上海地区苏云金芽孢杆菌的分离及其对草地贪夜蛾室内杀虫活性的测定  

Isolation of Bacillus thuringiensis in Shanghai and its indoor insecticidal activity against Spodoptera frugiperda

作　　者：陈诗杰 曹越平[1] CHEN Shijie;CAO Yueping(School of Agriculture and Biology,Shanghai Jiao Tong University,Shanghai 200240,China)

机构地区：[1]上海交通大学农业与生物学院,上海200240

出　　处：《上海农业学报》2022年第5期78-83,共6页Acta Agriculturae Shanghai

摘　　要：为进一步发掘国内苏云金芽孢杆菌(Bacillus thuringiensis,Bt)的菌株资源,在上海市若干地区搜集土壤样品,采用NaAc-抗生素筛选法从土样中分离野生Bt菌株,通过光学显微镜染色镜检、PCR-RFLP体系及SDS-PAGE电泳等方法对分离菌株的杀虫晶体蛋白形态、蛋白表达量及cry基因类型进行分析,并测定了分离菌株对草地贪夜蛾的室内杀虫活性。结果表明:在上海地区采集的165份土壤样品中共分离得到苏云金芽孢杆菌50株,总出菌率为8.20%;其伴孢晶体蛋白呈菱形、球形、椭球形、不规则形等晶体形态,以球形晶体为主;多数表达130 ku、90 ku、80 ku、50 ku蛋白;分离菌株所含cry基因类型丰富,包含cry1、cry3、cry8类等共16种基因类型,以cry32及cry8类基因居多;有6株菌株对草地贪夜蛾表现出高杀虫活性,72 h校正死亡率最高可达100%。上述分离菌株对后续开展草地贪夜蛾的绿色防控及抗性治理具有重要的实践意义。In order to further explore the resources of Bacillus thuringiensis(Bt)strains in China,soil samples were collected in several areas of Shanghai.The morphology,protein expression and cry gene type of the insecticidal crystal protein of the isolated strain were analyzed by optical microscope staining and microscopy,PCR-RFLP system and SDS-PAGE electrophoresis,and the insecticidal activity of the isolated strain against Spodoptera frugiperda(J.E.Smith)was determined in the laboratory.The results showed that a total of 50 strains of B.thuringiensis were isolated from 165 soil samples collected in Shanghai,with a total bacterial rate of 8.20%.The parasporal crystal proteins showed rhombic,spherical,ellipsoidal and irregular crystal shapes,mainly spherical crystals.Most of them expressed 130 ku,90 ku,80 ku and 50 ku proteins.The isolates contained rich cry gene types,including 16 types of cry1,cry3 and cry8,most of which were cry32 and cry8 gene types.Six strains showed high insecticidal activity against S.frugiperda,and the highest 72-hour corrected mortality was 100%.The above isolated strains have important practical significance for the subsequent development of green control and resistance management of S.frugiperda.

关 键 词：苏云金芽孢杆菌 伴孢晶体蛋白 CRY基因 Bt分离菌株 草地贪夜蛾 

分 类 号：S433.4[农业科学—农业昆虫与害虫防治] S476.8[农业科学—植物保护]