雅安周公山土壤苏云金芽孢杆菌菌株资源的筛选和初步鉴定

采集四川雅安市周公山土壤样品320份,利用醋酸钠-抗生素筛选法分离出苏云金芽孢杆菌132株。通过光学显微镜镜检发现,这些菌株均呈典型的苏云金芽孢杆菌形态,其伴孢晶体主要形态有长菱形、短菱形、方形、球形、不定型等。根据GeneBank中公布的cry基因序列,设计了14对杀虫cry基因的通用引物,对这132株分离菌进行的PCR-RFLP分析鉴定表明:在86株菌中分布有除cry18、cry27/29、cry34/35外的11种cry基因型,其中以cry1型最多,其次为cry2型。另46株菌未检测出cry基因。对这132株苏云金芽孢杆菌进行SDS-PAGE蛋白分析,发现杀鳞翅目、鞘翅目和双翅目的约130、80、65、75 kDa的蛋白带最丰富。

Screening and Identification of Bacillus thuringiensis from Mountain Zhou Gong of Yaan

In this study,132 Bacillus thuringiensis isolates were screened by Sodium acetate-antibiotic method from 320 soil samples collected from the primeval forest in Mt.Zhou Gong of Yaan Sichuan province.Several shapes of parasporal crystal such as bipyramidal,quadrate and spherical shape were observed by microscopic observation in the B.thuringiensis isolates.PCR-RFLP and fourteen pairs of primers designed were used to identify the cry gene-types of these 132 B.thuringiensis isolates.The results indicated that 86 isolates harbored 11 cry gene type except cryl8,cry27/29,cry34/35.The cry1 type was the most abundant,and then the cry2 type.Another 46 isolates were not identified any cry gene type.The SDS-PAGE assay of parasporal crystal showed that about 130,80,65,75 kDa proteins which were probably toxic for Lepidoptera,coleopteran and dipteran insects were the most abundant.