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Biotechnology,Research-for-Development at IITA

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Tissue Culture,Crop Genetic Transformation,Molecular marker applications,Capacity Building & partnerships

Tissue Culture,Crop Genetic Transformation,Molecular marker applications,Capacity Building & partnerships

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  • 1. BiotechnologyResearch-for-Development at IITA I Ingelbrecht IITA-Ibadan, 31 August 09
  • 2. Guiding principles Biotech complementary tool to conventional approaches Multidisciplinary approach (breeding, genetic resources, pathology, etc) Product oriented Partnerships: national research programs across SSA and advanced labs overseas Integrate capacity building with research
  • 3.  ~8 full time PhD scientists with additional scientists (breeding, virology, pathology, etc) Two main hubs: HQ in Nigeria + BECA, Kenya
  • 4. Biotechnology R-4-D ‘traditional’ ‘modern’ In vitro Tissue Culture Crop Genetic Molecular Marker Transformation Applications Capacity building
  • 5. I. In vitro Tissue Culture yam Banana, plantain cassava
  • 6. Activities In vitro micropropagation for distribution of elite cassava, yam and banana/plantain materials Disease clean up: production of certified, disease-free planting materials (with PQS) Training + technical backstopping of NARS in all aspects of in vitro micropropagation and set up of tissue culture facilities
  • 7. Impact  High demand by national programs across SSA  Technology well established: applications for breeding & genetic resource conservation (fairly) routine  Technology transferred to national programs, in some cases (high value crops) applied for commercial purposes In vitro tissue culture recently integrated with Genetic Resources Center at HQ, Ibadan, Nigeria
  • 8. II. Crop Genetic Transformation • GM crops developed and commercialized in industrialized as well as developing countries with demonstrated benefits for producers and environment • With proper identification of targets, GM technology can complement conventional breeding for intractable traits in SSA agriculture • For ‘orphan’ crops tools and products mostly in stage of development, some already available • Need to consider regulatory environment lacking in most countries in SSA; capacity building
  • 9. Ongoing projects Genetic transformation of cassava for resistance to the Cassava Brown Streak Disease, starch modification Genetic transformation of banana for bacterial blight resistance, DNA recombinant technology for construction of genes and analysis of transgenic plants Biosafety with application for CFT of GM crops, by national programs
  • 10. Example: cassava, bananaThree steps in genetic transformation protocol:1. In vitro shoot regeneration method2. Gene transfer method: Agrobacterium3. Selection and/or screening for transgenic shootsFocus on African genotypes
  • 11. Transformation of farmer-preferred cassavafor CBSD resistance Equatorial Guinea Uganda Kenya ROC DRC Tanzania Malawi Zambia Mozambique CBSD devastating CBSD damaging CBSD reported A transgenics approach for resistance to Potyviruses previously used and grown commercially: eg papaya resistant to Papaya ringspot virus
  • 12. Why target cassava landraces?Current transformation protocols are for ‘model’ genotypes, not usedby farmers or breeders in Africa: excellent research tool butlimited application in the fieldBottleneck since current protocols are genotype-dependent develop protocol for (African) farmer-preferred lines
  • 13. Summary cassava regeneration responses Explants; e.g. immature leaf lobes Embryogenesis +/- Shoot organogenesis Secondary SE * Primary SE ++++ ++++ Multiple shoots ++++ Embryogenesis ++ Cotyledonary Friable embryogenic stage SE * Callus * Adventitious shoots Embryos Protoplasts +/- Embryogenic +/- 0 - 40% suspensions Plantlets ++ 20 - 40%Fig. modified from Zhang et al. 2006 ++++ >90%
  • 14. Somatic embryogenesisSE produced for 9 genotypes: 4 landraces from ESA 4 landraces from WCA 1 IITA elite line cv Albert TME12
  • 15. Transgenic African cassava with reporter gene
  • 16. Transgenic African cassava with reporter gene a petiole 4 mm b Young stem 3 mm c Old stem 5 mm d Fibrous root 3 mm e tuber 1 cm Non transgenic Transgenic – 1st ratoon Transgenic – 2nd ratoon
  • 17. Status- Resistance genes developed and tested in N benthamiana: resistance is strain specific- Transformation of African cassava with resistance genes ongoing
  • 18. Banana transformation - status - Transgenic plants produced with resistance gene(s) for bacterial blight - Field trial planned for 2010 (Uganda)
  • 19. III. Molecular marker applicationsRationale• For characterization and management of germplasm and pathogens (diversity) and for diagnostics purposes• In marker assisted selection/breeder to accelerate breeding process; extensively used in cereals and other crops of developed world• As with GM technology, for ‘orphan’ crops tools are mostly in stage of development
  • 20. Ongoing projects Characterization of germplasm collections of cassava, yam, banana QTL mapping of virus (CMD & CBSD) resistance in cassava, drought tolerance in cowpea, maize and cassava MAS for striga resistance in cowpea Development of SSR and SNP markers for cassava & yam Diagnostic markers for bacterial blight + other pathogens Development of DNA microarray for gene discovery in cassava
  • 21. Example: EST-SSR marker development Total number of EST sequences investigated: 18,166 Number of unigenes used for in silico identification of SSRs: 8,577 Total number of unique SSR loci appropriate for primer modeling: 646 (3.3%) Number of candidate SSR investigated : 346 PCR successful: ~ 90% Failed PCR: ~ 10% PCR products with expected sizes Amplification of introns > 500 bp Eliminate Screen on diversity panel
  • 22. Example cassava- 3078 cassava accessions with minimal passport data held in trust in GRC DNA fingerprinting for effective management and use by breeding program
  • 23. New Cassava SSR markers M 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 1718 19 20 21 22 23 24 180 new SSR markersUsed for fingerprinting collection, mapping and variety identification
  • 24. Semi-automated genotyping using fluorescent labeling Different technologies can be used for diversity studies, QTL mapping
  • 25. Example: cassava microarray ~14,000 genes or 1/4 to 1/3 of all cassava genes
  • 26. - Hybridization and scanning- Identify differentially expressed genes for use in transgenic program or complement QTL mapping
  • 27. IV. Capacity Building & partnerships- Degree (MSc, PhD) and non-degree training of students or VS; mostly integrated with ongoing research activities; topical also possible if funded- Workshops: genetic transformation, molecular breeding, biosafety- Infrastructure: eg MARI, Tanzania for GM Advanced labs: DDPSC, USA; ETH, Switzerland; KU, Copenhagen, Univ Virginia, USA; others National research institutions and universities NRCRI, Nigeria; MARI, Tanzania, NABDA, Nigeria; many universities in Nigeria and other African countries for joint degree research.
  • 28. Development investors  USAID  RF  BMGF  Generation CP  AATF  Gatsby Charitable Foundation  Masterfoods

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