Bt杀虫基因向水稻内生细菌的转化研究  

Insecticidal Activity of Bacillus thuringiensis Expressed in Endophytic Bacteria of Rice

作　　者：刘云霞[1] 张青文[1] 周明 

机构地区：[1]中国农业大学植保系

出　　处：《农业生物技术学报》1997年第2期188-193,共6页Journal of Agricultural Biotechnology

摘　　要：本研究成功地构建以水稻体内定殖的优势细菌巨大芽孢杆菌为载体菌的工程杀虫内生细菌，这一内生工程杀虫细菌的建成是以水稻内生细菌的动态研究，定殖研究以及重组ＤＮＡ和细菌转化方法研究背景为基础，利用杀虫毒性强，表达苏云金芽孢杆菌δ－内毒素较高的重组质粒为供体，通过改进的ＰＥＧ原生质体转化法及新型高效电脉冲穿孔转化法完成。工程菌杀虫晶体蛋白的表达可不通过芽孢的萌发，电镜下可见营养生长期细胞内似苏云金杆菌晶体形状的毒蛋白产生。生测结果表明，工程菌液（１０５～１０６ｃｆｕ·ｍＬ－１）经超声波破碎浸水稻幼苗饲养二化螟幼虫，６天后校正死亡率均达８４．７％。In this paper reported for the first time establishment of pesticidal endophytic bacteria in rice. A transformed isolate of B.megaterium carrying a delta endotoxin gene from B.thuringiensis subsp. kurstaki strain HD 1 was successfully accomplished throgh introduction on plasmid using PEG mediated protoplast transformation and electroporation mediated transformation. By transmission electto microscopy techniques, the crystal protein was detected during vegetative growth in the transformed B.megaterium. In room experiments, insect bioassay showed that the transformants express pesticidal levels of the Bt. toxin protein.

关 键 词：苏云金芽孢杆菌 转化 杀虫内生细菌 水稻 

分 类 号：S511.035.3[农业科学—作物学]