Agrobacterium-mediated transformation of Arabis gunnisoniana |
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Authors: | Taskin Kemal Melik Turgut Kenan Gulhan Ercan A Scott Rod J |
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Institution: | (1) Department of Field Crops, Faculty of Agriculture, University of Akdeniz, P.O. Box 510, Antalya, Turkey;(2) Department of Biology and Biochemistry, University of Bath, Bath, BA2 7AY, UK |
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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, -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 21 medium supplemented with 50 mg l kanamycin. PCR analysis verified the presence of the nptII gene in the plant DNA isolated from kanamycin resistant shoots. |
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