摘要
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cry1、cry2、cry3、cry4、cry5、cry8、cry9、cry10、cry11、cry1I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cry1、cry2、cry1I3类基因的有21株菌,6株菌含有cry1、cry2类基因,4株菌含有cry1和cry1I基因,只含有cry1基因的有1株,cry2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、抗虫育种新的苏云金芽孢杆菌cry基因资源。
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 PCRRFLP and SDSPAGE. The results showed that 21 strains contained three crytype genes: cry1, cry2 and cry1; 6 strains contained both cry1 and cry2 genes, 4 strains contained both cry1 and cry1I type genes; 1 strain contained cry1 type gene, 4 strains contained cry2 type gene, 36 strains did not contain the ten crytype genes. Most cry1 genes encoded 130 kD protein that is toxic to Lepidopteron pest, and most cry2 gene encoded 60 kD protein. The isolates of containing cry1Aa, cry1Ac and cry2A 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.
出处
《林业科学研究》
CSCD
北大核心
2003年第1期19-25,共7页
Forest Research
基金
国家自然科学基金资助项目(NO.39970607)
