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Agrobacterium-mediated transformation of Arabis gunnisoniana

Taskin, K.M., Turgut, K., Ercan, A.G., Scott, R.J.
Plant cell, tissue, and organ culture 2003 v.72 no.2 pp. 173-179
benzyladenine, micropropagation, 2,4-D, callus, shoots, dose response, hypocotyls, Agrobacterium radiobacter, explants, Arabis, apomixis, naphthaleneacetic acid, in vitro regeneration, culture media, genetic transformation, gene transfer, Boechera stricta, kanamycin, cultured cells
An efficient method for adventitious shoot regeneration for Arabis drummondii and a transformation protocol for A. gunnisoniana from hypocotyl explants are described. Hypocotyl explants from 7-day-old aseptically grown seedlings were cultured on MS medium containing plant growth regulators (6-benzylaminopurine, 1-phelyl-3-(1,2,3-thiadiazol-5-yl) urea, α-naphthaleneacetic acid and 2,4-dichlorophenoxy-acetic acid). After 4 weeks in culture, high frequency of adventitious shoot regeneration was observed. Regenerated shoots were rooted on half-strength MS basal medium supplemented 1% (w/v) sucrose, with or without NAA. This protocol was then used to produce transformed Arabis gunnisoniana plants. A. gunnisoniana hypocotyl explants were co-cultivated with Agrobacterium tumefaciens strain GV3101 harbouring pBJ40. Transgenic shoots were selected on MS medium supplemented with 50 mg l-1 kanamycin. PCR analysis verified the presence of the nptII gene in the plant DNA isolated from kanamycin resistant shoots.