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

Abstract

In order to generate transgenic radish (Raphanus sativus L., cv. Jin Ju Dae Pyong), hypocotyl explants were cultured on Murashige and Skoog medium containing 4mg l-1 AgNO3, 5mg l-1 acetosyringone, 4mg l-1 6-benzyladenine, and 3mg l-1 α-naphthaleneacetic acid in addition to either 10 mg l-1 hygromycin or 100 mg l-1 paromomycin after cocultivation 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.