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Conquering gene pools in pigeonpea

  1. Conquering gene pools of pigeonpea [Cajanus cajan (L.) Millspaugh] Nalini Mallikarjuna Grain legumes Program International Crops Research Institute for the Semi-Arid Tropics, Patancheru, India. ICRISAT is a member of the CGIAR Consortium
  2. ICRISAT Collaborators ICRISAT is a member of the CGIAR Consortium
  3. 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 base ICRISAT is a member of the CGIAR Consortium
  4. How to tackle the brewing storm? Water Scarcity and water loggingICRISAT is a member of the CGIAR Consortium
  5. Gene pools of pigeonpea Primary Gene Pool Cajanus cajan and its land races. P S T Q Secondary Gene Pool C. cajanifolius, C. lineatus, C. lanceolatus, C. laticepalus, C. albicans, C. reticulatus, C. sericeus, C. scarabaeoides, C . trinervius, C. acutifolius,. Tertiary Gene Pool C. 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 Pool Flemingia, 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
  6. 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
  7. 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
  8. 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
  9. Helicoverpa armigera damage to pigeonpea ICRISAT is a member of the CGIAR Consortium
  10. And the result is…!!! ICRISAT is a member of the CGIAR Consortium
  11. 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
  12. Pod borer resistant Cajanus acutifolius Fig.A total of 166 C.platycarpus and 301C.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 ICRISAT is a member of the CGIAR Consortium
  13. High seed weight lines Fig. Helicoverpa armigera pod damage and high 100 seed wt. in a Cajanus acutifolius derived advanced progeny line 0 10 20 30 40 50 60 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 R- cont- icp- 332 S- cont- 87 progenies Poddamage(%) 0 2 4 6 8 10 12 14 16 18 20 (%) Dam 100 seed wt ICRISAT is a member of the CGIAR Consortium
  14. 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
  15. A5 CMS system A5 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
  16. Cajanus cajan Cajanus lanceolatus Traits of interest transferred: A9 CMS system, pod borer and bruchid resistance INTER-SPECIFIC CROSS THAT IS FERTILE Introgression of traits
  17. 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.
  18. 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
  19. Helicoverpa resistant Cajanus platycarpus
  20. 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 techniques BC2 [87.5%A+12.5%B] Screen for Helicoverpa ↓ [ May require growth regulator applications, embryo rescue techniques] Interspecific hybridization
  21. 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
  22. Molecular analysis DArT 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.
  23. Cajanus platycarpus derivatives are showing variation for plant growth, height, seed weight, disease and pest resistance C. platycarpus seeds
  24. Fig. A total of 166 C. platycarpus and 301C. 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
  25. A7 CMS system A7 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
  26. Meiosis Premeiotic Postmeiotic
  27. 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.
  28. 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
  29. Bruchid resistance 0 10 20 30 40 50 60 70 80 90 ICPW- 130 ICPW-66 ICPW-94 ICPW- 15671 ICPW-64 ICP- 15613 ICPW-68 Controls 85010 (Solidbars(%)seeddamage 0 10 20 30 40 50 60 70 80 90 Linegraphis(%)eggsunhatched Fig. 4. Bruchid damage in w ild relatives of pigeonpea 0 20 40 60 80 100 237-1 238-1 A4-10--7--1 A4-10--7--2 A4-10--7--4 A4-10--7--7 A4-10-7--19 A4-10-7--20 7018--40-26-7-1 Cont.ICPL- 85010 Solidbars(%)seeddamage 0 20 40 60 80 100 Smallbars(%)eggdensity& linegraphis(%)eggs unhatched Publication: Jadhav et al., 2012.
  30. 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**
  31. C. volubilis First plant: Hybrid Second plant: Cajanus cajan Third plant: C. volubilis
  32. SSR analysis of C. volubilis hybrid
  33. Dwarf plant type and early maturity in F2 onwards Srikanth et al., manuscript under preparation
  34. Gene pools of pigeonpea Primary Gene Pool Cajanus cajan and its land races. P S T Q Secondary Gene Pool C. cajanifolius, C. lineatus, C. lanceolatus, C. laticepalus, C. albicans, C. reticulatus, C. sericeus, C. scarabaeoides, C . trinervius, C. acutifolius,. Tertiary Gene Pool C. 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 Pool Flemingia, Rhynchosia, Dunbaria, Erisema Paracalyx, Adenodolichos, Bolusafra, Carissoa, Chrysoscias, Baukea. Fig. 1. Taken from Mallikarjuna et al., 2011
  35. Quaternary gene pool Recent Success : Succeeded in crossing pigeonpea with Rhynchosia species and hybrids were obtained.
  36. Crossing pigeonpea with R. bracteata
  37. SSR marker showing polymorphism
  38. Genomic in-situ hybridization
  39. Summary • Species from secondary, tertiary and quarternary gene pool crossed successfully with pigeonpea
  40. Summary Diversification of CMS base CMS 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)
  41. 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.
  42. Legume Cell Biology Staff • Deepak R Jadhav • V. Balakrishna • S. Satyanarayaya • K. Avinash • K. Lakshmi • A. Amrutha and the students Thank you Cell Biology staff for your help and understanding and sharing the passion to pre- breed ICRISAT mandate legumes.
  43. End of the pigeonpea story from me!! Would you like to listen to the grandoise groundnut ground-breaking research??
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