皂角豆象成虫触角转录组及嗅觉相关基因分析  

Analysis of the antennal transcriptome and olfaction-related genes in Megabruchidius dorsalis(Coleoptera:Chrysomelidae)adults

作　　者：彭启艳 潘秀奎 张苏芳 武承旭 杨茂发[3,4] PENG Qi-Yan;PAN Xiu-Kui;ZHANG Su-Fang;WU Cheng-Xu;YANG Mao-Fa(College of Forestry,Guizhou University,Guiyang 550025,China;Key Laboratory of Forest Protection of National Forestry and Grassland Administration,Research Institute of Forest Ecology,Environment and Nature Conservation,Chinese Academy of Forestry,Beijing 100091,China;Guizhou Provincial Key Laboratory for Agriculture,Pest Management of the Mountainous Region,Institute of Entomology,Guizhou University,Guiyang 550025,China;College of Tobacco Science,Guizhou University,Guiyang 550025,China)

机构地区：[1]贵州大学林学院,贵阳550025 [2]中国林业科学研究院森林生态环境与自然保护研究所,国家林业和草原局森林保护学重点实验室,北京100091 [3]贵州大学昆虫研究所,贵州省山地农业病虫害重点实验室,贵阳550025 [4]贵州大学烟草学院,贵阳550025

出　　处：《昆虫学报》2024年第8期1039-1049,共11页Acta Entomologica Sinica

基　　金：贵州省科技计划项目(黔科合基础-ZK[2022]一般120);贵州省林业局基础科研项目(黔林合(2021)06);贵州省特色林业产业科研项目(GZMC-ZD20202098)。

摘　　要：【目的】本研究旨在建立皂角豆象Megabruchidius dorsalis雌雄成虫触角转录组数据库,挖掘皂角豆象嗅觉相关基因。【方法】采用高通量测序平台Illumina HiSeq对皂角豆象雌雄成虫触角进行转录组测序、序列组装、功能注释及差异表达基因分析;通过qRT-PCR技术测定和验证皂角豆象雌雄成虫触角中部分高丰度表达基因的表达量。【结果】皂角豆象雌雄成虫触角转录组共获得42 053条unigenes, N50长度为2 066 bp。与NR, Swiss-Prot, Pfam, eggNOG, GO和KEGG六大公共数据库比对共注释到18 039条unigenes(占43.57%),其中注释到NR数据库的unigenes数量最多,为17 687条;注释到KEGG数据库的unigenes数量最少,为9 779条。依据注释信息分析,鉴定出183个皂角豆象候选嗅觉相关基因,包括25个气味结合蛋白(odorant binding protein, OBP)基因、 3个化学感受蛋白(chemosensory protein, CSP)基因、 126个气味受体(odorant receptor, OR)基因(包括125个典型OR基因和1个非典型OR基因)、 8个离子型受体(ionotropic receptor, IR)基因、 18个味觉受体(gustatory receptor, GR)基因和3个感觉神经元膜蛋白(sensory neuron membrane protein, SNMP)基因。通过比较皂角豆象雌雄成虫触角转录组,共筛选出357个差异表达基因,其中包含8个OR基因和1个IR基因;这357个差异表达基因中,152个基因在雌性触角中高表达,205个基因在雄性触角中高表达。此外,qRT-PCR验证结果显示,6个嗅觉相关基因(MdorCSP3,MdorIR2,MdorIR6,MdorGR10,MdorSNMP2和MdorSNMP3)在皂角豆象雌成虫触角中高表达,4个嗅觉相关基因(MdorOBP15,MdorOBP22,MdorORco和MdorGR8)在雄成虫触角中高表达。【结论】本研究获得了皂角豆象成虫触角转录组数据,并鉴定筛选出候选嗅觉相关基因,结果为进一步研究皂角豆象的基因功能及嗅觉感受机制奠定分子基础。【Aim】This study aims to establish the antennal transcriptome database of female and male adults of Megabruchidius dorsalis and explore olfaction-related genes.【Methods】The high-throughput sequencing platform Illumina HiSeq was used to perform antennal transcriptome sequencing,sequence assembly,functional annotation and differentially expressed gene analysis of female and male adults of M.dorsalis.The expression levels of some highly expressed genes in the antennae of female and male adults of M.dorsalis were assayed and confirmed by qRT-PCR.【Results】A total of 42053 unigenes with the N50 length of 2066 bp were obtained from the antennal transcriptome of female and male adults of M.dorsalis.A total of 18039 unigenes(43.57%)were annotated against six major public databases NR,Swiss-Prot,Pfam,eggNOG,GO and KEGG,among them the largest number of unigenes(17687)were annotated into the NR database,and the minimum number of unigenes(9779)were annotated into the KEGG database.By further functional annotation and identification,183 candidate olfaction-related genes of M.dorsalis including 25 odorant binding protein(OBP)genes,three chemosensory protein(CSP)genes,126 odorant receptor(OR)genes(including 125 typical OR genes and one atypical OR gene),eight ionotropic receptor(IR)genes,18 gustatory receptor(GR)genes and three sensory neuron membrane protein(SNMP)genes were identified.After comparison of the antennal transcriptome of female and male adults,357 differentially expressed genes including eight OR genes and one IR gene were screened.Of the 357 differentially expressed genes,152 genes were highly expressed in the antennae of females and 205 in the antennae of males.In addition,qRT-PCR results showed that six olfaction-related genes(MdorCSP3,MdorIR2,MdorIR6,MdorGR10,MdorSNMP 2 and MdorSNMP 3)were highly expressed in the female adult antennae of M.dorsalis,while four olfaction-related genes(MdorOBP15,MdorOBP22,MdorORco and MdorGR 8)were highly expressed in the male adult antennae.【Conclusion】In this stud

关 键 词：皂角豆象 触角转录组 高通量测序 嗅觉相关基因 QRT-PCR 

分 类 号：Q966[生物学—昆虫学]