Conquering gene pools of pigeonpea [Cajanus cajan (L.) Millspaugh]

Nalini Mallikarjuna

Grain legumes ProgramInternational Crops Research Institute for the

Semi-Arid Tropics, Patancheru, India.

ICRISAT is a member of the CGIAR Consortium

ICRISAT Collaborators

ICRISAT is a member of the CGIAR Consortium

Pigeonpea Domestication Bottleneck/s

One founder species i.e. Cajanus cajanifolius

Indian origin

Diploid cultivated pigeonpea

Farmer’s selection

Plant breeding

Pigeonpea with a narrow genetic baseICRISAT is a member of the CGIAR Consortium

How to tackle the brewing storm?

Water Scarcity and water loggingICRISAT is a member of the CGIAR Consortium

Gene pools of pigeonpeaPrimary Gene PoolCajanus cajan and its land races.

P

S

T

Q

Secondary Gene PoolC. cajanifolius, C. lineatus, C. lanceolatus, C. laticepalus, C. albicans, C. reticulatus, C. sericeus, C. scarabaeoides, C . trinervius, C. acutifolius,.

Tertiary Gene PoolC. goensis, C. heynei, C. kerstingii, C. mollis,C. rugosus, C. volubilis, C. platycarpus, C. niveus, C. gandiflorus, C. crassicaulis, C. rugosus, C. elongates, C. villosus, C . confertiflorus, C. visidus, C. aromaticus, C. crassicaulis, C. lanuginosus, C. pubescens, C. cinereus, C. marmoratus, C. mareebensis.C. lanuginosus, C. pubescens.

Quaternary gene PoolFlemingia, Rhynchosia, Dunbaria, Erisema Paracalyx, Adenodolichos, Bolusafra, Carissoa, Chrysoscias, Baukea.

Fig. 1. Taken from Mallikarjuna et al., 2011

ICRISAT is a member of the CGIAR Consortium

Primary gene pool:

• >13,600 accessions

• Sixty-six cultivars based on germplasm and improved breeding lines developed by ICRISAT have been released in several countries of Asia (38), Africa (22), Australia (3) and USA (3). Short and medium duration cultivars have made a significant impact in Asian countries. However, long and medium duration varieties with fusarium wilt resistance and consumer preferred large, cream colored seeds have created impacts in eastern and southern Africa.

• Variation for days to flower, maturity, yield, harvest index, shelling ratio with limited amount of disease and pest resistance etc.

• High yielding varietiese and useful genetic stocks

ICRISAT is a member of the CGIAR Consortium

Secondary gene pool

•Compatible wild relatives

•Traits discovered:

• C. acutifolius: A5 CMS system, pod borer, pod fly, bruchid resistance, phytophthora blight, dwarf plant type, high seed weight, beige seed color.

• C. lanceolatus: A9 CMS system, bruchid resistance.

ICRISAT is a member of the CGIAR Consortium

Cajanus acutifolius

• A wild relative from secondary gene

pool.

• Native of Auatralia

• Has a repertoire of disease and

pest resistance traits.

• One way cross more successful

than the reciprocal.

ICRISAT is a member of the CGIAR Consortium

Helicoverpa armigera damage to pigeonpea

ICRISAT is a member of the CGIAR Consortium

And the result is…!!!

ICRISAT is a member of the CGIAR Consortium

How does this happen?

Happy Helicoverpa but sad peas!!!!

Publications: Mallikarjuna et al., 2007, Mallikarjuna et al., 2012a, Mallikarjuna et al., 2012b

ICRISAT is a member of the CGIAR Consortium

Pod borer resistant Cajanus acutifolius

Fig. A total of 166 C. platycarpus and 301 C. acutifolius interspecific derivative lines with low Helicoverpa armigera damage

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Helicoverpa pod damage

C. platycarpus

C. acutifolius

ICRISAT is a member of the CGIAR Consortium

High seed weight lines

Fig. Helicoverpa armigera pod damage and high 100 seed wt. in a Cajanus acutifolius derived advanced progeny line

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1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 R-cont-icp-332

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ICRISAT is a member of the CGIAR Consortium

C. acutifolius lines with multiple disease resistance

ICPLNos FW SM PB SWT

ICPL2011146 12.5** 0** 8** 13.6**

ICPL2011147 11.1** 0** 2** 11.6**

ICPL2011148 0** 0** 1** 17.3**

ICPL2011150 0** 0** 2** 12.4**

ICPL2011160 14.3** 0** 2** 18.4**

ICPL2011164 12.5** 0** 7** 20**

ICPL2011168 14.3** 0** 3** 19.6**

ICPL2011169 16.7** 0** 3** 19.6**

ICPL2011183 11.1** 0** 2** 18.8**

ICPL2011170 11.1** 0** 3** 19.2**

ICPL2011161 0** 0** 2** 18.8**

ICPL2011203 12.5** 0** 1** 16.9**

Publications: Jadhav et al., 2012 a & b, Mallikarjuna et al., manuscripts under preparation

ICRISAT is a member of the CGIAR Consortium

A5 CMS systemA5 CMS system was developed utilizing C. acutifolius and with cultivated pigeonpea cytoplasm.

•First report by Mallikarjuna and Saxena (2005).

•The only system with cultivated pigeonpea cytoplasm

•Has post meiotic pollen abortion

•Plenty of restorers available

•Maintainers have been identified

•Handed it over to the breeders

ICRISAT is a member of the CGIAR Consortium

Cajanus cajan

Cajanus lanceolatus

Traits of interest transferred: A9 CMS system, pod borer and bruchid resistance

INTER-SPECIFIC CROSS THAT IS FERTILEIntrogression of traits

A9 CMS system

• A9 CMS system was recently discovered

by us.

• The beauty of the system is that it is developed on cultivated pigeonpea cytoplasm.

• It was possible to identify maintainers and restorers in pigeonpea cultivars

• Will be handed over to breeders shortly.

Tertiary gene pool

•Incompatible Cajanus species

•C. platycarpus: A7CMS, clasmogamy,pod borer, pod-fly, bruchid and Phytophthora blight resistance.

•C.volubilis: dwarf plant type, early maturity and high pod number.

ICRISAT is a member of the CGIAR Consortium

Helicoverpa resistant Cajanus platycarpus

Compatible species Incompatible species

Cultivated species wild species[100% A] [100% B]

 

F1

[50%A+50%B]  

BC1

[75%A+25%B]Screen for Helicoverpa

  

BC2

[87.5%A+12.5%B]Screen for Helicoverpa

  

BC3

[93.75%A+6.25%B]Screen for Helicoverpa

 

Cultivated species Wild species[100%A] [100%B]

[or vice versa] 

          Growth regulator applications

          Embryo rescue techniques F1

[50%A+50%B] 

          Growth regulator applications

          Embryo rescue techniques BC1

[75%A+25%B]Screen for Helicoverpa

 

          Growth regulator applications,

          Embryo rescue techniquesBC2

[87.5%A+12.5%B]Screen for Helicoverpa

 

[ May require growth regulator

applications, embryo rescue techniques]

BC3

[93.75%A+ 6.25%B]Screen for Helicoverpa

 

Interspecific hybridization

Mature seed set from BC2

onwards

Embryo abortion in BC1

F

Embryo abortion in F1

FM

Embryo rescue in recurrent backcrosses

Publications: Mallikarjuna et al, 2007, Mallikarjuna et al., 2011; Mallikarjuna et al., 2012

F

Molecular analysisDArT analysis showed the presence of C. platycarpus genome ranging from 2.0 to 4.8%. The presence of non-parental alleles, presumably due to recombination, ranged from 2.6 to 10.4%. These two categories have contributed to the variation observed in the progenies.

Cajanus platycarpus derivatives are showing variation for plant growth, height, seed

weight, disease and pest resistance

C. platycarpus seeds

Fig. A total of 166 C. platycarpus and 301 C. acutifolius interspecific derivative lines with low Helicoverpa armigera damage

0

50

100

150

200

250

300

11-10 11-20 20-30 30-40

Helicoverpa pod damage

C. platycarpus

C. acutifolius

A7 CMS systemA7 CMS system was developed utilizing C. platycarpus, a species from tertiary gene pool and incompatible with pigeonpea

• First reported by Mallikarjuna et al., 2011

• Has both pre-meiotic and post- meiotic pollen abortion.

• Handed over to pigeonpea breeders in 2012

Meiosis

Premeiotic

Postmeiotic

Important traits of A7 CMS system

• Clasmogamous flowers

• Pre and post meiotic anther abortion

• Non-dehiscent anthers

Publications: Cherian et al., 2007, Mallikarjuna et al., 2012.

Clasmogamous or open flower is an important trait in the development of CMS systems in pigeonpea. A7 CMS system had open flowers accompanied by sterile and non-dehiscent anthers, an important trait in this CMS system

Bruchid resistance

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Fig. 4. Bruchid damage in w ild relatives of pigeonpea

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Publication: Jadhav et al., 2012.

C. platycarpus lines with multi disease resistance ICPL NO FW SM PB SWT

ASHA 40.8 60.64 31.3 9.67

ICPL2011007 100.0** 0.00** 1.00** 8.2**

ICPL2011013 100.0** 0.00** 1.00** 7.7**

ICPL2011014 100.0** 0.00** 1.00** 7.3**

ICPL2011015 100.00** 0.00** 1.00** 8.2**

ICPL2011016 83.3** 16.7** 1.00** 7.6**

ICPL2011018 100.0** 0.00** 3.00** 7.6**

ICPL2011024 0.00** 100.0** 2.00** 12.0**

ICPL2011025 100.0** 0.00** 3.00** 8.7**

ICPL2011029 100.0** 0.00** 3.00** 8.7**

ICPL2011033 85.7** 0.00** 2.00** 8.4**

ICPL2011037 88.9** 0.00** 4.00** 8.7**

ICPL2011041 83.3** 16.7** 3.00** 8.5**

ICPL2011044 100.0** 0.00** 7.00** 7.8**

ICPL2011047 100.0** 0.00** 3.00** 11.1**

ICPL2011061 100.0** 0.00** 2.00** 8.1**

ICPL2011072 0.00** 100.0** 3.00** 8.6**

ICPL2011074 0.00** 0.00** 5.00** 7.8**

ICPL2011088 88.9** 0.00** 2.00** 6.6**

ICPL2011090 100.0** 0.00** 2.00** 7.8**

ICPL2011124 75.0** 12.5** 1.00** 8.2**

ICPL2011127 100.0** 0.00** 2.00** 7.3**

ICPL2011128 75.0** 0.00** 4.00** 8.00**

ICPL2011129 100.0** 0.00** 4.00** 7.9**

ICPL2011132 100.0** 0.00** 7.0** 12.4**

ICPL2011133 100.0** 0.00** 2.0** 8**

C. volubilis

First plant: Hybrid

Second plant: Cajanus cajan

Third plant: C. volubilis

SSR analysis of C. volubilis hybrid

Dwarf plant type and early maturity in F2 onwards

Srikanth et al., manuscript under preparation

Gene pools of pigeonpeaPrimary Gene PoolCajanus cajan and its land races.

P

S

T

Q

Secondary Gene PoolC. cajanifolius, C. lineatus, C. lanceolatus, C. laticepalus, C. albicans, C. reticulatus, C. sericeus, C. scarabaeoides, C . trinervius, C. acutifolius,.

Tertiary Gene PoolC. goensis, C. heynei, C. kerstingii, C. mollis,C. rugosus, C. volubilis, C. platycarpus, C. niveus, C. gandiflorus, C. crassicaulis, C. rugosus, C. elongates, C. villosus, C . confertiflorus, C. visidus, C. aromaticus, C. crassicaulis, C. lanuginosus, C. pubescens, C. cinereus, C. marmoratus, C. mareebensis.C. lanuginosus, C. pubescens.

Quaternary gene PoolFlemingia, Rhynchosia, Dunbaria, Erisema Paracalyx, Adenodolichos, Bolusafra, Carissoa, Chrysoscias, Baukea.

Fig. 1. Taken from Mallikarjuna et al., 2011

Quaternary gene poolRecent Success :Succeeded in crossing pigeonpea with Rhynchosia species and hybrids were obtained.

Crossing

pigeonpea

with

R. bracteata

SSR marker showing polymorphism

Genomic in-situ hybridization

Summary

• Species from secondary, tertiary and quarternary gene pool crossed successfully with pigeonpea

SummaryDiversification of CMS baseCMS system Wild species

A1 C. sericeus

A2 C. scarabaeoides

A3 C. volubilis

A4 C. cajanifolius

A5 C. acutifolius (Mallikarjuna & Saxena, 2005)

A6 C. lineatus

A7 C. platycarpus (Mallikarjuna et al., 2011)

A8 C. reticulatus

A9 C. lanceolatus (Srikanth et al, manuscript under preparation)

Summary

• Disease and pest resistant stable pre-breeding lines obtained from secondary gene pool species.

• Successfully crossed tertiary gene pool species and developed stable disease and pest pre-breeding lines.

• Dwarf and early plant type.

• High seed weight and number.

Legume Cell Biology Staff

• Deepak R Jadhav• V. Balakrishna• S. Satyanarayaya• K. Avinash• K. Lakshmi• A. Amruthaand the students Thank you Cell Biology staff for your help and

understanding and sharing the passion to pre-breed ICRISAT mandate legumes.

End of the pigeonpea story from me!!

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