豇豆胰蛋白酶抑制剂基因转化花椰菜的研究

Transformation of cauliflower (Brassica oleracea var.botrytis)with cowpea trypsin inhibitor gene

通过根癌农杆菌 Agrobacterium tumefaciens介导,将豇豆胰蛋白酶抑制剂(CpTI)基因导入花椰菜无菌苗的下胚轴和子叶中,卡那霉素(Kan)的筛选质量浓度为15 mg/L,抑制农杆菌生长的抗生素选用羧苄青霉素(carbencillin),质量浓度为500mg/L.对所获得的转基因植株进行PCR扩增,结果显示大多数为阳;PCR-Southern及Southern分子检测分析,结果证明CpTI基因已被整合到花椰菜植株的基因组中.转基因植株叶片的离体饲虫初步试验结果表明,对鳞翅目害虫菜青虫的生长发育有一定的抑制作用.

The cowpea trypsin inhibitor (CpTI) gene was transferred into the cotyledons and hypocotyls of cauliflower mediated by Agrobacterium tumefaciens. In the selective medium, the kanamycin concentration was 15 mg/L. Carbencillin (500 mg/L) was used to inhibit growth of agrobacterium. The regenerated transformant plants were assayed by PCR, PCR-Southern blot and Southern blot, and the target band was observed in most of the plants assayed. So the integration of the CpTI gene into cauliflower genome DNA was confirmed. In vitro leaf testing for evaluating resistance to cabbage worm (Pieris brassicae L.) was made, and the results showed that the transgenic plants were more resistant than non-transgenic plants.