Chaudhary Sarwan Kumar

Himachal Pradesh Krishi Vishvavidyalaya

Palampur- 176 062
Masters Seminar (GP 591) Write up

MALE STERILITY- ROLE IN HYBRID SEED

PRODUCTION WITH SPECIAL REFERENCE TO
VEGETABLE CROPS

Submitted to: Dr. HK Chaudhary

& Dr. RK Mittal
Submitted by:
Akhilesh Singh
(A-2014-40-024)

ABSTRACT
The trend of F1 hybrid seed usage in vegetable is increasing globally in term of
species, cultivars and volume of seed used. Despite the usefulness of F1 hybrids
worldwide, the rising cost of labour and materials necessary for seed production is
posing a serious problem. For which we need genetic mechanisms which can help
in reducing the cost of hybrid production and enhance the purity of the hybrid seed
produced. For example, reduction in labour costs can be achieved by the
elimination of manual emasculation process, as it represents about 40% of the total
expenditure in tomato (Yordanov, 1983) and pepper (Kumar et al., 2002). Pearson
(1933) using self-incompatibility mechanism in cabbage and Jones and Clarke
(1943) using cytoplasmic male sterility mechanism in onion, proposed the methods
to produce large scale pure hybrid seeds. F1 hybrid vegetable seed can be
categorized into hand-pollinated and gene control pollinated species. The genecontrolled species can be due to the effect of self-incompatibility genes or male
sterility genes (Tay, 2006). The major advantage of self-incompatibility over male
sterility for hybrid seed production in vegetable crops is that hybrid seed can be
collected from both the parental lines. However, in some crops male sterility has
been reported to be more stable than self-incompatibility. Therefore both methods
are useful tools in the economic hybrid seed production of vegetable crops.
Male sterility is also induced these days through genetic engineering and
intergeneric hybridization. Barnase/Barstar and Bcp1 gene expression are two
potential systems under genetic engineering. Erucastrum canariense and
Moricandia arvensis cytoplasms were recently reported to confer CMS in B.
oleracea following intergeneric hybridization (Chamola et al., 2013). A CMS line
of R. sativus with the cytoplasm of B. maurorum was produced (Bang et al., 2011).
Male sterility in vegetables is being commercially utilized to develop hybrids and
substantial progress has been made in understanding the mechanism governing it
(Prasanth et al., 2014). Molecular markers can also be used to identify male
sterility gene in certain vegetables. Shu et al., (2016) reported a generic SSR
marker closely linked to dominant genic MS gene (DGMs79-399-3) in broccoli.
Introduction

In F1 hybrid vegetable seed production, vegetables can be divided into two

groups: the hand-pollinated and the gene-control pollinated species. The genetic
control system can be due to the self-incompatible system where pollen of the
same plant or flower cannot pollinate itself or to the male sterile genetic system
where a female plant has no male organ, deformed organ or no functional pollen to
pollinate itself. When no such genetic control system is found or when it is not
introduced into inbred parental lines, tedious hand-emasculation and pollination
have to be used to produce F1 seed. In both the gene-control system and handpollinated species sufficient field or female flower isolation have to be maintained
to obtain high seed genetic purity. The vegetables with highly developed selfincompatibility system are those in the family Crucifereae. They include Brassica
oleracea (Brussel sprouts, cabbage, cauliflower, broccoli, kohlrabi and kale),
Brassica rapa (Chinese cabbage, turnip and a range of Asian leafy brassicas) and
Raphanus sativus. The genetics of the self-incompatibility system in the
cruciferous crops are so well developed that they consist of a series of genes (loci)
and alleles. Vegetable breeders have been very successful in using them for
decades in F1 hybrid seed breeding. Hybrid seed production of sweet corn, carrot
and onion are based on male sterility gene system and the genetic control can be
either just clear-cut male sterility genes or the interaction of a male sterility gene
with a cytoplasmic factor.
Most of the seed of F1 hybrid vegetables are produced by hand-pollination.
However, it is labor intensive and requires a team of skillful growers and many
dedicate pollinators with good eye-sight, gentle hands, a lot of patience and
commitment, and able to follow instructions accurately. The main task of a seed
producer is the management of the production system and business. To be costeffective, this system only works in species where a single pollination of a female
flower will produce many seeds. This is the case for all the solanaceous crops and
cucurbits. On the contrary, in legumes the small number of seeds per flower/pod
prevents hand-pollination to be efficient and thus no hybrid beans to date have
been produced. In this case the use of gene-control pollination has to be exploited.
Similarly, if a good gene-control pollination system is available in say tomato and
pepper their seed production could be transformed into less intensive large field
production system as in the brassicas and sweet corn. Ever since (since 1930s) the
discovery of male sterility (in onion) and self-incompatibility (in cabbage)
mechanisms and their proposed utilization in hybrid seed production, several

mechanisms and methods have been evolved for the development of experimental
and commercial hybrids. The commercial utilization many of these mechanisms
are not feasible, therefore, utilization of such mechanisms has been restricted to
develop only experimental crosses (Bassett, 1986; Kalloo and Bergh, 1993).
History:
Among the vegetables, first F1 hybrid of eggplant was released during 1924
in Japan (Nishi, 1967). Subsequently, hybrids of watermelon (1930), cucumber
(1933), radish (1935), tomato (1940) and cabbage (1942) were developed (Liedle
and Anderson, 1993). Pearson (1933) using self-incompatibility mechanism in
cabbage and Jones and Clarke (1943) using cytoplasmic male sterility mechanism
in onion, proposed the methods to produce large scale pure hybrid seeds.
In India, Choudhury and Singh in 1971 first developed the breeding of the F 1
hybrid in India. The species was bottlegourd (Lagenaria ciceraria). As F1 hybrids
came to be widely recognized, hybrid breeding was advanced by many researchers
and numerous F1 hybrids varieties were publicized, as follows:
1971 to 1980
bottlegourd, brinjal, chilli
1980s
tomato, brinjal, muskmelon, watermelon, sponge gourd
1990s
cauliflower, cabbage, summer squash, chilli, bitter gourd,
capsicum, cucumber, carrot
Status and importance:
Now a day, F1 hybrid breeding method is commonly utilized to exploit
heterosis in several economically important vegetables including tomato, eggplant,
hot and sweet peppers, onion, cabbage, cauliflower, other cole crops, radish, carrot,
melons etc. Vegetable breeders prefer to select hybrid breeding because it is
comparatively easy to incorporate resistant genes for biotic and abiotic stresses in
F1 hybrid and right of the bred variety is protected in terms of parental lines.
Moreover, despite high cost of hybrid seeds, there has been increasing concern of
the farmers on the cultivation of hybrids. This is because under optimum crop
production and protection management, crop raised from the seeds of F 1 hybrid has
several advantages like better yield, adaptability, uniformity and reactions to
certain stresses in comparison to crop raised from the seeds of improved pure line
or population.

Important genetic mechanisms:

Male sterility:
Male sterility is the failure of plants to produce functional anthers, pollen or
male gametes. Advances in plant science have led to better understanding and
control of male sterile systems in plants.
Male sterility has meanwhile been identified in over 150 plant species.
Today, two natural sources of male sterility present in the genetic diversity are
mainly used by breeders in their breeding programs:
Either male sterility is controlled by genes of the nucleus (Genic Male
Sterility) or by genes of the cytoplasm (Cytoplasmic Male Sterility) in the
organism, sometimes in interaction with the environment for the stability of its
expression. Numerous examples exist across crops: male sterility can thus be found
in maize, wheat, oilseed rape, sunflower, sorghum, sugar beet, rye, barley, rice,
cotton, flax, onion, spinach, carrot, asparagus, celery, cucurbits, tomato, pepper,
eggplant, leek, fennel, radish, cabbage, cauliflower, broccoli, turnip, chicory, etc.
Cytoplasmic Male Sterility (CMS) was commonly identified within the
genetic diversity of sexually compatible species. It is introduced in the crop either
by crossing or with the help of protoplast fusion. For example, in brassicas, CMS
was discovered in 1968 in a Japanese radish and introduced in cabbage by crossing
in 1974. In this specific case, protoplast fusion was later used to correct the
chlorophyll deficiency of cytoplasmic male sterile brassica plants (cabbage and
oilseed rape) by fusing a cabbage male-sterile cell without functional chloroplasts
with a cabbage cell that had functional chloroplasts.
Types:
Kaul (1988) classified male sterility in two major groups, viz., genetic
(spontaneous or induced) and non-genetic (induced) male sterility. On phenotypic
basis, genetic male sterility has been classified in three classes, i. e., sporogenous,
structural and functional. Similarly, non-genetic male sterility has been classified
as chemical, physiological and ecological male sterility. Further, on genotypic basis
genetic male sterility was grouped as genic, cytoplasmic and gene-cytoplasmic
male sterility (Kaul, 1988). Based on the location of gene(s) controlling genetic
male sterility, spontaneously isolated, artificially induced through mutagenesis,

artificially incorporated through protoplast fusion or genetically engineered male

sterility systems can be classified as (i) genic male sterility (gms; more precisely
nuclear male sterility) and (ii) cytoplasmic male sterility (cms; more precisely
cytoplasmic-nuclear male sterility).
Nuclear or Genic Male Sterility (gms)
As the name suggests, nuclear male sterility (earlier termed as gms) is
controlled by the gene(s) from the nuclear compartment. Pollen fail to self-fertilize,
either due to non-dehiscence of pollen or their special flower morphology e.g.
positional sterility in tomato (Atanassova, 1999) and functional male sterility in
eggplant (Phatak and Jaworski, 1989). The occurrence of predominantly recessive
male sterility clearly indicates that gms is the result of mutation in any gene(s)
controlling microsporogenesis (pollen development process), stamen development
or microgametogenesis (male gamete development process).
EGMS Line:
Certain gms lines are conditional mutants, meaning thereby in a particular
environment male sterile mutant plants turn into male fertile. After determination
of critical environment (usually temperature or photoperiod) for sterility and
fertility expression, such GMS mutants are classified under environmental
sensitive genic male sterile (egms) lines. In vegetable crops, mostly temperature
sensitive egms lines have been reported. From practical application viewpoint, it is
necessary to identify critical temperature or photoperiod for the fertility/sterility
expression in temperature and photoperiod sensitive genetic male sterility,
respectively.

Cytoplasmic (cms) Male Sterility

The expression of male sterility in cms plants is the result of incompatibility
between recessive nuclear gene (called maintainer gene; rf) and male sterile
specific cytoplasmic genome. Now it is well documented that specific open
reading frame (ORFs) in mitochondrial genome (mt-genome) are responsible for
the expression of male sterile trait (Kumar et al., 2000). Once dominant restorer
(Rf) gene (located in nuclear genome) responsible for pollen fertility of a
cytoplasmic male sterile line is identified, it is commonly known as cytoplasmicgenic male sterility (CMS). Therefore, those cytoplasmic male sterile lines for
which Rf gene(s) have been identified are widely known as genic-cytoplasmic male
sterility (g-cms) and more often treated as a separate class of male sterility system.
However, both cms and g-cms can be described under common head (i.e. cms)
because of the fact that in both these systems, expression of male sterility is due to
the defect in the cytoplasm (mt-genome). Based on mode of action of the pollen
fertility restorer (Rf) and mainainer (rf) alleles, CMS are of two types, viz., (i)
gametophytic and (ii) sporophytic. In gametophytic system, expression of restorer
allele is pollen specific, thus the pollen grains are the responding elements (e.g. Scytoplasm in corn, abortive cytoplasm in rice etc.). Therefore, a plant heterozygous
for maintainer-restorer locus (Rfrf) produces 50% aborted (rf) and 50% normal
(fertile) pollen (Rf). Pollen from such plant (Rfrf) crossed on to a sterile plant (rfrf),
will again produce plants with 50% each of aborted and normal pollen. In contrast,
all pollen are either fertile or sterile in sporophytic system, which is most common
(Pearson, 1981). A heterozygous restorer line (Rfrf) in this system produces all
fertile pollen and when crossed on to a sterile plant (rfrf), produces 50% absolute

male sterile and 50% absolute male fertile plants. Cytoplasmic male sterility may
originate from inter-generic or inter-specific crosses and may be artificially
induced through mutagenesis or antibiotic effects on cytoplasmic genes (Kaul,
1988). Cytoplasmic male sterile plants have also been developed in several
vegetables through protoplast fusion (Pelletier et al., 1995). In near future,
genetically engineered cytoplasmic male sterility may be available after
standardization of transformation technique for organelle genome.
Transgenic Male Sterility Systems
From the beginning of 1990s, new genetic approaches have been proposed
and implemented to develop male sterility systems through genetic transformation
(Mariani et al., 1992). The ability to design new molecular strategies and their
successful execution has been possible because of the isolation, cloning and
characterization of anther or pollen specific genes and promoter sequences. These
genes are expressed in pollen themselves (gametophytic expression) or cells and
tissues (sporophytic expression) that directly or indirectly support pollen
development, such as tapetum, filament, anther wall. Williams et al., (1997)
reviewed the reports on genetically engineered male sterility systems under
dominant male sterility, recessive male sterility, targeted gametocide and dual
method. Transgenic plants in which the Bcp1 gene is perturbed showed an
inheritable male sterile phenotype. Mature pollen grains lack cytoplasmic contents
and appear as empty, flattened exine shells. The transgenic plants are normal in all
other respects except their inability to produce functional pollen. The induction of
male sterility in the model plant Arabidopsis using Bcp1antisense RNA has
provided a new technology for the production of hybrid crop plants, that is
potentially applicable for the production of hybrid seed in Brassica crops.
However, based on mechanism of male sterility induction and fertility restoration,
all transgenic male sterility systems developed so far can be described under five
classes, viz.,
(i) abolition-restoration system,
(ii) abolition-reversible system,
(iii) constitutive-reversible system,
(iv) complementary-gene system and
(v) gametocide-targeted system.

Although in transgenic(s) developed within one system, mode of action of

trans-gene(s) remains the same, there can be variations in trans-gene constructs
including promoter, targeted site (depending upon the promoter used) and
methodology adopted within one system. All the transgenic male sterile lines
developed till date are gms, since they have been developed through
transformation of male sterility causing gene construct(s) inside the nuclear
genome. Regardless of the crop, all transgenic male sterility systems (except
constitutive- reversible) with the same trans-gene construct(s) may be utilized to
develop transgenic male sterile lines in those vegetables, where transformation and
regeneration protocols have been standardized.
Utilization of gms
Since gms is maintained through backcrossing, in hybrid seed production
field, 50% male fertile segregants (Msms) need to be identified and removed before
they shed pollen. In some gms lines, ms genes are tightly linked with the recessive
phenotypic marker genes. Such marker genes, especially which expresses at
seedling stage, are good proposition for the identification of sterile/fertile plants at
seedling stage. Hybrid seed production using EGMS line is more attractive because
of the ease in seed multiplication of male sterile line. Seeds of EGMS line can be
multiplied in an environment where it expresses male fertility trait while hybrid
seeds can be produced in other environment, where it expresses male sterility.
Because of more tedious maintenance process and non-availability of suitable
marker gene among the vegetable crops, utilization of gms is restricted only in few
vegetables. The identification of fertilizing cytoplasm for specific nuclear male
sterile gene (Horner and Palmer, 1995), is an interesting research area, which upon
success, may provide opportunity for most efficient utilization of gms lines, like
cms line.

Utilization of cms
The cms system is the most commonly utilized male sterility to produce
commercial hybrid seeds of several vegetables. The cms based hybrid development
is often term as three line method of hybrid breeding involving A line (male sterile;
S-rfrf), B line (maintainer; N-rfrf) and C or R line (restorer; S-or N-RfRf). As
mentioned, cms line without restorer male parent cannot be utilized in fruit
producing vegetables (e.g. chilli), but it can be utilized in vegetables where
vegetative part is of economic value (e.g. onion, cole vegetables, carrot, radish,
leafy vegetables etc.). The cms system though is the most commonly utilized, its
utilization is restricted in specific species because of the following limitations:
Non-availability of cms in many crops and their wild relatives.
Need of fertility restorer allele in fruit producing vegetables.
Undesirable pleiotropic effect of sterile cytoplasm on horticultural qualities.
Highly unstable sterile cytoplasm in several cases.
Poor cross pollination ability of flowers of plants with sterile cytoplasm due to
altered morphology.
Technical complexity involved in seed production and maintenance of parental
lines.
Besides, vulnerability of sterile cytoplasm to specific diseases is a major risk
due to monopolistic cultivation of hybrids derived from single source of sterile
cytoplasm. The devastation of corn hybrids derived from T-cytoplasm by

Helminthosporium blight in USA during 1970s (Levings, 1990), is a well-known

example of such risk.

Self-incompatibility:
Ever since the first discussion on self-incompatibility by Darwin, the
phenomenon has extensively studied in several plant families and now significant
amount of information is available on genes and gene products involved in the
expression of SI trait (Dodds et al., 1997).
Types:
There are two types of SI, viz., gametophytic and sporophytic. In
gametophytic system SI reaction of pollen and stigma is determined by the
genotype of the mother plant on which pollens are produced (e.g. tomato) while in
sporophytic system, pollen phenotype (SI reaction) is determined by the genotype
of the mother plant on which pollens are produced (e.g. cole vegetables). In
Brassicacae, sporophytic self-incompatibility (SSI) has been best characterized
and successfully utilized for the development of commercial hybrids (Pearson,
1983; McCubbin and Dickinson, 1997, Tripathy and Singh 2000, Singh 2000,
Singh et al., 2001).
SPOROPHYTIC SELF-INCOMPATIBILITY (SSI)
Sporophytic self-incompatibility (SSI) was first observed in radish (Stout,
1920), and its inheritance pattern was first demonstrated (Bateman, 1955). The
numbers of S allele at S-locus have been reported to be 34 in Raphanus sativus and
60 in B. oleracea.

Genetic and Molecular Basis of SSI

The complex genetics of SSI has been discussed by Dickson and Wallace
(1986). The compatible or incompatible reaction of pollen and stigma is not only
depends on the genotypes (homozygous/heterozygous) of male and female plant at
S locus, but also on the existence of any of the following four levels of interactions
between the two S alleles, which characterize the incompatibility/compatibility
specificity of the pollen and stigma: dominance (S1 <S2), co-dominance (S1 + S2),
mutual wakening (no action by either allele) and intermediate gradation (1100%
activity by each allele) (Dickson and Wallace,1986).The analysis of sequence data
of S-locus cDNA clones revealed that various S alleles exhibit high levels (up to
30%) of sequence divergence (McCubbin and Dickinson, 1997).
Utilization
For hybrid seed production both the parental inbreds should have two
different S alleles for strong self-incompatibility (in case of single cross hybrid).
One S.I. inbred is used as female parent and a good pollinator (an open pollinated
variety) as male to develop top cross hybrid, while four S.I. inbreds having
altogether different S alleles are used to produce double cross hybrids. Among the
cole vegetables like cabbage, cauliflower, broccoli etc., self-incompatibility
(sporophytic) mechanism is being utilized for hybrid seed production at several
places including India (Singh, 2000). Usually in cauliflower S.I. is weak and S.I
reaction is breaks at high temperature, resulting into selfing and sibling (brothersister mating) among the plants of female parent, thus deterioration in the genetic
make-up of F1 seeds. Following strategies are utilized to overcome sibling
problem:
(i)
use of parental lines with synchronized flowering,
(ii) use of parental lines with similar morphology
(iii) pollination by stored pollen and
(iv) use of male sterility as an alternative to S.I. inbreds.
Due to the inherent advantages, the last option i.e. use of cms line is being
getting more attention in the hybrid seed production of all cole vegetables.
The advantage of SI system is the possibility to produce hybrid seed using
two SI lines homozygous for different S alleles as parental components. Following
disadvantages can be considered: less reliable SI, which results in an undesirable
number of sibs in hybrid seeds; some difficulties connected with reproduction of

SI lines by means of high CO2 concentration (Jik 1985) or spraying with NaCl
solution (Kuera 1990).
Commercial utilization in vegetable crops:
In India vegetable hybrids based on CMS and CGMS system have been
limited. In India, genetic male sterility (GMS) has been exploited commercially
only in the cases of chilli and muskmelon to develop F 1 hybrid seed commercially.
Punjab Agricultural University (PAU), Ludhiana, India has released two chilli
hybrids (CH-1 and CH-3) and one muskmelon hybrid (Punjab Hybrid) based on
the GMS system. Similarly in tomato, work on GMS lines is in progress at PAU.
The CGMS system has been commercially exploited in chilli, onion and carrot. In
the recent past, chilli CGMS lines were introduced at the Indian Institute of
Vegetable Research (IIVR) from AVRDC, which are utilized directly or indirectly
to produce CMS-based hybrids, i.e. Kashi Surkh (CCH-2) and Kashi Early (CCH3). The Indian Institute of Horticultural Research (IIHR), Bangalore, India has also
released chilli hybrids based on the CGMS system, i. e. Arka Meghna (MSH-172),
MSH-149 and MSH-96. In carrot, the Indian Agricultural Research Institute (IARI)
regional station, Katrain (HP), India has developed one hybrid, 'Pusa Nayanjvoti',
which is based on petaloid CGMS and it was identified for release from Delhi state
seed subcommittee in 2009. In the tropical group of carrot, IARI, New Delhi India
has also reported CGMS system in different genetic backgrounds and evaluation of
different hybrid combinations is in progress. In onion, IIHR, Bangalore has
released two hybrids based on the CGMS system, i.e. Arka Kirtiman and Arka
Lalima, and IARI, New Delhi has developed two hybrids in onion (Hybrid-63 and
Hybrid-35) which are based on the same system. The CMS system has been
commercially exploited in cabbage, cauliflower and onion.
In tomato, approximately 19 male sterile based hybrids have been released,
17 of them are based on functional sterility controlled by gene positional sterility
(ps-2).
In pepper (Capsicum annuum; 2n = 24), The MS-12 (ms-10ms-10) line is
being utilized to produce hybrid seeds of CH-1 and CH-3 hybrids of hot pepper in
Punjab state. However, non-availability of Rf genes in most of the sweet pepper
genotypes is still a handicap in developing cms based commercial sweet pepper
hybrids.

Among cole vegetables (Brassica spp. 2n = 18) although gms based

experimental crosses have been developed, it has not been commercially utilized
mainly because of the difficulty in multiplication of male sterile seeds and
availability of self-incompatibility and cms systems. Similarly in many other
vegetable crops hybrids have been developed using these mechanisms.
Research efforts at IIHR, Bengaluru from past several years resulted in
development of male sterile lines in onion, chilli, okra, carrot, cauliflower, ridge
gourd and marigold for generating hybrids. Cytoplasmic male sterile (CMS) lines
were used in onion to develop F 1 hybrids namely Arka Kirtiman and Arka Lalima.
In chilli, the cytoplasmic genic male sterile (CGMS) lines were used to develop F 1
hybrids namely Arka Meghana, Arka Sweta, Arka Harita and ArkaKhyati.
Furthermore, GMS lines in okra, brown anther and petaloid types (CMS) in carrot,
CMS lines in cauliflower and petaloid types (CGMS) in marigold have been
developed. Besides, CMS lines in ridge gourd have been identified. Male sterile
lines developed in onion, chilli and okra are very popular and commercialized to
many private seed industries and public institutions.
So far majority of cruciferous hybrid cultivars have been created by means of SI
(Watanabe, Hinata 1999). The most widespread system used in Brassica oleracea
hybrid breeding is the improved Ogura CMS (Pelletier et al., 1983).
Future prospects:
Biotechnological approaches involving molecular markers and genetic
engineering have given a new dimension to utilisation of male sterility. Barnasebarstar system is one such development in genetic engineering of male sterility.
Recent molecular studies have reported association of male sterility with
chimeric mitochondrial open reading frames (ORFs) which are combination of
mitochondrial genes with ORFs created in plant mitochondrial genome due to their
high recombinogenic activity. The transcripts originating from these chimeras are
translated into unique proteins that appear to interfere with mitochondrial function
and pollen development. The physiological and in depth molecular aspects by
which the products of these chimers interfere with the formation of male
gametophytes are still the subject of intense research.
In recent years, brassica breeders are trying to use male sterility system
instead of the standard incompatibility system. Some of the difficulties
encountered were the reduction in nectary gland size and decreasing function of

these glands. Plants with male-sterility gene were therefore unable to attract insect
pollinators which are required for pollination.
Self-incompatibility and male sterility have proved instrumental in
commercialization of hybrids in cabbage followed by in other cole crops. Although
sporophytic self-incompatibility system was confirmed in cabbage by Adamson
(1965), but the methods of seed production by utilizing self-incompatible parental
lines had been suggested by Odland and Noll (1950) much earlier. The use of selfincompatibility involves the basic steps of identification of self-incompatible plants
in diverse genotypes, development of homozygous self-incompatible S-allele lines,
studying interallelic relationships of S-alleles, ascertaining stability of S-allele
lines in diverse environments, pinpointing the most heterotic S-allele parental lines
combination, maintenance of S-allele lines and their utilization in the production of
hybrid seed on commercial scale.
Currently at IARI Regional Station Katrain, sporophytic self-incompatibility(SSI)
and cytoplasmic male sterility (CMS) are being used in the development of
hybrids. KGMR-1 and KTCBH-81 based on SSI have already been identified for
release during 2005 and 2013 respectively and KTCBH-84based on CMS is being
evaluated in trials of All India Coordinated Research Project on Vegetable Crops.
At CSKHPKV Palampur, mild chill requiring genotypes viz., KGAT-I, II and III
have been developed through hybridization of temperate and tropical genotypes.

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