防治茄二十八星瓢虫Bt菌株的筛选及杀虫活性分析  

Screening of Bacillus thuringiensis Strains and Analysis of Their Insecticidal Activity Against Henosepilachna vigintioctopunctata

作　　者：马琪琪 胡宗伟 夏鹏亮 冯万祖 Natasha I T MBIZA 张建民[1] MA Qiqi;HU Zongwei;XIA Pengliang;FENG Wanzu;NATASHA I T Mbiza;ZHANG Jianmin(College of Agriculture,Yangtze University,Jingzhou,Hubei 434025;Enshi Tobacco Company,Enshi,Hubei 445000)

机构地区：[1]长江大学农学院,湖北荆州434025 [2]湖北省恩施州烟草公司,湖北恩施445000

出　　处：《北方园艺》2021年第14期31-38,共8页Northern Horticulture

基　　金：国家自然科学基金资助项目(31471783)。

摘　　要：以从湖北宜昌、襄阳、荆州、广东东莞、安徽安庆5个地区采集得到的52份土样为试材,分别采用温度筛选法、镜检法、PCR-RFLP法和SDS-PAGE等方法,分离了土壤中的苏云金芽孢杆菌,并鉴定了分离出的Bt菌株的基因型,通过茄二十八星瓢虫二龄幼虫的室内杀虫试验测定分离株的杀虫活性,同时分析了部分Bt菌株的伴孢晶体蛋白,以期挖掘新的Bt菌株资源,筛选出对茄二十八星瓢虫具有高毒力的Bt菌株。结果表明:分离得到10株含cry3基因的菌株,3株含cry7基因的菌株和2株含cry8基因的菌株,得到1株对茄二十八星瓢虫二龄幼虫有高毒力的Bt菌株,当其伴孢晶体悬浮液的浓度在40 mg·mL^(-1)时,校正死亡率最高可达40.42%;发现YC-2-20、GD-4-1均表达分子量约为67 kDa的晶体蛋白;GD-5-7表达分子量约为130 kDa和80 kDa的晶体蛋白;YC-12-6表达分子量约为130 kDa和70 kDa的晶体蛋白。试验筛选出的Bt菌株为农业生产中茄二十八星瓢虫的绿色防控提供了可应用的菌株及基因资源,同时对延缓害虫抗药性的产生也具有重要意义。Taking 52 soil samples as test materials, soil samples were collected from five areas: Yichang, Xiangyang, Jingzhou in Hubei Province;Dongguan in Guangdong Province;Anqing in Anhui Province.The temperature screening method, microscopic examination method, PCR-RFLP and SDS-PAGE were used as experimental methods.The Bt in the soil was isolated, and the genotype of the isolated Bt strain was identified.The insecticidal activity of the isolated strain was determined by indoor insecticidal test for the second instar larvae of Henosepilachna vigintioctopunctata,and the crystal protein of some Bt strains was analyzed.In order to discover new Bacillus thuringiensis strains resources.The results showed that a total of ten strains containing cry3 gene, three strains containing cry7 gene and two strains containing cry8 gene were isolated, and one strain of Bt strain with high virulent to the second instar larva of Henosepilachna vigintioctopunctata was obtained.When the concentration of its adjoint crystal suspension was 40 mg·mL^(-1),the corrected mortality rate could reach 40.42%.It was found that YC-2-20 and GD-4-1 both expressed crystal proteins with molecular weight of about 67 kDa.GD-5-7 expressed crystal proteins with molecular weights of about 130 kDa and 80 kDa.YC-12-6 expresses crystal proteins with molecular weights of about 130 kDa and 70 kDa.The Bt strains screened out in the experiment provided applicable strains and genetic resources for the green prevention and control of Henosepilachna vigintioctopunctata in agricultural production, and also had important significance for delaying the emergence of pest resistance.

关 键 词：苏云金芽孢杆菌 茄二十八星瓢虫 菌株筛选 CRY基因 杀虫活性 

分 类 号：S433[农业科学—农业昆虫与害虫防治]