绿眼赛茧蜂转录组分析及嗅觉相关蛋白基因的鉴定

Transcriptome sequencing and identification of the Zele chlorophthalmus olfactory related protein gene

【目的】绿眼赛茧蜂Zele chlorophthalmus是草地螟Loxoste gesticticalis幼虫重要天敌之一。本研究通过构建绿眼赛茧蜂触角转录组数据库,挖掘其与嗅觉相关的蛋白基因,为更好的利用绿眼赛茧蜂防治草地螟发挥其生防潜能提供理论依据。【方法】以Illumina Novaseq 6000高通量测序平台为基础,将绿眼赛茧蜂触角的基因进行转录组测序、组装序列,以及完成其生物信息学的研究分析,并对绿眼茧赛蜂触角的相关嗅觉基因做鉴定分析。【结果】成功构建绿眼赛茧蜂转录组数据库,数据库中的unigenes序列为65228条,N50为3882 bp。使用BLAST软件将绿眼赛茧蜂触角unigenes序列各自和Pfam、Swiss-Prot、NR、COG、KEGG、GO权威数据库进行对比,并完成基因功能相关注释,共注释基因数为18662条,占总数的28.61%。其中,NR数据库获得的注释最多,占总数的24.61%,为15863条,KEGG数据库获得的注释最少,为9612条(14.91%),其他依次为Pfam数据库注释数据库12164条(18.86%)、COG数据库注释15584条(24.17%)、GO数据库注释11634条(18.05%),Swiss-Prot数据库注释达到11634条,为总数的18.86%。借助GO数据库对unigenes的注释,其功能供分为三大类,可以细分49个分支,主要包括分子功能、细胞组分以及生物学过程。通过注释基因功能对嗅觉相关基因进行筛选,共发现151条和嗅觉有关的蛋白基因,包括3个感觉神经元膜蛋白基因、22个离子型受体基因、23个味觉受体基因、83个气味受体基因、6个化学感受蛋白基因、14个气味结合蛋白基因。【结论】成功收集了绿眼赛茧蜂触角转录组相关数据,并对与嗅觉相关的蛋白进行鉴定分析,为深入研究基因功能及嗅觉分子机制提供理论依据。

[Objectives]Zele chlorophthalmus is an important natural enemy of Loxoste gesticticalis larvae.Constructing the Z.chlorophthalmus antenna transcriptome database and mining this for olfactory-related protein genes provides a theoretical basis for optimizing the potential of this species as a biocontrol agent.[Methods]The Illumina Novaseq 6000 high-throughput sequencing platform,transcriptome sequencing and sequence assembly were used to sequence Z.chlorophthalmus antennae genes,after which bioinformatics research and analysis were used to identify associated olfactory genes.BLAST software analysis of antennal unigene sequences was compared with that of authoritative databases(Pfam,Swiss-Prot,NR,COG,KEGG and GO)and complete gene function related annotations.[Results]A transcriptome database was successfully constructed.A total of 65228,unigenes were sequenced with an N50 of 3882 bp.A total of 18662 genes were annotated,accounting for 28.61%of the total.Among these,the NR database had the most(15863)annotations,accounting for 24.61%of the total.The KEGG database had the least(9612)annotations comprising 14.91%of the total,the Pfam database had 12164(18.86%),the COG database had 15584(24.17%),the GO database had 11634(18.05%)and the Swiss-Prot database had 11634,18.86%of the total.Unigene functions could be divided into three categories based on GO database annotations,which could be further subdivided into 49 branches.These categories were mainly molecular functions,cellular components,and biological processes.Screening of olfactory-related genes by annotating gene function revealed a total of 151 protein genes related to olfaction,including 3 sensory neuron membrane protein(SNMP)genes,22 ionotropic receptor(IR)genes,23 gustatory receptor(GR)genes,83 odorant receptor(OR)genes,6 chemosensory protein(CSP)genes and 14 odorant binding protein(OBP)genes.[Conclusion]The Z.chlorophthalmus antenna transcriptome was successfully sequenced and antennal proteins related to olfaction identified and analyzed.These findings provide a theoretical basis for in-depth study of the function of olfactory genes and the molecular mechanisms underlying olfaction.