基于基因组与蛋白组分析挖掘高效杀虫菌株Bt S2480-1毒蛋白基因  

Exploration of insecticidal genes in Bt S2480-1: based on the analysis of genome and proteome

作　　者：周燕 杨天宝 潘甘霖 莫世栋 刘传顺 方宣钧 姜明国[1] ZHOU Yan;YANG Tianbao;PAN Ganlin;MO Shidong;LIU Chuanshun;FANG Xuanjun;JIANG Mingguo(Guangxi Key Laboratory for Polysaccharide Materials and Modifications,School of Marine Sciences and Biotechnology,Guangxi Minzu University,Nanning 530008,Guangxi,China;Institute of Life Science,Jiyang College,Zhejiang A&F University,Zhuji 311800,Zhejiang,China;Hainan Institute of Tropical Agricultural Resources,Sanya 572025,Hainan,China)

机构地区：[1]广西民族大学海洋与生物技术学院,广西多糖材料与改性重点实验室,广西南宁530008 [2]浙江农林大学暨阳学院生命科学研究所,浙江诸暨311800 [3]海南省热带农业资源开发利用研究所,海南三亚572025

出　　处：《微生物学报》2022年第9期3587-3596,共10页Acta Microbiologica Sinica

基　　金：广西民族大学2020年度校级引进人才科研启动项目(2020KJQD23);广西科技重大专项(桂科AA18242026);2022年度广西高校中青年教师基础能力提升项目(2022KY0159)。

摘　　要：【目的】鉴定苏云金芽孢杆菌野生型菌株Bt S2480-1的杀虫活性并挖掘该菌株含有的杀虫基因资源。【方法】采用Illumina Hiseq2000测序平台对Bt S2480-1菌株进行全基因组测序,通过生物信息学方法获得该菌株的基因组信息、基因预测以及毒蛋白基因预测识别;随后,利用LTQ-Obitrap nano-LC-MS/MS系统对该菌株的总蛋白进行了质谱分析;最后以致倦库蚊幼虫和甜菜夜蛾幼虫为靶标昆虫对该菌株进行了生物活性测定。【结果】Bt S2480-1基因组大小为6.2 Mb,GC含量为35.11%,拼接得到1个拟核和3个大质粒的可视化草图,预测编码基因6297个,其中含有12个预测毒蛋白基因。BtS2480-1菌株总蛋白在LTQ-OrbitrapMS/MS质谱分析中共有1 500个蛋白质获得鉴定,鉴定获得11个毒蛋白。Bt S2480-1菌株总蛋白对致倦库蚊幼虫表现出非常高的杀蚊活性,而对甜菜夜蛾幼虫杀虫活性相对偏弱,其LC50分别为27.636μg/m L (95%FL:12.559-61.707μg/m L)和496.833μg/mg (95%FL:320.134–776.964μg/mg)。【结论】Bt S2480-1菌株基因组分析显示共含有12个预测毒蛋白基因,LTQ-Orbitrap MS/MS鉴定获得11个毒蛋白,Bt S2480-1菌株总蛋白对致倦库蚊和甜菜夜蛾幼虫都具有杀虫活性。[Objective] To identify the insecticidal activity of wild-type Bacillus thuringiensis strain Bt S2480-1 and explore the insecticidal gene resources. [Methods] The whole genome of Bt S2480-1 was sequenced by Illumina Hiseq2000. The genome information, genes, and insecticidal genes of the strain were analyzed with bioinformatics methods. The total proteins of Bt S2480-1 were analyzed by LTQ-Obitrap nano-LC-MS/MS and the biological activity of Bt S2480-1 against Culex quinquefasciatus and Spodoptera exigua larvae was determined. [Results] The genome of Bt S2480-1 was 6.2 Mb, with GC content of 35.11%, and one nucleoid and three large plasmids were yielded through assembly, containing 6 297 coding genes with 12 encoding insecticidal proteins. Meanwhile, a total of 1 500 proteins of Bt S2480-1 were identified by LTQ-Orbitrap MS/MS, among which 11 were insecticidal proteins. The total proteins of Bt S2480-1 showed strong activity against C. quinquefasciatus larvae but relatively weak activity against S. exigua larvae, with LC50 of 27.636 μg/m L(95% FL: 12.559–61.707 μg/m L) and 496.833 μg/mg(95% FL: 320.134–776.964 μg/mg), respectively. [Conclusion] Bt S2480-1 contains 12 genes encoding insecticidal proteins and displays insecticidal activity against C. quinquefasciatus and S. exigua larvae.

关 键 词：苏云金芽孢杆菌 Bt S2480-1 毒蛋白 LTQ-Obitrap 杀虫活性 

分 类 号：S476[农业科学—农业昆虫与害虫防治]