An efficient Agrobacterium-mediated transformation method used in Chinese cabbage (Brassica rapa ssp. pekinensis)

摘要

An efficient method was developed for producing transgenic Chinese cabbage inbred lines Kenshin by co-culturing (Pqf1, Pqf2 and Pqf3). Vector pCAMBIA1301 which contains kanamycin and hygromycin resistance genes was used for transformation. The highest infection frequency was detected when cotyledonary explants were first infected by Agrobacterium for 15 min and subsequently co-cultivated with Agrobacterium for 2 days in co-cultivation medium supplemented with 50 mg/l acetosyringone. The media for shoot regeneration and selection contained 5.0 mg/l and 10.0 mg/l hygromycin, respectively. More than 60 putative transformants have been obtained from this method in Chinese cabbage inbred line Kenshin. The transformants were confirmed by GUS staining, PCR, Southern blot, Northern blot analysises and progeny test.