1. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Rani Lakshmi Bai Central Agricultural University
Jhansi -284003
Credit seminar on
Wide Hybridization in Chickpea
Sanjay H B
RLBCAU/AG/PG/0017/19
M.Sc. Ag. (Genetics and Plant Breeding)

2. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
 The present day high yielding varieties have narrow genetic
base as very few germplasm lines have been utilized in
development of these varieties (Kumar et al. 2008).
 Narrow genetic base results in higher vulnerability to biotic
and abiotic stresses.
 Hence, broadening of genetic base to reduce vulnerability
to stresses is very essential.
 The chickpea has low yield potential as compared to cereals
like wheat, rice and maize etc.
Wide Hybridization

3. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
 The reason for low genetic yield potential may be due to the
fact that genes responsible for higher yield might have been
eroded during long evolutionary process and natural selection
under harsh environmental conditions (dry area/rainfed) for
survival traits.
 This eroded gene can be regained by hybridizing the cultivated
species with wild relatives.
Wide hybridization: Is the process of crossing the individuals
belonging to two different species or genera, facilitating the
exploitation of useful genes from wild unimproved species for the
benefit of the cultivated species.
Wide Hybridization

4. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Importance of Wide Hybridization
 Distant hybridisation contributed immensely to crop
improvement, gene and genome mapping, understanding
chromosome behaviour, and to know about the evolution of
some crops.
 Ultimate goal of distant hybridisation is to transfer useful
genes from alien species to cultivated crop species.
 This helps to create the variability, stability, wide adoptability
in the cultivated species and high genetic gain under selection
among the germplasm can be achieved.
 Pre breeding lines, and parental lines can be developed
through wide hybridisation with multiple stress resistance,
desirable agronomic and yield attributing traits can be
introgressed from wild relatives.

5. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Chickpea
Primary centre of origin : South Eastern Turkey and Syria
Centers of diversity : Mediterranean, Central Asia, The near
Eastern centre and India.
Secondary centre of origin : Ethiopia
Source : Singh R et al., (2008)

6. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi Types of Chickpea
Desi type : Mainly grown in Indian Subcontinents , they have
greenish purple foliage with Purple coloured flower, Seeds are
angular shaped, rough surface, brown in colour.

7. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Kabuli type : Foliage is lite green, white coloured floweres,
Seeds are creamy coloured, owl head to pea shaped seed.
Types of Chickpea

8. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi Chickpea Gene Pool
Genera cicer consists of 9 annual and 34 perenial species.
Vander Maesen et al.,(2007) proposed recent classification
of chickpea gene pool.
Primary Gene Pool : It consists of cultivated and species
and its landraces. It includes all species that hybridizes
freely, show good chromosome pairing leading to gene
exchange and produce viable hybrids.
(Ohri D et al., 2016)
Ex: Cicer arietinum ( 2n=2X=16)
Cont.d

9. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Chickpea gene pool
Secondary Gene Pool : It includes species that can be used
as germplasm resource, however hybridisation with the
cultivated species is difficult because of genetic barriers or
chromosome alterations and some degree of sterility is
associated with the first generation hybrids. (Ohri D et al., 2016)
Ex: Cicer reticulatum (2n=2x=16)
Cicer echinospermum (2n=2x=16)
Tertiary Gene Pool : It consists of 6 annual species, and 34
perennial species that are not crossable with the cultivated
species. (Ohri D et al., 2016)
Ex: Cicer cuneatum
Cicer chorassanicum
Cicer yamashitae

10. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi Crossability potential
 The crossability of cultivars with wild species is a
prerequisite for alien gene introgression.
 Conventional crossing has been successful in producing
interspecific hybrids between Cicer areitinum and Cicer
reticulatum and between Cicer arietinum and C.
echinospermum.
 The availability of novel embryo rescue, tissue culture
technique circumventing crossing barriers has brightened
the prospects of transferring useful traits from the tertiary
gene pool. And as a result, hybrids were obtained
between Cicer pinnatifidum and C. bijugum.
Cont.d

11. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi Crossability potential
Pod set in the crosses between annual and perennial Cicer
species
Female parent Male parent Pod set %
C. microphyllum C. reticulatum 35
C. bijugum C. anatolicum 30
C. oxyodon C. reticulatum 24
C. microphyllum C. echinospermum 22
C. bijugum C. nursitanicum 20
C. bijugum C. oxyodon 15
C. pinnatifidum C. oxyodon 10
Crossing were carried out in 2006 at WSU, USA
Source : C.Kole (ed.), Wild crop relatives: Genomic and breeding resource, Legume
Crops and Forages, N Mallikarjuna et al.,(2011)

12. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi

13. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Barriers for production of Wide hybrids
Pre zygotic
barriers
Post zygotic
barriers
Sticky stigmatic secretion
Failure of pollen
germination, tube growth
Failure of fertilization
Embryo abortion
Inviability of F1 hybrids
Hybrid lethality and
weakness
Crossability
barriers

14. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi Strategy to over coming crossability barriers
Invitro techniques :
 Invitro techniques like embryo rescue, ovule culture
greatly reduce the barriers of wide hybridization in
chickpea.
 Clarke et al., (2006) suggested that the appropriate time
to rescue C arietinum × C bijugum hybrid is the early
globular stage of embryogenesis ( 2-7 days)
 In contrast, C arietinum × C pinnatifidum hybrids abort
later (15-20 days) at the heart shaped or terpedo stages
are easier to rescue invitro.
Cont.d

15. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Strategy to over coming crossability barriers
Use of growth harmones :
 Post pollination application of growth regulators like
gibberellic acid(120ppm), kinetin(15ppm), napthalene acetic
acid(30ppm), 2,4-D singly or in combination helpful in
maintaining the developing seeds by facilitating division of
hybrid zygote and endosperm. (Singh S et al.,2005)
 Mallikarjuna et al.,(2009) observed that application of
growth regulators (Zeatin + IAA) to pollinated pistils to
prevent initial pod abscission and to save the aborting
hybrids by embryo rescue technique in C. arietinum × C.
pinnatifidum
Cont.d

16. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Strategy to over coming crossability barriers
Reciprocal crossing :
1. Reciprocal differences in wide crosses are very common. If
disharmony between the genome of one species and
cytoplasm of other is a cause of a fertilization barrier,
reciprocal crosses can be successful in recovery of hybrids.
Ex: C. reticulatum × C. microphyllum was may not be
successful, its reciprocal cross C. microphyllum × C.
reticulatum produces successful hybrid with 34% seed setting.
2. In general using a female parent with higher chromosome
number is more successful.
Cont.d

17. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Strategy to over coming crossability barriers
Use of bridge species :
C. reticulatum and C. echinospermum could be considered
bridge species as means of transferring of genes from species
of the tertiary gene pool to the cultigen.
Ex: C. oxyodon carries resistant gene for Pea streak carlvirus
but it is cross incompatible with cultivated species of chickpea.
Hence a bridge species called C. reticulatum is used to transfer
that gene to primary gene pool.
C. oxyodon × C. reticulatum (24% seed set)
Progeny × C. areitinum
Progeny containing resistant gene for Pea streak carl virus

18. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Application of Wide hybridization in Chickpea
1. Disease resistance :
Ascochyta blight resistance, Fusarium wilt resistance,
Botrytis grey mould resistance, Phytophthora root rot
resistance gene is transferred from wild species to
cultivated species.
Sources of disease resistant traits identified in Cicer species.
Diseases Resistant wild species
Ascochyta blight C. judaicum, C. pinnatifidum, C. cuneatum
Fusarium wilt C. bijugam, C. reticulatum, C. echinospermum
Botrytis grey mould C. judaicum, C. pinnatifidum, C. bijugam
Rust C. reticulatum, C. reticulatum, C. echinospermum
Phytophthora root rot C. reticulatum, C. bijugam, C. pinnatifidum,
Stem rot C. yamashitae, C. judaicum, C. pinnatifidum
Source: Mohar Singh et al.,(2016)

19. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Application of Wide hybridization in Chickpea
Pest resistance :
Sources of pest resistant trait identified in wild species.
Pest Resistant wild species
Helicoverpa pod borer
(tolerance)
C. bijugam, C. reticulatum, C.echinospermum
C. microphyllum
Pulse beetle C. cuneatum, C. judaicum, C. bijugam,
Leaf miner C. judaicum, C. bijugam, C. reticulatum
Root lession
nematode
C.echinospermum, C. reticulatum
Bruchids C. reticulatum
Cyst nematode C. bijugam, C. pinnatifidum, C. reticulatum
Root knot nematode C. bijugam, C. judaicum
Source: Mohar Singh et al.,(2016)

20. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Application of Wide hybridization in Chickpea
Cold tolerance :
 C. reticulatum, C. bijugum, C. judaicum, C. pinnatifidum, are
the sources of cold tolerance identified in Cicer species used
for introgression in to elite genetic background of chickpea.
 FLIP87-82C is the pre breeding line for cold tolerance
produced from its wild species.
 Germplasm line ILC8262, the mutant ILC8617 also reported
as best source of cold tolerance.
Cont.d

21. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Application of Wide hybridization in Chickpea
Drought tolerance :
 Canci and Toker (2009) reported few accessions of C.
reticulatum, and C. pinnatifidum to perform better under
drought conditions and those lines could be considered as the
best available drought resistant sources.
 Dehydrin gene(cpdhn1) of C. pinnatifidum isolated from
cDNA, reported as Drought tolerant gene.
 C. stapfianum, C. subaphyllum and C. pungens have been
found to be drought resistant.
Cont.d

22. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Application of Wide hybridization in Chickpea
High yielding :
 Introgression of high yielding gene from C. reticulatum to
cultivatd variety L550, with interspecific derivatives
showing 6-17% yield advantage.
 A cross between Pusa 256 and C. reticulatum was made
and their F1 was again crossed with the wilt resistant
variety pusa 362 further selection conducted with
development of pusa 1103 which is high yielding and
multiple stress resistant.
 Singh and Ocampo transferred some genes from C.
echinospermum, C. reticulatum to cultivated chickpea and
observed up to 39% increase in seed yielding following the
pedigree method.
Cont.d

23. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Application of Wide hybridization in Chickpea
High protein content :
 C. pinnatifidum contain 268g/kg of seed and used as
donor parent for protein content.
 Chickpea cultivars were crossed
with C. pinnatifidum, C.reticulatum,
C. echinospermum on the premise
that recombination could result in
progenies with high yielding and
high protein content.
fig. C. pinnatifidum

24. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Breeding methods used in Wide hybridization
 The bulk method, variously modified is now the most
common selection method used after hybridization in
chickpea.
 SSD(Single Seed Descent ) method, a modification bulk
method is used for development of RIL for genome mapping.
 A population improvement method that involves inter
crossing of selected plants in F2s or F3s has been suggested
for legumes for enhancing the chances of recombination in
segregating generation.
 Van Rheenen et al.,(2009) proposed a method called polygon
breeding, where by segregating population and selection are
shared and exchanged between breeders.

25. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi Limitations of wide hybridization
 Incompatible crosses.
 F1 sterility
 Undesirable linkage
 Problems in transferring recessive oligogenes and
quantitative traits.
 Lack of flowering in F1
 Problems in using improved varieties in distant hybridization
 Presence of pre and post fertilization barriers.

26. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Achievements in Chickpea Wide hybridization
 Singh et al.,(2012) developed certain pre breeding lines
IPC71 (C. arietinum × C. judaicum) has been used as donor
parent under National Chickpea Crossing Program.
 Three cross combinations of Pusa 1103 × ILWC 46, Pusa 256
× ILWC239, shows higher variability for important yield
related traits.
 The most important varieties are chaffa, Dohad yellow,
BDN 9-3, Annegeri -1, JG315, JG 74, Pragati and BG 287 and
these selections are popular in many parts of India.
 Early flowering recombinants have also isolated from the
interspecific crosses C. arietinum × C. reticulatum
Cont.d

27. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Achievements in Chickpea Wide hybridization
 Jaiswal et al.,(2006) found 3 out of 5 F5 lines derived from the
crosses C. arietinum × c. reticulatum gave significant higher
yield than the superior chickpea check, with 1 line giving a
51.2% increase in yield.
 Singh and Ocampo reported numerous transgressive
segregrants for high yield up to 39% better than the locally
adopted parent in F2 population between C. arietinum × C.
echinospermum and C. arietinum × C. echinospermum.
 cpdhn 1 is the dehydrin gene identified in C. pinnatifidum and
it is used as donor parent for drought tolerance.

28. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Case study 1
Wide hybridization in chickpea for creating variability and
increasing yield via more number of primary branches per
plant. S.K.Chaturvedi et.al.,(2016)
In this investigation 5 diverse genotypes of chickpea were
utilized to develop 8 recombinations. 4 interspecific crosses
between C. arietinum × C. reticulatum and 4 intervarietal
crosses between Desi × Kabuli crosses were made to create
variability and number of primary branches per plant in
chickpea
Cont.d

29. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Results of the study
 The Diallele Selective Matting System (DSMS) is likely to
result in development of high yielding varieties in
chickpea.
 The mean value of hybrids indicated the presence of large
variation among crosses as evident from the coefficient of
variations for the trait studied.
 Among various traits, Primary branches per plant
exhibited maximum Coefficient of variation

30. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Case Study 2
• The material comprised three kabuli genotypes of chickpea,
and three wild species,C. cuneatum ( ICCW 47),
C. echinospermum (ICCW 47), and C. reticulatum (ICCW 48).
• The cultivated chickpea genotypes were crossed as females to
the wild species.
• Some of the true F1 hybrid plants obtained with C.
reticulatum were backcrossed twice to their respective female
parents. The pedigree method was followed to advance the
segregating populations from straight crosses (without
backcross), BC1 and BC2 generations to F5–F7.
Introgression from wild Cicer reticulatum to cultivated chickpea
for productivity and disease resistance.
Singh et al.,(2005)
Cont.d

31. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
 Over all, 22 promising uniform lines(14 desi, four kabuli and
four wild types) selected from
F6/F7/BC1F5/BC1F6/BC2F5/BC2F6 generations were
retained for preliminary evaluation for yield and yield
components.
 Finally, six lines (four desi and two kabuli) along with
standard check cultivars, GPF2 (desi) and L550 (kabuli), were
selected and evaluated
 These 6 lines possess a high degree of resistance to wilt, foot
rot and root rot diseases, and recorded a 6.1–17.0% seed
yield increase over the best check cultivars.
Results of the study

32. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi Future perspectives
 Most of the desirable gene complexes are present in non
crossable secondary and tertiary pools, and there is an urgent
need to augment germplasm collections in primary gene pool
and land races.
 An identification of area is required for further exploration
and targeted trait specific collections particularly from WANA
region.
 The evaluation and identification of wild Cicer species will
greatly aid in trait discovery.
 With the use of molecular tools and pre breeding activities,
trait of interest should be introgressed from wild species to
cultivars.

33. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Conclusion
 Wide hybridization causes broadening the genetic base and
bringing the characters like high protein content, early
maturity, high number of primary and secondary branches,
multiple stress resistance to the cultivated chickpea species.
 Crossability barriers during hybridization frustrate breeders
efforts in successful hybridization between Cicer species.
These barriers can be overcome through novel technologies.
 Among Cicer species C.reticulatum and C. echinospermum
are compatible with all other Cicer species as an female
parent so they can be utilized as bridge species.
 Crossing between species of same genera have contributed
immensely to crop improvement. Hence wide hybridization is
crucial for transferring useful genes from alien species to
cultivated species.

34. Rani
Lakshmi
Bai
Central
Agricultural
University
Jhansi
Thank You