FEASIBILITY OF HYGROMYCIN AS A SELECTION AGENT IN AGROBACTERIUM-MEDIATED TRANSFORMATION OF OILSEED RAPE (BRASSICA NAPUS L.)

Abstract

In this work the feasibility of the antibiotic hygromycin as a selection agent in Agrobacterium-mediated transformation of oilseed rape (Brassica napus L.) was evaluated. For this, two economically important commercial varieties Haydn and Hunter and tobacco as a model plant were subjected to Agrobacterium-mediated transformation. The 5-6 days-old oilseed rape hypocotyls and 4-6 weeks-old tobacco leaf segments were transformed with the binary vector pCambia1304. The T-DNA contained the reporter gfp:gus and the selectable marker htp genes. Regeneration of transformed cells was conducted under selection of 10 mg.l-1 (oilseed rape) and 30 mg.l-1 (tobacco) hygromycin. Putative transgenic plantlets were analysed by the mean of the histochemical GUS and PCR analyses. Transformation efficiency ranged from 1.0% (cv. Haydn) to 40.4% (tobacco). No transgenic shoots were detected for the cv. Hunter. It points out the oilseed rape cultivar specificity plays significant role in choice of suitable selection agent.