Expression of nptII marker and gus reporter genes and their inheritance in subsequent generations of transgenic Brassica developed through Agrobacterium-mediated gene transfer

摘要

NptII marker and gus reporter genes were introduced in Brassica juncea through Agrobacterium-mediated transformation showing a transformation frequency of 3-5% in the best responsive medium. Seventy-one per cent regeneration from hypocotyl explants was obtained on MS medium containing 0.01 mg/l 2,4-D and 2.0 mg/l BA supplemented with 20 #mu#M AgNO_3, and 0.7% agarose as the gelling agent. The presence of transgenes (ntpII and gus) ion T_0, T_1 and T_2 generations was confirmed by dot blot analysis, using nptII gene as the probe, and by histochemical assay for the gus gene, respectively. Mendelian inheritance of the transgene (nptII) was observed in the T_1 and T_2 generations. Both the marker and reporter genes co-segregated in the T_1 and T_2 generations. The kanamycin-resistant plants in the T_1 progeny were either homozygous or heterozygous for the transgene.