Regeneration of transgenic kale ( Brassica oleracea var. acephala), rape ( B. napus) and turnip ( B. campestris var. rapifera) plants via Agrobacterium rhizogenes mediated transformation

Abstract

Transgenic hairy root lines were developed for six Brassica cultivars: B. napus cv. Giant (rape), B. oleracea cvs. Kapeti, Midas, Rawara and Medium Stem (all kales) and B. campstris cv. Red Globe (turnip). Leaf explants or petioles of intact cotyledons were cocultivated with Agrobacterium rhizogenes strain A4T harbouring the binary vector pKIWI110, the T-DNA of which contains genes encoding β-glucuronidase (GUS), neomycin phosphotransferase II (NPTII) and chlorsulfuron resistance. Hairy root lines grew prolifically on hormone-free medium containing selective levels of kanamycin (50 or 100 mg/l) or chlorsulfuron (10 μg/l). Transgenic shoots were regenerated from hairy roots of all cultivars and plants have been successfully transferred to soil. Molecular analyses using polymerase chain reaction (PCR) and Southern blotting confirmed the trnasgenic nature of these plants.