Biofortification of highland bananas in Uganda - November 2012

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Presentation by Geoffrey Arinaitwe …

Presentation by Geoffrey Arinaitwe
Delivered at the B4FA Media Dialogue Workshop, Kampala, Uganda - November 2012
www.b4fa.org

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  • 1. Genetic modification of highland bananas: biofortification with PVA and iron G. Arinaitwe, H. Khana, P. Namanya, . Kiggundu, A W.K Tushemereirwe and J. Dale Research Framework National Research Laboratories - Kawanda
  • 2. Introduction Uganda has a population of 34 M people & is projected to be 53 M by 2025 Problems: Inadequate food quantities Imbalanced nutrients, containing mainly carbohydrates
  • 3. Banana is synonymous to food A B C
  • 4. Bananas in Uganda
  • 5. Banana weevils
  • 6. Nematode effect
  • 7. Banana bacterial wilt
  • 8. Black Sigatoka disease
  • 9. Micronutrient deficiencies in Uganda Micronutrient undernutrition The major micronutrient deficiencies in Uganda include: Vitamin A deficiency (VAD) Iron deficiency Anaemia (IDA) Iodine Deficiency Disorders (IDD) National Nutritional Surveys 1994-1999 31 out of 36 districts had VAD 50% of children have VAD 40% of children > 5 years have IDA 50% of pregnant women had IDA 30% of maternal deaths were attributable to IDA
  • 10. Sources of genes for biofortification psyB73 psy psy2 Psy2a crt I
  • 11. Constructs used in biofortification 1st Batch 2nd Batch (Opt series) 3rd Batch (GEN series) E1-ZmPsy1 E1-APSy2a E1-ZmPsy1Q60 E1-Crt1 Acs-ZmPsy1Q60+E1-S.Fer E1-APsy2a E1-APsy2a Acs-ZmPsy1Q60 BT4act-APsy2a E1-S.Fer E1-ZmPsy1Q60 Ubi-APsy2a 35S-Fro2 E1-ZmPsy1 Ubi-ced9 35S-IrtI Ubi-mced9 Ubi-AtBag4 Ubi-Gus E1-Gus
  • 12. Biofortified product for low and highlands Cultivars: ‘Nakinyika’ (EA-AAA) And ‘Sukali Ndizi’ (ABB) Cultivars: Hybrid M09 Tolerant to: Black Sigatoka, Nematodes and weevils
  • 13. ECSs work at Kawanda, Uganda 1. Male bud Genetic transformation 2. Inoculation stage 3. Embryogenic callus 4. Cell in suspension 5. Somatic embryos formed from suspensions 6. Embryo germination 7. Multiple shoot formation and rooting 8. Weaning and potting of plantlets
  • 14. Materials and methods Male bud Immature flower Embryogenic callus ECSs ECS initiation and transformation ECS infection Shoots from EC clones Selection Future activities Morphological analyses PCR analyses Southern blot analyses NARO-KARI Biotechnology Laboratory
  • 15. Transformation and selection Shoot regeneration improved 2 Weeks 1.5 Months 2.5 Months Shoot regeneration Control: 0mg GA3 0.5 mg/L GA3 1.0 mg/L GA3 1.5mg/L GA3
  • 16. Weaning and hardening
  • 17. First CFT at NARO, Uganda
  • 18. CFTs at NARO-Kawanda Several CFTs of transgenic lines containing disease resistance genes and micronutrient traits
  • 19. Phenotypes observed
  • 20. Carotenoid content at FG during storage BCE (µg/g DW) Cultivar %Water content Day1 Day7 Day 14 Mbwazirume 76.3 9.8 17.5 23.5 Nakinyika 73.9 76.9 75.1 74.5 58.2 65.2 13.5 19.6 9.1 4.3 18.2 0.8 21.0 23.4 18.0 11.0 22.3 1.7 26.5 28.3 18.8 16.8 23.1 2.2 Nakitembe Kisansa M9 Butobe Kayinja NAKINYIKA NAKITEMBE HYBRID M9 KAYINJA
  • 21. High carotenoid content in transgenic S. Ndizi 1 3 2 Control line 4 ZmPsy1 (12-39) line Cross section of the control and the ZmPsy1 (12-39) line; above: Chromatogram peaks for 12-39 fruit extract identified at λmax=450nm, peak 1, lutein at 6min; peak 2, α-carotene at 12.2; peak 3, trans-β-carotene 13.1min & peak4, cis β-carotene at 13.9min
  • 22. PVA retention in target cooking bananas Steamed Boiled
  • 23. FOC symptoms of the disease Microconidia entry to the corm is through wounded /cut root hairs Yellow leaves The conidia spreads up the xylem and blocks water and nutrient movement Discoloured pseudostem Death of plant releases the dormant chlamydospores back into the soil Reinfection is through running Discoloured water and propagating suckers corm
  • 24. FOC wilt in cultivar Sukali Ndizi ND control ND control ND 1 ND 1 ND 2 ND 2 ND 3 ND 3 ND 4 ND 4
  • 25. 12 12 55 72 83 NTC
  • 26. Acknowledgements (Program Leader) Dr. Wilberforce Tushemereirwe QUT NARO Rob Harding Doug Becker Marion Bateson Jason Geijskes Harjeet Khanna Anthony James Jen Kleidon Bulu Malazi A. Kiggundu G. Arinaitwe P. Lamwaka B. Magambo R. Ecodu P.Namanya Prof James Dale