MALE STERILITY

AND
ITS UTILISATION IN VEGETABLE CROPS
 MALE STERILITY

Male sterility refers to the condition

in which pollen is either absent or non
functional in flowering plants.
Caused due to pollen or anther
abortion.
 MANIFESTATIONS OF MALE STERILITY

• Absence or malformation of male organs (stamens) in

bisexual plants or no male flowers in dioecious plants.
• Failure to develop normal anthers.
• Abnormal microsporogenesis--deformed or inviable pollen.
• Abnormal pollen maturation; inability to germinate on
compatible stigmata.
• Viable pollen but non dehiscent anthers - sporophytic control.
Main features
Important out breeding device i.e., MS plants produce
seed only on cross pollination with functional pollen
of other plants.
Leads to heterozygosity.
Results from action of nuclear or cytoplasmic genes or
both.
Occurs in nature through spontaneous mutation or
may be induced artificially.
Can be observed in all diploid species.
 Utilisation

It is of special interest to the plant

breeder for production of hybrid seeds
more efficiently and economically.
HISTORY OF MALE STERILITY IN VEGETABLE CROPS
 Discovered for the first time in 1925 in Onion cultivar Italian Red in
California (Jones and Emsweller, 1936).
 Cytoplasmic male sterility in carrot (Welch and Grimball, 1947)
 First recessive male sterile gene ms-1, in muskmelon (Bohn and Whitaker,
1949).
 Jasmin (1954) first reported male sterility in eggplant.
 Cytoplasmic-genic male sterility (cgms) was first reported by Peterson (1958)
in an introduction of C. annuum from India (PI 164835).
 Watts (1962) reported the first genic male-sterile mutation in watermelon.
 Ogura (1968) reported gene-cytoplasmic male sterility (CMS) in radish.
 Two distinct recessive genes for male sterility were described by Shifriss and
Frankel (1969) .
 Functional male sterility in eggplant (Phatak and Jaworski, 1989), tomato
(Atanassova, 1999).
 Positional sterility in tomato (Atanassova, 1999).
 CLASSIFICATION OF MALE STERILITY

A. MORPHOLOGICAL

 “Pollen sterility” in which male sterile individuals differ

from normal only in the absence or extreme scarcity of
functional pollen grains (the most common and the only one
that has played a major role in plant breeding)

 “Structural or staminal male sterility” in which male

flowers or stamen are malformed, non functional or
completely absent

 “Functional male sterility” in which perfectly good and

viable pollen is trapped in indehiscent anther and thus
prevented from functioning (Kaul , 1988)
B. GENETIC

 Genic or Nuclear male sterility (GMS) : Genic/ genetic/ Mendelian,

male sterility is governed solely by one or more nuclear genes.

 Cytoplasmic male sterility (CMS): Sterility is governed by sterile

cytoplasm.

 Cytoplasmic-Genetic male sterility (CGMS): Sterility is governed

by sterile cytoplasm having fertility restorer nuclear gene. Also called
nucleoplasmic male sterility.

 Transgenic male sterility

C. NON GENETIC
Artificial induction through radiations and application of chemicals
(CHA).
 GENETIC OR NUCLEAR MALE STERILITY(GMS)

Controlled by nuclear genes.

Reported in Cucurbits, Tomato, Sugarbeet, etc
Originate through spontaneous mutation or can induced
artificially.
Usually governed by single recessive (ms) gene.
GMS lines produce 1:1 male sterile and male fertile plants.
Less frequently GMS has been controlled by :
 More than one recessive gene (Driscoll, 1986)
 Polygenes (Athwal et al, 1967)
 Dominant genes (Mathias, 1985)
SYSTEM
Two types of line:
A line: Genetic male sterile line (msms), used as female
parent in hybrid seed production.
B line: Heterozygous fertile (Msms) line, similar to A
line except for sterility, used to maintain sterile line.
A and B are isogenic lines with the difference of sterility/
fertility.
 Inheritance
msms x MsMs
(Male sterile) (Male fertile)

Msms
(Male fertile)

1MsMs, 2Msms, 1 msms Ratio 3:1

(Male fertile) (Male sterile)
Maintenance
A line is maintained by crossing recessive male sterile
plants (msms) with B line (Msms).
Crosses will yield 50% sterile and 50% fertile plants.
Male sterile plants are used as female parents in
hybrid seed production.
Fertile plants are rogued out.
Crosses are effected every year for mainatining male
sterility.
 Maintenance of MS line
msms x MsMs
Male sterile line A Male fertile line B

msms x Msms F1 (Msms) backcrossed with MS line A

Male sterile line A F1 Male fertile

1 msms : 1 Msms Maintained by sib mating

Male sterile x Male fertile Seed from MS plants only harvested
(Seed harvested) (Seed not harvested)

Maintained indefinitely
Disadvantages:
Less stable.
May be influenced by
 Temperature – Thermo sensitive MS (sterile at higher
temperature but exhibit normal fertiliy at low temperature).
 Day length- Photosensitive MS (sterile under long day
conditions, but exhibit normal fertility under short day
conditions).
Environmental sensitive male sterile mutants in vegetables
Vegetable Mutant Temperature
Cabbage TGMS, PGMS ≤ 100C
Brussels sprout TGMS ≤100C
Broccoli TGMS 10-110C
Pepper TGMS, TCMS ≤ 250 /170C
Carrot TGMS
Tomato TGMS, TCMS ≤300C /180C

Chive TCMS ≤ 240C

Chinese cabbage TCMS Low temperature
onion TCMS 14-220C

TGMS–Thermo sensitive genic male sterility

PGMS–Photoperiod sensitive genic male sterility (Mohammad and Ali, 2009)
TCMS–Thermo sensitive cytoplasmic male sterility
 Advantage of EGMS over GMS.

Ease in seed multiplication of male sterile line.

Two Line Hybrid Breeding.

Economic hybrid seed production.

Male fertility in hybrid crop is not affected.

 CYTOPLASMIC MALE STERILITY (CMS)

 Sterility is controlled by specific cytoplasmic gene or plasma

genes (S-genes).
 Not influenced by nuclear or other gene.
 Cytoplasm will be sterile only in the presence of specific S-
gene.
 May originate from inter-generic or inter-specific crosses and
induced through mutagenesis (Kaul, 1988) and protoplast
fusion (Pelletier et al, 1995).
 If a restore gene becomes available it is included in CGMS.
CMS

rr S
rr F

Male sterile Male fertile

rr x rr rr
S F S

MS MF MS
Features
Plants with particular cytoplasm are male sterile but
produce seed if pollinators are present. F1 seeds
(crossed seeds) produce only male sterile plants as
their cytoplasm is derived from female gamete.
System-
Aline- Male sterile line
B line- Male fertile line
A and B isogenic lines except for sterility.
Maintenance
CMS line mainatined by crossing A line with B line.
CMS can’t be utilised for hybrid seed production
without the use of restorer line as F1 seeds produce
only male sterile F1 plants.
The restorer line only can provide fertility in F1
hybrids.
CMS can be used for development of hybrids in
vegetatively propagated crops where seed is not
needed.
Not influenced by environmental factors, hence
stable.
Maintenance
rr) S x rr) F Cross of MF B line with MS A line

rr) S F1 is MS
 CYTOPLASMIC GENETIC MALE STERILITY (CGMS)
MS controlled by the interaction of cytoplasm and nuclear
genes.
A nuclear gene for restoring fertility in the male sterile line is
known.
System includes
 A line- MS line
 B line- isogenic with A line but Male fertile.
 R line- restorer line, restores fertility in F1 hybrid.
Fertility restorer gene ‘R’ is dominant found in certain
species, or may be transferred from related species.
Plant would be male sterile in the presence of male sterile
cytoplasm if maintainer nuclear gene is recessive “rr”.
 Various genotypes
rr) S : Male sterile
rr) F : Male fertile
RR) S : Cytoplasm sterile, nuclear
restorer gene in homozygous (RR)
condition
Rr) S : Effect of sterile cytoplasm
negated by restorer gene
Results of various crosses
rr) S MS x rr) F MF = rr) S MS

rr) S MS x RR) F/S MF = Rr) S MF

rr)S MS x Rr) F/S MF = 1 Rr) S MF, 1 rr) S MS

Maintenance
CMS line can be maintained by crossing the male
sterile cytoplasmic line with male fertile cytoplasmic
line.

(ms/ms) S x (ms/ms) F  (ms/ms) S

Male sterile Male fertile Male sterile
Merits and demerits
Widely used for hybrid seed production in seed and
vegetatively propagated crops.
Highly stable.
Not affected by environmental influences.
Requires labour and area as three lines are to be
maintained.
Poor agronomic performance.
 Mechanism of male sterility (GMS and CMS)

 Role of Tapetum : Aberration in tapetum cell development leads to the

failure of tapetum function, consequently failure of pollen development and,
therefore expression of male sterility. (Kaul, 1989 )
 Role of Callase : An enzyme required for breakdown of the callose that
surrounds the PMCs. Thus help in release of microspores (pollen) from
tetrad after meiosis. Mistiming callase activity resulting in male sterility.
(Mohammad and Ali, 2009 )
 Role of Esterase : Esterase enzyme helps in hydrolyses of the polymer
required for pollen formation. Decreased activity of esterase has adverse
effects on pollen development. (Sawhney, 1997)
 Role of PGRs : Endogenous PGRs play important role in stamen and pollen
development. Reduced level of Abscissic acid associate with seeds of GMS
and CMS plants indicates that both kind of male sterility.
 CHEMICALY INDUCED MALE STERILITY

A chemical hybridizing agent (CHA) is a chemical

that induces artificial, nongenetic male sterility in
plants.
First report was in maize by (Moor, 1950 and Naylor,
1950).
Chemicals sprayed on plants at a certain stage before
flowering.
It overcomes the lengthy time frame required to
obtain male sterile and restorer lines.
Any line can be used as a female parent.
Advantages
Rapid (one season) method of developing MS line
(Backcross method takes 4 to 5 years).
Less laborious and less expensive.
No need to maintain A,B, and R lines.
Genotype to be used as female parent in hybrid
development is treated with male gametocide to
suppress pollen formation.
Problems
 Unstable: Lack of effective male sterilizations, partial
female sterility and phytotoxicity.
 Non heritable
 If growth stage interval is too narrow, adverse weather
conditions may prevent a timely application of the
chemical.
 CHA causing male sterility in some vegetable crops
Vegetables Applied chemicals References
Tomato Gibberlin synthesis CCC Rastogi and Swahney,
inhibitors, ABA 1988
NAA McRae, 1985
Increased level of IAA Singh et al, 1992
Reduced level of cytokinins Sawhney, 1997
Eggplant Ethephon McRae, 1985

Pepper GA3 Sawhney, 1981

Cole crops Dalapon Hirose and Fujime, 1973

Pea Dalapon Brauer, 1959

Okra Mendok Dubey and Singh, 1968
MH, GA3 & etheral Deepak et al, 2007
Properties of an ideal gametocide
It should be selective in inducing male sterility.
It should not affect ovule fertility.
It should not be mutagenic.
It should give consistent results.
It should be economical.
Its method of application should be simple.
It should be safe for use and should have minimum
side effects on plant growth.
Drawbacks of present gametocides
Pollen abortion is incomplete and erratic.
Treatments are effective only at a specific stage of crop
growth and effect is short lived, hence require repeated
treatment.
Ovule fertility is also affected leading to low seed
setting.
Other side effects like deformation of leaf, stunting of
plant growth and sometimes wilting.
High cost of gametocides.
 Male Sterility Through Irradiations

Male sterile mutant plant T-4 was obtained via

gamma irradiation by treating dry seeds of tomato cv.
First (Masuda et al,
2000)
Male-sterility induced by gamma-ray irradiation of
African nightshade (Solanum nigrum L. ssp. villosum)
seed. (Ojiewo,2005)
 Genetically Engineered/ Transgenic Male
Sterility
Foreign gene (transgene) is also used for induction of
male sterility.
Generally gene responsible for inducing MS is obtained
from micro-organisms (e.g., Bacillus amiloliquefaciens).
Comes under genetic male sterility.
Heritable.
Genes are active only in the male reproductive organs and
prevent the emergence of male flower resulting in purely
female (“male sterile”) plants.
 Approaches for development of male sterility

1. Dominant Nuclear Male Sterility (Barnase-Barstar System)

 Barnase gene specific to anther tapetal cells, cause destruction of
tapetal cell layer by hydrolyzing the tapetal cell which results in
disruption of normal pollen formation and caused male sterility.

 To restore fertility, the barnase-specific RNase inhibitor, barstar,

was used (Mariana et al, 1992) and it was found that barstar gene
is dominant to the barnase gene.

 The barnase-barstar male sterility –fertility restoration system is

available in cauliflower and tomato (Banga and Raman, 1998)
2. Through Hormone Engineering

Induction of male sterility by manipulating endogenous

hormone levels (Spena et al, 1987)
Drastic change in endogenous levels of auxins have been
demonstrated to cause male sterility in tomato (Sawhney,
1997)
Such male sterile can be maintained by linking the gene
for herbicide resistance to the male sterilizing “rol C”.
Male fertile segregants can be chemically rouged out by
selective elimination in maintainer population.
Constraints Regarding Genetically Engineered Male Sterility

Unavailability of efficient gene construct.

Possible dispersion of transgene to other
related species.
Unavailability of efficient transformation
techniques.
Very high initial investment.
 UTILIZATION OF GENETIC MECHANISM FOR HYBRID
PRODUCTION
GENIC MALE STERILITY
Male Sterile Maintainer Block Hybrid Seed Production Block
Commonly utilized monogenic
msms (o) X Msms (+) ● + o ● o + ● o +
recessive GMS (msms) is
Male sterile Male fertile ● + + ● + + ● o o
maintained by back crossing it
● o o ● o o ● + o with heterozygous isogenic line
[ 50% msms (o) : 50% Msms (+) ] ● + + ● + o ● + o (Msms) for male sterility.
Male sterile : Male fertile ● + o ● o o ● o + Therefore, in hybrid seed
● + o ● + + ● + + production field, 50% male
Seeds collected from male sterile ● o + ● o + ● o o
fertile segregants (Msms) need
plants only (after identification) to be identified and removed
● o + ● + o ● + o
before they shed pollen
Male parent (●)
e.g., chilli, muskmelon, tomato
Male fertile plants (+) in the female rows
are removed after identification and seed
harvested from male sterile (o) plants are
hybrid seeds.
 UTILIZATION OF CMS
Male Sterile Maintainer Block Hybrid Seed Production Block

A line X B line x ● ● x ● ● x ● ●
Cytoplasmic-genetic
Smsms Nmsms x ● ● x ● ● x ● ●
male sterility can be
Male sterile Male fertile x ● ● x ● ● x ● ●
maintained by crossing
(Maintainer line) x ● ● x ● ● x ● ●
a cytoplasmic male
x ● ● x ● ● x ● ●
sterile line (Smsms) or A
Smsms (●) x ● ● x ● ● x ● ●
line with the male
All male sterile x ● ● x ● ● x ● ●
fertile line (Nmsms)
x ● ● x ● ● x ● ●
which is known as
N : Normal cytoplasm Male parent (x): male fertile restorer plant maintainer line or B line
S : Sterile cytoplasm (NMsMs or SMsMs)

ms : Maintainer allele Female parent (●): male sterile plant

Ms : Restorer allele (Smsms). Seed harvested from male sterile
(●) plants are hybrid seeds (i.e. fertile).
This CGMS system is used where seed is
of economic value.

Cytoplasmic male sterility can be utilized for producing hybrid seeds in those
vegetable where vegetative part is of economic value e.g. onion, carrot, radish,
cole crops etc.
 UTILIZATION OF CGMS

Male Sterile Maintainer Block Hybrid Seed Production Block

A line X B line x ● ● x ● ● x ● ● Cytoplasmic-genetic

Smsms Nmsms x ● ● x ● ● x ● ● male sterility can be
Male sterile Male fertile x ● ● x ● ● x ● ● maintained by
(Maintainer line) x ● ● x ● ● x ● ●
crossing a cytoplasmic
x ● ● x ● ● x ● ●
male sterile line
Smsms (●) x ● ● x ● ● x ● ●
(Smsms) or A line with
All male sterile x ● ● x ● ● x ● ●
the male fertile line
x ● ● x ● ● x ● ●
(Nmsms) which is
known as maintainer
N : Normal cytoplasm Male parent (x): male fertile restorer plant
S : Sterile cytoplasm (NMsMs or SMsMs) line or B line

ms : Maintainer allele Female parent (●): male sterile plant

Ms : Restorer allele (Smsms). Seed harvested from male sterile
(●) plants are hybrid seeds (i.e. fertile).
This CGMS system is used where seed is
of economic value.

For hybrid seed production, 2 to 3 rows of line A (Smsms) are alternated with 1 row
of line C, which is generally expected to NMsMs. e.g. Chilli, Onion, Carrot
 Identification of male sterile plant

On the basis of flower phenotype and flowering behaviour:

 In broccoli, Borchers (1966) recognized male sterile by their small
flowers, short stamens and shriveled anthers in GMS lines. Their
pollen grains were characteristically small, abortive, non-staining
and often clump together.
 Male sterile flower size is small than that of fertile flower in chilli
(Hundal and Khurana, 1993) and tomato (Sawhney, 1997)
 More than 50% male sterile plants started flowering in middle phase.
Screening period can be shortened by initiating identification of
male sterile plants towards the lateral part of early followed by
middle and initial part of late phase. (Dhatt and Singh, 1999)
Based on marker gene

Vegetable Marker gene for Reference

Broccoli Bright green hypocotyl Sampson, 1966

Tomato Potato leaf shape and green Kaul, 1988

stem colour
Absence of anthocyanin at Kalloo, 1995
seedling
Brown seed Bruzzone et al, 1983
Watermelon Delayed green seedling Zhang et al, 1996

Onion Brown seed Fustos, 1986

Chilli ms-10 gene linked with Dash et al, 2001

dark purple anther
Characteristics of male fertile and sterile plants in female parent

Character Male sterile plant Male fertile plant

Pollen (White Absent Present

powdery substance)
Anther color Purple or Yellow Light grey

Anther size Reduced to less than Normal

half
Anther bursting No bursting Anther burst to shed
pollen
Fruit set Low Heavy

Hundal and Dhall, 2005

 HYBRIDS DEVELOPED BY USING MALE STERILITY

CROP Name of hybrid Type of genetic Developing

mechanism Institution
Chilli CH-1, CH-3 GMS PAU, Ludhiana
Arka Meghna, Arka Sweta, CGMS IIHR, Bangalore
Arka Harita
CCH-2 CGMS IIVR, Varanasi
Muskmelon Punjab Hybrid, Punjab GMS PAU, Ludhiana
Anmol
Cabbage KCH-5 Ogura CMS IARI regional station,
Katrain
H-11, H-46 CMS ,,,
Onion Arka Kirtiman, Arka Lalima CGMS IIHR, Bangalore

Hybrid-63, Hybrid-35 CGMS IARI, New Delhi

Carrot Pusa Nayanjyoti Petaloid CGMS IARI regional station,
Katrain
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