海南五指山热带雨林区Bt菌株的分离及其新型cry基因鉴定  

Screening Bacillus thringiensis Isolates from Soil Samples of Wuzhishan Primitive Tropic Rainforest in Hainan and Identifying Novel cry-type Genes

作　　者：吴仲琦[1] 罗爽[1] 张捷[1] 杨毅然[1] 王苏环[1] 郭玲玉[1] 钤江朝[1] 黄楠楠[1] 徐志霞[1] 吴红萍[1] 王锐萍[1] 张文飞[1] 

机构地区：[1]海南师范大学生命科学学院,热带动植物生态学省部共建教育部重点实验室,海口571158

出　　处：《基因组学与应用生物学》2013年第1期46-52,共7页Genomics and Applied Biology

基　　金：海南师范大学博士启动基金;海口市重点科技计划项目项目<厌氧消化池中硫化氢形成及其控制>(2009030);海南省重点科技项目"EBPR系统高效聚磷菌菌群构建及产业化研究(ZDXM20100065)"资助

摘　　要：五指山原始热带雨林区位于海南岛中部,是海南岛地区面积最大的热带原始雨林。我们依据不同的海拔高度对五指山原始雨林区进行了系统的取样,共采集了234份土壤样品,采用醋酸钠-高温处理的方法分离出各类产芽孢杆菌886株,并通过显微镜观察鉴定出产伴胞晶体蛋白的Bt菌株21株,其Bt菌株分离率为2.3%。为了进一步挖掘五指山丰富的Bt杀虫基因资源,还利用PCR方法结合测序技术对21株Bt分离株的cry基因进行了鉴定,鉴定结果显示cry1、cry4、cry39、cry40、cry31和cry46等基因型存在,表明五指山热带Bt菌株含有丰富cry基因资源,这将为为进一步开展Bt杀虫基因的分离克隆打下基础。The Wuzhishan, located at the center of Hainan Island, is the largest Primitive Tropic Rainforest in Hainna island. In this study, we collected 234 soil samples from Wuzhishan at the different heights above sea level from 700 m to 1 650 m. Following the approach of selection culture medium with sodium acetate and treatment of high temperature, we obtained 886 Bacillus isolates and further detected 21 Bacillus thuringieusis （Bt） isolates through observation of microscopy, of which the isolating rate reached 2.3%. In order to mine the insecticidal genes from the identified Bt isolates, the PCR strategy with universal primer pairs, combined with DNA sequencing technology was used to detected cry genes. The result showed that the Bt isolates from Wuzhishan include cryI-like, cry4-1ike, cry39-1ike, cry40-like, cry31-like, cry46-1ike genes and so on, presenting that the obtained Bt isolates are abundant in novel cry genes, which may facilitate to clone novel Bt crystal toxin genes.

关 键 词：苏云金芽孢杆菌 菌株分离 CRY基因 五指山原始热带雨林 

分 类 号：S718.8[农业科学—林学]