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Agrobacterium-Mediated Transformation of Cry8db Gene in Vietnam Sweet Potato Cultivar

Journal of Life Sciences 2015年第6期9卷 262-271页

作者： Pham Bich Ngoc Vu Thi Lan Tran Thu Trang Nguyen Hoai Thuong Le Thu Ngoc Chu Hoang Ha Le Tran Binh Institute of Biotechnology Vietnam Academy of Science and Technology 18-Hoang Quoc Viet Cau Giay Ha Noi 10000 Vietnam College of Science Thai Nguyen University Tan Thinh Thai Nguyen 23000 Vietnam University of Science and Technology of Ha Noi 18-Hoang Quoc Viet Cau Giay Ha Noi 10000 Vietnam

Sweet potato [Ipomoea batatas （L.） Lam.] is an important food crop in the world as well as in Vietnam. It is well known as a recalcitrant crop for gene transformation and tissue culture because of its genotype dependent in vitro responses. In present study, Agrobacterium-mediated transformation of cry8Db from Bacillus thuringiensis into KB 1 sweet potato variety has been studied. The C58cv strain carrying a pBl 121 backbone which contained cry8Db delta-endotoxin gene regulated under 35 S CaMV prom oter, and the selection marker gene, neomycin phosphotransferase （npt11） gene, was subjected for plant transformation. Callus induced from shoot tips and leaf explants were inoculated and cocultured with A. tumefaciens. The selection occurred during callus producing and plant regenerating steps. A total of 201 transgenic putative plant lines were produced, and 21 transgenic lines were positively confirmed by PCR and finalized by Southern blot. Four putative transgenic lines confirming a single copy of the crySEIb gene were transferred into soil pots in greenhouse. Biological activity evaluation for the insecticidal capacity of these transgenic lines under controlled conditions showed that the level of infestation by sweet potato weevils （cylasformicarius. in untransformed plants was higher than that of transgenic lines.

关键词： Sweet potato Agrobacterium tumefaciens plant transformation cry8Db sweet potato weevil resistance cylas formicarius.

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