Agrobacterium-mediated transformation of hybrid poplar (Populus alba x P. grandidentata) and analysis of foreign gene expression Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/vm40xv32m

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  • A method for Agrobacterium-mediated transformation of hybrid poplar (Populus alba x P. grandidentata) suspension cultures and regeneration of transformed plants is described. The best protocol was one in which suspension cultures were inoculated with Agrobacterium tumefaciens to a density of 10⁷ cfu's/ml, cocultivated for 48 hours, plated to cellulose acetate filters at a density of 14 colonies/mm², and cultured on medium containing 1 mg/1 2,4-D. Although cefotaxime inhibited callus growth, it was used in the plating medium to suppress proliferation of Agrobacterium. Selection appeared to be more reliable using hygromycin as compared to kanamycin or geneticin (G418). Transgenic plants were regenerated by culturing the calli on media containing thidiazuron, but no shoots could be regenerated using BA.
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