杀虫晶体蛋白基因Cry1Ac在生防菌B-916中的表达  

Expression of Insecticidal Crystal Protein Cry1Ac Gene in Biocontrol Bacillus subtilis Strain B-916

作　　者：罗楚平[1] 陈志谊[1] 刘永锋[1] 刘邮洲[1] 聂亚锋[1] 

机构地区：[1]江苏省农业科学院植物保护研究所,江苏南京210014

出　　处：《江苏农业学报》2007年第4期306-311,共6页Jiangsu Journal of Agricultural Sciences

基　　金：国家"863"计划现代农业技术领域重大项目(2006AA10A211);江苏省高技术研究项目(BG2006326);江苏省农业科学院科研基金项目(6110540)

摘　　要：枯草芽孢杆菌生防菌B-916对水稻纹枯病和稻曲病具有良好的防治效果,为拓宽其防治范围,兼治病虫害,本研究采用B-916内源芽孢形成期特异表达启动子成功实现了杀虫晶体蛋白基因Cry1Ac在野生型生防菌B-916的高效表达,成功构建了兼治水稻病、虫害的工程菌E-916。经SDS-PAGE电泳分析,异源Cry1Ac基因在对数生长后期其表达产物杀虫晶体蛋白占到胞内总蛋白的28%。杀虫和抑菌试验结果表明构建的工程菌对水稻二化螟具有杀虫活性,且保持了其原有对水稻纹枯病的拮抗性能。构建的工程菌具有良好的分离稳定性,连续培养25h,稳定性高于95%;连续培养50 h稳定性高于70%;连续稀释培养50 h,质粒丢失频率为每代小于10-3个。Strain B-916 of Bacillus subtilis has a good efficacy on rice sheathblight and rice false smut. To broaden its control objects including insect pests, a genetically engineered strain （E-916） against insect pests and diseases in rice was constructed through the heterologous expression of insecticidal crystal protein gene crylAc in post-exponential-phase B, subtilis strain B-916. SDS-PAGE electrophoresis showed that the insecticidal protein in strain E-916 accounted for about 28% of the total protein. Strain＇E-916 not only kept the antagonistic action on rice sheathblight disease, but acquired the insecticidal activity to rice stem borer through genetic engineering. The recombinant plasmid segregated stably with segregation stability higher than 70% and the plasmid losing frequency less than 0. 000 1 per generation even after continuous culture for 50 h. Growth test revealed that the expression of gene crylAc had no negative effect on growth of B-916.

关 键 词：生物防治 枯草芽孢杆菌 杀虫晶体蛋白 工程菌 

分 类 号：S476[农业科学—农业昆虫与害虫防治]