铁硫簇组装蛋白在意大利蜜蜂耐受吡虫啉毒害中的应答分析  

Response Analysis of Iron-Sulfur Cluster Assembly Protein in Apis mellifera ligustica Tolerance to Imidacloprid Toxicity

作　　者：何彪 王悦弟 秦启倩 陈兰春 段佳欣 许金山[1] 李治[1] HE Biao;WANG Yuedi;QIN Qiqian;CHEN Lanchun;DUAN Jiaxin;XU Jinshan;LI Zhi(College of Life Sciences,Chongqing Normal University,Chongqing 401331»China)

机构地区：[1]重庆师范大学生命科学学院,重庆401331

出　　处：《重庆师范大学学报（自然科学版）》2021年第4期21-29,共9页Journal of Chongqing Normal University:Natural Science

基　　金：现代农业产业技术体系建设专项资金(No.CARS-44-KXJ21);国家自然科学基金(No.31101770,No.31302037);重庆市教育委员会科学技术研究项目(No.KJ1703051);重庆市前沿与应用基础研究计划项目(No.cstc2014jcyjA80039,No.cstc2015jcyjA80014);重庆师范大学自然科学基金项目(No.13XLB009)。

摘　　要：【目的】用生物信息学方法鉴定意大利蜜蜂(Apis mellifera ligustica)铁硫簇组装蛋白(AmIscA),分析它在意大利蜜蜂工蜂耐受杀虫剂吡虫啉毒性中的应答特征。【方法】多重序列比对分析AmIscA的氨基酸序列特征和结构域组成;在线预测该蛋白的结构以及互作蛋白网络,预测它的生物学功能;构建系统进化树分析该蛋白的分类和进化关系;构建半致死模型,实时荧光定量PCR分析AmIscA基因在工蜂耐受杀虫剂吡虫啉毒性0~48h中的表达特征。【结果】AmIscA基因cDNA序列全长514bp,编码130个氨基酸残基,计算出AmIscA的相对分子质量为14.1kDa,pI为9.06。AmIscA为疏水性蛋白,具有铁硫簇组装蛋白保守的半胱氨酸和天冬氨酸残基,并且不具备信号肽,定位于线粒体中。经48h的半致死剂量吡虫啉暴露后的工蜂存活率为45%±5%。AmIscA基因表达受吡虫啉胁迫而上调,且呈先升高后下降趋势。【结论】AmIscA积极响应吡虫啉导致的毒性胁迫,可能通过抗氧化作用潜在地参与了意大利蜜蜂对吡虫啉毒性的耐受。[Purposes]To bioinformatics characterizes the iron-sulfur cluster assembly protein of Apis mellifera ligustica(AmIscA)and analyzes its response in worker tolerance imidacloprid toxicity.[Methods]Multiple sequence alignment was used to analyze the sequence characteristics and domain composition of AmIscA.The structure,interaction network and biological function of AmIscA were predicted by using an On-line program.A phylogenetic tree was constructed to analyze the classification and phylogenetic evolutionary of AmIscA.The semi-lethal model was constructed,and the expression characteristics of AmIscAgene in worker bees tolerance to imidacloprid toxicity were detected by real-time fluorescence quantitative PCR after 0~48 hours exposure.[Findings]The full-length cDNA sequence of the AmIscAgene was 514 bp,which encodes 130 amino acids.This protein AmIscA is predicted to have a molecular weight of 14.1 kDa,a theoretical pI value of 9.06.The hydrophilic-hydrophobic analysis showed that this protein was hydrophobic with cysteine and aspartic acid conserved residues of classify iron-sulfur cluster assembly protein family.The signal peptide and subcellular localization prediction showed that this protein had no signal peptide and was localized in mitochondria.After being exposed to a half-lethal dose of imidacloprid for 48 hours,the bees\survival rate was 45%±5%.Realtime quantitative PCR analysis showed that the expression levels of AmIscAgene was up-regulated under imidacloprid stress and presented a tendency of upregulation first,then downregulation compared to the control group.[Conclusions]AmIscA actively responds to the toxic stress caused by imidacloprid and potentially participates in the resistance of bees to imidacloprid toxicity.

关 键 词：铁硫簇组装蛋白 吡虫啉 半致死 意大利蜜蜂 

分 类 号：Q966[生物学—昆虫学]