Transgenic Brassica campestris Plants Expressing the gfp Gene

A protocol has been worked out for efficient regeneration and genetic transformation of summer rape (Brassica campestris L. var. oleifera). Cotyledons of the 5-day-old seedlings were transformed with Agrobacterium tumifaciens strain AGL cells comprising a binary vector with a selectable neomycin phosphotransferase II gene and a reporter gene encoding green fluorescent protein (GFP). Explants were cultured on a regeneration MS medium supplemented with ABA, and transformants were selected on the same medium (minus ABA and plus 70 mM AgNO3 and 15 mg/l kanamycin). The frequency of shoot regeneration on explant petioles was 30–40%. Transgenic plants were identified by GFP fluorescence and by polymerase chain reaction and Western blotting analysis. The transformation efficiency was as high as 75% of the total number of regenerated shoots.