我国不同森林立地带土壤中苏云金芽孢杆菌cry基因资源研究

利用PCR－RFLP鉴定体系和SDS－PAGE表达蛋白的分析方法，分析了来自我国不同森林立地带（寒温带，中温带，暖温带，北亚热带，中亚热带，南亚热带，高原亚热带，热带）自然保护区森林土壤中分离的72株苏云金芽孢杆菌的cry 1，cry 2，cry 3，cry 4，cry 5，cry 8，cry 9，cry 10，cry 11，cry 1I杀虫晶体蛋白基因类型，表达蛋白和杀虫活性的生物测定。研究表明：同时含有cry 1，cry 2，cry 1I3类 有21株菌，6株菌含有cry 1，cry 2类基因，4株菌含有cry 1和cry 1I基因，只含有cry 1基因的1株，cry 2基因的4株，36株菌不含所鉴定的10类基因。同时证明：绝大多数含有cry1基因的菌株表达了130kD蛋白，含有cry2基因的菌株表达了60kD的蛋白。对不同农，林害虫棉铃虫，杨扇舟蛾，舞毒蛾，马尾松毛虫，黄粉甲，榆兰叶甲，落叶松叶蜂等幼虫的杀虫活性进行了生物测定。进一步证明了苏云金芽孢杆菌cry基因，表达蛋白及杀虫活性三者的相关性。为生产和科研提供了生物治虫，抗虫育种的苏云金芽孢杆菌cry基因资源。

Studies on Resources of Cry-type Genes of Bacillus thuringie-nsis from Different Soil of Forest Site Zone in China

cry genes, express protein and bioassay of 72 isolations of Bacillus thuringiensis from different forest site zones(cool temperate zone, temperate zone, warm temperate zone, north subtropical zone, subtropical zone, south subtropical zone, high plateau subtropical zone, tropical zone) in China were analyzed by the methods of PCR RFLP and SDS PAGE. The results showed that 21 strains contained three cry type genes: cry1, cry1 and cry1; 6 strains contained both cry1 and cry1 genes, 4 strains contained both cry1 and cry1I type genes; 1 strain contained cry1 type gene, 4 strains contained cry1 type gene, 36 strains did not contain the ten cry type genes. Most cry1 genes encoded 130 kD protein that is toxic to Lepidopteron pest, and most cry1 gene encoded 60 kD protein. The isolates of containing cry1Aa, cry1Ac and cry1A type genes had higher toxicity to Helicoverpe armigera, Clostera anachoreta, Lymantria dispar, Dendrolimus punctatus larvae, but isolations of containing one or no cry gene had low toxicity to these pests.