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Agrobacterium-mediated transformation of Arabis gunnisoniana

Author:
Taskin, K.M., Turgut, K., Ercan, A.G., Scott, R.J.
Source:
Plant cell, tissue, and organ culture 2003 v.72 no.2 pp. 173-179
ISSN:
0167-6857
Subject:
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
Abstract:
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.
Agid:
673112