硼酸诱导的大豆子叶节转化新方法

Boric Acid Mediated Transformation of Soybean(Glycine max) Cotyledonary Node

采用农杆菌介导法,以苏云金芽孢杆菌(Bacillus thuringiensis)毒性蛋白基因B t为目的基因,研究了不同硼酸浓度处理对大豆子叶节转基因的影响。结果表明:200 mg·L-1硼酸浓度诱导培养基诱导的大豆丛生芽长势最好,而且转基因效率高达3.3%。对获得大豆转化幼苗的根和叶片采用PCR、逆转录PCR和Southern blotting等方法进行筛选与鉴定,结果均有2株幼苗呈阳性。

The agrobacterium-mediated transgenic technique combined with the treatment of boric acid was studied for the transformation of foreign gene Bt to soybean cotyledon nodes.The results showed that shoots regenerated from the explants were multiplied best at 200 mg·L^-1 of boric acid concentration,and the transformation ratio reached 3.3%.Regenerated seedlings were screened by genomic PCR amplification and two seedlings on inductive medium of 200 mg·L-1 boron acid was obtained and proved to be positive by reverse PCR and Southern blotting.It was demonstrated that the target gene had been transformed into the soybean genome.