斜纹夜蛾表皮蛋白基因克隆及表达谱分析  

Cloning and Expression Profiling of Cuticular Protein Genes of Spodoptera litura

作　　者：赵鹏 张帅 赵晨晨 胡方梅 崔金杰[2] 李绍勤[1] Zhao Peng;Zhang Shuai;Zhao Chenchen;Hu Fangmei;Cui Jinjie;Li Shaoqin(Hubei Insect Resource Utilization and Sustainable Pest Management Key Laboratory/College of Plant Science and Technology,Huazhong Agricultural University,Wuhan 430070,China;Institute of Cotton Research of Chinese Academy of Agricultural Sciences/State Key Laboratory of Cotton Biology,Anyang,Henan 455000,China)

机构地区：[1]华中农业大学植物科学技术学院/湖北省昆虫资源利用与害虫可持续治理重点实验室,武汉430070 [2]中国农业科学院棉花研究所/棉花生物学国家重点实验室,河南安阳455000

出　　处：《棉花学报》2020年第4期292-304,共13页Cotton Science

基　　金：国家现代农业产业技术体系——棉花产业技术体系(CARS-15-20)。

摘　　要：【目的】分析斜纹夜蛾不同发育阶段14个表皮蛋白(Cuticular protein, CP)基因的表达方式,为深入探究CP在斜纹夜蛾生长发育过程中的功能提供参考。【方法】利用聚合酶链反应(Polymerase chain reaction, PCR)扩增斜纹夜蛾表皮蛋白基因cDNA序列,分析其核苷酸序列和相应的氨基酸序列,并使用MEGA5.2软件构建了鳞翅目系统发育树。采用逆转录PCR(Reverse transcription PCR, RT-PCR)技术检测分析表皮蛋白基因在3~6龄幼虫各组织和2日龄成虫生殖器官中的表达。【结果】克隆了14个斜纹夜蛾表皮蛋白基因(Insect cuticular protein gene, ICPG),序列分析表明斜纹夜蛾表皮蛋白具有35~36个氨基酸残基保守基序,属于几丁质结合蛋白,它们与棉铃虫的CP具有高度的序列同源性。RT-PCR结果表明,14个ICPG在3~6龄幼虫和2日龄成虫中均有表达,其相对表达量在高龄期幼虫的表皮中较高,在低龄期幼虫的脂肪体中也较高。此外,所有14个ICPG均在睾丸和卵巢中低水平表达。在蛹期,SlICPG-11和SlICPG-14相对表达量最高。【结论】克隆了14个斜纹夜蛾CP基因的cDNA。其表达表现出组织和发育阶段特异性,暗示它们在斜纹夜蛾发育中的功能不同,为控制斜纹夜蛾靶标位点的选择奠定了基础。[Objective]The expression pattern of fourteen cuticular protein(CP)genes(ICPG)in Spodoptera litura were analyzed in this study,which provided reference for further study on the function of cuticular proteins in the growth and development of S.litura.[Method]Polymerase chain reaction(PCR)was used to amplify the cDNAs of the genes encoding CPs of S.litura.Their nucleotide sequences and corresponding amino acid sequences were analyzed,and lepidopteran phylogenetic tree was constructed using MEGA5.2 software.The expression of CP genes was analyzed in the third to the sixth instar larvae and 2-day-old adult genital organs by the reverse transcription PCR(RT-PCR)technique.[Result]Fourteen cDNAs coding 14 CPs of S.litura were obtained.The sequence analysis showed 14 CPs harbor the conserved motif of 35-36 amino acids that is a typical feature of chitin-binding proteins and share high sequence homology with CPs of Helicoverpa armigera.RT-PCR results showed that these CP genes were expressed in the third to the sixth instar larvae and 2-day-old adults.In particular,their relative expression levels are higher in the cuticulars of older larvae and lower in the fat bodies of younger larvae.In addition,the relative expression of 14 CP genes was low in testes and ovaries.The highest relative expression of SlICPG-11 and SlICPG-14 in the pupal stage.[Conclusion]In this study,the cDNA of 14 S.litura CP genes were cloned.Their expression exhibited tissue and developmental stage specificity indicating their putative roles in S.litura development.It lays the foundation for target site selection for the control of S.litura.

关 键 词：斜纹夜蛾 表皮蛋白 基因克隆 逆转录聚合酶链反应(RT-PCR) 组织表达 发育阶段 

分 类 号：S433.4[农业科学—农业昆虫与害虫防治]