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Ye, X; S. Al-Babili, A. Kloti, J. Zhang, P. Lucca, P. Beyer, and I. Potrykus. 2000. Engineering the pro-vitamin A (beta-carotene) biosynthetic pathway into (carotenoid-free) rice endosperm. Science 287 (5451): 303. Presented by Igy Martinez. VITAMIN A DEFICIENCY.
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Ye, X; S. Al-Babili, A. Kloti, J. Zhang, P. Lucca, P. Beyer, and I. Potrykus. 2000. Engineering the pro-vitamin A (beta-carotene) biosynthetic pathway into (carotenoid-free) rice endosperm. Science 287 (5451): 303. Presented by Igy Martinez
VITAMIN A DEFICIENCY • 124 million children affected worldwide • 1 to 2 million deaths per year among children • Major cause of seeing disorders • Other afflictions • Prevalent in 26 countries where rice is a major food staple
RICE (ORYZA SATIVA) • Primary food source in over ½ the world population • Poor source of essential nutrients • Vitamin A deficient • Synthesizes GGPP
Rational • Combat malnutrition • Find alternatives to deliver vitamin A in underdeveloped countries • Supplement rice endosperm with vitamin A
Three rice groups tested Control group with no transformation Single group that underwent one transformation carrying a combination of three vectors Double group that underwent two transformations each carrying one vector Vector Sequencing Three vectors in single transformation group combined sequencing for phytoene synthase from daffodil and phytoene desaturase from Erwinia bacterium One vector in the co-transformation group sequenced phytoene synthase and phytoene desaturase while the second vector sequenced lycopene beta cyclase Recombinant DNA Technology
“Golden Rice” • Single transformant line revealed 3:1 colored/uncolored ratio. • Co-transformant line were all variably colored. • Vitamin A synthesized in both lines. • Transgenic rice plants showed normal vegetation and were fertile.
HPLC Results • Both transformed lines detected beta carotene, lutein, and zeaxanthin • None of the transformed lines detected lycopene beta cyclase
Relevance • GGPP in rice endosperm can be successfully converted to pro-vitamin A. • The detection of pro-vitamin A in both transformed groups indicates that the enzymes necessary to convert lycopene into pro-vitamin A are already expressed in normal rice endosperm.