用接合转移方法构建杀虫防病荧光假单胞菌  

Construction of Genetically Engineered Strains of Pseudomonas fluorescens against Plant Pathogen and Pest by Conjugal Transfer

作　　者：张杰 彭于发 赵建周[1] 陈彩层 黄大 

机构地区：[1]中国农科院植保所植物病虫害生物学国家重点实验室,中国农科院蔬菜与花卉研究所

出　　处：《农业生物技术学报》1995年第2期75-81,共7页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金;八六三计划青年基金

摘　　要：以经过改造的含有转座子Ｔｎ５的自杀质粒ｐＳＵＰ２０２１为载体，通过接合转移将苏云金杆菌杀虫蛋白基因ｃｒｙＩＡ（ｃ）片段插入生防细菌荧光假单胞菌Ｐ３０３菌株染色体组。Ｓｏｕｔｈｅｒｎ和Ｗｅｓｔｅｒｎ印迹分析分别证实杀虫基因的导入和杀虫蛋白的表达。室内生物测定结果表明新构建的ＰＴ２１０、ＰＴ２１２等荧光假单胞菌工程菌菌株不仅保持了野生型自然菌株对小麦全蚀病良好的抑菌活性，而且表现出对小菜蛾和玉米螟显著的毒杀作用，校正死亡率分别达８５．２％和９６．６％以上。The cry I A(c )gene from Bacillus thuringiensis was cloned into the modified suicide plas-mid pSUP2021 harboring Tn5 fragment. This vector was introduced into a strain P303 of Pseudolnonas flu-orescens by conjugation. Southern blot hybridization demonstrated that the cry I A(c)gene was integratedinto the genome of tht baacterium. Western blot analysis conformed the expression of B. t toxin protein. Re-sults of bioassay showed that PT210,PT212and other genetically engineered strains were not only inhibito-ry to the plant pathOgen Gaeulnannomyces graminis var. tritic,as the same as P303.but also insecticidal tothe larvae of diamond back moth and corn borer.The corrected mortality for above two insects were morethan 85.2%and 96.6%respectively.

关 键 词：苏云金芽孢杆菌 荧光假单胞菌 接合转移 

分 类 号：S476.12[农业科学—农业昆虫与害虫防治]