花椒窄吉丁触角转录组及嗅觉相关基因分析  

Analysis of the antennal transcriptome and olfaction-related genes of Agrilus zanthoxylumi(Coleoptera:Buprestidae)

作　　者：巩雪芳 谢寿安[1] 杨平 郭莉 陈迪 车显荣 王延来 吕淑杰[1] GONG Xue-Fang;XIE Shou-An;YANG Ping;GUO Li;CHEN Di;CHE Xian-Rong;WANG Yan-Lai;LÜShu-Jie(College of Forestry,Northwest A&F University,Yangling,Shaanxi 712100,China)

机构地区：[1]西北农林科技大学林学院,陕西杨陵712100

出　　处：《昆虫学报》2020年第10期1159-1170,共12页Acta Entomologica Sinica

基　　金：国家公益性行业(林业)项目(201504324);云南省林业科技推广示范项目“花椒主要病虫害绿色防控技术推广与应用示范”(云[2019]TG01号);陕西省第二批特支计划领军人才项目。

摘　　要：【目的】建立花椒窄吉丁Agrilus zanthoxylumi成虫触角转录组数据库,挖掘嗅觉相关基因,为今后研究其触角的化学感受机制及生物防控提供理论支撑。【方法】采用高通量测序平台Illumina NovaSeq 6000对花椒窄吉丁雌雄成虫触角进行转录组测序,用Trinity软件对获得的高质量reads进行序列拼接与组装;使用BLAST软件将触角转录组数据比对NR,NT,Swiss-Prot,GO,KEGG,BLASTX,eggNOG,Pfam,TmHMM,SignalP,KO,Map,BLASTP和RNAMMER公共数据库;基于初步筛选到的花椒窄吉丁候选气味结合蛋白(odorant binding protein,OBP)和化学感受蛋白(chemosensory protein,CSP)以及其他鞘翅目昆虫的同源蛋白的核苷酸序列,利用MEGA软件进行系统进化分析。运用RPKM(reads per kilobase per million mapped reads)值对嗅觉相关基因表达量进行分析。【结果】花椒窄吉丁雌雄成虫触角转录组测序共获得36209条基因和90982条转录本,其N50分别为2103和2523 bp,组装完整性较高。注释到NR数据库的基因最多(41.62%),其中与赤拟谷盗Tribolium castaneum相似基因所占比例最高(19%)。在GO数据库中比对到11614个基因,按功能分为细胞组分、分子功能与生物学进程三大类57个分支,其中分子功能大类中的结合(70.57%)与催化活性(45.51%)相关基因占比最多;KEGG代谢途径分析表明,7427条基因参与了5类代谢通路,其中涉及信号转导的基因最多,为815条;筛选到7个候选OBP基因和5个具有全长开放阅读框的CSP基因,其编码蛋白均具有化学感受蛋白家族的典型特征。系统进化分析表明,花椒窄吉丁OBPs和CSPs分别与苹果小吉丁A.mali的OBPs和CSPs氨基酸序列一致性最高。RPKM值表明,嗅觉相关基因AzanOBP1和AzanOBP2在雌成虫触角中不表达,在雄成虫触角中微量表达;AzanOBP3在雄成虫触角中高丰度表达。【结论】首次获得了花椒窄吉丁成虫触角转录组数据,筛选到了花椒窄吉丁OBP,CSP,Or,IR和SNMP等的嗅觉相关�【Aim】To establish the antennal transcriptome database of Agrilus zanthoxylumi adult and to explore target genes related to olfaction,so as to provide the theoretical basis for the future study of the chemosensory mechanism of its antenna and its biological control.【Methods】High-throughput sequencing platform Illumina NovaSeq 6000 was used for transcriptome sequencing of the antennae of female and male adults of A.zanthoxylumi.Trinity software was used for sequence splicing and assembly of the high-quality reads obtained.BLAST software was used to compare the antennal transcriptome data with public databases NR,NT,Swiss-Prot,GO,KEGG,BLASTX,eggNOG,Pfam,TmHMM,SignalP,KO,Map,BLASTP and RNAMMER.Phylogenetic analysis was carried out using MEGA software based on the nucleotide sequences of the preliminarily screened candidate odorant binding proteins(OBPs)and chemosensory proteins(CSPs)of A.zanthoxylumi,and homologous proteins from other coleopteran insects.The RPKM(reads per kilobase per million mapped reads)value was used to evaluate the expression levels of olfaction-related genes.【Results】By sequencing of the antennal transcriptome of female and male adults of A.zanthoxylumi,a total of 36209 unigenes and 90982 transcripts were obtained with the N50 of 2103 and 2523 bp,respectively,indicating the high completeness of assembly.Most unigenes(41.62%)were annotated to the NR database,with the highest proportion(19%)of genes similar to those of Tribolium castaneum.In the GO database,11614 unigenes were annotated and divided into 57 branches of three functional categories,cellular component,molecular function and biological process,among which binding(70.57%)and catalytic activity(45.51%)related unigenes in the category of molecular function were the most.The KEGG metabolic pathway analysis indicated that 7427 unigenes are involved in five categories of metabolic pathways,with the most unigenes(815)involved in signal transduction.Seven candidate OBP genes and 5 CSP genes with full-length open reading frame were

关 键 词：花椒窄吉丁 触角转录组 基因注释 气味结合蛋白 化学感受蛋白 

分 类 号：S763.38[农业科学—森林保护学]