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Genomics for converting 3-lines to 2-lines hybrid system in pigeonpea By Dr Lekha T Pazhamala, ICRISAT

  1. Genomics for converting 3-lines to 2-lines hybrid system in pigeonpea Lekha T Pazhamala DST Women Scientist
  2. Pigeonpea (Cajanus cajan) ICPH 2671 Local check Food legume with diversified uses such as food, feed, fodder and fuel India : Largest producer, consumer, importer Commodity crop Myanmar, Tanzania, Kenya, Malawi, Uganda, Mozambique Pigeonpea hybrids World’s first commercial legume hybrid system Hybrids yielding upto 40% over varieties
  3. Exploring two-lines hybrid system Environment-sensitive male sterile line Restorer lineCytoplasmic male sterile line Restorer line
  4. 8 Environment-Sensitive Male Sterile (ESMS) lines Further evaluation in two years in fields Phenotypic studies Transcriptomic studies What is happening at the cellular level? What genes are involved in fertility reversion? What factors cause fertility reversion? Three lines evaluated under controlled conditions Cytological studies Evaluation & characterization of ESMS lines • Photoperiod • Day temperature • Night temperature • Light intensity • Humidity
  5. Key environmental factor: Day temperature 30°C day 16°C night 13 h day length 11 h day length 8 h day length Sterile Photoperiod Night temperature Light intensity Humidity 30°C day 13h day length 16°C night 10°C night 8°C night 12°C night Sterile 12°C night 11h day length 30°C day 25°C day 18°C day Sterile Fertile
  6. Critical day temperature: 24 °C 0 10 20 30 40 50 60 70 80 90 100 107/1 107/2 107/3 107/4 <24°C Sterility% > 24° C a Critical threshold temperature 30°C 29°C 28°C 27°C 26°C 25°C 24°C 23°C 22°C 21°C 20°C 19°C 18°C SterileFertile Plant 1 Plant 2 Plant 3 Plant 4
  7. Critical stages: Tetrad & Microspore
  8. Genes involved in fertility reversion RNA Sequencing 337.93 million reads 283.84 million reads Fertile Sterile
  9. A systems biology approach Transcriptome analysis Metabolic pathways analysis F1 F2 F3 F4 F5 S1 S2 S3 S4 S5 F1 0 37 340 464 476 6 14 141 283 445 F2 23 0 197 338 396 28 39 79 213 416 F3 71 35 0 5 100 103 81 38 45 182 F4 94 78 3 0 76 115 112 44 9 103 F5 59 46 27 46 0 75 79 50 30 10 S1 9 84 356 470 496 0 11 158 301 498 S2 17 61 392 517 516 11 0 138 322 521 S3 38 12 91 186 338 45 26 0 70 369 S4 79 76 19 10 89 98 89 11 0 105 S5 92 93 88 107 5 97 104 65 45 0 Gene ontology analysis
  10. Role of auxin in reverting sterility With Auxin treatment Without Auxin treatment
  11. Possible molecular mechanism >24 °C <24 °C
  12. Summary • Molecular mechanisms for fertility reversion deduced • A stable thermo-sensitive male sterile line identified • 24 °C as a threshold day temperature • Tetrad and microspore as critical stages • Application of auxin in reverting sterility validated • This breakthrough research will pave the way for two line hybrid breeding system Next steps • Evaluation of identified line in targeted temperature zones for deployment in two-lines hybrid breeding system • Candidate genes/genomic region responsible for fertility reversion
  13. Rajeev Varshney Rachit Saxena Anu Chitikineni Prasad Bajaj KB Saxena CV Sameer Kumar Anupama Hingane Acknowledgements Blue Sky Research Initiative from ICRISAT Thank You
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