in soybean seed. Low levels of infection (1 to 4%) could be
detected in extracts from seed lots.
Serologically Specific Electron Microscopy (SSEM). This method and a few others
comprise the category of methods known as immunoelectron microscopy. Essential steps in
the SSEM procedure include: (1) the copper specimen support or grid is covered with a
parlodion film, (2) the filmed grid is floated on antiserum diluted 1: 100 to 1:5000 in tris
buffer, (3) after 30 minutes the grid is washed repeatedly to remove unabsorbed serum
proteins, (4) the grid is floated on an extract of virus-infected tissue from 10 minutes to 24
hours, (5) the grid is washed repeatedly to remove cellular debris and salts, (6) the grid is
then stained with 5% uranyl acetate for 1.5 minutes or longer; (7) the grid is washed with
distilled water or 95% ethanol, and fmally (8) the grid is blotted dry and examined in the
electron microscope. The procedure is shown in Figure 16.8. For seed diagnosis, dry seed is
ground by mortar and pestle or Wiley mill, or soaked seed is homogenized. The seed
preparation is applied to an electron microscope grid containing an antiserum specific for the
virus in question.
Seed Pathology and Pathological Testing 373

Figure 16.6. The agar double diffusion test. (A) Schematic showing the preparation and serological
analysis of barley embryo extracts. Antigen from the virus-infected embryo (ViE) formed a
precipitate (lens-shaped area) with antibody (Ab) molecules. (B) Quadrant petri dish with white
precipitates due to positive viru5-antibody reactions. Precipitate lines occur only between infected
embryos on the peripheral disks and the center antisera disks A, B, and C [Reproduced by
permission of the American Phytopathological Society from (Carroll /980)).
374 Seed Pathology and Pathological Testing

Figure 16.7. The latex jlocculation test (AJ Schematic of latex jlocculation method, (B) Two 100-111
pipettes containing suspensions of antibody-sensitized latex. Virus presence in the upper pipette
caused the suspension to becomejlocculated or aggregated Absence of virus in the lower pipette
was indicated by the dispersed appearance of the latex suspension [Reproduced by permission of
Zeitschr. Pflkrankh. Pjlshutz. From Lundsgaard (1976)].
Seed Pathology and Pathological Testing 375

/\

c
Figure 16.8. The serologically specific electron microscopy (SSEM) method (A) Principle of the
SSEM method, (B) schematic of the SSEM method, (C) electron micrograph showing presence of the
spherical particles of tobacco ringspot virus (Reproduced by permission of the American
Phytopathological Society from Brlansky and Derrick 1979).
376 Seed Pathology and Pathological Testing
Samples from infected seed lots reveal the presence of virus particles characteristic for
the suspect virus. According to Carroll (1979), when tobacco ring spot and soybean mosaic
viruses were tested, each virus could be detected in a singly infected soybean seed. The
SSEM method was sensitive enough to detect the two soybean-infecting viruses in a mixture
of 1 infected seed per 1000 healthy seeds. The method is simple for trained personnel, and it
is rapid, selective, and sensitive. However, it does require the use of a transmission electron
microscope.

Questions

1. Why isn't pathological testing of commercial seed lots considered more often as a routine
test by seed testing laboratories?
2. How and why has the international commerce in seeds influenced the need for testing for
seedborne diseases?
3. Do you consider the testing for seedborne diseases to be more difficult or complicated
than other kinds of seed quality tests? Why or why not?
4. Name some crops grown in your state, province, or country for which pathological
testing is very important.

General References

Agarwal, V. K.. and J. B. Sinclair. 1987. Principles of Seed Pathology. Vol. 1 and 2. Boca
Raton, FL.
Agarwal, V. K. and J. B. Sinclair. 1997. Principles of Seed Pathology, 2nd edition, Lewis
Publishers, Boca Raton, FL.
American Pathological Society. 1973. A compendium of corn diseases. 1973. St. Paul, MN:
American Phytopathological Society.
American Pathological Society. Compendium of soybean diseases. 1975. St. Paul, MN:
American Phytopathological Society.
Bacon, C. W. 1983. The fungal endophyte and tall fescue. In: Proceedings of Tall Fescue
Toxicosis Workshop, Atlanta, GA. pp. 34-42.
Baker, K. F. 1962a. Principles of heat treatment of soil and planting material. Journal of
Australian Institute ofAgricultural Science 28: 118-126.
_ _ . 1962b. Thermotherapy of planting material. Phytopathology 52:1244-1255.
_ _ . 1969a. Seed pathology-concepts and control. Journal of Seed Technology 4(2):57-67 .
. 1969b. Aerated steam treatment of seed for disease control. Horticultural Research
9:5973.
_ _ . 1972. Seed pathology. In: Seed Biology, ed. T. Kozlowski, 2:317-416. New York:
Academic Press.
Barnett, N. L., and B. B. Hunter. 1972. Illustrated Genera of Imperfect Fungi, 3rd ed.
Minneapolis. MN: Burgess Publishing Company.
Bolesky, D. P., J. J. Evans, and S. R. Wilkinson. 1985. Amino acid composition of tall fescue
seed produced from fungal endophyte (Acremonium coenophealum)-free and infected plants.
Agronomy Journal 77:796-798.
Booth, C. 1971. The Genus Fusarium. Kew. Surrey, England: Commonwealth Mycological
Institute.
Seed Pathology and Pathological Testing 377
Brlansky, R. H., and K. S. Derrick. 1979. Detection of seedborne plant viruses using
serologically specific electron microscopy. Phytopathology 69:96-100.
Carpenter, P. L. 1977. Microbiology. 2d ed. Philadelphia: W. B. Saunders Company.
Carroll, T. W. 1979. Methods of detecting seedborne plant viruses Journal of Seed
Technology 4(2):82-95.
_ _ . 1980. Barley stripe mosaic virus: Its economic importance and control in Montana.
Plant Disease 64: 138-140.
Carroll, T. W., P. L. Gossel, and D. L. Batchelor.1979. Use of sodium dodecyl sulfate in
serodiagnosis of barley stripe mosaic virus in embryos and leaves. Phytopathology 69:12-14.
Chidambaram, P., S. B. Mathur, and P. Neergaard. 1973. Identification of seedborne
Drechslera species. Friesia 10: 165-207.
Christensen, C. M., and R. A Meronuck. 1986. Quality Maintenance in Stored Grains and
Seeds. Minneapolis, Minn.: University of Mitmesota Press.
Christensen, C. M., and D. B. Sauer. 1982. Microflora. In: Storage of Cereal Grains and
Their Products, 3rd ed. C. M. Christensen, pp. 219-240. St. Paul, Minn.: American
Association of Cereal Chemists.
Chualprasic, C., S. B. Mathur, and P. Neergaard. 1974. The light factor in seed health
testing. Seed Science and Technology 2:457-475.
Clark, M. F., and A N. Adams. 1977. Characteristics ofthe microplate method of enzyme-
linked immunosorbent assay for the detection of plant viruses. Journal of General Virology
34:475-483.
Clay, K. 1987. Effects of fungal endophytes on the seed and seedling biology of Lotium
perenne and Festuca arundinacea. Oecologia 73:358-362.
Coleno, A, A. Tregalet, and B. Digat. 1976. Detection des lots de semances cantamines par
une bacterie phytopathogene. Annales de Phytopathologie 8:355-364.
Cooper, V. C., and D. G. A Waldey, 1978. Thermal inactivation of cherry leaf roll virus in
tissue cultures of Nicotiana rustica raised from seeds and meristem tips. Annals of Applied
Biology 88:273-278.
Cross, J. E. 1979. Importance of seed pathology in seed trade quality control programs.
Journal of Seed Technology 4(2):99-102.
DeBattista, J. P., C. W. Bacon, R. Severson, R. D. Plattner, and J. H. Bouton. 1990. Indole
acetic acid production by the fungal endophyte oftaH fescue. Agronomy Journal 82:878-880.
Derrick, K. S., and R. H. Briansky. 1978. Assay for viruses and mycoplasma using
serologically specific electron microscopy. Phytopathology 65:815-820.
Echandi, E., and M. Sun. 1973. Isolation and characterization of a bacteriophage for the
identification of Corynebacterium michiganense. Phytopathology 63: 1398-1401.
Ellis, M. B.1971. Dematiaceous Hyphomycetes. Kew, Surrey, England: Commonwealth
Mycological Institute.
Funder, S. 1961. Practical Mycology. 2d ed. New York: Hainer Publishing Company.
Guthrie, J. W. 1979. Routine methods for testing and enumerating seedborne bacterial plant
pathogens. Journal of Seed Technology 4(2):78-81.
Guthrie, J. W., D. M. Huber, and H. S. Fenwick. 1965. Serological detection of halo blight.
Plant Disease Reporter 49:297-299.
Hamilton, R. I., and C. Nichols.1978. Serological methods for detection of pea seedborne
mosaic virus in leaves and seeds of Pisum sativam. Phytopathology 68:539-543.
378 Seed Pathology and Pathological Testing
Hepperly, P. R., and J. B. Sinclair. 1978. Quality losses in Phomopsis-infected soybean
seeds. Phytopathology 68:1684-1687.
Heydecker, W., J. Higgins, and Y. J. Turner.1975. Invigoration of seeds? Seed Science and
Technology 3 :881-888.
Hutchins, J. D. and J. C. Reeves (eds.). 1997. Seed Health Testing. CAB International:
Wallingford. 263 pp.
Ilyas, M. B., O. D. Dhingra, M. A. Ellis, and J. B. Sinclair. 1975. Location of mycelium of
Diaporthe phaseolorum var. sojoe and Cercospora kikuchii in infected soybean seeds. Plant
Disease Reporter 59:17-19.
Jeffs, K. A., ed .. 1978. Seed Treatment. Cambridge, U.K.: Heffers Printers Ltd.
Kulik, M. M., and J. F. Schoen. 1977. Procedures for the routine detection of seedborne
pathogen fungi in the seed testing laboratory. Journal of Seed Technology 2(1):23-39.
Letham, D. B. 1977. Seed treatment research. New South Wales Nurserymen 2(9):7.
Limonard, T. 1965. Ecological Aspects of Seed Health Testing. Wageningen: International
Seed Testing Association.
Lister, R. M. 1978. Application of the enzyme-linked immunosorbent assay for detecting
viruses in soybean seed and plants. Phytopathology 68:l393-1400.
Lister, R. M., S. E. Wright, and J. M. Kloots. 1978. Sensitive detection of barley stripe
mosaic virus in barley seed and embryos by ELISA. Phytopathology News 12: 198.
Lundsgaard, T. 1976. Routine seed health testing for barley stripe mosaic virus in barley
seeds using the latex-test. Zeitschrijt Fur ZenkranAheiten Pjlsarzzen Pathologie 83:278-283.
Malone, J. P., and A. E. Muskett. 1964. Seedborne fungi. Proceedings of the International
Seed Testing Association 29:179-384.
Maude, R. B. 1996. Seedborne Diseases and Their Control. CAB International:
Wallingford. 280 pp.
McGee, D. C. 1979. Epidemiological aspects of disease control. Journal ofSeed Technology
4(2):96-98.
McGee, D. C. 1988. Maize Diseases: A Reference Source for Seed Technologists. St. Paul,
Minn.: APS Press.
McGee, D. C. 1992. Soybean Diseases: A Reference Source for Seed Technologists. St.
Paul, Minn.: APS Press.
Mink, G. 1., and 1. L. Parsons. 1978. Detection of pea seedborne mosaic virus in pea seed by
direct-seed assay. Plant Disease Reporter 62:249-253.
Nath, R., P. Neergaard, and S. B. Mathur. 1970. Identification of Fusarium species on seeds
as they occur in the blotter test. Proceedings of the International Seed Testing Association
35:121-144.
Naumova, N. A. 1972. Testing of Seeds for Fungus and Bacterial Infection. 3rd ed.
Washington, D.C.: U.S. Department of Agriculture and the National Science Foundation.
Neergaard, P. 1977. Seed Pathology, Vol. t, New York: John Wiley and Sons.
Neergaard, P. 1977. Seed Pathology, Vol. 2. New York: John Wiley and Sons.
_ _ .1978. Detection of seedbome pathogens by culture tests. Seed Science and Technology
1:217-254.
Noble, M. 1965. Introduction to series 3 of the handbook on seed health testing. Proceedings
of the International Seed Testing Association 30:1045-1121.
Noble, M., and M. J. Richardson. 1968. An Annotated List of Seedborne Diseases. 2d ed.
Wageningen, Netherlands: International Seed Testing Association.
Seed Pathology and Pathological Testing 379
Parker, M. c., and L. L. Dean. 1968. Ultraviolet as a sampling aid for detection of bean seed
infected with Pseudomonas phaseolicola. Plant Disease Reporter 52:534-548.
Phatak, H. C. 1974. Seedborne plant viruses-id(~ntification and diagnosis in seed health
testing. Seed Science and Technology 2:3-155.
Powell, C. C., Jr., and D. E. Schlegel. 1970. The histological localization of squash mosaic
virus in cantaloupe seedlings. Virology 42: 123-127.
Ralph, W. 1978. Enhancing the success of seed thermotherapy: Repair of thermal damage to
cabbage seed using polyethylene glycol (PEG) treatment. Plant Disease Reporter 62:406-
407.
Rice, J. S., B. W. Pinkerton, W. C. Stringer, and D. J. Undersander. 1990. Seed production
in tall fescue as affected by fungal endophyte. Crop Science 30: 1303-1306.
Richardson, M. J. 1979. An annotated list of seedborne diseases, 3rd ed. Commonwealth
Mycological Institute Phytopathological Papers 23:1-320.
Rodriguez-Marcano, A., and 1. B. Sinclair, 1978. Fruiting structures of Colletotrichum
dematium var. truncala and Phomopsis sojae in soybean seeds. Plant Disease Reporter
62:873-876.
Schaad, N. W. 1978. Use of direct and indirect immunofluorescence tests for identification of
Xanthomonas campestris. Phytopathology 68:249-252.
Schaad, N. W., and W. C. White. 1974. A selective medium for soil isolation and
enumeration of Xanthomonas campestris. Phytopathology 64:876-880.
Schneider, R. W., O. D. Dhingra, 1. F. Nicholoson, and 1. B. Sinclair. 1974. Colletotrichum
lrunea/um borne within the seedcoat of soybeans. Phytopathology 64: 154-155.
Schuster, M. L. 1972. Leaf freckles and wilt of corn incited by Corynebacterium
nebraskense Schuster, Hoff. Mandel, Lazar. Nebraska Research Bulletin 270: 1-40.
Shepard, 1. F. 1972. Gel-diffusion methods for the serological detection of potato viruses X,
S, and M. Montana Agricultural Experiment Station Bulletin 662.
Shepherd, R. 1. 1972. Transmission of viruses through seed and pollen. Principles and
Techniques in Plant Virology, ed. C. J. Kado and H. O. Agrawal, pp. 267-292. New York:
Van Nostrand Reinhold Company.
Sheppard, 1. W. 1979. Methods for routine detection of seedborne fungal pathogens. Journal
of Seed Technology 4(2):74-77.
Sinclair, 1. B. 1977. The microcosm of the soybean seed. Illinois Research 19(1):12-13.
_ _ _. 1979. The seed: A microcosm of microbes. Journal of Seed Technology 4(2):68-73.
Slack, S. A., and R. J. Shepherd. 1975. Serological detection of seedborne barley strip
mosaic virus by a simplified radial-diffusion technique. Phytopathology 65:948-955.
Taylor, 1. D. 1970. The quantitative estimation of the infection of bean seed with
Pseudomonas phaseolicofa. (Burkh.) Dowson. Annals ofApplied Biology 66:29-36.
 17
Seed
Marketing
For centuries, fanners used their own seed. This approach offered the advantage of
immediate availability of supply. However, it suffered from the disadvantage that farmers
were oriented more toward production of grain or forage and their harvesting and storage
facilities generally were not suitable for maintenance of optimum seed quality. Therefore, in
some years, fanners experienced stand failures from planting inferior seed. Moreover, this
process did not encourage the planting of seeds with improved genetic potential. The modem
seed industry has filled this void with its emphasis on varietal development and increased
yields which have economically benefitted the farmer. It is for these reasons that most farmers
now recognize that the purchase of seeds from a professional seed producer or reputable seed
company is in their best interest.
Seed marketing is the process through which seed moves from the farm where it is
produced to the consumer who plants it. Depending on the type of seed and the proximity of
its production to the site of its use, the marketing process may be simply a farmer exchange
or it may be a complicated series of transactions involving several mid-transactions and a
highly organized seed industry. Figure 17.1 illustrates the seed marketing cycle in its most
complicated form. Much of the seed produced in the world today, especially that which is
produced outside its area of use, follows this kind of marketing system.
Successful seed marketing requires a demand for seed, a mechanism to supply the seed
in a timely fashion, and an organizational structure to ensure that high-quality seed is
produced.

DEMAND

One of the first questions a seed company must address when considering seed
marketing is the size of the demand for seed. The detennination of a potential market is based
on cultivated acreage, seeding rates, and renewal rates. While the first two factors are
relatively easy to determine, the renewal rate is more difficult to calculate because it is based
on the return of the farmer to purchase seed on an annual or seasonal basis. When improved

L. O. Copeland et al., Principles of Seed Science and Technology

© Kluwer Academic Publishers 2001
Seed Marketing 381

\
E
J
Who1esa1er(s)

Figure 17.1. A generalized seed marketing scheme.

varieties are not available, farmers commonly set aside a portion of the harvested product for
the next year's seed. Since the varietal purity and yield potential of high quality seed of self-
pollinating varieties in some instances may be satisfactorily maintained for several
generations, replacement of seed may not be higher than 25-30% on an annual basis. This is
particularly true in developing countries and is one of the reasons why the establishment of
seed industries in lesser developed countries is slow to evolve. In contrast, the sale of hybrid
crops is more stable since the renewal rate of hybrid seed is 100%.
Another important component of seed mark~~ting is the creation of a demand for high
quality seed of improved varieties. Farmers tend to be conservative and must be convinced
that new varieties or related cropping techniques (cultivation, fertilizer application, pest
control, etc.) are superior to their present production practices, particularly when increased
costs and extra effort are involved. Therefore, most seed companies provide local field
demonstrations and yield trials to demonstrate the superiority of their newer, higher-yielding
varieties. They also provide an opportunity for public relations and a chance to promote their
public image to farmers.

SUPPLY

Successful seed marketing requires that seed be supplied at the right time in the correct
quantity, at the place where it is used, and in appropriate packaging. Planting of seed
generally occurs in an intense period of only a few weeks at the beginning of the growing
season. If satisfactory quantities of seed are not available prior to planting, the seed will not
382 Seed Marketing

be sold. To facilitate timely supply, farmers are encouraged to preorder their seed so that the
seed company can anticipate the level of demand. Because many farmers tend to delay this
important decision until varieties and costs of seed are compared, inventory management in a
seed company is a difficult process. While seed can be delivered directly to the farmer, this
often does not occur. Instead, most farmers obtain the product directly from a retailer.
Therefore, the seed must be packaged in a manner which enables ease of transport and
handling. Often, this means that seed is packaged in 50-lb paper, plastic, or burlap bags.
More recently, many suppliers have began supplying seed in large bulk containers so that
there is less handling of numerous bags and the time-consuming process of opening smaller,
individual bags is avoided.

ORGANIZATION

Seed production often is centered in one area of a country. This is because certain
locales have better soil and climatic regimes conducive to production of high-quality seed.
Yet, the demand for the seed may be national and international. As a result, marketing
requires an organizational infrastructure where there is a system of distribution of seed from
a central production point to the customer. This often entails the use of brokers, wholesalers,
and retailers to ensure satisfactory seed distribution (Figure 17.1).
Seed marketing is central to the success of any seed operation. The most efficient seed
company will be organized so that all operations of the company flow through a central
marketing office (Figure 17.2). This allows the marketing manager to forecast farmer
demand, plan production dates and location of seed stock needs (inventory control of varieties
and amounts), determine the level of sales promotion, and to communicate the status of seed
supply with retail outlets (seed dealers). Such an organizational structure results in fewer
management crises and permits smoother operation of the company.
Various organizations market seed. When no reliable and effective marketing system
exists (e.g., in many developing countries), national governments often establish seed
companies that are operated by the government or its agency. This allows the government to
encourage the development of improved varieties and to educate farmers about their benefits
in producing higher yields. The rate at which a new variety is accepted by farmers depends on
a number of factors. The first is agronomic superiority. The increased yield or improvement
in other aspects of quality must cover the extra price for the seed and any other production
inputs. In many cases, the government reduces the cost of the seed to the farmer to minimize
the alternative use of feed grains which are often provided at below-market costs. Eventually,
farmers understand that their profits are greater when they purchase improved seed rather
than using their own home-grown seed. When demand increases, most governments
encourage privatization of the seed industry. This can occur by groups of interested seed
purchasers establishing cooperatives where the integrated marketing and distribution
operations are shared by a central authority. More often, however, private seed companies
recognize that profits are better in a stable marketplace with a continuing demand for high-
quality seed from a better educated clientele. When this occurs, a private seed industry
quickly develops.
Seed marketing schemes in the United States and Canada for various kinds of seeds,
including small-seeded grasses and legumes, hybrid corn and sorghum, cereal grains and
soybean, and miscellaneous crops are described below.
Seed Marketing 383

PRESIDENT

I I I I
PRODUCTION PROCESSING MARKE11NG FINANCIAL PERSONNEL
MANAGER MANAGER MANAGER MANAGER MANAGER

I I
MANAGER GENERAL
MARKETING RESEARCH. PLANS SALES MANAGER
FACILITATING SERYICES

1 J
FIELD SALES ORGANIZATION
MARKET PLANNING
ADVERTISING SALES OFFICE CONTROL
SALES ANALYSIS. BUDGETING CUSTOMER SERVICE AND
FORECASTING, PROMOTION PRODUCT SERVICE
INVENTORY CONTROL
PRODUCTION SCHEDULING
DISTRIBUTION QUOTAS &
TERRITORIES

Figure 17.2. Effective, market-oriented seed program organization which can focus all activities
toward identifoing, developing, and satisfYing farmers ' demandfor improved seed. (From Law et al.
1971)

Small-Seeded Grasses and Legumes

Since most of the forage and turf grasses and legumes is produced outside its major area
of use, almost none is sold directly to the user; instead it is sold to local seed houses and
enters the marketing chain similar to that illustrated in Figure 17.1. The seed buyer may be an
independent seed producer or part of a large seed company with a national or even
international organization. The seed may be sold after it has been cleaned in the producer's
farm cleaning plant or it may be taken "in the rough" directly from the field; in either
instance, however, the grower is usually paid on a dean seed basis.
Although seed of the small-seeded grasses andllegumes may be produced and sold on the
open market, much of it is produced under contra(;t with local seed dealers. These contracts
are usually made before the seed field is established, and they specifY the terms of the price to
be paid, the quality expected, and responsibilities of both the grower and the contractor. Such
contracts protect the seed producers by ensuring them a supply of seed at a reasonable,
predetermined price. Contracts tend to stabilize seed prices, acting as a hedge against short
supplies and rising prices. They provide growers with a reasonable margin of profit above
their production costs and protect them against an unstable market. The contracts often
384 Seed Marketing

specifY the services of a technical representative who advises the producer on cultural
practices needed for maximum seed production.
If growers elect to produce seed without a contract, they may do so; however, the price
they receive depends on the market at the time they are ready to sell. Sometimes this may
work in their favor, although the odds may favor them if they choose to produce under
contract. The success of contract production and the mutual satisfaction of both the seed
producers and growers are reflected in the large amounts of the seed of small-seeded grasses
and legumes produced under contract.
In Chapter lOwe learned that there is still some grass and legume seed produced within
the area of utilization. Most of this is wholesaled to local seed outlets on the open market or
grown under contract similar to that discussed above; however, because of the proximity and
accessibility to the consumers, some may be retailed directly by the seed grower to the
farmers who uses it.

Hybrid Corn and Sorghum

Most of the seed of hybrid corn and sorghum is produced by selected growers under
contract with a few large seed companies, who mostly market the seed through farmer-dealers
to their customers. However, there is an increasing trend to sell through local elevators or
other seed outlets who, in turn, retail it to local farmers. Hybrid corn seed is also sold by
numerous small seed companies who produce and market it throughout the corn belt.

Other Small Grains and Soybeans

Most seed of small grains and soybeans is marketed through local elevators or seed
dealers. Seed ofthese crops is usually produced without contracts and is sold at the prevailing
market price; however, contract production is becoming more common, especially by larger
seed handlers, who wish to ensure themselves adequate supplies of seed for their customers at
a reasonable cost.

Flower and Vegetable Seeds

Flower and vegetable seeds are produced almost entirely under contract and under rigid
control of private seed companies, who usually determine the varieties to be grown, the
quantity of seed needed, and the price of seed to the producer as well as the ultimate
consumer. Rigid control is possible because the varieties are usually privately developed,
have secret pedigrees, and are needed only in carefully regulated quantities.

Field Bean Seed

The marketing of field bean seed depends on its area of production. Perhaps one-half of
the seed of field beans is produced in the western United States, where it is usually grown
under contract with seed companies; however, field bean seed is also produced in the
commercial bean areas, particularly in North Dakota, Michigan, Ontario, and New York. In
these areas, most seed is sold without contracts to local elevators at the prevailing market
Seed Marketing 385
price. However, many of the elevators, commerciall bean companies and even some large seed
growers arrange for contract production to hedge against price increases and ensure adequate
supplies.
In contrast to field beans, seed of garden beans is grown almost exclusively in the
western United States-mostly under contract with local seed dealers who may represent
independent businesses or parts of larger seed companies. Many of the independent seed
producers also handle field bean seed as well.

Cotton Seed

In the southern United States, the great majority of cotton seed is controlled by a few
large companies who contract with dependable farmers to produce the seed under certain
restrictions and the state certification standards. In the West, most of the cotton seed is
produced by grower-owned seed organizations who contract with farmers for the seed
production.

MARKETING SEED OF PUBLIC VERSUS PRIVATE VARIETIES

Traditionally, access to seed of new crop varieties that have been developed and released
from public experiment stations or university plant-breeding programs has been free and
equally available to all growers and seed producers wishing to produce, promote, and market
the seed. The initial seed of new varieties has traditionally been released through foundation
seed organizations for increase and subsequent availability as certified seed. Thereafter, the
seed enters the public domain and is made available to all growers and farmers desiring it.
Consequently, no individual or seed company is able to monopolize the seed production and
marketing of publicly developed varieties.
In contrast, the seed production and marketing of crop varieties developed by privatr:
seed companies is tightly controlled. This system has been relatively easy for varieties of
vegetables, where the commercial product grown from the seed is also under company
control. It has also worked well for corn and sorghum hybrids, for which seed stocks are
under company control and loss of hybrid vigor incurred by replanting the progeny from seed
comprises a type of biological varietal protection. Prior to passage of the Plant Variety
Protection Act of 1970, no legal protection was available to owners or developers of varieties
of sexually propagated crops. This Act, discussed more fully in Chapter 18, provides
protection for sexually propagated crop varieties and provides legal protection to their owners
in controlling the production and marketing of seed.
In recent years, considerable numbers of publicly developed varieties, particularly
forage and turf varieties, have been released to private seed companies. Such exclusive
releases are usually covered by formal memoranda of understanding setting forth the specific
terms of the agreement. Normally, the exclusive ndease does not circumvent the usual role of
foundation and certified seed organizations, but merely gives the company the right to control
seed production and marketing of the variety. Some agreements stipulate that only certified
seed may be sold under the variety name.
Exclusive releases seem to be justified for varieties for which: (I) the area of seed
production is outside the area of commercial use, (2) seed production is a difficult or
hazardous specialty (e.g., tobacco and sugar beet seed), or (3) the volume of seed needed of
386 Seed Marketing

certain specialty crops is not large enough to justifY participation of many different parties. In
such cases, it is felt that no individual company or person can be expected to incur the
expense of seed production, promotion, and marketing of unproved varieties without a
guarantee that their competitors will not be able to capitalize on their efforts. Many superior,
badly needed varieties have "died on the vine" because they were released openly and no one
company would risk investing in seed production and promotion.
Since the passage of the Plant Varietal Protection Act, many public institutions are
obtaining varietal protection on their new varieties. Such varietal protection obviously
benetits the recipient company in the case of exclusive release by assuring them that another
company cannot legally violate their assigned rights by selling seed of the same variety.
With the advent of variety protection, owners of protected varieties may sell ownership
of their varieties or collect royalties from others whom they allow to produce and sell such
seed.

THE SEED INDUSTRY

In a survey of retail seed outlets in New York and New Jersey, 81 % of the respondents
indicated that the retail seed business was a sideline rather than a main enterprise. Aside from
the farmer-dealer marketing of com and sorghum hybrids and seed of certain other specialty
crops, this is probably a fairly accurate reflection of the marketing of most field seeds
throughout North America. Most crop seed is sold by local elevators or cooperatives as a
sideline to the grain, feed, fertilizer, and agricultural chemicals industry. While much of this
is purchased wholesale from local seed growers, a substantial portion, particularly of turf,
forage, and vegetable seed, is purchased from wholesale seed dealers or brokers who trade
almost exclusively in seeds.
Aside from the type of seed outlet described above, many companies exist that specialize
in wholesaling seed. Much of their business is concerned with trading in seeds produced
outside their marketing area, although they may handle locally produced seed as well. Most of
the turf and forage seed produced in the western United States is eventually sold in the
consuming states through these types of wholesale outlets. In tum, the wholesalers supply the
seed to local elevators, cooperatives, and lawn and garden stores, who ultimately retail it to
farmers, homeowners, sod growers, and hobbyists. Many seed wholesalers of this type are
equipped with extensive conditioning facilities; therefore, they can purchase seed in bulk from
larger seed companies or western seed outlets and reclean, blend, package, and prepare the
seed according to the needs of their customers and their own specifications. Because of the
competitive nature of this type of business, many of these companies have an aggressive sales
program, which includes salespeople who regularly service and attend to the needs of their
customers.
Another type of seed business is characterized by large seed companies that are
organized on a regional, national, or even international basis. These may be active in the
production as well as in the consuming areas; some marketing is done through elevators and
cooperatives, and some seed is sold through their own retail outlets in the consuming area.
The companies may be specialized in scope, such as seed of tield and garden beans,
vegetables, and flowers; others are highly diversified, with different divisions handling seeds
of field crops, vegetables, turfgrass, flowers, and some specialty crops. These companies are
active in the international seed trade, and have highly organized domestic seed production and
marketing programs.
Seed Marketing 387
SEED TRADE ORGANIZATIONS

Like most professions, the seed trade is highly organized. Seed trade organizations
provide their members an opportunity to meet and associate on a professional and social level
with persons having mutual interests. More importantly, the organization gives the seed trade
identity and power in influencing public opinion as well as state and national seed legislation.

United States - American Seed Trade Association (www.amseed.com)

The seed trade within the United States is organized into state, national, regional, and
specific interest associations. While most states have their own seed dealers associations, the
national seed industry is organized into the Ameriican Seed Trade Association (ASTA). The
AST A was organized in 1883. Today it is divided into five regional associations representing
the Pacific, Western, Northern, Atlantic, and Southern regions. It is divided by commodity
interests into five divisions-the farm seed division, garden seed division, hybrid corn and
soybean division, lawn and turf division, and flower seed division. Each division has its own
staff of officers and committees. The ASTA has a board of directors, and officers are elected
annually, but a permanent full-time executive vice president is employed to give continuity to
its administration.
The activities of AST A are many and varied and, of course, geared to the interests of the
seed industry. The AS T A has been highly effectiv,e as a lobbying organization in influencing
state and federal seed legislation. An example of its effectiveness was the passage ofthe Plant
Variety Protection Act of 1970, for which the ASTA was largely responsible. In addition,
ASTA sponsors educational meetings, such as its annual Farm Seed Conference and the
Hybrid Corn, Sorghum and Soybean Research Conferences. It also sponsors a national crops
judging contest for agronomy students. Through a special organization, known as the
American Seed Research Foundation, it sponsors seed research projects at public institutions.
The results of this research are published and made available throughout the industry, and are
doing much to broaden our knowledge of seeds.

Canada - Canadian Seed Trade Association (www.cdnseed.org)

The seed industry of Canada is organized into the Canadian Seed Trade Association
(CSTA). It was organized in 1923, following a small gathering of seed trade representatives
called by the Seed Commissioner of Canada to appoint trade representatives to the Advisory
Board under the new Canadian Seeds Act, which was soon to become effective. Since that
time, it has continued to be active as a lobbying agency in the interest of the seed trade of
Canada; it has also fostered professional association within the Canadian seed industry and
with that of the United States and other parts of the world.
CST A maintains a full staff of officers-a president, first vice president, and full time
executive secretary. All officers assume responsibility for contacting members of the trade in
their area and passing on information to their members in other parts of the country through
the executive secretary's office.
388 Seed Marketing
Federation Internationale du Commerce des Semances (FIS) (www.worldseed.org)

The Federation International du Commerce des Semances is an international seed trade

organization. Its purpose is to foster cooperation among the nations of the world in respect to
facilitating the international commerce in seeds. It acts as a voice of the international trade in
seeds to discourage national policies restricting the free movement of seeds across national
boundaries. Although it has no policy-making powers, it is effective as an educational force
in influencing national and international policies affecting the international movement of seed.

Questions

1. What are the advantages and disadvantages of growing seed under contract?
2. How is the system of marketing hybrid corn seed different from that of grass seed
crops?
3. How much seed is retailed directly to the ultimate consumer by seed growers in your
state?
4. Do you think it is good business for soybean seed producers in the Midwest to sell part
of their production directly to consumers and to wholesale part of it through elevators
and seed dealers? Why or why not?
5. Do you think that exclusive release of publicly developed varieties is justified? If so,
when and under what circumstances?
6. What class or species of seed generally follows the marketing cycle illustrated in Figure
17.1? How does the marketing of other kinds of seed deviate from this?
7. Do you feel the sale of seed in the local supermarket promotes high-quality seed?
8. If you wanted to establish a home lawn, where would you purchase your seed?

General References

Douglas, J. E., ed. 1980. Successful Seed Programs: A Planning and Management Guide.
Boulder, CO: Westview Press.
Gregg, B. R. 1983. Seed marketing in the tropics. Seed Science and Technology 11: 129-148.
Law, A. G., B. R. Gregg, P. B. Young, and P. R. Cherty. 1971. Seed Marketing. New Delhi,
India: National Seeds Corporation.
Thompson, J. R. 1979. An Introduction to Seed Technology. New York: John Wiley and
Sons.
U.S. Department of Agriculture. 1961. Seeds: The Yearbook of Agriculture. Washington,
D.C., U.S. Government Printing Office.

I. Carter, W. B., and E. P. Bugbee, Jr. How we get seeds of vegetables and flowers, pp.
493-499.
2. Christensen, D. K., Earl Sieveking, and J. W. Neely, Handling seed of the field crops,
pp. 499-506.
3. Heckendorn, W., and R. A. Edwards. Jr. The four types of seed trade associations, pp.
517-521.
4. Kuzelka, T. J., and W. H. Youngman. Statistics and trends, pp. 521-530.
5. McCorkle, C. O. Jr., and A. D. Reed. The economics of seed production, pp. 530-540.
Seed Marketing 389
6. Schiffman, 1. F., and R. W. Schery. The responsibilities of the seedsman, pp. 514-517.
7. Schery, R. W. Grass seeds for lawns and turf, pp. 507-513.

Van Gastel, A. J. G. 1986. Seed marketing. In: Seed Production Technology, eds. J. P.
Srivastava and L. T. Simarski, pp. 232-236. Aleppo, Syria: ICARDA.
Wagner, K. P., H. F. Creupelandt, and W. H. Verburgt. 1975. Seed marketing. In: Cereal
Technology ed. W. P. Feistritzer, pp. 108-129. Food and Agriculture Organization, Rome.
Wheeler, W. A., and D. D. Hill. 1957. Grassland Seeds. New York: D. Van Nostrand
Company.
 18
Seed Legislation
and
Law Enforcement
Thou shalt not sow thy fields with mingled seed-(Lev. 19: 19)

Seed laws are designed to aid in the orderly marketing of seed. They establish regulations
governing the sale of seed, thereby providing legal protection to both buyers and sellers. No
country can expect to have a well-developed, effective seed industry without seed control
regulations. In the United States, seed legislation exists at both the state and federal levels.
The basic purpose of both state and federal seed law is truth-in-Iabeling. Although some
seed laws are designed to protect even the uninformed consumer, most require only that the seed
be completely labeled for quality. It is assumed that buyers will read the label and select seed
lots that meet their criteria. In many states, it is legal to sell even poor-quality seed if it is
properly labeled.
The truth-in-labeling concept was developed to avoid the type of caveat emptor, or "let the
buyer beware" marketing philosophy prevalent in the early English markets. There,
unscrupulous vendors sometimes used clever gimmicks to pass on virtual trash as agricultural
seed to unsuspecting customers. There were even cases of "factories" established for the purpose
of preparing so-called seed for sale. Often this was done by coloring small gravel and other inert
material to substitute for, or mix with, the seed offered for sale. Another fraudulent scheme was
to place sweepings, chaff, or other seed types in the center of seed containers to increase their
bulk, a practice sometimes called "stove piping."

FEDERAL SEED LEGISLATION

History of Federal Legislation

Federal seed legislation in the United States dates back to 1905, when the Annual
Appropriations Act was passed, giving the USDA authority to purchase seeds on the open
market, test them for adulteration or mislabeling, and publish the test results together with the

L. O. Copeland et al., Principles of Seed Science and Technology

© Kluwer Academic Publishers 2001
Seed Legislation and Law Enforcement 391
names of the persons offering the seed for sale. Under the provisions of this Act, between 1912
and 1919 approximately 15,000 samples were tested, and 20% of the samples were found to be
adulterated or mislabeled.
The second major federal seed legislation was the Seed Importation Act of 1912 restricting
the importation of principal forage crops that were below minimum purity specifications and
above maximum weed seed content. It was first amended in 1916 to require a minimum live seed
requirement for imported seed, amended again in 1926 to require coloration of imported alfalfa
and red clover, and further amended in 1926 to prohibit the shipment in interstate commerce of
falsely or fraudulently labeled seed. The first amendment was added because much of the alfalfa
and clover seed imported was unsuited for production in the United States, and coloration was
intended to inform buyers of the possible lack of adaptation of crops grown from the seed.

The Federal Seed Act

Beginning in about 1936, discussions took place between several interested agencies with
respect to further changes in the seed laws, which resulted in the enactment of the Federal Seed
Act in 1939. This Act is the single most important piece of seed legislation in United States
history. It applies to all agricultural and vegetable seeds, to imported seeds, and to that sold in
interstate commerce. A major improvement over the Seed Importation Act, it provided more
detailed labeling requirements and did not require proof of intent to defraud in cases of
mislabeling. The Act was amended in 1956 to allow civil prosecution for complaints of
violations and was amended again in 1960 to require labeling of pesticide-treated seed. Further
amendments have been made as needed.
The Federal Seed Act is a truth-in-Iabeling law that governs the sale of seed in interstate
commerce and seed imported into the United States. The aim of the Act is to provide detailed
regulations covering sale of seed on a national basis. Normally, it has no jurisdiction over seed
produced and marketed within state boundaries. The federal and state seed laws contain
somewhat similar requirements. If seed is labeled to comply with the Federal Seed Act and is
shipped in interstate commerce, it will normally comply with the labeling requirements of the
state into which it is shipped. Thus, the Act helps maintain the integrity of each state seed law;
however, no state may set standards for seed moving into the state from another below the
minimum required by the Federal Seed Act. A state may have standards below those of the
Federal Seed Act covering seed sold within the state's boundaries. However, it is a violation of
the Federal Seed Act to move seed into a state in which it does not comply with the state's
noxious weed seed restrictions, even though the seed may otherwise comply with the
requirements of the federal law.
Though the Federal Seed Act is seemingly detailed and complex, most of its requirements
are simple and should be well understood by everyone buying or selling seed. In general, the
following information is required on the label (see Figure 18.1):

1. The name of the kind or kinds and variety for each agricultural seed component present in
excess of 5% of the whole and the percentage by weight of each. If the crop is one that is
regarded by the Secretary of Agriculture as generally labeled as to variety, the label shall
bear either the name of the variety or the statement "Variety Not Stated."
392 Seed Legislation and Law Eriforcement

r- ""

/ ANALYSIS
TAG
~
Perfection Seed Co.
Middletown, U.S.A. •
Vernal
Variety & Kind Alfalfa
Lot. No: 307·31

Pure Seed: 98.90%

Inert Matter: 01.05%
Other Crop Seed: 00.00%
Weed Seed: 00.05% !
Noxious Weeds: 00.00% II

Germination 90.00%
Hard Seed 05.00% '
Date Tested: Jan. 1994
Net Weight: 601bs.

Figure 18-1. A typical labeling tab.

2. Lot number or other identification.

3. Origin, if determined by the Secretary of Agriculture that the crop is one in which the
origin is important from the standpoint of crop production. If the origin is unknown, that
fact shall be stated.
4. Percentage by weight of weed seeds, including noxious types.
5. Kind and rate of occurrence (per lb. or oz.) of noxious weed seed, labeled in accordance
with the law of the state into which the seed is shipped.
6. Percentage by weight of agricultural seed present other than those named in No. 1 above.
7. Percentage by weight of inert matter.
8. Germination percent for each agricultural seed in excess of 5% and percentage of hard
seeds, and the calendar month and year the test was completed to determine such
percentages.
9. Name and address of the seller, or the person to whom the seed is sold, together with a
code designating the seller.
10. The year and month beyond which an inoculant, if shown on the labeling, is no longer
claimed to be effective.

STATE SEED LAWS

The Federal Seed Act is important because it applies to all seed marketed on a national
basis across state boundaries. However, it has no jurisdiction on seed sold produced and sold
within state boundaries. As a result, states have established their own seed laws.
Seed Legislation and Law Enforcement 393
Each of the 50 states has its own seed law that regulates the sale of seed within the state.
Seed laws vary among states, since each is constructed to meet the unique needs of a particular
state. All are primarily truth-in-labeling laws, although some are more protective and specify
minimum quality standards that seed must meet to be eligible for sale.
In most states, seed laws are administered by the state department of agriculture. If a state
does not have a department of agriculture, another state agency is responsible for seed law
enforcement. For example, in Indiana seed law eJllforcement is the responsibility of Purdue
University, which designates a faculty member as the Commissioner of Agriculture. Regardless
of the specific arrangement, the responsible agency must maintain a staff of seed inspectors, who
actually go to locations where seed is marketed and check for compliance with the seed law.
Representative seed samples are routinely and randomly collected from seed dealers, elevators,
railroad cars, lawn and garden stores (see Figure 18.2), and even supermarkets to be tested for
compliance with the seed law. Violations are reported, and appropriate action taken to correct
the violation or to stop sale ofthe seed. A stop sale is the action by a state seed control agency
that prohibits further sale of seed until violations are corrected. Often, the state seed law requires
that a notice of all violations be periodically published and made available to the public.

The Need for Standardization

A major goal of the American seed industry has been to promote uniformity among state
seed laws. Uniformity among laws is an advantage to the seed trade in that it simplifies interstate
commerce in seed. Encouragement toward uniformity has also come from seed testing
associations as well as federal seed control officials. The first suggested uniform state seed law
was developed by the Association of Official Seed Analysts (AOSA) and the seed trade in 1907-
1917. After the Federal Seed Act of 1939 was enacted, the USDA promoted the Recommended
Uniform State Seed Law patterned after suggestions made by the AOSA and the American Seed
Trade Association. The movement for uniformity among seed laws was aided in 1949 by the
formation of the American Association of Seed Control Officials (AASeO), an organization of
state and federal seed control officials. This organization has as its major goal the uniformity
in seed laws, in rules and regulations, and in the uniform administration of laws pertaining to
the sale and distribution of seeds.
In 1957, the AASCO published a "Recommended Uniform State Seed Law" (RUSSL) with
the approval of its various member agencies. This law has no officialjurisdiction, but is merely
a model for states that desire to revise their seed laws. It provides both form and wording for
revisions that can be made to promote uniformity among state seed laws. Although this law has
been useful, it is recognized that each state has its own unique situations and its own interests
to protect; consequently, no move toward complete uniformity is anticipated.
FEDERAL-STATE COOPERATION
Enforcement of the Federal Seed Act is the responsibility of the Seed Branch of the
Livestock, Meat, Grain, and Seed Division, Agricultural Marketing Service of the USDA. To
help carry out this responsibility, a federal seed laboratory is located at Beltsville, Maryland.
This laboratory has historically assumed the additional responsibility for helping to standardize
seed-testing activities and procedures throughout the United States. Federal-state cooperative
agreements have been worked out through which apparent violations of the Federal Seed Act
394 Seed Legislation and Law Ef'!forcement

Figure 18.2. Sampling garden and flower seed at the point ofsale.

are brought to the attention of Seed Branch officials by various state officials. When an apparent
labeling violation occurs, seed samples are exchanged and tested by both the federal and state
laboratories. If a violation is established, the federal seed officials may make further
investigations and send all information and evidence to the Seed Branch headquarters in
Beltsville, MD. Depending on the nature and seriousness of the violation, several alternatives
can be followed. A warning letter may be sent to the violator about the apparent violation so that
appropriate steps may be taken to prevent such action in the future. Another option is a cease
and desist proceedings, which is similar to a court order to cease from such violations. In
addition, the violator may be fined or even prosecuted under the criminal section of the act. Also,
seed may be seized by the courts to prevent further sales. When seed is seized through court
procedure, the court may permit the sale of seed after the violation has been corrected, or may
permit its sale in another state where the seed would not be in violation of state laws. In extreme
cases, the court may even order the seed destroyed. As with many state seed laws, the Federal
Seed Act requires that a list of violations be maintained showing violators' names, the nature of
the violations, and the penalty imposed.
Seed Legislation and Law Enforcement 395

PROVISIONS OF STATE AND FEDERAL SEED LAWS

Farmer Seed Exchange

Exchanges of seed between farmers are exempt from labeling laws; however, the laws
carefully define such exemptions. Ifthe seed is advertised in any way, the seed must be labeled.

Current Germination Tests

Since seed will deteriorate over time, the month of the test on which the germination is
based must appear on the label. Under the Federal Seed Act, no more than five months may
elapse between the last day of the month in which the test was completed and the date of
transportation and delivery in interstate commerce. If the seed is in hermetically sealed
containers, a period of 24 months is permitted. State seed laws vary in the length of time
permitted since the last germination test.

Labeling Vegetable Seed Containers

Minimum germination standards are outlined lor vegetable seeds in the Federal Seed Act.
If seed being sold or shipped in interstate commerce has a germination as good or better than
the minimum standard for that particular kind of setxi, the germination of seed in packets of one
pound or less does not have to be indicated on the label. If the germination is below the
minimum standard, the statement "Below Standard" and the germination percentage must be
given on the label.

Transport for Conditioning

If consigned to a seed cleaning or conditioning establishment, seed is exempt from labeling

laws provided the statement, "Seed for Conditioning" is on the invoice and other shipping
records and is clearly indicated on the seed container.

Disclaimers Not Allowed

Statements disclaiming responsibility for the information on the seed label are disallowed
by all federal and state seed laws.

Collection of Damages

No damages may be collected through the seed laws even though a violation is established.
Damage must be collected through separate action in a civil court.

Proof of Intent not Needed

Original seed laws were often ineffective because even when violations occurred, the state
had to prove intent to defraud, which was hard to do. After the 1956 amendment to the Federal
Seed Act, violators could be penalized without proof of intent or carelessness.
396 Seed Legislation and Law Enforcement

Coloration and Labeling of Treated Seed

A Federal Food and Drug rule requires that seed that has been treated with a chemical
pesticide must be dyed a color that contrasts with that ofthe seed. Usually red or green dyes are
used. In addition, under federal and state seed laws the seed container must bear a statement that
the seed has been treated, along with the commonly accepted chemical or abbreviated chemical
name of the substance; if the chemical is harmful to humans or animals in the amount applied,
a statement such as "Do not use for food or feed or oil purposes" must also be included on the
tag, along with an antidote to be used ifthe chemical is taken internally.

Noxious Weed Seeds

In interstate trade, it is a violation of the Federal Seed Act to ship seeds containing noxious
weed seeds that are in excess of that allowed by the receiving state. Since each state seed law
has established a list of prohibited and restricted noxious weeds (designated as primary and
secondary noxious weeds in some states), the requirements vary among states. The sale of seed
lots containing primary noxious weed seeds is prohibited. The sale of seed lots containing
secondary noxious weed seeds is permitted, but the name and number (per ounce or pound) must
be stated on the label. Some states limit the number of secondary noxious weed seeds that may
be present per pound. Table 18.1 shows the classification of noxious weeds of Michigan.
Because of the importance of accurate identification of weed seeds, the Association of
Official Seed Analysts has developed Handbook 25 on the Uniform Classification of Weed and
Crop Seeds. This handbook was first published in 1952 and was subsequently revised in 1964,
1977, 1993 and 1998 because of continuing changes in scientific terminology. The purpose of
the handbook is to (I) develop a better system of classifying contaminating species as other crop
or weed seed, (2) provide a comprehensive listing and classification of over 2,000 species, (3)
bring the scientific nomenclature of these species up to date, and (4) provide a comprehensive
reference source to other scientific nomenclature systems. Handbook 25 is a valuable resource
for standardizing the classification of noxious weed seeds found in a seed lot. Without this,
incorrect evaluations could be made that would make perfectly acceptable seed lots unacceptable
for sale because they would be labeled to contain weed seeds.

Imported Seeds

Special regulations of the Federal Seed Act apply to seeds imported into the United States.
Screenings of any seed are not permitted to be imported except for screenings of wheat, oats,
rye, barley, buckwheat, field corn, sorghum, and certain other edible crops that are not imported
for seeding purposes and are declared for cleaning, conditioning, or manufacturing purposes.
Screenings of other kinds of seeds may not be imported.
Imported seeds that contain 10% or more of seeds of alfalfa, red clover, or both must be
stained to indicate that such seed is of foreign origin. When foreign-grown seed is mixed with
domestically produced seed, each component must contain at least 10% stained seed. If a seed
lot contains any specified noxious weed seeds or more than 2% by weight of any weed seeds or
less than 75% pure live seed (with certain exceptions), it is considered unfit and ineligible for
import into the United States.
Seed Legislation and Law Enforcement 397
Table 18.1. Noxious Weeds of Michigan

Prohibited Noxious
(Prohibited in all seed offered for sale in Michigan)
1. Bindweed (Convolvulus arvensis) 5. Russion knapweed (Centaurea picris)
2. Canada thistle (Cirsium arvense) 6. Leafy spurge (Euphorbia esula)
3. Perennial sow thistle (Sonchus arvensis) 7. Quack grass (Agropyron repens)
4. Whitetop (Lepidium draba) 8. Horse nettle (Solanum carolinense)
Restricted Noxious
(Restricted occurrence in all seed offered for sale in Michigan)
1. Dodder (Cuscuta spp.) 6. Wild carrot (Daucus carota)
2. Fan weed (Thlaspi arvense) 7. Wild onion (Allium spp.)
3. Wild mustard (Brassica kaber, juncea 8. Giant foxtail (Setaria faberii)
and nigra) 9. Yellow rocket or wintercress (Barbarea
4. Hoary alyssum (Bmeroa incana) vulgaris)
5. Buckhorn plantain (Plantago lanceolata)

Keeping of Records
State and federal seed laws require that comlPlete records for each lot of seed sold be
maintained for varying lengths of time. Complete records refer to information relating to origin,
treatment, germination, or purity of each lot of agricultural seed handled. The following
information must also be maintained: declarations, labels, seed samples, records of purchases,
sales, cleaning and bulking, handling, storage, analysis, and tests. Each person shipping seed
in interstate commerce must retain a sample representing each lot of agricultural seed shipped.
Federal regulations provide that any seed sample may be discarded one year after the entire lot
represented by such sample has been sold.

Definition of Sale or Offer for Sale

Seed legislation is intended to cover regulations for seed sold or offered for sale. The term
offer for sale includes representations of the salesperson, oral or written advertisements,
including radio and television advertising, and price lists, newspapers, catalogs, and pamphlets
that pertain to the seed for sale.

Definition of Seed Quality Terminology

The definition for seed quality terminology used in state and federal seed laws relies on the
Rules for Testing Seeds of the Association of Official Seed Analysts (AOSA 2000). For
example, the Federal Seed Act defines the term germination to mean the percentage of seeds
capable of producing normal seedlings under favorable conditions, which is the definition in the
Rules for Testing Seeds of the AOSA. Many other terms, such as pure seed, other crop seeds,
inert matter, also were adopted from the AOSA rules.

Labeling
The Federal Seed Act and most state seed laws define the term label as the display or
disp lays of written, printed, or graphic matter upon or attached to the container or seed. For
398 Seed Legislation and Law Enforcement

example, any of the printing on the bag or on any cartons or on any tags attached to bags is
considered to be a part of the label. The term label is not confined to the information on the tag
attached to the bag.

Amending Seed Laws

In establishing seed laws and regulations in the United States, state and federal officials
give due consideration to the interest of the seed industry, for which many of the laws and
regulations are intended. From the beginning of seed legislation in the United States, the seed
industry and government officials have worked together in preparing legislation. Any regulations
that are passed by state and federal seed laws, or any changes to these regulations, are adopted
only after a public hearing at which any person in the state or United States is free to make
comments in person or in writing. This makes certain that the views of interested
parties-whether in industry, in seed law enforcement, or among consumers are given due
consideration. However, the final decision for the new law or changes in the law must be made
by the duly constituted legislative bodies, relying largely on the advice of officials of the agency
responsible for enforcing the act.

THE PLANT VARIETY PROTECTION ACT-

LEGAL PROTECTION FOR CROP VARIETIES

The U. S. Plant Variety Protection Act, signed into law in 1970, provides legal protection
in the production and sale of seed by owners or developers of new varieties of sexually
propagated crops. Developers of varieties of asexually propagated (budding or grafting, etc.)
crops have received protection since 1930 through the U.S. Patent Office. This protection has
enab led them to control the propagation of their varieties and to collect a royalty for any use and
propagation of their varieties by other parties. Prior to the Plant Variety Protection Act of 1970,
such protection had not been available to originators of sexually propagated crop varieties.
The Plant Variety Protection Act is administered by the Plant Variety Protection Office
within the U.S. Department of Agriculture. The office receives applications and evaluates each
candidate on the basis of the following criteria: (1) novelty, (2) uniformity, and (3) stability.
Novelty is described as distinctiveness (i.e., whether it can be distinguished from all known
varieties of the same crop on the basis of morphological, physiological, or cytological
characteristics). Uniformity requires that all variations within the population must be
describable, predictable, and commercially acceptable. Stability requires that its essential and
distinctive characteristics remain unchanged throughout successive generations of seed increase.
The owner of a protected variety may assign hislher rights to others, or even sell them.
Protection lasts for 17 years, at which time the variety becomes public property. In cases of
emergency, or when needed to supply the country with sufficient food, fiber, or feed, the
Secretary of Agriculture may declare a variety available to the public; however, in such rare
instances, however, the owner would be compensated for its public use.
The Act provides two avenues of protection to the owner of a variety. First, it provides
exclusive rights for the propagation and use of a protected variety. In cases of infringement,
however, the owner is responsible for pursuing or defending these rights in a civil court. Second,
it gives the owner the right to stipulate in the application that the variety name be protected
through seed certification. It is a violation of Title V ofthe Federal Seed Act to sell by variety
name uncertified seed of varieties protected under this stipulation.
Seed Legislation and Law Enforcement 399

A special exemption for farmers grants them the right to produce seed of a protected
variety for their own use. Plant breeders may also use protected varieties for breeding purposes
since the use of a protected variety for breeding purposes does not constitute infringement of the
ownership rights under the Act.

CANADIAN SEED LAWS

The Canadian Seed Control Act was enacted in 1905 to set minimum standards for pure
seed, common and noxious weed content, and germination. It was amended in 1911 to establish
more definite requirements. In 1923, a grading system was introduced for all seeds in commerce.
The Seed Branch of Agriculture Canada (the Canadian Department of Agriculture) provides the
seed inspection work for law enforcement.
The philosophy of Canadian seed legislation provides truth-in-Iabeling for seeds in
commerce; however, the Canadian seed law is considerably more protective than the U.S.
Federal Seed Act. Crop varieties in Canada are lic<msed to protect the seed user or consumer
against losses that can occur from the purchase of unknown and inferior seed. Licensing is also
intended to prevent deception from the sale of seed under modified or false variety names. The
Canadian law provides a system whereby only registered (licensed) varieties that have been
tested and found agronomically and economically desirab Ie for Canadian agriculture are allowed
to be sold, advertised, or imported. The licensing of varieties is administered by the Plant
Products Division of Agriculture Canada and covers all agricultural and vegetable crop seeds.
Seeds of root and vegetable crops, other than seed potatoes, are exempt.

EUROPEAN SEED LAWS

In contrast to the truth-in-Iabeling seed laws in the United States, most European seed laws
appear to be based on the assumption that the gov~rnment knows what is best for the farmer.
Many European seed laws strictly control what can be sold within or imported into a country.
This is done in various ways.
In France, seeds of varieties of most cereals, herbage crops, peas, vetches, horse beans,
white clover, and the most important grasses cannot be sold unless the variety name appears on
the official list. Only certified seed of these varieties can be sold. Field trials for approval of
varieties are conducted by an official government agency. Five years of testing are required for
approval. France has one of the most restrictive seed laws in effect, and its seed law is often
used as a model by other countries.
England's varietal protection law has a varietal indexing system, official trials for testing
all varieties submitted, and a list of acceptable varij~ties based on their performance. Although
participation in the official tests is voluntary, seed cannot be sold by varietal name unless it has
been entered into the testing program.
Most other European countries have rather restrictive seed laws similar to those in France
and England. Until recently, Denmark had no seed law but relied almost entirely on an education
approach to consumer protection. However, due to their membership in the European Common
Market, they have recently drafted and adopted seed control legislation similar to that in other
common market countries.
400 Seed Legislation and Law Enforcement

SEED LEGISLATION IN DEVELOPING COUNTRIES

Fanners in most developing countries generally save their own seed from year to year or
exchange seed with their neighbors. This system of agriculture involves almost no commerce in
seed and has little need for seed control legislation. However, as improved varieties become
available and agricultural development occurs, trade in seed naturally follows, leading to the
necessity for seed control legislation.
The first seed laws in developing countries are usually truth-in-labeling laws, requiring that
seed be labeled as to quality, and that such labels truthfully represent the actual seed quality.
For such laws to be effective, seed testing laboratories in which quality tests can be performed
must be available, and farmers and seed dealers must be encouraged to use these services.
Strong research, seed certification, and extension programs also promote effectiveness of seed
control legislation.
As agriculture and the seed trade gain sophistication and become more complex, seed
legislation tends to become more complex and restrictive. Most countries contemplating seed
legislation seek advice from other countries that have long histories of successful seed
legislation.

Questions

1. Can you list several noxious weeds in your state? What is meant by the term caveat
emptor? How does this concept apply to seed labeling today?
2. What is meant by the concept of truth-in-labeling?
3. In State A, the minimum germination of agricultural seed allowed for sale is 60%. If a lot
of alfalfa seed was shipped from State B into State A where it was offered for sale,
sampled by state seed control officials, and found to germinate 45%, would the seed lot
be in violation of the seed law in State A? The Federal Seed Act? Both? Why? Assume
that a part of the same seed lot was shipped into State C, which has no minimum
germination requirement. Would it still be a violation of the Federal Seed Act? Why or
why not?
4. What information is required on the seed label in your state?
5. What is the difference between a stop sale and a seizure?
6. What procedure must a seed regulation undergo to be amended?
7. Is it illegal to offer seed that has not been tested for gennination and purity to a
neighboring farm?
8. Is it illegal to sell untested seeds in a retail outlet or store?
9. What is a disclaimer and what is its legality?
10. What is the law in your state pertaining to the labeling of treated seed?
11. What is the philosophical difference between seed laws in the United States compared to
those in most European countries? Why do you think this difference exists?
12. Assume that a certified seed grower conditions his/her own seed, obtains a seed analysis,
and labels the seed accordingly. The seed is then sold to a wholesaler, who in tum sells
it to a local elevator where it is resampJed and found to be mislabeled. However, the
grower's name is still listed on the label with the labeling infonnation. Who is responsible
for the mislabeled information according to the seed law?
Seed Legislation and Law Enforcement 401
13. What is the basic purpose (or philosophy) of seed laws in the United States?
14. What kind of plants are covered by the Plant Variety Protection Act of 1970? What crops,
if any, are excluded from coverage under this Act?

General Refer4mces

Association of Official Seed Analysts. 2000. Rules for testing seeds. Lincoln, NE. AOSA
Publication.
Douglas, 1. E., ed. 1980. Successful Seed Programs: A Planning and Management GuMe.
Boulder, Colo.: Westview Press.
Larsen, A. L., 1. H. Wiersema, and T. Handwerker. 1993. Uniform Classification of Weed and
Crop Seeds. 137 pp. Contribution No. 25 to the Handbook on Seed Testing. Association of
Official Seed Analysts, Lincoln, NE.
Thompson, J. R. 1979. An Introduction to Seed Technology. New York: John Wiley and Sons.
U. S. Department of Agriculture. 1961. Seeru: The Yearbook ofAgriculture. Washington, D.C.:
U.S. Government Printing Office.

1. Clark, E. R. Sometimes there are frauds in seeds, pp. 478-482.

2. Clark, E. R., and C. R. Porter. The seeds in your drill box, pp. 474-478.
3. Crispin, W. R. Seed marketing services, pp. 470-474.
4. Davidson, W. A. What labels tell and do not tell, pp. 462-469.
5. Rollin, S. F. and F. A. Johnston. Our laws that pertain to seeds, pp. 482-492.
 Glossary
AASCO. American Association of Seed Control Officials representing state and federal seed
law enforcement officials throughout the United States.
Abnormal seedling. A seedling (in a germination test) that does not have the essential
structures indicative ofthe ability to produce a normal plant under favorable conditions.
Absorption. The uptake of moisture into the tissm:s of an organism (e.g., seed).
Achene. A small, dry, one-seeded fruit with a thin, dry wall that does not split open at
maturity (e.g., sunflower seed).
Acorn. The fruit of an oak; see the definition of a nut.
ADP. Adenosine-diphosphate, a complex sugar-phosphorus compound formed as the result
of expenditure of energy and the loss of a phosphate group from the energy-rich ATP
(adenosine-triphosphate) compounds.
Adsorption. The accumulation and adhesion of a thin layer of water (or gases) on the surface
of another substance.
Adventitious embryony. A condition in seed in which the embryo arises from somatic (body)
rather than reproductive tissue. This condition is common in certain grasses and often results
in multiple embryos.
After-ripening. A term for the collective changes that occur in a dormant seed that make it
capable of germination. It is usually considered to denote physiological changes.
Agamogony. A type of apomixis in which c,ells undergo abnormal meiosis during
megasporogenesis, resulting in a diploid ~mbryo sac rather than the normal haploid embryo
sac.
Agamospermy. A type of apomixis in which sporophytic tissue is formed, ultimately leading
to seed development.
Aggregate fruit. Fruit development from several pistils in one flower, as in strawberry or
blackberry .
Air screen cleaner. The basic piece of equipment for cleaning seed, utilizing air flow, and
perforated screens (also called a fanning mill).
Albuminous seed. A seed having a well-developed endosperm or peri sperm (nucellar origin).
Aleurone layer. The layer of high-protein cells surrounding the storage cells of the
endosperm. Its function is to secrete hydrolytic enzymes for digesting food reserves in the
endosperm.
Ambient conditions. The outside conditions (e.g., relative humidity and temperature) that
exist at any given time and place.
404 Glossary

Amino acid. Organic acid containing one or more amino groups (-NH 2), at least one carboxyl
group (-COOH), and sometimes, sulfur. Many amino acids are linked together by peptide
bonds to form a protein molecule. Proteins are a fundamental constituent of living matter.
Amphitropous ovule. A type of ovule arrangement in which the ovule is slightly curved so
the micropyle is near the funicular attachment.
Amylase. The enzyme responsible for catalyzing the breakdown of starch into sugars. It may
be active in one of two forms: a-amylase and p-amylase.
Amylopectin. A type of starch mo lecule com posed of long, branched chains of glucose units
(a polysaccharide).
Amylose. A type of starch molecule made up of glucose units in long, unbranched chains (a
polysaccharide ).
Anatropous. A type of ovule arrangement in which the ovule is completely inverted, having
a long funiculus with the micropyle adjacent to the base of the funiculus.
Androecium. Collectively, the stamens of a flower.
Angiosperm. A kind of plant that has seeds formed within an ovary.
Annual. The type of plant that normally starts from seed, produces its flowers, fruits, and
seeds, then dies within one growing season.
Annuoline. The fluorescent protein pigment exuded by the roots of ryegrass seedlings. The
fluorescent nature of this material makes it useful in distinguishing annual and perennial
ryegrass.
Anther. The saclike structure of the male part (stamen) of a flower in which the pollen is
formed. Anthers normally have two lobes or cavities that dehisce at anthesis and allow the
pollen to disperse.
Anthesis. The period of pollenation, specifically the time when the stigma is ready to receive
the dispersed pollen.
Antipodal nuclei. Three of the eight nuclei that develop from the megaspore by mitotic cell
divisions within the developing megagametophyte (embryo sac). They are usually located at
the base of the embryo sac and have no apparent function in most species.
AOSA. Association of Official Seed Analysts, the organization of state and federal seed
analysts from the United States and Canada.
AOSCA. Association of Official Seed Certifying Agencies, the organization composed
mostly of certification agencies from the United States and Canada-formerly (prior to 1968)
known as the International Crop Improvement Association (lCIA).
Apical placentation. A type of free-central placentation in fruit where the seeds are attached
near the top of the central ovary axis.
Apogamy. A type of apomixis involving the suppression of gametophyte formation so that
seeds are formed directly from somatic (body) cells of the parent tissue.
Apomixis. Seed development without the benefit of sexual fusion of the egg and the sperm
cells.
Archesporial cell. The cell ofthe nucellus that differentiates and gives rise to cells ultimately
destined to undergo meiosis and produce the megaspore mother cell.
Glossary 405

Aril. A loose, papery appendage in some seeds (e.g., elm) originating as an extension (or
proliferation) from the outer integument.
Aspirator. An air-blast separator. A seed conditioning (cleaning) machine that uses air to
separate seeds according to specific gravity (weight) and resistance to air flow.
Asexual reproduction. Reproduction by vegetative means without the fusion of two sexual
cells.
ASTA. American Seed Trade Association.
Astered embryo type. A type of embryo classification in which the terminal cell ot the
proembryo divides by a longitudinal wall and both the basal and terminal cells contribute to
embryo development.
ATP. Adenosine-triphosphate, an energy-rich complex sugar phosphorus compound which
provides energy for many metabolic reactions.
Auxins. A group of growth regulators that may stimulate cell growth, root development, and
other growth processes, including seed germination.
Awn. A slender appendage often associated with seeds, such as the "beards" of wheat or
barley.
Axile placentation. The type of ovule attachment within a fruit in which the seeds are
attached along the central axis at the junction of the septa.
Bacteriocide. A chemical compound that kills bacteria.
Bacteriophage. A virus that infects specific bacteria and usually kills them. Specific phages
are used to identify certain bacterial plant pathogens.
Basal placentation. A type offree-central placentation in which the seeds are attached at the
bottom of the central ovary axis.
Berry. A simple, fleshy, or pulpy and usually many-seeded fruit that has two or more
compartments and does not burst open to release its seeds when ripe (e.g., blueberry).
Biennial. A kind of plant that produces only vl~getative growth during its first growing
season. After a period of storage or overwintering lOut-of-doors, flowers, fruits, and seeds are
produced during the second year and the plant dies (i.e., a plant that requires two years to
complete its life cycle).
Biological seed treatment. Use offungi or bacteria to control soil and seed pathogens instead
of synthetic chemical seed treatments.
Bitegmic testa. A testa (seed coat) composed of two integumentary layers.
Blend. A term applied to seed mixtures of different crop varieties (or species) that have been
mixed together to fulfill a specific agronomic purpose.
Brand. A legal trademark registered by a particular company or distributor for its exclusive
use in marketing a product such as seeds or plants.
Breeders' rights. Varietal protection-the legal rights of a breeder, owner, or developer in
controlling seed production and marketing of crop varieties.
Brick grit test. A type of seedling emergence (vigor) test utilizing uniformly crushed brick
gravel through which seedlings must emerge to be considered vigorous.
406 Glossary

Buckhorn machine. A machine that mixes water and sawdust with seed lots containing seeds
of buckhorn plantain. The mucilagenous seed coat of the buckhorn seed attracts the water and
sawdust and changes its size and specific gravity, allowing it to be separated by the air screen
machine or gravity separator.
Bulb. An enlarged, fleshy, thick, underground part of a stem surrounded by thickened, leafy
scales and shortened leaves. Roots develop at the base of a bulb (e.g., wild onion).
Bulbil. A small bulb or bulblet produced above the ground, as in wild garlic.
Bumper mill. A machine designed to clean timothy seed by a continuous bumping action on
an inclined plane. The uncleaned seed is metered onto the plane, which is continuously
bumped by sets of knockers. The cylindrical timothy seeds are rolled into separate grooves
while noncylindrical contaminants are jarred off the end of the inclined plane and separated.
Callus. A hard or thickened layer at the base of certain grass florets.
Calyx. A collective term for all the sepals surrounding a flower; it forms part of the covering
of some seeds.
Campylotropous ovule. A type of ovule arrangement in which the ovule is slightly curved
and the micropylar end is pointed slightly downward so the funiculus and micropyle are close
together on the mature seed on opposite sides of the hilum, as in legumes.
Capsule. A dehiscent fruit with a dry pericarp usually containing many seeds.
Carpel. Female reproductive organ of flowering plants. One or more carpels may be united
to form the pistil.
Caruncle. A fragile appendage or outgrowth of the outer integument of the seed of some
species (e.g., leafy spurge).
Caryopsis. A dry, indehiscent one-seeded fruit (as in grasses) in which the pericarp and
integuments are tightly fused.
Catalase. An enzyme that catalyzes the degradation of hydrogen peroxide to water and the
oxidation by hydrogen peroxide of alcohols to aldehydes during seed germination.
Catalyst. A substance that can induce or accelerate a chemical reaction without undergoing
any change itself.
Catkin. A spike in fluorescence with a single unisexual flower arising from the peduncle, as
in Alnus rubra (red alder).
Caveat emptor. A Latin expression meaning "let the buyer beware," often applied to seed
marketing prior to the consumer protection era.
Cellular endosperm. A type of endosperm in which the early development is characterized
by cell wall formation accompanying each nuclear division.
Cellulose. A long-chain complex carbohydrate compound (polysaccharide) with the general
formula (C 6 H 120 6 )n. It is the chief substance forming cell walls and the woody parts of plants.
Certified seed. Seed produced under an officially designated system of maintaining the
genetic identity of, and including provisions for, seed multiplication and distribution of crop
varieties. It also refers to the class of certified seed which is the progeny of registered or
foundation seed. It is identified by a blue tag; thus, it is sometimes called "blue-tag" seed.
Glossary 407
Chaffy grass divider. A subsampling device used to divide a sample of chaffy grass seed into
a working sample.
Chalaza. The part of an ovule where the integuments originate. In orthotropous ovules the
chalaza is directly underneath the funicular attachment. In other types of ovule arrangement
it can sometimes be distinguished on the outside of the seed near the hilum (e.g.,
campylotropous (legumes)).
Chromosome. A rodlike-bearer of hereditary material (genes) inside the nucleus of all cells.
Chenopodiad embryo type. A type of embryo classification in which the terminal cell of the
pro-embryo divides by a transverse wall and both the basal and terminal cells contribute to
embryo development.
Circadian rhythm. See endogenous rhythms. A lype of rhythmic plant or animal growth
response which appears to be independent of external stimuli.
Circinotropous ovule. A type of ovule arrangement in which the funiculus is very long and
completely encircles the ovule, which otherwise has an orthotropous (straight) arrangement.
Circumscissle capsule. A capsule which at maturity splits open at the middle so that the top
comes off like a lid (e.g., plantain).
Clone. A group of individuals (plants) of common ancestry that have been propagated
vegetatively, usually by cuttings or by multiplication of bulbs or tubers.
Cold test. A type of stress (vigor) test that tests th,~ performance of seeds in cool, moist soil
in the presence of various soil microorganisms. The test is conducted by planting the seeds in
moist, unsterilized field soil, exposing them to cool (5-10°C) temperatures for about a week,
then allowing them to germinate in the same soil at warmer temperatures.
Coleoptile. A transitory membrane covering the shoot apex of certain species that protects the
plumule as it emerges through the soil. The coleoptile is photosensitive and stops growth when
exposed to light, allowing the plumule to break through and continue growth.
Coleorhiza. A transitory membrane covering the emerging radicle (root apex) in some species.
It serves the same function for the root as the coleoptile does for the plumule.
Color separator. A machine that separates seeds on the basis of their color differences.
Coma. A tuft of hairs attached to a seed (e.g., "brush" on wheat).
Complete flower. A flower that has pistils, stamens, petals and sepals.
Complete hybrid. A legal designation for a seed lot indicating that at least 95% ofthe seed
represents hybrid seed.
Compound cyme. A determinate inflorescence where there is secondary branching, and each
ultimate unit becomes a simple cyme (e.g., Sapanoria officinales).
Conditioning. The term used to describe the process of cleaning seed and preparing it for
market; formerly called processing.
Conductivity test (of seed leachates). An electrical conductivity test that associates the
concentration of leachates from seeds, after soaking in water, to their quality.
Constancy of destination. The theory of em bryo development that suggests that each part of
the mature embryo inevitably arises from predetermined cells of the proembryo.
408 Glossary

Corymb. An indeterminant inflorescence in which the lower pedicels arising from the
peduncle are successively longer than the upper ones, giving a rounded or flat-topped
appearance (e.g., Prunus emarginata).
Cotyledon. Seed leaves of the embryo. In most dicotyledon seeds they are thickened and are
storage sites of reserve food for use by the germinating seedling.
Coumarin. A chemical growth inhibitor that has germination-inhibiting capability.
Crassinucellate. The nuclear condition in which the embryo sac originates and develops deep
(several cell layers) under the nucellar epidermis.
Cristae. The inner folds of the mitochondrial membranes where many enzymes catalyzing
metabolic processes are located.
CSTA. Canadian Seed Trade Association.
Crucifer embryo type. A type of embryo classification in which the terminal cell of the
proembryo divides by a longitudinal wall and the basal cell plays only a minor part (or none)
in subsequent embryo development.
Cultivar. A variety of a cultivated crop.
Cutin. A complex fatty or waxy substance found on the surface of certain seeds or leaves,
often making them impermeable to water.
CVPO. Certification of seed on the basis of varietal purity only, referring to a type of seed
certification that certifies seed as to genetic purity without specific criteria for purity,
germination, and other aspects of mechanical seed quality.
Cyme. A type of inflorescence in which the main axis ends in a flower. Further growth is by
lateral branches, which may also terminate in a flower.
Cytoplasm. The contents of a cell between the nucleus and the cell wall.
Damping-off. A condition in which young seedlings are attacked (parasitized) and killed by
soil-borne fungi immediately following germination. This condition is sometimes mistaken
for poor seed quality.
Day-neutral plants. Plants that have no daylength requirements for floral initiation.
Desiccant. A chemical applied to crops that prematurely kills their vegetative growth; often
used for legume seed crops so the seed can be harvested prior to normal plant senescence.
Dehiscence. The splitting open at maturity by pods or capsules along definite lines or sutures.
Detasselling. Artificially removing (cutting or pulling) the tassel of the female parent to
prevent selfing during hybrid seed corn production.
Determinant flower. A floral axis which terminates as a flower rather than a vegetative bud.
Dicot. An abbreviated name for dicotyledon which refers to plants having two seed leaves.
Dioecious (diecious). Refers to plants having stamens and pistils on different unisexual plants.
Therefore, both sexes must be grown near each other before seed can be produced (e.g.,
American holly).
Diploid (2n). Refers to two sets of chromosomes. Germ cells have one set and are haploid;
somatic cells have two sets and are diploid (except for polyploid plants).
Diploid apogamety. A type of diplospory (apomixis) in which seed develops from some cell
other than the egg but does not require the stimulus of pollination.
Glossary 409
Diploid parthenogenesis. A type of diplospory (apomixis) in which seed develops from the
unfertilized egg of the embryo sac without the need for pollination.
Diploid pseudogamy. A type of diplospory (apomixis) in which the stimulus of pollination
is needed for seed development.
Diplospory. A type ofagamospermy(apomixis) in which a diploid embryo sac is formed from
archesporial origin.
Disclaimer. A statement (legally invalid) on a seed container or label disavowing
responsibility for the information contained on the label.
Disinfectant. A chemical treatment used to disinfect seed for planting. It is especially useful
for surface-borne pathogens.
Diverticulae. The tendrill ike forks projecting from the ends of the haustoria (adsorptive arms)
of the developing embryo or endosperm.
DNA. Deoxyribonucleic acid, a component of the nucleus (chromosomes) and the basic
building blocks of genes. It carries the hereditary information of a cell.
Dormancy. A physical or physiological condition of a viable seed that prevents germination
even in the presence of otherwise favorable germination conditions.
Double cross hybrid. The first generation progcmy of a cross between two single-cross
hybrids.
Drupe. One-seeded, stone fruit (e.g., cherry, peach, plum).
Dry-bulb temperature. The temperature measured with an ordinary thermometer.
Ecotype. A strain within a given species adapted to a particular environment.
Equilibrium moisture content. The moisture content (e.g., of a seed) at which there is no
further tendency to gain or lose moisture at any given relative humidity.
Electrophoresis test. A method for separating and mapping protein bands from homogenized
plant (or seed) preparations. The separations are made within a gel preparation across an
electrical field. The test may have potential use in variety identification and tests for varietal
purity.
Electrostatic separator. A machine that separates seed on the basis of their ability to accept
and retain an electrical charge.
Elite seed. The class of Canadian pedigreed seed c:orresponding to the foundation seed class
in the United States.
Embryo. The generative part of a seed that develops from the union of the egg cell and sperm
cell and during germination becomes the young pliant.
Embryo sac. The female sexual spore of the ovule; also known as the mature female
gametophyte or megagametophyte.
Embryogeny. Embryo growth and development.
Endogenous rhythm. A type of rhythmic plant or animal response or growth capacity that is
not affected by external stimuli (see circadian rhythm).
Endocarp. Inner layer of the fruit wall (pericarp).
410 Glossary

Endosperm. The tissue of seeds that develops from sexual fusion ofthe polar nuclei of the
ovule and the second male sperm cell. It provides nutrition for the developing, growing
embryo.
Enzyme. A catalyst produced in living matter. Enzymes are specialized proteins capable of
promoting chemical reactions without themselves entering into the reaction; consequently,
they are not changed or destroyed.
Epicotyl. The portion of the embryo or seedling above the cotyledons.
Epidermis. The outer layer of cells in plants that protects them against drying and mechanical
injury.
Epigeal germination. A type of germination in which the cotyledons are raised above the
ground by elongation of the hypocotyl.
Epistase. The development of a well-defined nucellar on integumentary tissue (or plug) in the
micropylar or chalazal area of an ovule.
Ergot. Dark spur-shaped sclerotium that develops in place of a healthy seed in a diseased
(fungus-infected) inflorescence. Ergot sclerotia are toxic to both man and livestock and were
an original source of the hallucinatory drug LSD.
Exalbuminous seeds. Seeds with only small amounts of endosperm.
Excised embryo test. A quick test for evaluating the growth potential of a root-shoot axis that
has been detached from the remainder of the seed.
Exhaustion test. A type of vigor test that measures the ability of seeds to grow rapidly under
rigidly controlled conditions of high temperature, relative humidity, and moisture content in
continuous darkness.
Exocarp. Outermost layer of the fruit wall (pericarp).
F) hybrid. Denotes the first generation offspring from the mating of two parents.
F2 seed. The second generation progeny from the mating of two parents.
Fanning mill. The air-screen machine that utilizes air flow and sieving action in separating
and cleaning seeds.
Far-red light. The radiant energy in the long wavelength range of the visible spectrum
between 700 and 760 nanometers.
Fat. An ester of three fatty acids and glycerol (or another alcohol) found in plants or animals.
When they exist in liquid form, they are frequently called oils.
Fatty acid. Organic compound of carbon and hydrogen that combines with glycerol to make
a fat.
Fermentation. Chemical change in sugar induced by the activity of the enzyme systems or
microorganisms under anerobic conditions. For example, in the brewing industry, yeast
enzymes produce carbon dioxide and alcohol from sugar by fermentation.
Field burning. The practice of burning the stubble and plant residue of seed fields after
threshing. It is a cultural practice for aiding insect and disease control as well as stimulating
tiller production and delaying the onset of sodbinding in perennial sod-forming grasses
Filament. The stalk that supports the anther in the stamen (male part) of a flower.
Film coating. Application of seed additives in a "sticky" polymer directly to the seed in one
to multi-layers that increase the seed's weight 1 to 10% to improve seed performance.
Glossary 411

Firm seeds. A term sometimes applied to grass caryopses that are dormant due to seed coats
that are impervious to water or gases.
First generation hybrid. An F 1 hybrid.
FIS, or (ASSINSEL). Federation International du Semances-an international federation of
the seed trade.
Floral induction. The physiological changes in n::sponse to external stimuli (light quality,
daylength, etc.) that occur in vegetative meristems and subsequently allow them to become
reproductive meristems and undergo floral initiation.
Floral initiation. The morphological changes in the development of a reproductive meristem
from a vegetative meristem.
Floret. The smallest unit of a flower. In grasses it consists of the lemma, palea, stamens, and
pisti I.
Florigen. The universal hormone that supposedly causes plants to change from the vegetative
to the reproductive state.
Fluid drilling. Pregermination of seeds in gels sw;h as hydroxyethyl cellulose followed by
furrow planting to enhance germination, stand establishment, and seedling growth.
Follicle. A fruit with a simple (single) pistil that at maturity splits open along one suture (e.g.,
milkweed, larkspur).
Foundation seed. The progeny of breeder seed; used as planting stock for registered and
certified seed.
Free central placentation. The type of ovule attachment within a fruit that bears seeds along
a free central axis with no separations (septa).
Fruit. A mature ovary and any associated parts.
Fungicide (seed). A chemical that disinfects the se1ed and/or protects it from soil-borne fungi
during germination.
Funiculus. The stalk that connects an ovule (seed) to the placenta of the ovary wall.
Gamete. A sex cell that unites with another sex cell to form a zygote.
Gamet precision divider. A type of mechanical halving device for subdividing a large seed
sample to obtain a smaller working sample for germination or purity analysis. It has an
electrically operated rotating cup into which the seed is funneled to be spun out and into one
of two spouts.
Gametophyte. The part of the flower that produces gametes or sex cells.
Gene(s). Units of inheritance located in linear ord,er on chromosomes.
General seed blower. A precision seed blower us,~d to aid in separating light seed and inert
matter from heavy seed.
Genetic drift. A gradual (or sometimes abrupt) change in the germplasm balance of a cross
pollinated variety causing a change in its characteristics. Usually applies to grass or legume
varieties when seed is produced outside their area of adaptation. The shift may be caused by
selective differences in plant mortality or flowering habit under the different environment.
Genetic purity. Trueness to type or variety, usually referring to seed.
412 Glossary

Genotype. The hereditary makeup ofa plant (or variety) which determines its inheritance.
Germ. A term for the embryo of some seeds, especially the cereal grains.
Germplasm. An expression used in a broad sense to denote the hereditary properties of an
individual plant or plant population that are transmitted from one generation to another.
Germ tube. The tube that grows out from the pollen grain, usually into the stigma, down the
style and into the ovary to perm it sexual fusion.
Germination. The resumption ofactive growth by the embryo culminating in the development
of a young plant from the seed.
Gibberellic acids. A group ofgrowth promoting substances first discovered in Gibberella spp.
which regulate many growth responses and appear to be a universal component of seeds as
well as other plant parts.
Glomerule. A very compact cyme (e.g., Saxilfaga integrifolia).
Glumes. The pair of chafty bracts that occur at the base of a grass spikelet, often completely
closing it.
Gravity separator. A machine utilizing a vibrating porous deck and air flow to separate seeds
on the basis of their different specific gravity.
Growth regulator. A synthetic compound produced in the laboratory which controls growth
responses in plants and seeds.
Gymnosperm. A kind of plant that produces seeds but no fruits. The seeds are not borne
within an ovary and are said to be naked (hence the name).
Gynoecium. The female part of a flower or pistil formed by one or more carpels and
composed ofthe stigma, style, and ovary.
Hard seed. A seed that is dormant due to the nature of its seed coat, which is impervious to
either water or oxygen.
Haploid (IN). A term indicating one-half the normal diploid complement of chromosomes.
Haustoria. In seeds, a type of armlike absorptive organ sometimes projecting from the
developing endosperm or embryo into other seed parts to gather nutritive support.
Head. An inflorescence in which the floral units on the peduncle are tightly clustered and
surrounded by a group of flowerlike bracts called an involucre (e.g., sunflower).
Helobial endosperm. Intermediate between the nuclear and cellular endosperm types in which
development is characterized by free nuclear division as well as cell wall formation in some
areas.
Hemianatropous ovule. A type of ovule arrangement in which the straight ovule axis
orientation is perpendicular to that of the funiculus.
Hemicellulose. Complex cell wall constituent that is similar in appearance to cellulose but
more easily broken down to simple sugars. Common forms include xylan, mannans, and
galactans.
Hesperidia. Berrylike fruit with papery internal separations (septa) and a leathery, separable
rind (e.g., citrus fruits).
Hilum. The scar remaining on the seed (ovule) at the place of its detachment from the
seedstalk (funiculus).
Glossary 413

Hormone. A chemical substance that is produced in one part of a plant and used in minute
quantities to induce a growth response in another part.
Hybrid vigor. The increase in vigor of hybrids over their parental inbred types; also known
as heterosis.
Hydrogen peroxide (H202) test. Quick test to determine seed viability. In response to a H2 0 2
soak, viable seeds elongate their roots through a cut in the seed coat; a commonly used quick
test for conifer seeds.
Hygroscopic. In seeds, the high tendency to take up moisture, even as water vapor.
Hypha. A thread of a fungus mycelium. Plural-hyphae.
HypocotyI. The part of the embryo axis between the cotyledons and the primary root which
gives rise to the stalk of the young plant.
Hypogeal germination. A type of germination in which the cotyledons remain below the
ground while the epicotyl grows and emerges above the ground.
lelA. International Crop Improvement Association which in 1968 was renamed the
Association of Official Seed Certifying Agencies (AOSCA).
Imbibition. The initial step in seed germination involving the uptake of moisture by
absorption from the germination media and hydration of the seed tissue.
Imperfect flower. Unisexual flowers; flowers lacking either male or female parts.
Inbred. A plant from successive self-fertilizations of parents throughout several generations.
Inclined draper. A device for separating seeds using an inclined endless belt onto which
seeds
are metered; seeds are separated on the basis of their different tendencies to roll down the
plane or to catch and be carried up and into a separate discharge spout.
Incomplete flower. A flower that lacks any of the four basic parts (pistils, stamens, sepals,
petals).
Increase (seed). To multiply a quantity of seed by planting it, thereby producing a larger
quantity of seeds which may also be called an increase.
Indehiscent. Not splitting open at maturity.
Indent cylinder separator. A seed separator utilizing a rotating indented cylinder through
which seeds are passed for cleaning. It lifts shorter seeds from longer seeds, thus separating
them.
Indent disk separator. A seed separator utilizing multiple rotating disks inside a cylinder
through which seeds are moved. It lifts shorter seeds from longer seeded types, thus separating
them.
Indeterminate flower. A flower which terminates in a bud which continues to be
meristematic throughout the growing season, resulting in flowers of different maturity within
the same inflorescence.
Inert matter. One of the four components of a purity test; it includes both non seed and seed
material that is classified as inert according to the Rules for Testing Seeds.
Inhibitor. A chemical substance that retards or prevents a growth process such as
germination.
414 Glossary

Indexing. The process used to test vegetatively reproduced planting stock for freedom from
virus diseases. Disease evaluations are based on greenhouse or field plot growout tests.
Inflorescence. The flowering structure of a plant (e.g., umbel, spike or panicle).
Inoculant. A preparation containing a specific nitrogen-fixing bacteria that is added to legume
seed prior to planting to assure that the resulting crop will have nitrogen fixation ability.
Inoculum. Any material such as spores, bacteria or fungus bodies that serve as a means of
propagating or spreading a pathogenic disease. In legume seed inoculation, the inoculum is
the bacterial inoculant (see above).
Integumentary tapetum. The layer of nucellar cells with radial elongation and two nuclei
that immediately surround and provide nutritive support to the developing embryo sac of some
species. Later it becomes hardened and provides a protective layer to the mature seed.
Integuments. The tissues covering or surrounding the ovule, usually consisting of an inner
and outer layer which comprises the seed coat of the mature ovule.
Interagency certiflcation. The certification of seed through the cooperation of two different
agencies. Usually the term indicates that the field inspection is performed by one agency and
the seed inspection is made by a second agency which completes the certification process and
issues tags.
Intermediate day plants. Plants initiate flowers best under intermediate day-lengths.
Involucre. A close collection of bracts surrounding an inflorescence or flower.
ISTA. International Seed Testing Association.
Isolation requirement. The spatial separation required between a seed field and other sources
of mechanical and genetic contamination, especially between cross pollinated varieties.
Isotherm (absorption). A graph showing the curve ofthe relationship between seed moisture
content and its equilibrium relative humidity; also called moisture equilibrium curve.
Keel (flower). The two fused anterior petals of a legume flower.
Legume. A member of the pea family characterized by having dry, multiseeded pods that
dehisce along two sutures at maturity.
Lemma. One of two bracts of the grass floret; it is located on the side nearest the embryo and
opposite the rachilJa.
Lignin. The extremely complex strengthening or deposition material in plants that tend to
make them hard and woody. Chemically, lignin shows both phenolic and alcoholic
characteristics.
Locule. The cavity of an ovary.
Loculicidal capsule. A type of capsule that at maturity splits open through the midrib of the
carpel into the locules (e.g., iris, tulip).
Long day plants. Plants that initiate flowers best under long day (short night) regimes.
Longevity. Length of life or viability of organisms; often used in terms of seed longevity.
Macrogametophyte. A name sometimes applied to the megagametophyte (see
megagametogenesis, below).
Magnetic separator. A machine that separates seeds on the basis of their magnetic
characteristics (whether they are attracted or repelled by a magnet).
Glossary 415

Male sterile. Producing no functional pollen.

Malpighian layer. A protective layer of cells in the coats of many seeds characteristically
comprised of close-packed, radially positioned, heavy-walled, columnar cells without inter-
cellular spaces. They are usually heavily cutinized or lignified and relatively impervious to
moisture and gases.
Matriconditioning. Hydrating seeds in low water potential solid carriers such as clays or ver-
miculite followed by subsequent drying to enhance germination, stand establishment, and
seedling growth (also known as solid matrix priming).
Megagametogenesis. The development of the female gametophyte (megagametophyte) from
a functional megaspore.
Megaspore. One of the four cells (of archesporial cell origin) formed in the ovule of higher
plants as a result of meiosis, or sexual cell reduction division. One of these later undergoes
mitosis to give rise to the female gamete (megagametophyte, or embryo sac).
Megasporogenesis. The development of the megaspore from the archesporial cell.
Meiosis. Cell division during which homologous chromosomes pair; one member of each pair
separates and passes to daughter cells, each having one-halfthe original chromosome number.
Also called reduction division.
Mericarp. One-half of a two-sectioned fruit known as a schizocarp. Characteristic of the
carrot family.
Meristem. Undifferentiated tissue located at the tips or growing points of vegetative or
reproductive organs capable of undergoing cell division and elongation.
Meristematic cells. Undifferentiated cells in plant meristems which are capable of undergoing
cell division.
Mesocarp. Middle layer of the fruit wall (pericarp).
Messenger RNA. The form of ribonucleic acid that conveys the genetic messages coded in
the deoxyribonucleic acid of the nucleus to the ribosomes of the cytoplasm where proteins are
formed.
Metabolism. The chemical changes within a living cell.
Microgametogenesis. The development ofthe microgametophyte (pollen grain) from a micro-
spore.
Microgametophyte. A mature pollen grain, or male gamete.
Microspore. The male spore in the anther from which the male gametes develop.
Microspore mother cell. One of many cells in the microsporangium (anther) which undergoes
microsporogenesis to yield four microspores.
Microsporogenesis. The development of micros pores from the microspore mother cell.
Micropyle. The integumentary opening of the ovuldhrough which the pollen tube enters prior
to fertilization.
Middlings. The seed from the middle portion of the gravity separator. This portion usually
contains a mixture of light and heavy seed and is recirculated for further conditioning.
Mitochondria. Microscopic rod-shaped or spherical organelles present in living cells. They
contain the enzyme systems active in the respiration process.
416 Glossary
Mitosis. Normal cell division in which each daughter cell has exactly the same chromosome
number as the mother cell.
Monocot. An abbreviated name for monocotyledon, referring to plants having single seed
leaves (cotyledons). Examples are bamboo and com.
Monoecious (monecious). Having stamens and pistils in different unisexual flowers on the
same plant (e.g., com).
Monogerm seed (sugar beet). A sugar beet "seed" (botanically a fruit) containing only one
ovule in contrast to a mUltigerm "seed" which represents aggregate fruit containing several
ovule units.
Mother cells. Special cells in the anther and ovule that give rise to pollen or egg cells.
Multigerm seed (sugar beet). An aggregate fruit containing several ovules.
Mucilages. The gummy (sticky when wet) complex carbohydrate substances which cover the
seeds, bark or stems of some plant species (e.g., buckhorn plantain seeds).
Multiline. A composite (blended) population of several genetically related lines of a self-polli-
nated crop.
Multiple fruit. Developed from a cluster of flowers on a common base (e.g., fig).
National Seed Storage Laboratory. The laboratory at Ft. Collins, Colorado, that is operated
by the USDA as a permanent repository for storing germplasm of all kinds of plants under
controlled temperature and relative humidity conditions.
Nick. In hybrid seed production, the condition existing when two inbred plants flower and are
ready for sexual crossing at the same time (i.e., the majority of the pollen is ready when the
flowers of the other parent are ready to be fertilized).
Nonrecurrent apomixis. A type of apomixis in which a haploid embryo sac is produced as
a result of apomeiosis (abnormal cell division during megasporogenesis). It results in seed
with a haploid embryo and thus haploid plants that are usually sterile.
Noxious weed. A weed species that is defined by law as being noxious; usually highly
objectionable when found in crop seed lots.
Nucellus. The tissue of the ovary wall in which the archesporial cell arises and where
megasporogenesis, megagametogenesis, and ovule development occurs.
Nuclear endosperm. The type of endosperm in which the early development is characterized
by rapid cell enlargement accompanied by nuclear division without cell wall formation.
Nucleic acid. A highly complex organic molecule found in the nucleus of cells; believed to
be the substance that determines heredity and governs the behavior of all cells.
Nucleus. The part of the cell bearing the chromosomes.
Nut. A dry, indehiscent, one-seeded fruit with a hard, woody shell.
Nutlet. A small, dry, indehiscent fruit composed of one-haifa carpel enclosing a single seed;
developed by folding and splitting the carpel into a compound pistil.
OECD. Organization for Economic Cooperation and Development, an international agency
which, among other things, has developed specifications, procedures, and standards for an
international seed certification scheme.
Glossary 417
Open pollinated variety. A heterogeneous variety of a cross pollinated crop that is allowed
to interpollinate freely during seed production. In contrast to hybrids, representing controlled
crosses, open pollinated varieties are not common in modern crop production.
Operculum (seed). A type of epistase (integumentary proliferation) that is deposited inside
the ovule, forming a tight-fitting micropylar or chalazal plug in the mature seed; contributes
to water impermeability and hard seed coat dormancy.
Orthotropus ovule. The simplest type of ovule arrangement in which the ovule is erect, with
the micropyle at one end and the funiculus at the other.
Osmoconditioning. Soaking seeds in aerated, low water potential osmotica such as
polyethylene glycol or salts followed by subsequent drying to enhance germination, stand
establishment, and seedling growth (also known as priming).
Other crop seed (percentage). One of the four components of a purity test; the total
percentage (by weight) of seed of all crop species each comprising less than 5% of the seed
lot.
Ottawa seed blower. A type of seed blower developed by C.W. Leggitt of the Canadian
Department of Agriculture that has a slender metal blowing tube used for small seeded crops.
Ovary. The part of the pistil containing the ovule.
Ovoid. Egg-shaped.
Ovule. The structure within the ovary of the flower that becomes the seed following
fertilization and development.
Ovum. An egg cell.
Palea. One ofthe thin bracts of a grass floret enclosing the caryopsis that is located on the side
opposite the embryo.
Palisade layer. In seeds, this term is used interchangeably with malpighian layer.
Panicle. An inflorescence in which the lateral branches arising from the peduncle produce
flower-bearing branches instead of single flowers (e.g., Avena sativa).
Paper-piercing test. A stress test for seedling vigor utilizing sand covered by filter paper
through which the seedlings must emerge to be considered vigorous.
Pappus. A tuft of delicate fibers or bristles such as the feathery appendage on a ripe dandelion
seed representing a modified calyx.
Parietal cell. The sister cell of the megaspore mother cell originating from the division of the
archesporial cell. It is nonfunctional and usually degenerates.
Parietal placentation. A type of placentation in which the seeds are attached in the ovary near
the outer ovary wall; usually associated with vestiges of septa rather than along the ovary axis
as in other types of placentation.
Parthenocarpy. Production of fruit without seeds as in bananas and some grapes.
Partial hybrid. A legal designation for seed lots representing at least 75 but less than 95%
hybrid seed.
Pathogen. Any organism capable of causing disease by obtaining its nutrition either partially
or wholly from its diseased host.
Pedicel. The stalk of a floret.
418 Glossary

Pelleted seeds. Seed that are commercially prepared for precision planting by pelleting them
inside a special preparation to make them more uniform in size. Sometimes special nutrient
or growth-promoting substances are placed in the pellets to aid in seed germination and
growth.
Pepo. A fruit with a hard rind without internal separations or septa (e.g., cantaloupe, water-
melon, cucumber).
Perennial. A plant which survives and produces vegetative growth and flowers year after year
without being replanted.
Perfect flower. A flower having both staminate (male) and pistillate (female) organs.
Perianth. A collective term for all the petals of a flower.
Pericarp. The ovary wall. It may be thin and fused with the seed coat as in com, fleshy as in
berries, or hard and dry as in pods of legumes.
Perisperm A type of endospermlike storage tissue in a mature seed that develops from the
nucellus of the parent plant-thus it has the 2n chromosome number. Examples of species
with well-developed peri sperm tissue include beet and pigweed.
Petal. A unit of the inner perianth whorl or corolla.
Petiole. The stem of a leaf.
Phage-plaque. A clear area caused by a bacteriophage in a bacterial colony and caused by
dissolving of specific bacterial cells.
Photoperiodism. The response (e.g., flowering, germination) of organisms to the relative
length of daily periods of light and darkness.
Physiological dormancy. Seed dormancy caused by internal physiological conditions that
prevent germination. Often referred to as epicotyl or embryo dormancy.
Physiological maturity. The maturity of a seed when it reaches its maximum dry weight. This
usually occurs prior to the normal harvest date.
Phytase. An enzyme that catalyzes the breakdown of phytin, the source of inorganic
phospnorus in seed metabolism.
Phytochrome. The bluish photoreversible protein pigment responsible for the photoperiodic
control of flowering and seed germination. It exists in two forms in plants, the biologically
active PFR (receptive to far-red light) and the biologically inactive PR (receptive to red light).
Phytosanitary certificate. A certificate issued by a legally constituted authority offederal or
state government stating that a seed lot has been inspected and found to be free of disease
infestation. These certificates are frequently used in international seed trade agreements to
prevent the spread of seedborne diseases among countries.
Pipered embryo type. A type of embryo classification in which the second waIl ofthe zygote
(fertilized egg) is longitudinal, or nearly so.
Pistil. The female, or seed-bearing organ of the flower. It is composed of the ovary, style, and
stigma.
Placentation. The method of attachment of the seeds within the ovary.
Plastid. Small cytoplasmic organelles containing pigments (e.g., chloroplasts, which give the
green color to plant leaves).
Glossary 419

Plenum chamber. The chamber, or air chest, associated with (usually underneath) a crop
(seed) dryer into which air is moved and allowed to distribute immediately prior to its entry
into the drying bed.
Plumule. The major leaf bud of the seed or seedling. That part of the embryonic plant axis
above the cotyledons. Also known as epicotyl.
Pneumatic conveyor. A method of conveying seed within a conditioning plant utilizing air
as the driving force.
Pod. A fruit that is dry and nonfleshy when ripe, and splits open at maturity to release its
seeds.
Polar nuclei. Two nuclei of the female gametophyte (sex cell) that unite with one ofthe sperm
cells to form the endosperm of a developing seed.
Pollen. The small, almost microscopic, yellow bodies that are borne within the anthers of
flowers and contain the male generative (sex) cells. The mature microgametophyte.
Pollen tube. A microscopic tube that grows down the stigma from the pollen grain through
which the sperm cells are deposited into the embryo sac.
Pollination. The process by which pollen is transferred from the anther where it is produced
to the stigma of a flower.
Polyembryony. The condition in which an ovule has more than one embryo. This condition
is common to certain grasses.
Polymer. A large molecule formed by joining together many small identical molecular units
(e.g., starch formed by long chains of glucose).
Polyploid. An individual (plant) that carries two or more complete sets of homologous (pairs)
chromosomes.
Pome. A fruit in which the floral cup forms a thick outer fleshy layer and that has a papery
inner peri carp layer (endocarp) forming a multi-seeded core (e.g., apple, pear).
Poricidal capsule. A capsule that at maturity splits open at pores near the top, releasing
mature seed (e.g., poppy).
Prechill. The practice of exposing imbibed seeds to cool (5-1 OUC) temperature conditions for
a few days prior to germination at warmer conditions. See definition of stratification.
Prehydration. Soaking seeds in water or gels prior to planting to enhance germination, stand
establishment, and seedling growth.
Priming. Soaking seeds in aerated, low water potential osmotica such as polyethylene glycol
or salts followed by subsequent drying to enhance germination, stand establishment, and
seedling growth (also known as osmoconditioning).
Primorida. Organs in their earliest stage of development as a leaf primordia or meristem.
Processing. The term formerly used to describe the technology of cleaning seed and preparing
it for the market. See conditioning.
Proembryo. The young embryo in its early stages of development.
Progeny. Offspring.
Propagule. Any type of plant part used to reproduce another individual plant asexually.
420 Glossary

Protein. An essential constituent of all living cells. Proteins occur naturally and are complex
combinations of amino acids linked by peptide bonds.
Protoplasm. The essential complex living substance of cells on which all vital functions of
nutrition, secretion, growth, and reproduction depend.
Pseudocarpic fruit. A fruit consisting of one or more ripened ovules attached or fused to
modified bracts or other nonfloral structures (e.g., sandbur).
Pseudogamous apogamety. A type of diplospory (apomixis) in which the seed develops from
some cell of the diploid embryo sac other than the egg, but one in which the stimulus of
pollination is required before development will begin.
Pseudogamy. A type of apomixis in which the diploid egg cell develops into the embryo
without fertilization of the egg cell, although only after fertilization of the polar nuclei with
one of the sperm cells from the male gamete to form a normal triploid (3n) endosperm.
Pubescence. A covering of short, soft hairs.
Pure line variety. A variety (cultivar) ofa self-pollinated species derived from a single plant.
Pure live seed (PLS). The percentage of pure seeds in a seed lot that have the ability to
germinate. The percentage of PLS is determined by multiplying percent germination by
percent pure seed and dividing by 100.
Pure seed content (percentage). The percentage of each crop species that comprise five
percent or more (by weight) of a seed lot.
Quick test (seed testing). A type of test for evaluating seed quality more rapidly than by
standard laboratory tests.
Quicker method (of purity testing). The method of purity testing that distinguishes between
seed and inert matter purely on the basis of physical characteristics. See stronger method.
Quiescence. The absence of growth, usually inferring the absence of environmental conditions
favoring growth; although dormant seeds are quiescent, quiescence is distinguished from
dormancy, which implies the inability to germinate even in the presence of environmental
conditions favoring growth.
Raceme. A type of inflorescence in which the single-flowered pedicels are arranged along the
sides of a flower shoot axis.
Rachilla. The central axis of a grass floret.
Rachis. The main axis of a flower (or leaf).
Radicle. The rudimentary root of the seed or seedling that forms the primary root of the young
plant.
Raphe. A ridge (seam), sometimes visible on the seed surface, which is the axis along which
the ovule stalk ( funiculus) joins the ovule.
Receptacle. The basal structure to which the flower parts are attached, sometimes forming
part of the mature fruit, as in apple.
Registered seed. A class of certified (generic sense) seed which is produced from foundation
seed and planted to produce certified (blue-tag) seed. It is identified by a purple tag.
Release. A crop variety (or germplasm) that is released and designated to be reproduced,
marketed, and made available as seed for public use.
Glossary 421
Renovation (seed production). Usually refers to the mechanical removal of plants from a
very dense, unproductive, or sodbound stand for the purpose of revitalizing its productivity.
Respiration. The metabolic process by which a plant (or animal) oxidizes its food and
provides energy for assimilation of breakdown products.
Restorer line. An inbred line that perm its restoration of fertility to the progeny of male sterile
lines to which it is crossed.
Rhizome. A nonfieshy, more or less horizontal, underground stem.
RNA. Ribonucleic acid, a component of the nuchms, and to some extent, the cytoplasm,
which relays the genetic messages coded in the DNA of the nucleus to the ribosomes of the
cytoplasm, which in tum form amino acids into proteins.
Rogue. An off-type plant. When used as a verb it refers to the act of removing such plants.
Rotary dryer. A seed or grain dryer utilizing forced air in a revolving drum, into which seed
flows in and out in a continuing pattern.
RST. Registered Seed Technologist, a designation for a private (commercial)
seed analyst who has passed tests and met other professional and academic requirements to
merit a seal and the designation of "Registered Seed Technologist."
Rudimentary. Incompletely developed.
RUSSL. Recommended Uniform State Seed Law, a model for state seed laws prepared by the
American Association of Seed Control Officials (AASCO).
Samara. An indehiscent, winged fruit in which the seed coat is loose inside the peri carp (e.g.,
maple, ash).
Sampling. The method by which a representative sample is taken from a seed lot to be sent
to a laboratory for analysis. It is most commonly accomplished using triers, or seed probes,
although hand methods and mechanical sampling methods are also used.
Scalper. A screening or air blast machine used to s~~parate the very large, very small, or very
light contamination from a seed lot-a rough cleaner.
Scopoletin. The growth promoting substance reported to stimulate seed germination of some
plant species.
Scarification. The process of mechanically abrading a seed coat to make it more permeable
to water. This process may also be accomplished by brief exposure to strong acids (e.g.,
sulfuric acid).
Schizocarp. A dry, two-seeded fruit of the carrot family that separates at maturity along a
midline into two mericarps. Each mericarp has a dry, indehiscent peri carp enclosing a loose
fitting ovule.
Sclerotium (pI. sclerotia). Compact mass offungus hyphae usually with black outer surface
and white inner surface. Capable ofremaining dormant for long periods of time and eventually
giving rise to fruiting bodies.
Scorpioid cyme. A determinate inflorescence in which the lateral buds on one side are sup-
pressed during growth, resulting in a curved or coiled arrangement.
SCST. Society of Commercial Seed Technologists, an organization of commercial and private
registered seed technologists and seed analysts ofthe United States and Canada.
422 Glossary

Scutellum. A shield~shaped organ of the embryo of grass. It is often viewed as a highly

modified cotyledon in monocotyledons.
Secondary dormancy. A type of dormancy imposed by certain adverse environmental condi-
tions in previously nondormant seeds, or seeds in which primary dormancy has been broken.
Seed. A mature ovule consisting of an embryonic plant together with a store of food, all sur-
rounded by a protective coat.
Seed borne. Carried on or in seeds.
Seed coat. The protective covering of a seed usually composed of the inner and outer integu~
ments. Also called the testa.
Seed coating. Application of substances such as fungicides, insecticides, safeners, micronutri~
ents, etc. directly to the seed that do not obscure its shape.
Seedling. A young plant grown from seed.
Seed pellet. Obscuring the shape of the seed with an amalgam of fillers and cementing
additives (sometimes containing other substances such as plant growth regulators, innoculants,
fungicides, etc.) to a specific size to enhance mechanical planting and seed performance.
Seedstocks. Seed used as a source of germplasm for maintaining and increasing seed of crop
varieties.
Seizure. Legal court action that takes seed offthe market and makes it subject to court~ordered
disposition by law enforcement officials. It usually results from serious seed law violations.
Seminal organs. Pertaining to the seed or germ, or those already developed in the embryo
within the seed (i.e., seminal roots are roots arising from the embryo compared to adventitious
roots that arise later).
Sepals. A floral part-the outer whorl, referred to collectively as the calyx.
Septicidal capsule. A capsule that at maturity splits along the septa and releases the seeds.
Septum. A partition, as between the locules of a fruit.
Short day plants. Plants that initiate flowers best under short day (long night) regimes.
Sillique (sillicle). A fruit, characteristic of the mustard family, which has two valves that at
maturity split away from a persistent central partition. If it is several times longer than wide
it is termed a sillique. If it is broad and short it is called a sillicle.
Simple cyme. The simplest branched determinant inflorescence where the lateral flowers de~
velop later than the terminal flower (e.g., mouse-eared chickweed).
Simple fruits. Developed from a single pistil or ovary that may be simple or compound (e.g.,
berry, as in blueberry).
Single cross hybrid. A hybrid between two inbred lines.
Sodbinding. A condition of older grass seed fields where high plant population (interplant
competition), thatch build-up, and certain unknown factors create conditions in which seed
production decreases. This condition can often be relieved by fertilization or mechanical
renovation.
Solanad embryo type. A type of embryo classification in which the terminal cell of the
proembryo divides by a transverse wall and in which the basal cell plays a minor part (or no
part) in subsequent embryo development.
Glossary 423

Solid matrix priming. Hydrating seeds in low water potential solid carriers such as clays or
vermiculite followed by subsequent drying to enhance germination, stand establishment, and
seedling growth (also known as matriconditioning).
Solitary flower. The simplest expression of a determinant inflorescence.
Somatic cells. Pertaining to cells of the plant body other than reproduction tissue.
South Dakota blower. A popular type of seed blower used in purity testing of seeds. Air is
passed through plastic tubes to help in the separation of seeds according to their specific
gravity and resistance to air flow.
Sperm cell. The male generative cell that fertilizes the egg cell and unites with the polar
nuclei.
Spadix. A special type of spike with a fleshy inflorescence axis.
Spathe. A large bract surrounding an inflorescence, especially a spadix.
Spike. A basic type of inflorescence in which the flowers arising along the rachis are
essentially sessile ( stalkless).
Spikelet. The unit ofthe grass flower that includes the two basal glumes subtending one to
several florets.
Spiral separator. A type of seed separator with no moving parts. The seeds enter at the top
and slide or roll down an inclined spiral runway. The speed of seed movement and centrifugal
force allows separation ofthe heavier, round, fast-moving seeds from those that move slower.
It is sometimes called a vetch separator.
Spore. In seed plants, the spore is the first cell of the gametophyte generation. The two kinds,
microspore and megaspore, produce male and female gametes, respectively.
Stamen. The part of the flower bearing the male reproductive cells composed of the anthers
on the filament (stalk).
Stigma. The upper part of the pistil that receives the pollen.
Stock seeds. See seedstocks.
Stoner. A modification ofthe gravity separator especially constructed to separate stones from
crop seeds such as beans.
Stop-sale. An official administrative action by a state seed control agency which prohibits
further sale of seed. It is issued when evidence of mislabeling or a seed law violation is found.
Stratiflcation. The practice of exposing imbibed seeds to cool (S_lOOC) temperature
conditions for a few days prior to germination in order to break dormancy. This is a standard
practice in germination testing of many grass and woody species.
Strain. A term sometimes used to designate an improved selection within a variety.
Strophiole. A rare appendage arising from the seed coat of some species near the hilum area.
It may be variable in shape and has no apparent function.
Style. The stalk of the pistil between the stigma allld the ovary.
Subsampling. The procedure (usually by halving methods) by which a smaller representative
working sample is obtained from the larger sample submitted for seed analysis.
Sun drying. The use of direct sunlight and temperature to remove moisture from seed, usually
on a flat (concrete, asphalt, or wooden) surface.
424 Glossary
Suspensor. The group or chain of cells produced from the zygote that pushes the developing
proembryo toward the center of the ovule in contact with the nutrient supply.
Swath. A windrow. A row of cut or pulled crop usually waiting for drying or curing before
further harvesting.
Synergid nuclei. Two of the eight cells ofthe embryo sac-usually remaining nonfunctional.
Syn-l Syn-2. The first and second generation progenies, respectively, of a synthetic variety
after the individual lines are composited.
Syngamy. Sexual fusion of the sperm and egg cells.
Synthetic seeds. Seeds (often a somatic embryo surrounded by a synthetic encapsulation)
produced from vegetative tissue (usually by tissue culture) that are clones possessing identical
genotypes.
Synthetic variety. A variety composed of an interbreeding population of several cross-polli-
nated plant lines.
Tailings. Partly threshed material that has passed through the coarse shakers or "straw
walkers" and is eliminated at the rear of a threshing machine.
Tenuinucellate. The nucellar condition in which the embryo sac originates and develops only
one cell layer beneath the nucellar epidermis.
Testa. The outer covering ofthe seed; the seed coat.
Tetrad. A group (quartet) or tetrad offour spores formed by division ofthe same mother cell,
as in a tetrad of microspores.
Tetrazolium (TZ). Indicates a class of chemicals that have the ability to accept hydrogen
atoms (and undergo reduction) from dehydrogenase enzymes during the respiration process
in viable seeds. This is the basis of the tetrazolium test during which the tetrazolium chemical
undergoes a color change, usually from colorless to red.
TZ test. Quick test to determine seed viability (and sometimes vigor) using tetrazolium
solution.
Thermoperiod. A period of the proper temperature to elicit a specific growth or
developmental response-as in flowering or seed germination.
Three-way hybrid. A hybrid between an inbred line and single cross hybrid.
Tiller. A branch arising from the base of a monocot plant, especially in the grass family.
Tolerance. The amount by which a second test may differ from a first test without being
attributed to an actual difference in seed quality.
Trier. A hand manipulated probe for sampling seeds.
Umbel. A type of inflorescence in which the minute flower units are arranged into flat or
umbrella-shaped heads as in carrots and celery.
Unitegmic testa. A testa (seed coat) made up of only one integument.
Unsaturated fatty acid. A fatty acid that has a double bond between the carbon atoms at one
or more places in the carbon chain; hydrogen can be added at the site of the double bond.
Utricle. A small, thin walled, one-seeded fruit in which the seed is only loosely attached to
the pericarp.
Glossary 425
Varietal protection. Legal protection (breeders rights) to developers or owners of crop
varieties giving them exclusive right to control seed production and marketing.
Vegetative. Referring to asexual (stem, leaf, root) development in plants in contrast to sexual
(flower, seed) development.
Vermiculite. A porous form of mica, a mineral, which makes a good rooting medium for seed
germination because of its capacity to retain moisture and permit aeration.
Vernalization. Bringing into a spring condition. In reference to flowering, it is the process by
which floral induction is promoted. It is sometimes (perhaps erroneously) applied to seeds to
indicate stratification in order to break dormancy, t:nabling them to germinate.
Viable (viability). Alive. Seed viability indicat€~s that a seed contains structures and
substances including enzyme systems that give it the capacity to germinate under favorable
conditions in the absence of dormancy.
Vibrator separator. A machine utilizing a vibrating deck for separating seeds on the basis
of their shape and differing surface textures.
Vigor. The AOSA has defined vigor as "those seed properties which determine the potential
for rapid uniform emergence and development of normal seedlings under a wide range offield
conditions.
Vitascope. A commercial device for helping speed up the tetrazolium test utilizing a
mechanical seed slicing mechanism, a timer, and a controlled temperature bath.
Viviporous. In seeds, it denotes the condition in which they are able to germinate while still
attached to the mother plant.
Volunteer plants. Unwanted plants growing from residual seeds of previous crops.
Wagon-bed dryer. A seed or grain dryer utilizing forced air through a shallow drying
chamber over a similar-sized plenum chamber.
Wavelength. The distance between two corresponding points on any two consecutive waves.
For light, it is very small and is measured in nanometers (nm), which equal about 0.04
millionths of an inch.
Weed. Any plant in a place where it is a nuisance might be considered a weed. The term is
usually used to denote unwanted, noncultivated plants growing in fields, lawns, gardens, or
other areas used by man.
Weed seed (percent). The total percentage (by wl~ight) of a seed lot which is composed of
seed of plants considered to be weeds. One of the four components of a purity test.
Wet-bulb temperature. The temperature measured with a thermometer covered with a wet
wick.
Windrow. A loose, continuous row of cut or uprooted plants usually allowed to dry in place
until harvested. A swath.
Wing. A membrance, or thin, dry expansion or appendage of a seed or fruit.
Xenia. The direct, visible effects of the pollen on endosperm and related tissues in the
formation of a seed (e.g., seed color). It results in hybrid characteristics of form and color.
Zygote. A fertilized egg.
Index
2, 3, 5-triiodobenzoic acid, 4
2, 4-dichlorophenoxyacetic acid, 4
Abnormal seedlings, 77, 134,329,349
Abscisic acid (ABA), 34, 53, 94, 104, 150-152, 194, 221,
Absorption isotherms, 197, 199
Acacia, 61
Accelerated aging test, 168, 175-176, 182-183, 313-214
Acetone, 145
Acetylene, 4
Achene, 13
Acorn, 40
Acorus, 10
Adenosine triphosphate, 99, 101, 173, 218-219, 221
Advantage of climatic factors, 233,239
Adventitious embryony, 19
Aeration fans, 269
After-ripening, 79, 107, 110, 149, 151-154,156
Agamospermy, 18, 19
Agar, 359-363, 365-366, 368, 371-373
Agar testing for fungi, 359-361
Aggregate fruit, 12
Agrostis tenuis, 86
Air, influence on germination, 77-78
Air screen machine, 254-255
Albizia, 200
Albumin, 51
Albuminous endosperm, 26
Alcohol, 145
Aldehydes, volatile, 217
Aleurone layer, 26-27, 45, 51-52, 88, 95-96, 97, 99, 108
Alfalfa hard seed of, 141, 144
seed storability, 200-202
Alkali bees, 240, 244-245
Alkaloids, in seeds, 52
Allelopathy,67
Index 427

Almond, 47, 52
Alpha oxidation, 50, 99-100
Amaranthus
endogenous rhythms in, 144, 148, 155
germination photoperiod of, 85
Ambient conditions, 269-270, 275
Ambrosia, 67
Amending seed laws, 398
American Association of Seed Control Officials (AASCO), 393
American Seed Trade Association CASTA), 387
Amino acids, 29, 50, 100-10 I, 219
Amylase, 44-46, 95-96, 98-99, 106, 132, 214, 219
Amylopectin, 44-46, 98
Amylose, 44-46, 98
Anemochory, 61, 63, 65
Anemophily, 17
Annuoline,335
Anther, 4, 9
Anthesis, 11
Antipodal cells, 8, 19
Apical placentation, 6
Apomixis, 18
Apospory, 19
Apple
ABA content, 150-152
ABA decrease during stratification, 150, 152
Archesporial cell, 6, 8
Aril,10,65
Artemesia tridentata, 154
Asclepias, 63
Ascorbic acid, 53
Asexual reproduction, 18
Ash, 63, 65, 151
ABA, 141, ISO-lSI,
ABA decrease during stratification, ISO, 152
Aspergillus, 202, 356, 366-367
Association of Official Seed Analysts (AOSA), 125-126, 128, 167-168, 182, 184,317, 323,
329,331,335,349,393,397
Association of Official Seed Certifying Agencies, 298-299, 303-306, 311
Asteraceae, 9
Asterad embryo development, 23
ATP,99, 101, 173,218-219,221
428 Index
Autochory, 61, 63,
Auxins, effect on gennination, 3, 53, 103-104, 108-109
effect on floral induction, 3
Axile placentation, 6
Azalia, 13
Bacillus subtilis, 282, 357
Bacteria on seeds, 351-357
Bacterial pathogens on seeds, 367-369
bacteriophage testing for, 368-369
plant injection test for, 368-369
serological testing for, 368
testing for, 368-369
Bacteriophage testing for seedborne bacteria, 368
Bahia grass, 201
Barley, 26-29, 34,41,44-45, 102-103, 108, Ill, 147-149, 157,170,205,214,220,248,254,
279,300,356,359,362-363,367,369,371-372,373,396
chemical changes during seed development, 27-30
chemical content, 27, 42, 45
conditioned storage, 205
donnancy, 147-149, 151-152, 157
hybrid vigor in, 169
hydrogen peroxide, 108
kernel weight in, 170
physical changes during seed development, 26-29
photoperiod of, 3
radicle protrusion, 92, 102
secondary donnancy in, 157
Basal placentation, 6
Basin wildrye, 75
Bean
ABA effect, 34
bacterial pathogens in, 367
bleaching effect on vigor of snap beans, 171
carbohydrate storage in, 44
chemical composition, 39, 42, 44, 51
chilling injury, effect on respiration of, 182
chilling injury in, 81
chilling injury in lima bean, 81-82, 182
certification of, 300
color separation, 260
conditioned storage of, 205
conditioning, 257
conductivity test, 178
Index 429

cotyledon unfolding, 82
drying with wagon bed dryer, 270
effect of excess moisture on germination, 76
fat and oil content of, 47
flower type, 14
fruit type, 13
genetic effect on storability, 201
genotype effect on germination of, 113
gibberellin content of, 107
gravity separation-upgrading, 257
hard seed, 141-142
hypogeal germination pattern, 73
light induced epicotyl unfolding, 3, 82
mechanical damage of, 112-113, 170
micropyle role in moisture absorption, 169
mineral composition, 39
mobilization of nutrients during germination, 98
moisture equilibrium in lima bean, 198
moisture stress on protein content, 42
photoperiod of, 3
radicle growth during germination of, 102
seed coat bleaching in lima bean, 170
seed coat impermeability to water, 141
seed production, 233
seed storability of, 207
seed storage of, 205
seedborne diseases of, 358
seedborne viruses in, 369
seedling injection test of, 370
seed marketing, 384-385
segregant selection for seed coat color, 170
storability of, 201
tannin content of, 51
temperature effect on seed development, 31
winged seeds in, 63
Beech
chilling injury in, 81
conductivity test, 177
fat and oil content of, 47
fruit type, 13
hard seed, 140-146
hypogeal germination of, 73
430 Index

light induced epicotyl unfolding, 3, 82

lima
chilling injury in, 81-82, 182
moisture equilibrium content in, 198
seed coat bleaching in, 170
mechanical damage of, 112-1l3, 169
moisture equilibrium content in, 198
photoperiod of, 3
radicle growth during germination of, 102
seed coat impermeability to water, 141
seed storage, 205
storability, 20 I
tannin content of, 51
Beet
garden beet, 198
sugar. 2, 3, 9, 31, 34, 43,76,136,151,198,201,296,365,385
Belt conveyors, 263
Benefits of certified seed, 307
Bentgrass, 20 I
Bermudagrass, 77, 201, 239
Berry, 12
Beta oxidation, 50, 99
Beta limit dextrin, 46
Big bluestem, 201
Biennial, 2, 59-60, 68
Biological seed treatments, 282-283, 277
Biotin, 53
Birch, 55, 84, 157
Birdsfoot trefoil, 31, 201
Blackberry, 12
Blotter testing for fungi, 361-362, 363
Bluegrama, 201
Bluegrass, Kentucky
effect of maturity on germination, 149
endogenous rhythms in, 155-156
germination under alternating temperature, 80
germination under moisture stress, 75
radiation effect on germination, 112
seed storability, 201
Bluegrass, big, germination under moisture stress, 75
Bouncing bet, 14
Brazil nut, 47
Index 431

Breakdown of storage tissues during germination, 89, 95, 96-101

Breeder seed, 299-301, 303, 332
Brick grit test, 166, 181, 183
Broccoli, 201
Bromegrass, 201
Buckeye, 44
Buckhorn, 46-47
Buckwheat, 40, 47, 201, 300
Buffalograss, 201
Bumble bees, 240, 244
Burdock, 12
Cabbage, 3, 152, 197,201
Cacao, 40, 52
Calcium, 4, 41, 43,52,101,172,203,284,290-29\
Calyx, 4
Campanu/a, 63, 79
Canada thistle, 73, 75
Canadian Seed Trade Association (CSTA), 387-388
Canary grass, 3, 201
Canna, 200
Cantaloupe, 47, 49
Capsicumjrutescens, 43
Capsule, 13
Carbohydrate composition in seed, 40-41, 43
metabolism of, 98-99,102-103, 107
storage in seed, 44
Carbolic acid, 335
Carbon dioxide
requirement for germination, 77-78
effect of embryo puncture on dormancy of, \42
effect on germination, 77-78
evolution during deterioration, 215
influence on germination, 77-78, 144,208
permeability of nucellar membrane, 144
Carpels, 5
Carpetgrass, 201
Carrot
environmental effects on seed development, 31, 33-34
equilibrium moisture content, 198
dormancy in, 148
floral induction in, 2
flower type, 14
432 Index

fruit type, 13
inflorescence type, 16
influence of oxygen on germination of, 78
maturation, 202-203
seed storability, 201
Caruncle, 10
Caryophyllad embryo type, 23
Caryopsis, 13
Cassia, 200
Castalia, 10
Castorbean, 47
Catalase, 132, 152, 214
Catkin, 14
Cattail, 77
Cease and desist order, 394
Celery, 108, 148, 220, 279, 290
Cellular endosperm, 25-26
Certification, 297-3 15
ancillary programs, 311-312
application, 303
Association of Official Seed Certifying Agencies (AOSCA), 298-299,302-305,306,311
benefits, 307
blends, 310
breeder seed, 299-301, 303, 332
certification today, 298
certified seed (the class), 298
changing concepts and services, 308-312
competition and survival, 314
conditioning, 304
eligibility for, 302, 310
field inspection, 297, 303, 308, 312
foundation seed, 299-302
future speculation, 312-314
generation scheme, 299
genetic standards, 305
harvesting, 304
history, 297-298
identity preserved programs, 312, 314
interagency, 307-308
marketing, 306-307
new dimensions; new horizons, 314
nongenetic standards, 306
Index 433

past contributions, 312

performance and recommendation criteria, 310
phytosanitary, 311
planting stock, 297, 303, 308
procedure, 303-307
quality assurance, 312
registered seed, 299
sampling, 304
seed inspection, 304, 308, 310
select seed, 300
sod,310
tagging, 306
tree seed, 311, 231
USA certification, 311
varietal purity only certification, 308, 3 10, 312
Certified seed (the class), 299
Chaetomium, 282, 366
Chain conveyors, 263
Chalaza, 8, 11, 19,25-26,87,142
Chalazosperm, 26
Chemical composition, effect on vigor, 170
Chemical changes during seed germination, 102-103
Chemical tests, 335-337
Chemistry of seeds, 39-57
Chenopodiad embryo development, 23
Chenopodium, 67, 79, 147,369
Cherry
changes during stratification, 152
fruit type of, 13
increase in protein content of, 42
inflorescence type of, 14
mechanical restriction of germination in, 144
radicle growth during germination of, 103
rudimentary embryo, 149
Chestnut, 13, 44, 52
Chickpea, 41
Chickweed, 14,68
Chilling injury, 81-82, 89, 111
Chromatographic tests for variety testing, 334, 337
Chromosome test, 334-337
Chrysanthemum, thermoperiodism of, 2
Circadian rhythms, 149, 155-156
434 Index
Circumscissle fruit, 13
Citrus, 61, 194
Classification of grasses, 234
Cleaning equipment, 253-261
Clover
alsike, 33-34, 200
berseem, 201
crimson, 112, 141,201,261
Jadino, 73, 285
red, 75, 244,345,391, 396
subterranean, 141, 146, 201
sweet, 142,201,254,316
white, 141-143,399
Coatings, 277, 282-286
Cocklebur
impermeability to gases, 144
embryonic thrust of germinating seeds, 145
influence of oxygen on germination of, 78
mechanical restriction, 141, 145
photoperiod of, 2
Coconut, 13,47,49,53,63, 194
Coffee, 3, 52,143, 194
Cold test, 174-177, 182-183
Coleoptile, 73, 76-77,91, 104
Coleorhiza, 24
Color separator, 260
Colza, 47
Commercial Seed Analysts Association of Canada (CSAAC), 350
Complete flower, 6
Compound lipid, 49
Conditioning sequence, 253-254, 264
Conditioning plants, design of, 264
Conditioning and handling, 252-267
affinity for liquids, 253, 261
air screen machine, 254-255
certified seed, 304
cleaning equipment, 253-261
color separator, 260
dimensional sizing equipment, 257
electrostatic separator, 260-261
gravity separator, 255-257
inclined draper, 259
Index 435

indent cylinder separator, 257, 259

indent disk separator, 257-258
length separators, 257
precleaning equipment, 254
principles, 252-253
spiral separator, 260
surface texture separators, 257-262
timothy bumper mill, 261
velvet-roll machine, 257-259
vibrator separator, 261
width and thickness separators, 257
Constancy of destination theory, 23
Conveyors, 263
Cool gennination test, 175, 178
Cool-season grasses, 234-235
Corn cockle, 14
Corn
amylase activity in, 47
chemical changes during gennination of, 97, 101-102
chemical composition of, 40-43, 45-46, 89, 102
cold test of, 176-177, 183
fast green test, 134
fat and oil content of, 47
fatty acid content of, 49
gennination, 73-74,76-77,82-83,85,89-90,101-104
gennination under moisture stress, 76
hybrid vigor effect on germination, 169
imbibition, 90
influence of chemical composition on imbibition, 89
integuments, 10
low temperature on gennination, 99
photoperiod of, 3
prolamine content, 52
radiation effect on germination, 112
radicle growth during gennination, 82
respiration test, 182
seed storability of, 200-201, 205
starch composition of, 46
sweet, 201
vigor testing, 178
Corn (hybrid seed), marketing of, 382, 384-387
Corolla, 4
436 Index

Corymb,14
eastus, 10
Cotton
chemical composition of seed, 43, 48
chilling injury, 81
conductivity tests for seed vigor of, 177
cool germination test, 178
delinting and storability, 202
fat and oil content of, 47
fatty acid content of, 48
hardseededness in, 141-142
increase in free fatty acids during deterioration of, 215
influence of preharvest environment on vigor of, 170-173
photoperiod of, 3
seed storability, 201
Cotton seed, marketing of, 385
Cottonwood, 194
Cotyledon, 13,46,53,73,94-95,98,101-102,106-107,134,172,203, 213, 215-216
Cotyledonary knee, in onion, 216
Coumarin, 53, ISO-lSI
Cowpea
Aspergillus invasion of seed, 356
chemical changes during germination of, 102
conductivity testing of, 178
free fatty acidity test, 134
fruit abscission, 156
Fusarium infestation, 364
hybrid seed production problems, 287
intended for feed, 356
mineral composition, 39
nondelinted,322
seed marketing, 385
seed size, 34
seed storability, 201
structure of seed coat, 143
varietal eligibility for certification, 302
Cress, garden, 151
Crimson clover, 112, 141
Cross fertilization, 17
Crucifer embryo development, 23
embryo type, 23
Cucumber, 144,201,214
Index 437

Cultural practices on seed chemistry, 44

Curly dock, 78
Current germination tests, 395
Cyme, 14
Cynodon dactylon, 77
Cypsela, 63-64
Cytochrome oxidase, 214
Cytokinins, 4, 53
effect on germ ination, 107-108, 220
interaction with gibberellin, 152
Cytological tests for variety testing, 342
Dalligrass, 201
Dandelion, 13
Day length, influence on germination, 85, 155
Dehiscent fruits, 13
Dehydrogenase, 92,99, 128, 132-135, 178,214,219
activity tests, 133
Delphinium, germination of, 79
Derived lipid, 49
Dermatogens, 4
Design of conditioning plants, 264
Deterioration
conversion of ADP to ATP during deterioration, 221
color changes, 213
concepts of, 194-195
decrease in seed performance, 215-216
due to breakdown of germination triggering mechanisms, 220
due to enzyme degradation and inactivation, 219
effect of immaturity, 196, 202
effect of mechanical damage, 202
factors influencing, 203, 216
formation and activation of hydrolytic enzymes, 200, 214, 219
from accumulation of toxic compounds, 221
from depletion of food reserves, 220
from genetic degradation, 220
genetic influences, 200-201
inability of ribosomes to dissociate, 219
increase in free fatty acids, 215, 219
increase in seed leachates, 215
influence of microflora, 202, 214-215
lipid peroxidation, 216
loss of enzyme activity, 214, 219-221
438 Index

moisture influence, 194, 196, 200,202-203,205-206,208-209,214-216,219, 221

performance symptoms, 215-216
possible causes, 216-221
predicting, 203
reduced respiration, 214-215
symptoms of, 210-216
ultrastructure changes, 219
Determinate flowers, 14
Development of seeds, 19-38
Dextrin, 45-46
Digitalis, 63
Dimensional sizing equipment, 257-260
Dioecious, 6
Diploid
megaspore mother cell, 6
parthenogenesis, 20-21
pseudogamy, 20-21
Diplospory, 19
Disclaimers, 395
Disease resistance tests for variety testing, 331, 342-343
Dispersal of seed, vii, ix, x, 46
Diverticulae, 26
DNA
changes during seed development, 28
changes during seed germination, 107
content of seed, 50
role in protein synthesis, 50
Dock, 78, 86
Dormancy, 3, 53, 58, 61, 65-68,,78,81,88,91,104,107-108,127-128,130,135-136,140-
164
Dormant seeds, 125, 127, 130, 135, 143~ 144, 148-149, 157,327,329,331
Dormin, 53, 150
Double fertilization, 17
Double diffusion test for seed borne viruses, 371
Drought, 30, 41, 181-182, 239
Drupe, 13
Dry edible bean~disease-free seed production 358
Dry weight changes during germination, 90, 95, 101~103, 106
Ear~corn seed dryers, 269, 275
Effectiveness of inoculation of legume seed, 343
Effectiveness of seed treatments, 343
Egg apparatus, 7-8,17
Index 439
Eggplant, 198
Elaisome, 61, 63
Electrophoresis, 332, 334, 349-350
Electrostatic separator, 260
Eligibility for certification, 302-303
Elm, 63, 84, 194
Elymus, IO
Embryo excision, 135
Embryo development in Hordeum, 22
Embryo growth, during germination, 89-90, 96-99, 101-102
Embryo sac, 7, 14, 19,24
monocot and dicot, 24
Embryogeny, 19, 23
Encapsulations, 289-291
Endocarp, 12
Endogenous dormancy, 140, 147-149, 153-156
Endosperm, 9, 17, 19,23-29,34,46,53,90-91,93,95-97,99,101,106-107,130, 134, 157,
170, 178, 188, 210, 221
degradation, 95
development, 19, 23-24
haustoria, 26
nucleus, 17
types, 25
Endozoochorous plants, 61
Endozoochory, 61
Energy relations in the seed, 96-97, 99, 101
Enhancements for seed, 277-296
Entomophily, 17
Environment, 29, 31, 34, 39, 41, 43-44, 58-61, 65-68, 72, 75-76, 85,92,124,127,134,140-
141,147-148,150-151,155,166-167,169,170-175,182, 184, 196,201,203-206,208,215,
231-234,238-239,262,277,281-282,286-288,355-358, 366, 369
effect on seed maturity, 172
effect on seed vigor, 170-171
influence on seed development, 30
postmaturation-preharvest effect on vigor, 16'9, 171-172
Environmental influences on seed chemical content, 39, 41, 43-44
Enzyme activation during germination, 75, 89,95-97
Enzyme activity
loss during deterioration, 214
tests for estimating viability, 124-136
Enzyme degradation and inactivation, 219
440 Index

Enzyme-linked immunosorbent assay (ELISA) test, 339-340, 368, 372

Epigeal germination, 68, 70, 73
Epistase, 10
Epizoochorous plants, 61
Equilibrium moisture content, 197, 199-200,202,205,210-212,268,279,340,356-357
Ethanol toxicity, 77
Ethylene
effect on flowering, 4
effect on germination, 53, 108, 128, 220
evolution of, 104, 147
Ethylene chlorohydrin, 4
Euphorbia esuia, 9-10
Exalbuminous endosperm, 26
Excised embryo test, 135-137, 157
Exocarp, 12
Exogenous dormancy, 140-141, 145, 149,155
Fabaceae, xi
Farmers exemption, 395
Fast green test, 134
Fat and oil content of seeds, 39, 42, 44, 47-49
seed storability, 201
Fat degradation during germination, 100
Fatty acid content of seed, 44, 48-50, 99, 215-217
Fatty acids, 44, 48-50, 99, 133, 200, 215-217, 219
Federal Seed Act, 391-392
Federation Internationale du Commerce des Semances (FIS, or ASSINSEL), 388
Ferric chloride test, 134
Fertilization, 17
Fescue
Chewing's, 200
hairy, 126
hard,126
meadow, 126
red,75
tall, 201
Feterita,47
Field bean, 1
capacity, 75
Field bean seed, marketing of, 384-385
Field burning, 235, 237-238
Field inspection, 297, 303, 308, 312
Filament, 4
Index 441

Filbert, 13
Fir, Douglas, 155, 163
Firm ungerminated seeds, 329
Flax chemical content of, 31, 40, 43
Equilibrium moisture content, 197-200,202,205,210-212,268-269,279,340,356-357
Flora taxonomy, 14
Floral induction, 1-2, 4, 13-14, 53, 82, 85
Floral initiation, 4, 53
Floral morphology, 4
F1origen,3
Flower seed, marketing of, 384, 386-387
Follicle, 13
Food reserves depletion, 220-221
Formazan, 128, 178
Foundation seed, 232, 299-301
Foundation seed production, 300-302
Fraxinus, 143, 149
Free central placentation, 6
Free fatty acidity, 50, 99, 200, 215, 219
test for, 133
Free radicals, 216-218
Freezing injury, 111-112
Fruit development, 11
Fruit types, 12-14
Fungal endophyte, 366-367
testing for, 360, 367
Fungi on seeds, 355-357, 359-366
agar testing for, 359-361
blotter testing for, 359-362
classification of, 363-366
identification of, 363-366
methods of detection, 359-363
noncultural tests for, 362-363
saprophytic fungi, 366
sporulation of, 361
virulence testing for, 362
Funiculus, 8, 27
Fusarium, 139, 181-282,356,364-365
Gametophytic apomixis, 18
Gaps, in vegetation, 66
Gas impermeability, 143-144
Genetic drift, 61
442 Index

Genetic effects on seed deterioration, 200-201, 217, 220

Genetic influence on seed chemical content, 39-40
Genetic standards for certified seed, 303-305, 311
Geranium, 34
Germination, 72-123
biochemical mechanism, 104-106
chemicals, influence of, 67, 107-111
definition, 72
dry weight changes during, 90, 95, 101-103, 106
environmental factors, 72, 75-78
freezing during, 111-112
gas influence on, 77
light influence on, 82-89,104, 107-110
morphology of, 73
of seed banks, 58-59
osmotic pressure effect, 89, 92, 110
pattern of, 89-104
pH influence on, 91, 111
photoreversibility of, 84
requirements, 73-80
temperature influence on, 78-82
water influence on, 75-77
Germination speed, 111-112, 166, 179-181
Germination terminology, 125
Germination testing, 124- I 28, 165,326-329
abnormal seedlings, 134, 172, 179,329,349
breaking dormancy, 329-330
dormant seeds, 65,67,72,78,89,108,125,127,130,132,135,143-146, 148-151, 157,
234,237,327,329,357
evaluation of germination, 328-329
firm ungerminated seeds, 329
hard seeds, 329-330
normal seedlings, 8, 72,124-125,129,167,178-179,181-182, 220, 328-329,338-349,
397
prechilling, 329-331
procedures, 125-326
Gibberellins effect on floral induction, 4
effect on germination, 53, 88, 95, 98-99, 106-108
in dormancy interactions, 150, 152-153
in phytochrome interactions, 88
in seeds, 4, 52-53
role in physiological dormancy, 150, 152
Index 443

Globulin, 51
Glomerule, 14
Glucose, 44-46,52,92,97-99, 101
Glucosides, 52
Glutamic acid decarboxylase, 214, 219
Glutelin, 51
Glycerol, 48-50, 99, 181-182, 215
Glycolysis, 99
Glyoxylate cycle, 85, 99
Goodia,200
Gooseberry, 12
Grapefruit, 13
Grass seed production, 233-259
Grasses, 234-239
Grasses, marketing of, 383-384
Gravity separator, 255-256
Great plains grasses, 234-235, 237-239
Grow-out tests, 334, 343, 369
Gynoecium, 4
Gypsophila, 63
Haploid cell, 7
Haploid parthenogenesis, 19
Hard seeds, 91-92, 136, 141, 169-170, 196, 200, 202, 254, 329-330,
Harding grass, 201
Harvest maturity, 27, 33, 169
Hatch Act, 231
Haustoria, 25-26
Hazel, 13, 152
Head, 14
Heliotropicum curassavicum, 14
Helobial endosperm, 25
Hemicellulose, 44, 46-47
Hemlock, 157
Hemp, 47
Hesperidia, 13
Hilum, 8, 11,46,74,94, 141-142, 169
History of certification, 298-299
Hordeum distichum paimella, 22
Hormones, in seeds, 52-53
Horse chestnut, 52
Huckleberry, 12
Humidity, 76,90,92,127,142,170,172-173,175-176,183, 196-197, 199-200,202-203,205-
444 Index

206, 208-214, 218,

Humidity control, 211-212
Hybrid alfalfa, 249
Hybrid corn, 246-249
Hybrid seed production, 246-249
alfalfa, 249
corn, 246-249
sorghum, 248
wheat, 248
Hybrid sorghum, 248
Hybrid wheat, 248
Hydrochory, 61, 63
Hydrogen peroxide, effect on germination, 108, 134, 145
Hyoscyamus niger, 3
HypocotyJ, 68, 73~ 74, 102
Hypogeal germination, 68, 70, 73~ 74
Hysteresis, 199
Identity preserved programs, 312, 314
Imbibition, 81-82, 85-86, 88-92, 95, 110
Imbibition period, influence on light sensitivity, 85
Imbibitional chilling injury, 81-82, 89, 111,
Impaction, 142, 145
Impatiens, 126, 148
Imperfect flower, 6
Impermeable seed coat, 91, 141 ~ 142, 151, 183, 194, 200, 254, 286, 329
Imported seeds, 355, 391, 396,399
Inclined draper, 258-259
Incomplete flower, 6
Indehiscent fruits, 13
Indent cylinder separator, 257-259
Indent disk separator, 257, 259
Indeterminate inflorescences, 14
Indian ricegrass, 155, 157
Indigo carmine, 132-133
Indole acetic acid, 3, 103, 108
IndoxyJ acetate test, 134
Inhibitors in seeds, 28, 51, 53, 67, 78, 80,104,109,144,147,149-153,158,194,221
Integumentary tapetum, 10
Integuments, 9, 19
Interagency certification, 307
International Seed Testing Association (1STA), 130, 318, 340, 346-348, 350
Iris, 13
Index 445

Irrigated production, 233-234, 239, 258

!sopyrum biternatum, 149
Isotherm, 90, 197, 199
Juvenile, 1
Kafir, 40, 47
Kinins, 4, 53, 107-108, 150, 220
Kreb's cycle, 99, 101
Labeling requirements, 390-392
Labeling vegetable seed containers, 395
Larkspur, 13
Latex flocculation test for seedborne viruses, 372, 374
Laws of embryony, 23
Leaching, 111, 144, 151-152, 183
Leaf-cutting bees, 240, 243-244
Leafy spurge, 9
Leek, 155
Legislation and law enforcement, 390-398
Legume fruit, 13
Legume
seed production, 233, 239-246, 307,384
insect pollination, 233, 239-244
in area of use, 233
outside the area of use, 233-234
Lemma, 10
Lemon, 13, 151
Length separators, 257
Lentil, 141,213,300
Lepidium, 85-86, 151
Lespedeza, 201, 254
Lettuce, 44,52,77-78,83-88,101-102,107-108,110,136,148,155, 157, 183,200,213,215,
279-281,285,287,369
Leucadendron daphnoides, 108
Lifespan of seeds, 192-194
Lift trucks, 263
Light
influence on germination, 82-85
intensity, influence on germination, 82, 83
quality influence on germination, 83-85
requirement for breaking dormancy, 154-155
Light quality, 66, 82-83
influence on germination, 83-85
Lima bean, 81
446 Index

Lime, 13
Linoleic acid, 42, 48, 216
Lipase, 49,96,99, 101, 132, 152,215,219
Lipids, 47-50,82,92,96,99, 101,208,213-219
classification of, 49
environmental effects on synthesis, 49-50
hydrolysis of, 46, 48-50
metabolism during germination, 99
peroxidation, 208, 214, 216-218,
storage in seeds, 47-50
Lobelia, 10
Locule,5
Loculicidal fruit, 13
Locust, 13
Long-lived seeds, 192-193
Lotus, 192, 200
Low temperature effect on germination, 79, 86, 110-112
Lupin, 31, 68, 192, 200-201
Lupinus, 68, 200
Lysine content in seed, 51
Madia saliva, 47
Magnesium, 41,52
Maleic hydrazide, 4, 53
Maltose, 45-46, 97-98, 101
Mannitol, 85, 110, 151, 181-182,278
Maple
ABA decrease during stratification, 150
cytokinin influence on dormancy, 108
possession of wings, 63
short-lived seed of river maple, 194
stratification requirement for, 154
type of fruit, 13
Marketing certified seed, 306
Marketing of seeds, 380-399
cotton seed, 385
field beans, 384-385
flower and vegetable seeds, 384
hybrid corn and sorghum, 382, 384
public vs. private varieties small grains and soybeans, 385-386
small seeded grasses and legumes, 383-384
Matriconditioning, 279
effect on vegetable seed performance, 280
Index 447

Maturity, 1,8,11,13-14,26-27,33,43,72,75,82,111-113,124,148, 167, 172, 175, 184,

194,196,202,307,317,331,334,356
effect on vigor, 167, 172
Meadow saffron, 52
Meadowfoam, 44
Mechanical damage
effect on germination, 112
effect on seed storability, 202
excised embryo test, 136
fastgreen, 134
ferric chloride detection of, 134
indoxyl acetate test for, 134
influence of moisture content, 113
sodium hypochlorite for, 134
Mechanical injury, sources, 113
Mechanical restriction to germination, 141, 144
Megagametogenesis, 7
Megagametophyte, 7
Megaspore, 7
Megaspore mother cell, 6, 19
Megasporogenesis, 6
Meiosis, 7, 19
Membrane degradation, 173,215,218
Mericarp, 14
Meristematic cells, starvation of, 221
Meristems reproductive, 1
vegetative, 1
Mesocarp, 12
Mesocotyl, 104, 150-151
Metabolic inhibition of germination, 150-151
Microflora and seed deterioration, 202
Microgametogenesis, 11
Microgametophyte, 11
Micropyle, 10-11, 17-19,22, 141-142, 169
Microspore, 11
Microspore mother cell, 12
Microsporogenesis, 11
Milkweed, 13, 75
Millet, 201
Mirabilis, 78
Mistletoe, 110
Mitosis, 7
448 Index

Moisture effect on deterioration, 196-197

Moisture equilibrium, 197-199, 268-269, 279, 340, 356-357
Moisture stress, effect on germination, 75
Monocarpic, 60, 68
Monoecious, 6
Morphology of germination, 73
Mucilages, 44, 46-47, 61, 91,95
Multiple fruit, 12
Mummy seed, 192
Mustard, 40, 52, 110
Mutation, 217, 221
Myrmechory, 61
Naphthaleneacetic acid, 3
National Seed Storage Laboratory, 210
Natural selection, 61
Needlegrass, green, 81
New Zealand browntop, 83, 109
Nitrate, 67, 108-110, 127,331
Nitrogen, 4, 27-30, 41, 43-44, 50-52, 67,77,101-105,111,148,172,203,206,208
Noncultural tests for fungi, 362-363
Nongenetic standards for certified seed, 306
Nonirrigated production, 233-234
Normal seedling, 124, 129, 165, 167, 178-179, 181-182, 204, 220, 329, 338, 349
Noxious weed seed examinations, 326-327
Noxious weed seeds, 240, 258,304,320-321,326,331,346,391,392,396-397,399
Nucellar membrane, 143-144
Nucellus, 6, 8, 19
Nuclear endosperm, 25-26
Nucleic acids, 50, 101-103, 106-107
Nucleoproteins, 50
Nucleus, 17-19, 22, 24
Nut, 13
Nutlet, 14
Oak,14,43,48,52
Oats
chemical composition of, 41
conditioned storability of, 205-206
effect of irrigation during seed development of, 41
effect of low temperature in germination, 112
effect of prechilling on dormancy of, 153
effect of puncturing hull on dormancy of, 146
equilibrium moisture content, 198
Index 449
inflorescence type, 14
photoperiod of, 3
Oatgrass,75
OECD certification, 297, 308-309, 311, 314
Oil content of seed in relation to protein, 44
Okra, 198
Oleic acid, 42, 48
Olive, 13
Onion, 77, 155,201,216,218,279,285
cotyledonary knee, 216
moisture equilibrium in, 201
photoperiod of, 3
Operculum, 10
Orchardgrass, 31, 130, 201, 287
tetrazolium testing of, 128-13 2
Organic acids, inhibition of germination, 151
Organizations for seed testing, 346-350
Association of Official Seed Analysts, 346, 348-349
Commercial Seed Analysts Association of Canada, 350
International Seed Testing Association, 347-348
Society of Commercial Seed Technologists, 349
Orobanche, 63, 110
Orthodox seeds, 194-195, 206
Osmoconditioning, 111
Osmotic inhibition, 150-151
Osmotic pressure, effect on germination, 110, 151, 182
Osmotic stress test for vigor, 182
Outside the area of use, 233-234, 380, 383, 385
Ovary, 4-5, 11
Ovule, 8, 11
Ovule primordia, 6
Oxidase test for viability, 132-133
Oxygen
effect on dormancy, 143-144, 149, 152
effect on germination and viability, 77-78, 110-111, 127, 134
effect on longevity, 203, 208, 215-217,
influence on germination, 77-78
Pappus, 63-64
Parthenium, 67
Pattern of seed germination, 89-104
Pea
boron deficient seeds of, 172
450 Index

chemical composition of, 41, 43

deterioration, 214
effect of light on seed size, 31
effect of short days on seed size, 33
environmental effects on seed development, 4) , 43
equilibrium, 203
fat and oil content of, 47
fruit development in, 11
fruit type, 13
germination, 73
imbibition of heat-killed seed, 91
marsh spot in, 203
seed storability, 208, 214
Peach
embryo excision of dormant seed, 157
fruit type, 13
mechanical restriction of germination of, 144
metabolic inhibition, 150
Peanut
chemical composition of, 40-41, 43
effect of radiation, 99, III
effect of soil fertility on abnormal development, 172
equilibrium moisture content, 203
ethylene on germination, 108
farty acid content of, 48
seedling vigor classification of, 179
storability of seed, 205, 213-214
Pear, 13
Pectic compounds, 47, 92
Pedicel,9
Peduncle, 9
Pelleting, 277, 283-286, 325
composition of, 284
Penicillium, 202
Pennycress, 14
Pepo, 13
Pepper, moisture equilibrium, 198
priming, 279
Peppergrass
germination of imbibed seeds in response to red light, 86
germination under mineral deficient conditions, 203
imbibition period, 85
Index 451
increase in sensitivity to red light in response to temperature, 86
interaction of light and temperature on germination, 86
osmotic inhibition of germination in sugar bet:t juice, 151
photoreversibility, 85-86
Peptide bond, 50, 100
Perfect flower, 6
Pericarp, 5, 11,
Perisperm, 9, 26
Permanent wilting point, 75
Peroxidase, 96, 99, 132, 144, 152,214
Persicaria, swamp, 14
Petals, 4
pH, effect on germination, 57, 91, 94, Ill, 215
Phacelia,87
Phenol tests, 335-336
Phenols, 150
Phleum arenarium, 154
Phospholipid, 49, 101, 214-216, 218-219
Phosphorus, 29, 41, 43, 52, 101, 103,203
Photodormancy, 158
Photoperiod, 2-4, 33, 85, 147, 154-155,
Photoreversibility of germination, 84-85
Physiological maturity, 26-27, 33-34, 73, Ill, 124, 157, 169, 194, 196,202
Physiological dormancy, 149-150, 152, 155
Phytase, 52, 96, 101
Phytin, 52, 101
Phytochrome, 3, 85-89, 108, 111, ISS
Phytosanitary certification, 3 10
Pigwet:d, 78, 144, 155
Pine
cell division and elongation during germination, 103
Chinese red, 153
eastern white, 86
Japanese black, 153
light quality, 71-72
loblolly, 85-86
morphology of germination, 73
red, 181
sugar, 153
ultrastructural changes during deterioration, 213
Virginia, 86, 155
Pineapple, 12
452 Index

Pipered embryo development, 22-23

Pistil, 4
Placenta, 6
Plant injection testing for seedbome bacteria, 368-369
Plant Variety Protection Act (PVPA), 385, 387, 398-399
Plantago, 61, 144, 149
Plantain, 13
Plenum chamber, 256, 272
Plum, 52
Plumule, 73, 102, 107, 182
Pneumatic conveyors, 263
Poaceae, xi
Polar nuclei, 7, 17-18,23-24
Pollen grain, 4
sterility, 4
tube, 10, 17-18,30
Polyethylene glycol (PEG), 110, 151, 175, 181
Polygonaceae, ovule arrangement in, 11
Pome,13
Poplar, seed longevity of, 194
Populus, 63
Poppy,52
Poricidal fruit, 13
Potassium, 43, 52
Potassium nitrate
effect on germination, 331, 109
use of, 331
Potato, 3
Prechilling influence on germination, 81,127,153,329-330,331
Precleaning equipment, 254
Precocious germination, 76, 288
Predation, 63, 68
Prehydration, 278
Presoaking, effect on germination, 82, 111
Primary dormancy, 107, 140-157
interactions, 155-157
Primary noxious weed seeds, 326, 396
Priming, 218, 278-281, 286
factors affecting, 281
physiological mechanical responsible for, 280-281
seed matrix priming, 279
Primordia, 1
Index 453

Princess tree, 87
Proembryo, 22-23, 27
Prohibited noxious weeds, 326, 396
Prolamine, 51
Promotors in seeds, 53, 149
Protea compacta, 108
Protein content of seeds, 26-27, 40-41, 43-44,48, 50-52
metabolism of, 100-101
storage in seeds, 40, 50-51
Pseudocarpic fruit, 12
Pseudogamy, 19
Pseudomonasfluorescens, 282-283
Pumpkin, 47, 201
Pure live seed, 253
Purity testing, 285, 321-326
philosophy of, 323
procedures, 323-326
Pyrola, 63
Pythium, 282
QIO,90
Quality assurance, 312-314
Quiescence, 72, 75, 95, 140,290
Quince, 13
Raceme, 14
Radial diffusion test for seedborne viruses, 371<172
Radiation, effect on germination, 84-87, 89, 112, 182
Radicle, growth during germ ination, 72, 82, 102··104, 106-107
Radish, 3,
Ranunculus, 11, 149
Rape
aged seed, 20 I, 220
chemical composition of, 40, 42, 44, 48, 54, 89
Raphanus, 68
Raphe, 8,
Raspberry, 144
Recalcitrant seeds, 66, 194, 195
Receiving, elevating and conveying equipment, 263
Recommended Uniform State Seed Law (RUSSL), 393
Records, 395, 397
Red cotyledon malfunction, of lettuce, 215
Red clover, 3
Redtop, 198
454 Index

Registered seed, 299, 303

Relativehumidity,90, 127, 142, 173, 175-176,183,196-200,202-203,205-206,208,210-214
Reproductive Meristems, 1
Respiration, 53, 77-78,80,95,99,106, 108, 196,214-215,221
Respiration test, for vigor, 182
Respiratory quotient, 215
Restricted noxious weeds, 326, 396
Rhizobia, 325
Rhizoctonia, 282
Ribonuclease, 95, 96, 106
Ribosomes, 219
Rice
chemical composition of, 30, 40
fat and oil content of, 47
germination of, 77, 109
lipid composition of, 47
moisture equilibrium in, 201
oryzenin in, 51
photoperiod of, 3
radiation injury to, 112
seed storability, 198
starch composition of, 47
Ricegrass, [ndian, 155, 157
RNA, 28-29,50,96,101,106-107,150,173,219
changes during seed development, 28-29
Rose, 150, 153
Rudimentary embryo dormancy, 149
Rules for germination testing, 125, 127-128, 153, 196
Rumex obtusifolius, 86, 108
Russian pigweed, 78
Russian thistle, 13
Rye
chemical composition of, 40
cold requirement for floral induction, 2
deterioration, 213
equilibrium moisture content, 201
example of short-lived seed, 198, 200
fat and oil content, 47
germination, 99
photoperiod of, 3
prolamine in seed of, 51
protein of seed, 42
Index 455

short life-span of seeds, 200

Ryegrass
cold requirement for floral initiation, 2
developmental changes in seed of, 106
environmental effects on seed development, 30-31, 33
photoperiod of, 3
seed storability of, 198
Salix, 63
Salvia, 61, 65
Samara, 13
Samples, 261, 318, 321
mailing, 320
subdividing, 318, 321
Sampling,
from bags, 318
from bulk lots, 3 18
from small containers, 318
submitted sample, 318
the sampling process, 318
Sampling certified seed, 304
Saponins, 144
Saxifrage, 14
Scales, 264
Scarification, 145, 151-152, 155
chemical, 155
mechanical, 151-152
Schizocarp, 14
Screw conveyors, 263
Scutellum, 24, 45, 52, 95-97, 99, 108
Secaie cereaie, 1
Secondary donnancy, 107, 117, 140, 148, 157-158
Secondary noxious weed seeds, 326, 396
Sedum, 63
Seed bank, 58-60, 65-67, 140, 148
Seed blowers, 316, 325
Seed coat, 8,10,13,27,43,51,62,67,72,87,89,91-95,103,106, 112, 133-136, 140-145,
]47,149,151-152,154-155,157,170,181,200-202,213,2]6
impermeabi lity to water, 141-142
penneability, effect on imbibition, 91
radicle growth through, 72, 103
scarification of, 145-146, 151-152, 155
structure of, 74
456 Index

water entry through, 91-92

Seed coatings, 283-287
Seed deterioration
concepts, 194-195
prediction, 203
symptoms, 210-216
Seed development, 19
under high night temperatures, 43
Seed dispersal, 61, 65-67, 78, 81, 88,91,104, 107-108, 127-128, 131, 135, 136, 140-164,
Seed dormancy, 53, 58, 61, 65-67, 81, 86, 91,104,107,108,127,130,135-136,140-164,183,
194
Seed drying, 268-276
bin batch-dryer, 270
bin layer-dryer, 270
column batch-dryer, 273
continuous-flow crossflow dryer, 273
drying methods, 269-276
drying principles, 268
ear-com drying, 275
modified bin batch-dryer, 276
rotary dryers, 273-275
sun drying, 270
wagon batch dryer, 270
Seed enhancements, 277-296
Seed formation, 17
Seed handling equipment, 263-264
belt conveyors, 263
chain conveyors, 263
conveyors, 263
lift trucks, 263
receiving, elevating and conveying equipment, 263-264
receiving pit, 263
pneumatic conveyors, 263
screw conveyors, 263
vibrating conveyors, 263
Seed hydration, 278-281
Seed industry, 277, 314, 380,382,386
Seed industry, development of, 231-232
Seed inspection of certified seed, 304
Seed laws, 390-401
Seed legislation, 390-401
amendments of, 391
Index 457

Canadian seed laws, 399

cease and desist order, 394
collection of damages, 395
coloration and labeling of treated seed, 391, 396
definition of sale, 397
European seed laws, 399
Federal Seed Act, 391-399
federal seed legislation, 393-394
federal-state cooperation, 393-394
in developing countries, 400
offer for sale, 397
state seed laws, 392-397
Seed longevity, 192, 196,200-203,206
Seed matrix priming, 278-279, 281
Seed maturity
influence on storability, 202
influence on vigor, 170, 172-173
Seed moisture content, 27,30,33, 76, 85,95, 111-113, 127, 141-142, 148, 153, 183, 194-200,
202-206,208-209,214-215,219
Seed moisture tests, 344-346
dry weight, 325, 344, 346
wet weight, 344
Seed pelleting, 277, 283-285, 356
Seed production, 231-251
advantage of climatic factors, 233-234
cool-season grasses, 234-239
grasses, 234-239
great plains grasses, 234-235,237-239
hybrid alfalfa, 249
hybrid corn, 246-248
hybrid sorghum, 248
hybrid wheat, 248-249
in area of use, 232
irrigated production, 233, 239, 358
legumes, 239-246
nonirrigated production, 233-334
outside the area of use, 233-234
southern grasses, 234, 237
warm-season grasses, 234, 237, 239
western legumes, 239-244
Seed quality, 27, 41, 43
Seed rain, 60, 65
458 Index

Seed shadows, 60-61

Seed size, xiii, 29-31, 34, 60, 63, 68, 95, 167, 169, 172, 175-176, 195
Seed-soil contact, 95
Seed storage conditioned, 205-206, 210
containerized, 210
cryogenic, 204, 206, 208
hermetic, 204, 208, 216, 218
Seed testing, 316-353
Seed testing tolerances, 346
Seed treatment, 261-263
formulations and equipment, 262-263
history of, 262
Seedbome diseases, control of, 358-359
postharvest control of, 358-359
preharvest control of, 358
Seedling
emergence of, 72, 89,102,125,165-167,169,172-173,176-177,181-182,184
establishment, 58, 60, 65-68
growth rate test, 178, 183
morphology, 68, 165, 194
vigor classification test, 178-179, 181-182
Select seed, 300
Selenite viability testing, 133
Self fertilization, 17
Sepals, 4
Septa, 6
Septicidal fruit, 13
Serological testing for seedbome bacteria, 368-369
Serological tests for seedbome viruses, 369-376
Serologically specific electro microscopy (SSEM) test, 372, 375-376
Sesame, 48, 206
Shepherdspurse, 84
Short-lived seeds, 194,200
Sibling competition, 61
Silica gel, 210,
Sillique (sillicle), 13
Simple lipid, 49
Simple fruit, 12
Sisymbrium,67
Skunk cabbage, 14
Small grains and soybean seed, marketing of, 384
Smooth bromegrass, 73
index 459

Society of Commercial Seed Technologists, 331,346,349-350

Sod certification, 283, 310-311
Sodium hypochlorite test, 135
Soil fertility, 29-30,41, 43, 170, 172
effect on seed vigor, 170, 172
Soil moisture, effect on seed vigor, 170
Soil water, 92, 95
Solanad, 22-23
Solanad embryo development, 23
Solid matrix priming, 278-279, 281, 287
Solidago, 67
Somatic embryogenesis, 287, 289,291
Somatic embryos, 277, 287-291
Sorbus aucuparia, 151
Sorghum
conditioned storage requirements of, 205
germination, 77, III
high lysine, 40
imported seed, 396
inhibitors in, 67
marketing, 382, 384-386
moisture equilibrium in, 201
nitrogen content, 41
priming, 279
radiation influence on seed germination of, III
resistance to freezing injury, 100
respiration test for seed vigor of, 182
seed development of, 31, 33
seed production, 248
seed storability of, 205, 207
storage of, 207
variety release, 302
variety tests, 335
Southern grasses, 234, 249
Sowthistle, 73, 75
Soybean
accelerated aging test of, 168
breakdown of storage tissues, 98
chemical composition of, 4]
cold test for, 176
composition influence on imbibition of, 90
conditioned storage of, 206
460 Index

conductivity test of, 178

deterioration of, 206, 213-214
environmental influence on seed development of, 30-31,33
equilibrium moisture content in seed, 90
fat and oil content of, 47
fatty acid content of, 48-49
fruit type, 13
germination of, 73, 77
hydrolysis of protein bodies, 101
hypocotyl growth, 104
imbibitional injury, 82
indoxyl acetate test, 134
leakage of solutes through seed coat, 92
lipid composition of, 42
lipoxygenase, 217
low temperature injury to, 82, III
mechanical damage, 170-172
mineral content, 39
moisture equilibrium in, 90, 201
photoperiod of, 2
presoaking injury to, 110
protein content of, 40, 42-44, 48, 50
role of seed coat in delaying moisture imbibitional injury, 92
seed coat thickness, 141
seed development (fill), 141
seed maturity, effect on vigor of, 172
seed size, 30-31, 33-34, 160, 172
seed storage of, 198, 206, 215
seed vigor, test of, 176, 178
soaking injury, 111
sodium hypochlorite test, 134
state of moisture in seeds, 89
storability, 206
vigor, index of, 180-181, 184
vigor testing, 168, 176, 178, 184
Spadex, 14
Special quality tests, 331-346
Sperm cells, 17
Spherosomes, 50
Spike, 14
Spikelet, 18
Spinach, 3,
Index 461

Spiral separator, 260

Spirea, fruit type, 13
Spruce, tigertail, 153
Spurge, fat and oil content of, 47
Squash
fatty acid content of, 49
fruit of, 13
moisture equilibrium of, 201
Stamens, 4
Starch, 27, 44-46,51,54,90-91,98-99,101,106,132,208
content of seed, 27, 44, 46
hydrolysis of, 46, 99
metabolism of, 45
storage in seeds, 44-46
State seed laws, 391, 392-393
cease and desist order, 394
need for standardization, 393
Recommended Uniform State Seed Law (RUSSL), 393
stop sale, 393
Stigma, 4
Stop sale, 393
Storability, genetic influence, 200-201
Storage, 192-230
conditioned, 204-206, 210
containerized, 204, 210
cryogenic, 204, 206-208
degradation of functional structures during, 218-219
effect on vigor, 173,205
fungi, 202, 208, 210
hermetic, 204, 208, 216, 218
National Seed Storage Laboratory, 210
principles, 269
Stratification, 152-154, 157
breaking physiological dormancy, 81, 86, 110, 127, 150-151, 152-154, 157
influence on germination, 150
influence on inhibitor/promoter balance, 152
influence on physiology of dormant seed, 140
removing secondary dormancy, 157
Strawberry, 12
Striga, 110
Strophiolar cleft, in sweet clover, 142
Strophiole, 10
462 Index

Strychnos nux vomica, 52

Style,S
Stylophorum, 10
Subdividing samples, 31S-3 20, 321
Subsampling, 321-322
importance, 321
techniques, 321
Sucrose, 27
Sugar maple, lOS, ISO
Sugar beet, 2,9,31,34,43, 76, 136, 151
environmental effects on seed development, 31, 34,43
floral induction in, 2
germination of, 76
leaching, 151
osmotic inhibition, 151
peri sperm, 9
photoperiod of, 3
seed storability, 201
tern perature, 3 1
x-ray testing of, 136
Sugar beet seed germination, osmotic inhibition of, 151
leaching inhibitors from, 151
Sugarcane, 3
Sulfuric acid chemical scarification with, 145
moisture equilibrium with, 211
Sunflower
allelopathic substances in plant residue, 67
beta oxidation during germination, 99
chemical composition of, 40, 47
environmental influence on oil quality, 43
ethylene influence on breaking seed dormancy, 108
ethylene influence on seed germination, lOS
fat and oil content of, 42, 44
fatty acid content of, 42-43
flower type, 14
fruit type, 13
germination of, 77-78
inflorescence of, 16
membrane change during deterioration, 214
mineral composition, 39
oxygen influence on seed germination, 77
phospholipid decrease during deterioration, 214
Index 463
photoperiod of, 2
seed storability, 201, 214
Surface texture separators, 257
Suspensor, 23
Switchgrass, 235
Symplocarpus, 10
Synergids, 7-9
Syngamy, 17, 19
Synthetic seeds, 277, 282, 287-291
celery seeds, 290
challenges, 288
encapsulations, 290-291
principles, 287
somatic embryogenesis, 291
Synzoochorous plants, 61
Synzoochory,63
Tagging certified seed, 306
Tannins, 51, 92
Taraxacum kok-saghys, 22
Temperature
effect of imbibition temperature, 85
effect of low temperature on germination, 110
effect on chemical content, 41-42
effect on dormancy, 147-150, 152, 157-158
effect on germination, 66, 78-82, 86, 127
effect on hard seededness, 91
effect on imbibition speed, 92
effect on light requirement, 84, 86
effect on moisture equilibrium, 199-200
effect on photoperiodic response, 85
effect on radiation injury, 112
effect on seed chemistry, 41-42
effect on seed deterioration, 196, 200, 203-204
effect on seed development, 31
effect on seed storability, 196
floral induction, 1-2
high temperature lipid autoxidation, 216
high temperature loss of germination, 195
interaction with KN03 on germination, 110
interaction with moisture in influencing storability, 200
low temperature injury, 110
low temperature substitution for gibberelJin, 107
464 Index

need for cold temperature for germination, 104

requirement, role in dormancy, 152-154
resistance to low temperature injury, 112
susceptibility to freezing injury, 194
Testa, 8,
Tetrazolium test, for seed viability, 128-132, 175, 178, 180, 182-183
Thermo-dormancy, 158
Thermoperiodism, 2
Thiamine, 53
Thiourea, effect on germination, 110
Thistle
Canadian, 73, 75, 397
Russian, 13
Timothy
photoperiod of, 3
seed, CO2 requirement for germination, 78
seed, media toxicity test, 125
seed storability, 201
Timothy bumper mill, 261
Tobacco
alkaloids in, 52
germination with KN03, 110
photoperiod of, 3
sensitivity of seed germination to light, 73, 82-89
Tolerances for seed testing, 346
Tomato
breaking of physiological dormancy, 151
decrease in phospholipids during deterioration, 214
effect of carbohydrate nutrition on flowering, 4
environmental influence on seed development of, 30
floral induction, 2
fruit, 6, 12
hydrogen peroxide, 108
inhibition of germination by organic acids, 151
osmotic inhibition of seed germination, 151
seed, equilibrium moisture content, 198
seed storability, 201
thermoperiodism of, 2
Touch-me-not, 148
Transient seed banks, 65
Transport for conditioning, 395
Treated seed, labeling of, 396
Index 465
coloration of, 396
Tree of heaven, 13
Tree seed, certification of, 3 II
Trefoil, 31, 75
Tricarboxylic acid (TCA) cycle, 99, 101
Trichomes, 65
Trifolium, 200
Triglycerides, 48-49
Trillium, 10
Tube cell, 17-18
Tube nucleus, 17
Tulip, 13
Tung, 52
Turnip, 193,198
Ultraviolet tests, 334-335
Umbel, 14, 148
USA certification, 311
Utricle, 13
Varietal purity only (VPO) certification, 310
Variety
chemical tests, 335
chromatographic tests, 334, 337
chromosome counts, 334-335, 337-338
cytological tests, 342
definition of, 302
disease resistance tests, 342-343
electrophoresis, 332, 334, 339-342
grow-out tests, 334
phenol tests, 335-336
release, 302
review boards, 302-303
testing, 332, 335, 342-343
ultraviolet tests, 334-335, 337
Vegetative
apomixis, 18-20
meristems, 1-2,4
proliferation, 18
propagation, 18
Velvet-roll machine, 240, 257-258
Vernalization, 1
Vetch, 40, 103
466 Index

Viability, 43, 65, 108, 111-112, 124

Viability testing, 124-136
Vibrating conveyors, 263
Vibrator separator, 261
Viburnum, 149
Vigor, 165-191
definition of, 166-167
factors influencing, 169-1 73
history of, 166-168
post maturation, preharvest maturation, effect on, 170, 172-173
tests for, 173-184
Vigor tests characteristics of, 175
classification of, 174-175
standardization of, 166, 168, 175-176, 178-179, 181-183
use of, 183-184
Viola, 64, 68
Virulence testing for fungi, 362
Viruses on seeds, 357-358, 369-376
double diffusion test for, 371
enzyme-linked immunosorbent assay (ELISA) test for, 372
latex flocculation test for, 372, 374
radial diffusion test for, 371-372
serological tests for, 368-376
Vital coloring methods, for viability testing, 132
Vitamins, in seeds, 53
Vivipary, 18
Volatile aldehydes, 217
Walnut fat and oil content, 47
fruit type, 13
influence of stratification on seed chemical content, 152
Warm-season grasses, 234, 237-239
Water availability, influence on imbibition, 92-95
effect on chemical composition, 41, 89, 92
effect on germination, 75-76, 85, 89-90, 95, 111-113
imbibition, 92-95
impermeability,141
potential, 89, 92, 95, 104
types of water in seeds, 196-199
Watercress, 43
Watermelon, 142,201
Wheat
accumulation of toxic compounds in, 221
Index 467
chemical content, 30-31, 33-34, 43-44, 46
cuticle, 93
cytokinins, 107
depletion of food reserves, 220-221
deterioration, 213 -215, 221
dormancy, 147, 151, 157
effect of nutrients on seed chemical composition, 43-44
embryos, 221
equilibrium moisture content, 201
genetic influence on deterioration, 200-201
germination, 75, 77, 111
high lysine, 40
inflorescence type, 14
influence of protein content on vigor, 172
life span, 200
nitrogen content, 41, 44
permeability to water, 92
photoperiod of, 3
protein content, 41, 43, 51
radiation injury to, 111
requirements for conditioned storage, 205
respiration test for vigor in, 182
sprouting, 76
Wheatgrass,75
Width and thickness separators, 257
Wild rice, 194
Wild rye, 75
Wild water pepper, stratification requirements of, 154
Willow, 52, 194
Witchweed, 110
Xanthium, 144
X-ray tests, for seed quality, l35-l36
Yucca, l3
Zeatin, 53
Zoochory, 63
Zygote, 17, 19,22-24,26