High-frequency non-selective transformation method in Brassica napus for obtaining marker-free plants
Non-selective transformation was investigated as a method for obtaining marker-free transgenic plants. The hypocotyl explants of Rapeseed (Brassica napus cv Westar) were co-cultivated with a general Agrobacterium tumefaciens strain LBA4404 containing a pBI121-NPTII plasmid in which there is only GUS gene as an interesting gene in the T-DNA and the NPTII gene is deleted. Co-cultivation was carried out under the condition of no culture medium, which mimics natural process of Agrobacterium tumefaciens infecting plants and greatly improves the transformation frequency. According to the X-Gluc histochemical staining,GUS was expressed in 63-89％ of the harvested callus and 27-51％ of the harvested plants. Furthermore, this transformed nature was confirmed by PCR and Southern blot analysis. These results suggest that not only the isolation of transformants without a selective marker gene is feasible, but high-efficiency could be reached. This method may provide a simple and useful experimental tool to obtain marker-free transgenic plants in rapeseed.
Non-selective transformation Marker-free transgenic plants Brassica napus Agrobacterium tumefaciens
WANG Jingqiao CHEN Wei LI Jingfong ZHANG Guojian DONG Yunsong LI Genze
Institute of Cash Crops, Yunan Academy of Agricultural Sciences, Yunnan Kummimg P.R.China 650205 Vocational-technical College of Agriculture in Yunnan, Yunnan Kummimg P.R.China 650031
国际会议
第十二届国际油菜大会( The 12th International Rapeseed Congress)
武汉
英文
487-491
2007-03-26（万方平台首次上网日期，不代表论文的发表时间）