桃小食心虫触角气味结合蛋白和化学感受蛋白基因的鉴定与进化分析

Identification and Phylogenetic Analysis of OBPs and CSPs of Antennae Carposina sasakii Matsumura(Lepidoptera:Carposinidae)

借助转录组测序技术发掘桃小食心虫(Carposina sasakii)触角中的气味结合蛋白(odorant binding protein,OBPs)和化学感受蛋白(chemosensory protein,CSPs)的基因,为后续研究以上两类嗅觉基因的功能提供基因资源保障。利用Illumina HiSeq 4000测序仪对桃小食心虫成虫触角cDNA文库进行测序,利用昆虫OBPs和CSPs的特征序列鉴定桃小食心虫的OBPs和CSPs基因,并对OBPs和CSPs编码氨基酸序列的相似性进行比对,通过构建邻接树对桃小食心虫OBPs、CSPs与其他鳞翅目昆虫同类基因的进化关系进行分析。结果表明,此次转录组测序共产生67060329个clean reads,从头拼接后得到82426个转录本,去冗余后最终获得63327个独立基因Unigenes。通过关键词搜索和PSI-BLAST比对,在桃小食心虫触角中共鉴定到26种OBPs和16种CSPs基因。序列比对发现桃小食心虫触角中的不同OBPs和CSPs的序列相似性较低,分别介于10.00%~78.57%和20.47%~75.20%之间;进化分析表明桃小食心虫OBPs和CSPs分化程度较高,绝大多数基因聚类至不同的进化分枝。本研究获得了桃小食心虫触角中大量的OBPs和CSPs基因,为进一步研究该虫嗅觉蛋白外周编码的分子机制提供了基础保障。

The genes of odorant binding protein(OBPs)and chemosensory protein(CSPs)were identified by transcriptome sequencing of the antennae of Carposina sasakii,which provided the genetic resources for the further study on the functions of these two kinds of olfactory proteins.The cDNA libraries of C.sasakii antennae were sequencing using an Illumina Hiseq 4000 platform.The OBPs and CSPs genes were identified by comparing them with the characteristic sequences of OBPs and CSPs from the other insects.The evolutionary relationships between the OBPs and the CSPs of C.sasakii and those of the other Lepidopteran insects by constructing neighbor joining trees.The study obtained 67060329 clean reads.82426 transcripts were generated after the de novo assemblies of the clean reads data,and finally generated 63327 unigenes after removed redundant transcripts.Using keyword searching and PSI-BLAST comparing,26 OBPs and 16 CSPs genes of C.sasakii antennae were identified.The results of sequence alignment showed that the sequence similarities among different OBPs or different CSPs of C.sasakii antennae were very low,ranging from 10.00%to 78.57%and 20.47%to 75.20%,respectively.The results of phylogenetic analyses showed highly differentiations among different OBPs or among different CSPs.Most of the genes were clustered into different evolutionary branches.In this study,we obtained a large number of OBPs and CSPs genes in the antennae of of C.sasakii,providing a basic guarantee for further study on the molecular mechanisms of peripheral olfactory proteins of this insect.