Golden bananas for Africa D/Prof James Dale Centre for Tropical Crops and Biocommodities, Queensland University of Technology, Brisbane, Australia & National Agricultural Research Organisation, Kampala, Uganda
The Top 10 Food Crops Centre for Tropical Crops and Biocommodities Ranking Crop Production mt Also animal feed 1 Corn/maize 823 +++ 2 Wheat 690 + 3 Rice 685-4 Potatoes 314-5 Cassava 232-6 Soybeans 230 +++ 7 Banana/plantain 122-8 Sweet potato 110-9 Sorghum 65 +++ 10 Yams 51 -
Worldwide production Top 10 countries Production (million tonnes) Population (million) India 27.6 1333 China 12.4 1384 Uganda 9.5 40 Philippines 8.6 102 Brazil 6.9 210 Ecuador 6.0 16 Colombia 5.4 48 Indonesia 5.4 261 Cameroon 5.2 24 FAOSTAT, 2013 data including plantains
The problem for bananas The vast majority of domesticated bananas are selections from the wild, not bred Most cultivated bananas are triploids but there still exist edible diploids Domesticated bananas are very low male and female fertility: essentially sterile
Micronutrient deficiencies o third most important public health problem worldwide after HIV/AIDS and malaria (WHO) o disproportionately affects developing countries o vitamin A, iron, zinc, iodine and folic acid
Vitamin A deficiency 670,000 kids die of VAD every year Another 350,000 go permanently blind VAD: mortality, blindness, night blindness, impaired immunity system, impaired brain development
Human Nutrition Challenges in Uganda Micronutrient undernutrition The major micronutrient deficiencies in Uganda are: Iron deficiency anaemia (IDA) Vitamin A deficiency (VAD) Demographic and Health Survey 2006 15-32% of children < 5 years had VAD 13-31% of women had VAD 50-80% of children < 5 years had IDA 32-64% of women had IDA Uganda banana consumption > 0.5kg/person/day Centre for Tropical Crops and Biocommodities
Biofortification of staple crops Grand Challenge in Global Health No.9: Create a full range of optimal, bioavailable micronutrients in a single staple plant species: 2005 Banana21 BioCassava Plus Super Sorghum Golden Rice Centre for Tropical Crops and Biocommodities
The project To alleviate vitamin A deficiency and iron deficiency anaemia in Uganda through the enhancement of micronutrients in the staple food of Uganda, bananas
QUT Australia Cavendish Genes NARO Uganda EAHB M9 Transformation Analysis Continuous technology transfer Transformation Analysis Field Trials Pro-vit A Iron Field trials Feeding trials Stewardship Pro-vit A Iron Made in Uganda
Our banana biofortification target Estimated Average Requirement (EAR) Bioconversion of -carotene to retinol 20 g/g dwt -carotene equivalents Daily consumption (children v adults) Processing losses (cooking: steaming)
Genetic modification of banana Timeline from explant to harvest Banana flower Harvesting Banana flower explants Embryogenic cell suspensions Rooted plantlets Multiplied & acclimatised First harvest 9 months 12 months 6 months 12 months 6 months Transformation Planted in field Harvesting Months 9 21 27 39
GM field trials in Australia Two field trials (AFT1 and AFT2) Started in 2009 48 combinations of transgenes and promoters 14 plantings with close to 2000 lines Banana varieties Dwarf Cavendish Centre for Tropical Crops and Biocommodities
A selection of transgenes and promoters ZmPsy1 = phyotene synthase gene from maize MtPsy2a = phyotene synthase gene from Asupina (banana) CrtI = CrtI gene from Erwinia ACO = ACC oxidase promoter from banana (fruit expressing) Ubi = polyubiquitin promoter from maize (constitutive)
The Gold Standard Golden Rice 2 Centre for Tropical Crops and Biocommodities ZmPsy1 Maize B73 CrtI
Carotenoid content of selected ripe raw banana cultivars (μg/g dwt edible portion) Genotype α-carotene (ug/g dwt) β-carotene (ug/g dwt) β-carotene equivalents (ug/g dwt) Asupina (PNG) 41.2 117 137.6 Cavendish 1.4 1.0 1.7 Lady finger 0.8 0.4 0.8 East African Highland 10.2 6.2 11.3 Hybrid M9 7.3 2.2 5.9 Plantain 10.9 24.2 29.5 Target: 20 g/g dry weight -carotene equivalents Centre for Tropical Crops and Biocommodities
Golden bananas Centre for Tropical Crops and Biocommodities MtPsy2a Asupina
β-carotene equivalents (µg/g DW) β-carotene equivalents (µg/g DW) Centre for Tropical Crops and Biocommodities ACO-ZmPsy1 PC-FG PC-FR SC-FG SC-FR RC1-FG RC1-FR RC2-FG RC2-FR 50 40 30 20 10 0 FT467 FT475 FT479 FT483 ACO-MtPsy2a PC-FG PC-FR SC-FG SC-FR RC1-FG RC1-FR RC2-FG RC2-FR 50 40 30 20 10 0 FT497 FT504 FT508 FT511 FT518
β-carotene equivalents (µg/g DW) β-carotene equivalents (µg/g DW) Centre for Tropical Crops and Biocommodities ACO-MtPsy2a PC-FG PC-FR SC-FG SC-FR RC1-FG RC1-FR RC2-FG RC2-FR 50 40 30 20 10 0 FT497 FT504 FT508 FT511 FT518 Ubi-MtPsy2a PC-FG PC-FR SC-FG SC-FR RC1-FG RC1-FR RC2-FG RC2-FR 90 80 70 60 50 40 30 20 10 0 FT294 FT295 FT324 FT330 Variation between crops and seasons
Selected individual lines in AFT-2 Line Construct βce range (μ/g dwt) βce average (μ/g dwt) 584 ACO>ZmPsy1 + Exp1>CrtI 14-44 27 504 ACO>MtPsy2a 18-23 20 246 Exp1>MtPsy2a 14-24 18 479 ACO>ZmPsy1 19-30 26 294 Ubi>MtPsy2a 26-31 29 324 Ubi>MtPsy2a 40-74 55 162 Wildtype 4-5 5
Elevated provitamin A Wildtype Cavendish High PVA Cavendish Line with gene from banana
The project in Uganda o Two constructs transferred to NARO: ACO>MtPsy2a and Ubi>MtPsy2a o Transformed into two cultivars - Nakitembe (an EAHB clone) - M9 (a conventionally bred cooking banana with high disease resistance) Centre for Tropical Crops and Biocommodities
M9 fruit phenotype 41.86 µg/gdw, 16Kg; Ubi>MtPsy2a 5.4 µg/g DW; 20 Kg
FT12364 FT12350 Centre for Tropical Crops and Biocommodities Nakitembe top lines 71 µg/g DW; 12.5 Kg ; ACO>MtPSY2a WT 84.86 µg/g DW; 11 Kg, Ubi>MtPsy2a
Timelines 2005-2009 2010-2012 2013-2017 2018-2021 Phase 1 Phase 2 Phase 3 Phase 4 QUT: Australia Gene and promoter discovery; transformations First field trial: assessment Sucker field trial; transgene expression and stability; molecular characterisation Molecular characterisation; biosafety; deregulation documentation NARO: Uganda Transformation: model Transformation: East African Highland banana and M9 Elite line selection field trial Multi-location regulatory field trial; deregulation submission
Technology Phenotype Nutrition The Product Bananas with high levels of PVA and iron released in Africa Stewardship Impact Communication
Some key points: Our target country in Africa is Uganda initially and then potentially, Kenya, Rwanda, Tanzania, Burundi and Congo; India commenced August, 2012 The population in Africa in these countries is 125 million people Our planned release date is 2021 Collaboration, technology transfer and education are the key components
Our researchers and supporters James Dale Rob Harding JY Paul Bulukani Mlalazi Anthony James Jen Kleidon Jimmy Tindamanyire Wilberforce Tushemereirwe Jerome Kubiriba Stephen Buah Priver Namanya Moses Matuvo Sarah Nanyiti Doreen Amumpaire Joan Marunga James Kawuma Annette Magala Financial Support Jimmy Tindamanyire Field trials (in Australia) Jeff Daniels (DAF) Department for International Development Queensland university of Technology National Agricultural Research Organisation