双委夜蛾3个信息素结合蛋白基因的分子鉴定及组织表达分析  

Molecular identification and tissue expression analysis of three pheromone binding protein genes in Athetis dissimilis

作　　者：闫祺 陈严 刘晓龙 李柏桦 廖辉 张小晴 董双林[1] YAN Qi;CHEN Yan;LIU Xiaolong;LI Baihua;LIAO Hui;ZHANG Xiaoqing;DONG Shuanglin(College of Plant Protection/Key Laboratory of Monitoring and Management of Crop Diseases and Pest Insects, Ministry of Agriculture and Rural Affairs,Nanjing Agricultural University,Nanjing 210095,China;School of Forestry and Landscape Architecture,Anhui Agricultural University,Hefei 230036,China)

机构地区：[1]南京农业大学植物保护学院/农业农村部作物病虫害监测与防控重点开放实验室,江苏南京210095 [2]安徽农业大学林业与园林学院,安徽合肥230036

出　　处：《南京农业大学学报》2018年第6期1037-1044,共8页Journal of Nanjing Agricultural University

基　　金：国家重点研发计划项目(2017YFD0200900)

摘　　要：[目的]本文旨在鉴定重要农业害虫双委夜蛾(Athetis dissimilis Hampson)的信息素结合蛋白(pheromone binding protein,PBP)基因,并分析其序列和组织表达特征,为进一步研究双委夜蛾的嗅觉感受机制奠定基础。[方法]基于双委夜蛾触角转录组数据获得3个PBP基因(Adis PBP1、Adis PBP2、Adis PBP3)的全长序列,通过RT-PCR对3个基因进行克隆和验证,利用RT-qPCR对3个基因在雌、雄成虫的组织表达谱进行分析。[结果]从双委夜蛾触角中克隆3个PBP基因的c DNA序列,3个基因Adis PBP1、Adis PBP2及Adis PBP3分别编码166、169和164个氨基酸,并具有6个保守的半胱氨酸、N端1个信号肽序列等该类基因的典型特征。聚类分析表明:这3个基因分别被聚到3个不同的PBP分支中,且均与同属害虫二点委夜蛾(Athetis lepigone)的相应PBP最近。表达谱分析显示:3个基因均仅在雌、雄虫触角中高表达; Adis PBP1在雄性触角的表达量显著高于雌性触角,而Adis PBP2和Adis PBP3在两性触角中的表达量相似; 3个基因雄、雌蛾触角的表达量差异(雄∶雌)分别为1.84、1.14和0.95,暗示其在性信息素及其他气味感受中的不同作用。[结论]克隆了双委夜蛾3个PBP基因的c DNA全长序列,明确了3个基因的组织表达谱及雌、雄表达差异,对3个PBP基因的可能功能进行了讨论。[Objectives]This study aims to identify the pheromone binding protein(PBP)genes and clarify their tissue expression pattern,provide basis for further study on the olfactory mechnism of Athetis dissimilis.[Methods]The full-length cDNA of three PBP genes(AdisPBP 1,AdisPBP2,AdisPBP3)from A.dissimilis were obtained by analyzing the antennae transcriptomic data and further verified by RT-PCR experiments.The tissue expression patterns of three PBP genes in male and female adults were analyzed by RT-PCR and RT-qPCR.[Results]Three PBP genes were cloned from the antennae of A.dissimilis adults,which were named AdisPBP 1,AdisPBP2,AdisPBP3,respectively.The cDNA sequence of the three PBP genes encoded 166,169 and 164 amino acids,respectively.The encoded proteins of the three genes had typical characteristics of PBP,including an N-terminal signal peptide and six conserved cysteines.Phylogenetic analysis revealed that the three genes were clustered into three distinct PBP groups and had the highest homology with the PBP of Athetis lepigone.Expression patterns investigation revealed that the three genes were highly expressed in adult antennae of both males and females.The relative expression levels of AdisPBP 1,AdisPBP2 and AdisPBP 3 in the antennae of male adults were 1.84,1.14 and 0.95 times as high as those in the antennae of female adults,respectively.[Conclusions]Three A.dissimilis PBP genes were identified,their tissue and sex expression patterns were determined,and their functions were discussed.

关 键 词：双委夜蛾 夜蛾科 信息素结合蛋白 组织表达谱 

分 类 号：S965.8[农业科学—水产养殖]