SELF INCOMPATIBILITY

Dr. L.K.Gangwar
 SELF INCOMPATIBILITY
“It refers to the failure of pollen from a flower to
fertilize the same flower or other flowers on the
same plant.”
“The prevention of fusion of fertile (=functional) male
and female gametes after self-pollination.”(Gowers,
1998, in Banga and Banga, 1998)
“Lack of seed set on self-pollination.”
SI was first reported by Koelreuter in middle of
eighteenth century.
More than 300 species belonging to 70 families of
angiosperms show SI.
Self Incompatibility may arise due to any one
or in combination of the following reasons :
Pollen grains fail to germinate on the
stigma of the flower that produced them.
If some pollen grains do germinate, pollen
tubes fail to enter the stigma.
In many species, the pollen tubes enter in
the style, but they grow too slowly to effect
fertilization before the flower drops.
Sometimes, fertilization is effected, but the
embryos degenerate at a very early stage.
Self Incompatibility appears to be a
biochemical reaction, but the precise
nature of these reactions is not clearly
understood.
On the basis of the interaction between
pollen grains and pistil, self-
incompatibility is classified into the
following two types:
1.Complementary system of SI
2.Oppositional system of SI
 1.Complementary system of System of
Self-Incompatibility
This system is also called stimulatory type of self-
incompatibility.
In this system, pistil and pollen together provide
substances, which stimulate pollen germination and
growth of pollen tube.
If the pollen grains differ in SI genotype from that of
the pistil; the germination and growth of pollen
having similar genotype is not stimulated.
The complementary type of self-incompatibility
occurs in Dendrobium, where incompatibility leads to
flower abscission.
 1.Oppositional system of System of Self-
Incompatibility
This system is also known as inhibitory type of self-
incompatibility.
In this system, pollen and pistil produce such
substances, which prevent pollen germination and/or
pollen tube growth if the pollen has the same SI
reaction as the pistil.
However, germination and growth of pollen differing in
SI reaction is not inhibited.
Almost all cases of self-incompatibility are of this type.
The genetic control of oppositional type of self-
incompatibility reactions is relatively simpler.
 Classification of Self-
incompatibilty a/c to Lewis (1954)
1. Heteromorphic system
2. Homomorphic system
1a. Gametophytic control
1b. Sporophytic control
 Heteromorphic System
In this system, flowers of different incompatibility
groups are different in morphology.
For example, in Primula there are two types of
flowers, pin and thrum.
Pin flowers have long styles and short short stamens,
while thrum flowers have short styles and long
stamens.
This situation is referred to as distyly.
Pin and Thrum flowers are produced on different
plants.
The only compatible mating is between pin and thrum
flowers.
 Heteromorphic System Contd…
This characteristic is governed by a single locus s;
Ss produces thrum, while ss produces pin flowers.
The incompatibility reaction of pollen grains is
determined by the genotype of the plant
producing them.
The gene governing SI reaction has two alleles S
and s; allele S is dominant over s.
In some species, two genes, each having two
alleles, control the SI reaction.
The incompatibility system, therefore, is
heteromorphic-sporophytic.
 Heteromorphic System Contd…
The pollen grains produced by pin flowers,
would all be s in genotype as well as
incompatibility reaction.
The pollen produced by thrum flowers would be
of two types genotypically, S and s, but all of
them would be S phenotypically.
The mating between pin and thrum plants would
produce Ss and ss progeny in equal frequencie.
In some plants, a more complex situation occurs.
E.g., in Lythrum salicaria, styles are of three
different lengths; long, medium and short.
 Homomorphic System
In the homomorphic system, the system
found in the majority of self-incompatible
species, incompatibility is not associated with
morphological differences among flowers.
The incompatibility reaction of pollen may
be controlled by the genotype of the plant on
which it is produced (sporophytic control) or
by its own genotype (gametophytic control).
 Gametophytic System
Gametophytic incompatibility was first described by East
and Mangelsdorf in 1925 in Nicotiana sanderae.
The incompatibility reaction of pollen is determined by its
own genotype, and not by the genotype of the plant on
which it is produced.
This is because the biochemical substance involved in SI
reaction of the pollen is produced after meiosis.
In style, both the S alleles express themselves, i.e., show
co-dominance.
The incompatibility reaction may be controlled by one or
two genes; on this basis, gametophytic self-incompatibility
is classified into the following two groups : a)
Monofactorial, b.) difactorial systems.
 Sporophytic System
Sporophytic incompatibility was first reported by
Hughes and Babcock in 1950 in Crepis foetida, and by
Gerstel in Parthenium argentatum in 1950.
In the sporophytic system, self-incompatibility is
governed by a single gene, S, with multiple alleles;
often the number of alleles is 50 or more.
In general, the number of S allele is considerably
larger in the gametophytic than in the sporophytic
system.
The incompatibilty reaction of pollen is governed by
the genotype of the plant on which the pollen is
produced, and not by the genotype of the pollen.
 Mechanism of Self-Incompatibility
The mechanism of SI is quite complex
and poorly understood. The various
phenomenon observed in Self-
Incompatible mating are grouped into
three categories:
1. Pollen-stigma interaction
2. Pollen tube-style interaction
3. Pollen tube-ovule interaction.
 Pollen-Stigma Interaction
 These interactions occur just after the pollen
grains reach the stigma and generally prevent
pollen germination.
 At the time they reach stigma, pollen grains
generally have two nuclei in the
monofactorial gametophytic system, while
they have three nuclei in the difactorial
gametophytic and sporophytic systems.
 This was once considered to be the basis for
the gametophytic and sporophytic SI systems.
 Pollen-Stigma Interaction Contd…
Similarly, the structure of stigmatic surface once
appeared to be definitely involved in the differences
between the two systems.
In the gametophytic system, the stigma surface is
plumose having elongated receptive cells and is
commonly known as ‘wet’ stigma.
Incompatible pollen grains generally germinate on
reaching the stigma; the incompatibility reaction occurs
at a later stage within the stigma.
There are clear-cut serological differences among the
pollen grains with different S genotypes; such
differences have not been observed in the sporophytic
system.
 Pollen-Stigma Interaction Contd…
In the sporophytic system, the stigma is papillate and dry, and is covered
with a hydrated layer of proteins known as ‘pellicle’.
There is evidence that the pellicle is involved in incompatibility reaction.
There are striking differences in the stigma antigens related to the S
allele composition.
Within few minutes of reaching the stigmatic surface, the pollen releases
an exine exudate, which is either protein or glucoprotein in nature.
This exudate seems to induce immediate callose formation in the papillae
(which are in direct contact with the pollen) of incompatible stigma.
Often callose is also deposited on the young protruding pollen tubes
preventing any further germination of the pollen.
Thus in the sporophytic system, stigma is the site of incompatibilty
reaction; once the pollen tube crosses the stigmatic barrier, there is no
further inhibition of pollen tube growth.
 Pollen-Stigma Interaction Contd…
However, the difactorial gametophytic
system shows the various features of
sporophytic system, but pollen SI reaction
shows gametophytic control.
It has, therefore, been concluded that
pollen and stigma morphology, and pollen
cytology appear to be correlated with the
site of inhibition rather than with the
mode of genetic control of SI reaction.
 2. Pollen Tube-Style Interaction
In most cases of the gametophytic system, pollen grains
germinate and pollen tubes penetrate the stigmatic surface.
But in incompatible combinations, the growth of pollen
tubes is retarded within the stigma, e.g., in Oenothera, or a
little later in the style, e.g., in Petunia, Lycopersicon, Lilium,
etc.
In the latter case, there is a cessation of protein and
polysaccharide synthesis in the pollen tubes, which leads to
the degeneration of tube wall and the bursting of pollen
tube.
It has been suggested that SI reaction mimics the sperm
ejection stimulus of fertilization so that the pollen tubes
burst prematurely.
2.Pollen Tube-Ovule Interaction
In some cases, e.g., Theobroma cacao,
pollen tubes reach the ovule and effect
fertilization.
However, in incompatible combinations,
embryo degenerate at an early stage of
development.
Generally, such species have hollow
styles, e.g., Lilium, Ribes, Narcissus, etc.
 RELEVANCE OF SELF-INCOMPATIBILITY
Self-incompatibility effectively prevents self-pollination.
As a result, it has a profound effect on following breeding
approaches and objectives:
1. In self-incompatible fruit trees, it is necessary to plant two
cross-compatible varieties to ensure fruitfulness. Further,
cross-pollination may be poor in adverse weather conditions
reducing fruit set. Therefore, it would be desirable to develop
self-fertile forms in such cases.
2. In case of pineapple, commercial clones are self-incompatible.
As a result, their fruits develop parthenocarpically and are
seedless. Self-compatible clones would produce fruits
containing hundreds of very hard inedible seeds; such fruits
would not be acceptable to the consumer. Therefore, at least in
one case, i.e., pineapple, commercial success depends on the
clones being self-incompatible.
 RELEVANCE OF SELF-INCOMPATIBILITY Contd….
3. Some breeding schemes, e.g., development of hybrid varieties, etc.,
initially require some degree of inbreeding. Although, sib-mating
leads to inbreeding, but for the same degree of inbreeding it takes
twice as much time as selfing. Further, for the maintenance of inbred
lines selfing would be necessary.
4. Self-incompatibility may be used in hybrid seed production. For
this purpose:
i. Two self-incompatible, but cross-compatible, lines are
interplanted; seed obtained from both the lines would be hybrid
seed.
ii. Alternatively, a self-incompatible line may be interplanted with a
self-compatible line. From this scheme, seed from only the self-
incompatible line would be hybrid.
iii. Schemes for the production of double cross and triple cross
hybrids have also been proposed and their feasibility has been
demonstrated in the case of brassicas.
 RELEVANCE OF SELF-INCOMPATIBILITY Contd….
 The gametophytic system has been used, to a limited extent, for
hybrid seed production in clover, Trifolium (Leguminosae).
In Solanaceae, the cultivated species are generally self-fertile,
and self-incompatibility is confined to wild species.
 The sporophytic system has been exploited for hybrid seed
production in brassicas (Cruciferae), primarily by the
Japanese seed companies.
In Compositae, another economically important family
showing sporophytic self-incompatibility, the cultivated
varieties are generally self-fertile.
PROBLEMS IN USING SELF-INCOMPATIBILITY FOR HYBRID SEED PRODUCTION
1. Production and maintenance of inbred lines by hand pollination is tedious
and costly.
2. This raises the cost of hybrid seed.
3. Continued selfing leads to a depression in self-incompatibility, and it
unintentionally, but unavoidably, selects for self-fertility.
4. In the gametophytic system, continued inbreeding gives rise to new
incompatibility reactionss, which may limit the usefulness of such inbreds as
parents.
5. Environmental factors, e.g., high temp. & humidity etc., reduce or even
totally overcome self-incompatibility reaction leading to a high (30% or
more) proportion of selfed seed.
6. Bees often prefer to stay within a parental line, particularly when the
parental lines differ morphologically. This, in turn, increases the proportion
of selfed seed.
7. Transfer of S alleles from one variety or, more particularly, species into
another variety or species is tedious and complicated.
 These above problems has prevented the use of self-incompatibility in hybrid
seed production in Solanaceae and Compositae.
Recent investigations suggest that easy, reliable and economical
methods for multiplication of inbred lines may become available in the
near future. Some of the promising approaches are followin:
1. Inbreds could be multiplied in greenhouse; self-
compatibility is produced bt a temperature of 300C or
above.
2. Inbreds could be multiplied in suitable polythylene
tunnels. The CO2 level of the tunnels could be elevated,
e.g., by placing in them CO2 tablets. Pollination could be
effected by hand or by a suitable insect pollinator, e.g.,
blow flies.
3. Field multiplication of inbreds can be done using a 5-10%
sodium chloride spray for 3-5 days. This method has been
successful with B. napus, B. oleracea and B. campestris.
This is the only approach applicable at the field level.
 ELIMINATION OF SELF-
INCOMPATIBILITY
In many cases, self-fertile forms will be highly
desirable and, in such cases, it would be useful to
eliminate self-incompatibility.
1. In the case of single-locus gametophytic system,
incompatibilty may be eliminated by doubling
the chromosome number, e.g., in potato
diploidization leads to self-incompatibilty.
2. Isolation of self-fertile(sf) mutations is a very
useful tool in the elimination of self-
incompatibilty.
 ELIMINATION OF SELF-INCOMPATIBILITY Contd..
Flower buds are generally irradiated at the PMC
stage, and pollen from these buds is used to pollinate
flowers with known S alleles.
Generally, selection for Sf alleles is much more
complicated in the sporophytic system than in the
gametophytic system due to the temporary loss in
incompatibility and pseudofertility in the case of the
former. In Oenothera, Sf mutations occur
spontaneously at the rate of 10-8, and the rate of
induction with X-rays is 1.6 x 10-8/r unit.
3. Self-compatibility alleles may be transferred from
related species or varieties of the same species., if
available, through a backcross programme.
TEMPORARY SUPPRESSION
OF SELF-INCOMPATIBILITY
In many situations, e.g., during the production of
inbreds for use as parents in hybrid seed
production, it is essential that temporary self-
fertility is achieved in a manner so that self-
incompatibilty is fully functional in the selfed
progeny.
Such self-fertility is known as pseudofertility and
is achieved by temporarily suppressing the
incompatibility reaction using one of the
techniques given in next slides :
 TECHNIQUES USED FOR TEMPORARILY
SUPPRESSION OF SELF-INCOMPATIBILITY
Bud Pollination
Surgical Techniques
End-of-Season Pollination
High Temperature
Increased CO2 Concentration
High Humidity
Salt (NaCl) Sprays
Irradiation
Grafting
Double Pollination
Other Techniques : CO, Immunosuppressants, electric potential,
phytohormones, Protein synthesis inhibitors, steel brush
pollination.
 Bud Pollination
Bud pollination means application of mature pollen to immature
non-receptive stigma, generally 2-4 days prior to anthesis.
This is the most practicable and successful method both in the
gametophytic and sporophytic systems.
Obviosly, SI system does not become active until shortly before
anthesis.
In some cases, lack of stigma receptivity may be a problem, e.g.,
in Lilium longiflorum.
Seed set varies considerably in different inbred lines.
Bud pollination is adequate for the production of inbred lines,
but for their multiplication, other methods need to be employed.
In some cases, application of the fluid from mature stigmas may
improve the success of bud pollination.
 Surgical Techniques
Removal of the stigmatic surface, the whole of stigma
or a part or whole of the style may permit an
otherwise incompatible mating.
Removal of the stigma is very useful in the
sporophytic system, e.g., Brassica oleracea, but it does
not work in B. campestris.
In B. napus, stigma removal may be more effective
than bud pollination, while removal of the style may
be removed and the pollen grains may be directly
dropped on to the ovules in the ovarian cavity.
 End-of-Season Pollination
In some species, the degree of incompatibility is
reduced towards the end of flowering season or in
mature plants.
But there are controversial reports on the usefulness
of this technique.
 High Temperature
In some species, e.g., Trifolium, Lycopersicon, Brassica,
Oenothera, etc., exposure of pistils to temperature upto
600C induces pseudo-fertility.
In some lines, temperature of 300C or more induce seed
set.
But in many cases, it is a genotype-dependent response.
For example, in Trifolium hybridum a single dominant
gene produces self-incompatibility at 32oC.
In B. campestris, polygenes are reported to influence
this response.
In addition, high temperatures may reduce the strength
of self-incompatibilty.
 Increased CO2 Concentration
Increased CO2 concentration is reported to overcome
self-incompatibilty in the sporophytic system.
This approach has promise for use on a large scale for
multiplication of inbred lines.
Plants of B. napus and B. oleracea were grown in
polythene tunnels (2m high) with CO2 pellets; a small
fan was used for air circulation.
Pollination was effected by either hand or blowflies,
and a good seed set was obtained.
 High Humidity
High humidity can be created simply by enclosing the inflorescence
within a suitable bag, e.g., polythene bags.
High seed set in Brussels sprout was obtained by covering inflorescence
with cellophane bags and introducing within them blowflies as
pollinators.
This could have been due to increased CO2 levels due to blowfly activity
or, possibly, due to increased humidity.
Covering of the inflorescence with polythene bags results in a rapid
increase in humidity inside the bags.
This also results in seed set from similar lines of Brassica sp.
A combination of high humidity and high CO2 concentration is very
effective in promoting high seed in cauliflower and B. napus; this type of
gas mixture is obtained by breathing out.
Good seed set was obtained in B. napus when the inflorescence was
enclosed within a polythene bag and its air was displaced with the
breathed-out gas mixture simply by blowing into it.
 Salt (NaCl) Sprays
Chinese workers have developed a salt spray technique to
overcome self-incompatibility in B. napus.
In this technique, the flowers are sprayed with a 5-10% sodium
chloride solution for 3 to 5 days.
The method is easy, economical and as effective as bud
pollination.
It has also been shown to work with B. campestris and B.
oleracea.
However, it is not known if it will work with the gametophytic
system as well.
This approach, in any case, has the promise to develop into a
simple and reliable methods of field multiplication of self-
incompatibile inbred lines and of hybrid seed production,
particularly in the case of complex hybrid.
 Irradiation
In the single-locus gametophytic system, e.g., in
Solanaceae, acute irradiation with X-rays or gamma-
rays induces a temporary loss of self-incompatibility.

Grafting
Grafting of a branch onto another branch of the same
plant or of another plant is reported to reduce the
degree of self-incompatibility in Trifolium pratense.
There is only one report on this phenomenon, and the
mechanism of this reduction is not known.
 Double Pollination
In some species, self-incompatible matings become possible
when incompatible pollen is applied as a mixture with a
compatible pollen, or it is applied after pollination with a
compatible pollen.

Other Techniques
A number of other technique have been tried with varying
degrees of success, but they are not commonly used.
These techniques are, treatment of flowers with carbon
mono-oxide, injecting styles with immunosuppressants,
application of electrical potential difference of about 100V
between the stigma and pollen grains, treatment of pistil with
phytohormones and with protein synthesis inhibitors, and
steel brush pollination.