苏云金芽胞杆菌BRC-HZP7杀虫晶体蛋白多样性分析

ICPs polymorphism of Bacillus thuringiensis BRC-HZP7

分离自武夷山土壤的苏云金芽胞杆菌BRC-HZP7菌株对小菜蛾和甜菜夜蛾幼虫有较好的毒杀作用.对该菌株进行了生理生化测定、cry基因的PCR-RFLP检测,以及通过SDS-PAGE检测晶体蛋白,并对蛋白点进行肽段指纹图谱鉴定.结果表明该菌株含有cry1Aa、cry1Ba、cry1Ia、cry2Ab、cry2Ac、cry2Ae等6种cry基因.从选取的8个蛋白条带肽段指纹图谱可以看出:标记3、6、7、8的4个蛋白点都是杀虫晶体蛋白,蛋白点6、7、8分别鉴定为Cry2Aa、Cry1Aa、Cry1Ba,蛋白点3可能是Cry1Ba的部分片段.表明该菌株有丰富的ICP资源,可作为杀虫应用的候选菌株.

Bacillus thuringiensis strain BRC-HZP7 was isolated from the soil of Wuyi mountain,which exhibited the high activity against Plutella xylostella larvae and Spodoptera exigua larvae.Biochemical reactions were tested for its biological characters.The cry gene types and their encoded proteins of BRC-HZP7 were identified by the methods of PCR-RFLP,SDS-PAGE and PMF.The results showed that cry1Aa,cry1Ba,cry1Ia,cry2Ab,cry2Ac and cry2Ae existed in BRC-HZP7.Eight interesting protein bands were picked out to analyze further by MALDI-TOF/MS.PMF matching analysis results showed that band 3,6,7,8 were ICPs.The band 6,7,8 were identified as Cry2Aa,Cry1Aa,Cry1Ba,respectively.The band 3 was possible partial fragment of Cry1Ba.The strain with abundant ICPs would be a good resource in pest biocontrol.