Agrobacterium-mediated genetic transformation of radish (Raphanus sativus L.)

概要

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In order to generate transgenic radish (Raphanus sativus L., cv. Jin Ju Dae Pyong), hypocotyl explants were cultured on Murashige and Skoog medium containing 4 mg l−1 AgNO3, 5 mg l−1 acetosyringone, 4 mg l−1 6-benzyladenine, and 3 mg l−1 α-naphthaleneacetic acid in addition to either 10 mg l−1 hygromycin or 100 mg l−1 paromomycin after co-cultivation with disarmed Agrobacterium tumefaciens harboring a plant expression binary vector. Explants co-cultivated with A. tumefaciens GV3101 harboring pCAMBIA1301 and A. tumefaciens EHA101 harboring pPTN290 produced putative transgenic adventitious shoots at frequencies of 0.26% and 0.18%, respectively. Northern blot analysis revealed the gus gene transcript was detected in 8 regenerated plants which confirmed their genetic transformation. The transgenic plants were grown to maturity after vernalization in a greenhouse and appeared morphologically normal. Progeny analysis of independent transgenic plants demonstrated that the gus gene was transmitted in a Mendelian pattern in 3 lines, indicating a single copied gene was incorporated into the genome.
日本植物細胞分子生物学会の論文
2008-03-01