一株具有烟草甲幼虫毒杀作用苏云金芽孢杆菌的遗传多样性分析及cry基因鉴定  

Genetic Analysis and cry Gene Profiling of a Strain of Bacillus thuringiensis with Pathogenicity to Lasiodiodema serricome

作　　者：胡逸超 雷薇 孙建生[1] 李季刚[1] 邹克兴[1] 苏赞[1] 梁伟[1] 蔡联合[1] 陈琦 邹承武[3] HU Yi-chao;LEI Wei;SUN Jian-sheng;LI Ji-gang;ZOU Ke-xing;SU Zan;LIANG Wei;CAI Lian-he;CHEN Qi;ZOU Cheng-wu(Guangxi Zhongyan Industrial Co.,Ltd.,Guangxi Nanning 530001,China;Nanning Goldtech Biotechnology Co.,Ltd.,Guangxi Nanning 530001,China;Agricultural College of Guangxi University,Guangxi Nanning 530004,China)

机构地区：[1]广西中烟工业有限责任公司技术中心,广西南宁530001 [2]南宁国拓生物科技有限公司,广西南宁530001 [3]广西大学农学院,广西南宁530004

出　　处：《西南农业学报》2020年第2期319-324,共6页Southwest China Journal of Agricultural Sciences

基　　金：广西自然科学基金项目(2016GXNSFAA380027);广西中烟工业有限责任公司项目(GXZYCX2017B005);2019年度广西中央引导地方科技发展专项(桂科计字【2019】183号)。

摘　　要：【目的】明确一株具有烟草甲幼虫毒杀作用苏云金芽孢杆菌在蜡样芽孢杆菌群中的进化关系,并对该菌株所携带的cry基因进行鉴定,为其在农业上的开发应用提供参考依据。【方法】对苏云金芽孢杆菌GXZY032菌株的16S rRNA和gyrB基因序列进行PCR扩增测序,并进行相应的进化树分析;使用PCR技术对GXZY032菌株基因组所包含的cry杀虫基因进行鉴定。【结果】菌株GXZY032的16S rRNA序列经PCR扩增得到1.5 kb片段,在进化树中以较低的置信值与多个BtII群菌株聚到一起。gyrB基因序列经PCR扩增并测序得到1.2 kb片段,所构建的进化树有6个置信值较高的分支,构成BtI、BtII、B.cereus I、BtIII、Bt+Bw和Bt+Ba 6个群,其中菌株GXZY032的序列与BtII群聚集一起,置信值为99%。对8种cry基因进行PCR鉴定,结果表明,菌株GXZY032包含cry1、cry2和cry10 3种类型cry基因。【结论】苏云金芽孢杆菌菌株GXZY032属于BtII群,具有cry1、cry2和cry10 3种类型cry杀虫基因。【Objective】The present paper aimed to analyze the genetic relationship of a strain of Bacillus thuringiensis with pathogenicity to Lasiodema serricome in B.cereus,and cry gene in it was identified to provide reference for its development in agriculture.【Method】The sequence of 16S rRNA and gyrB gene in GXZY032 strain was amplified and sequenced to construct phylogenetic tree for analyzing the genetic distance,respectively.The profiles of cry genes in GXZY032 strain were obtained by PCR.【Result】The 1.5 kb sequence of 16S rRNA was amplified from strain GXZY032 by PCR,which is clustered with some sequences of BtII in the phylogenetic tree under low confidence value.The 1.2 kb sequence of gyrB gene was obtained by PCR and sequen-cing.The phylogenetic tree of gyrB gene have 6 clusters with high con-fidence value,which were attributed as BtI group,BtII group,B.cereus I group,BtIII group,Bt+Bw group and Bt+Ba group.Sequence of strain GXZY032 was clustered with sequences from BtII group under confidence value of 99%.Using PCR profiling,cry1,cry2 and cry10 were found in strain GXZY032.【Conclusion】B.thuringiensis strain GXZY032 belongs to BtII group and has three types of cry gene(cry1,cry2 and cry10).

关 键 词：苏云金芽孢杆菌 进化关系 16S RRNA序列 cry基因鉴定 

分 类 号：S476.1[农业科学—农业昆虫与害虫防治]