苏云金芽孢杆菌晶体杀虫蛋白Cry1Ac基因在虫生真菌球孢白僵菌中的表达  

Expression of Cry1Ac gene in entomogenous fungus Beaveria bassiana

作　　者：刘振邦[1] 任小芳[1] 宋文婧[1] 朱虹[1] 黄勃[1] 李增智[1] 

机构地区：[1]安徽农业大学微生物防治省重点实验室,合肥230036

出　　处：《安徽农业大学学报》2011年第5期742-747,共6页Journal of Anhui Agricultural University

基　　金：国家"863"项目(2006AA10A212);安徽省高校省级自然科学研究重点项目(TD200708)共同资助

摘　　要：依据苏云金芽孢杆菌晶体杀虫蛋白Cry1Ac基因序列设计引物进行PCR扩增,纯化产物与载体pbarGPE1连接,构建真菌表达载体pBARGPE1-Cry1Ac,通过芽生孢子转化法转入野生型球孢白僵菌中,获得多拷贝转化子;毒力测定结果表明转化子对马尾松毛虫的LC25比出发株降低5.4倍,LD25减少7.1倍,LT25缩短2.2 d,毒力明显提高;喂食处理和虫体喷雾喂食结合处理与单纯虫体喷雾处理相比,累计致死率分别提高了55.6%和63.0%,致死中时分别缩短了4.5 d和5.3 d。结果表明,苏云金芽孢杆菌晶体杀虫蛋白Cry1Ac基因的表达显著提高了球孢白僵菌的毒力。这是Cry1Ac基因首次在虫生真菌中表达。Bt delta-endotoxin Cry1Ac gene was cloned by PCR method and ligated with plasmid pbarGPE1 to construct a recombinant plasmid pbarGPE1-Cry1Ac.This recombinant plasmid pbarGPE1-Cry1Ac was then transferred into a wild-type strain of Beauveria bassiana by blastospore transform,resulting in a multicopy trans-formant.The results of bioassay against the Masson’s pine caterpillar,Dendrolimus punctatus,showed that LC25 of the transformant was reduced by 5.41 times,LD25 was decreased by 7.1 times,and LT25 shortened by 2.2 days,indicating a substantial virulence enhancement.Compared to spray treatment,the cumulative mortality of feeding treatment and feeding with spray treatment were increased by 55.6% and 63.0%,and LT50 shortened by 4.5 days and 5.3 days.These data indicated that the stomach toxicity of Bt delta-endotoxin gene enhanced the virulence of B.bassiana greatly.And this is the first expression of Cry1Ac gene in the insect fungi.

关 键 词：BT毒蛋白 CRY1AC基因 球孢白僵菌 表达 

分 类 号：S476.12[农业科学—农业昆虫与害虫防治]