苏云金芽孢杆菌cry1Aα核心毒素基因转化烟草及其杀虫活性  

Transformed tobacco with Bacillus thuringiensis cry1Aα active toxin gene and its entomocidal activity

作　　者：韩蓓[1,2] 袁志明[1] 陈士云[1] 

机构地区：[1]中国科学院武汉病毒研究所,武汉430071 [2]中国科学院研究生院,北京100039

出　　处：《高技术通讯》2005年第9期62-66,共5页Chinese High Technology Letters

基　　金：国家自然科学基金（39770170）资助项目.

摘　　要：将苏云金芽孢杆菌cry1Aα的C端编码区截短，获得编码N端609个氨基酸约1.8kb的核心毒素基因，将其构建到植物表达载体转化烟草，研究直接表达的活性ICP核心毒素被昆虫取食后的杀虫效果。T1代种子发芽的抗生素抗性分离和PCR检测证实cry1Aα核心毒素基因整合到了烟草基因组，Western杂交检测表明转化烟草能够正常表达大小为75．6kD的核心毒素蛋白。生物测定结果表明，转化烟草对初孵棉铃虫平均有高于60％的致死率，对初孵甜菜夜蛾平均达到70％的致死率，最高致死率均可达到90％以上，并对昆虫的生长发育有明显的抑制作用。The N terminal truncated sequence encoding 609 amino acids of the active toxin of the cry 1Aα gene from Bacillus thuringiensis was cloned by PCR, and was used for tobacco transformation. Twenty independent transgenic tobacco lines with hyromycin resistance were obtained via Agrobacterium-mediated transformation. T1 progeny seeds were germinated on medium with the segregation of hygromycin resistance and further analyzed by PCR to confirm the integration of the transgenes into tobacco genome. Western blotting analysis of T1 plants indicated that the cry 1Aα active protein with the size of 75.6kD was produced in transgenic tobacco leaf tissues. The entomocidal bioassay with the neonate larvae of exigua and Helicoverpa armigera showed that the transformed plants reached more than 60 % average mortality against Helicoverpa armigera larvae, and 70 % average mortality against Spodoptera exigua larvae, with the highest mortality rate of 90%. The transgenic plants also showed inhibition to the growth and development of neonate larvae of both insects.

关 键 词：苏云金芽孢杆菌 cry1Aα 核心毒素 烟草 杀虫活性 基因转化 毒素基因 Western 植物表达载体 PCR检测 抗生素抗性 

分 类 号：S476.11[农业科学—农业昆虫与害虫防治] S511[农业科学—植物保护]