大蜡螟触角转录组及嗅觉相关基因分析
Analysis of the antennal transcriptome and olfaction-related genes of the greater wax moth Galleria mellonella (Lepidoptera: Pyralidae)
杨 爽；赵慧婷；徐 凯；郭丽娜；杜亚丽；李新宇； 苏文婷；冯宇佳
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DOI：10.7679/j.issn.2095-1353.2019.139
作者单位：云南省农业科学院蚕桑蜜蜂研究所，蒙自 661101；山西农业大学动物科技学院， 太谷 030801；山西农业大学生命科学学院，太谷 030801
中文关键词：大蜡螟；触角转录组；高通量测序；基因注释；嗅觉相关基因；差异表达基因
英文关键词：Galleria mellonella; antennal transcriptome; high-throughput sequencing; gene annotation; olfaction-related genes; differentially expressed genes
中文摘要：【目的】 大蜡螟Galleria mellonella Linnaeus是世界范围内蜂群中普遍存在的害虫，给世界养蜂业带来了巨大的危害。本研究旨在建立大蜡螟触角转录组数据库，挖掘大蜡螟嗅觉相关基因，为今后利用嗅觉基因靶标防治大蜡螟提供分子生物学信息数据。【方法】 利用高通量测序平台（Illumina HiSeqTM 2500）对大蜡螟触角进行转录组测序和组装，通过筛选转录组数据库，鉴定出嗅觉相关基因。【结果】 成功构建了大蜡螟触角转录组，经序列拼接获得188 278条unigenes，平均长度为781 bp，N50为1 161 bp。使用BLAST软件将大蜡螟触角unigenes序列与7大公共数据库比对，共注释到108 047条unigenes，其中NR数据库注释的unigenes最多，为78 746条，占41.82%，且NR注释的大蜡螟unigenes中与家蚕Bombyx mori类似基因最多，为18.4%。将unigenes与GO数据库比对发现，共有69 794条unigenes注释到GO数据库中，按照其所参与的生物过程、细胞组分和分子功能对其进行GO功能分类，共含有55个亚类，其中参与结合和催化功能及代谢过程的unigenes比例较大；KEGG代谢途径分析表明，14 699条unigenes形成231条代谢通路，其中被注释在信号传导通路中的unigenes最多，为2 401条，占16.33%。进一步基因注释分析，鉴定得到226个候选嗅觉相关基因，包括52个气味结合蛋白（Odorant binding proteins，OBPs）基因，36个化学感受蛋白（Chemosensory proteins，CSPs）基因，80个气味受体（Odorant receptors，ORs）基因，56个离子型受体（Ionotropic receptors，IRs）基因和2个感觉神经元膜蛋白（Sensory neuron membrane proteins，SNMPs）基因；通过比较雌、雄大蜡螟转录组鉴定出114个差异表达基因，包括5个嗅觉相关基因（OBP8、OBP17、CSP3、CSP34和OR15）；114个差异表达基因中，66个基因在雌蛾触角中高表达，48个基因在雄蛾触角中高表达。【结论】 本研究获得了大蜡螟触角转录组数据，并鉴定出候选嗅觉相关基因，结果为进一步研究大蜡螟的基因功能及嗅觉感受机制奠定分子基础。
英文摘要：[Objectives] The greater waxmoth (Galleria
mellonella) is a ubiquitous pest of honeybee colonies and causes
significant economic losses to the global apicultural industry. The objective
of this study is to establish the antennal transcriptome database of G. mellonella and explore
olfaction-related gene data, which
could provide information useful facilitating the prevention and control of G. mellonella using olfaction-related gene targeting. [Methods] The antennal
transcriptome of G. mellonella adults
was sequenced using an Illumina HiSeqTM 2500 platform, analyzed
bioinformatically and successfully
assembled. [Results] In total, 188 278 unigenes with a mean
length of 781 bp and an N50 length of 1 161 bp were obtained. Of these unigenes,
108 047 unigenes were annotated using a BLAST search based on seven public
databases. 78 746 unigenes (41.82%) were annotated to the NR database. G. mellonella unigenes had the highest
similarity (18.4%) to those of Bombyx
mori. 69 794 unigenes were divided into 55 branches and three categories
(biological processes, cellular components and molecular function) using Gene
Ontology (GO), and many were found to be related to binding, catalytic activity
and metabolic processes. In the KEGG database, 14 699 unigenes were assigned to
231 known metabolic pathways. Among these, 2 401 unigenes (16.33%) are involved
in the signal transduction pathway. By further analyzing transcriptome data,
226 olfaction-related genes of G.
mellonella including 52 odorant binding protein (OBP) genes, 36
chemosensory protein (CSP) genes, 80 odorant receptor (OR) genes, 56 ionotropic
receptor (IR) genes and 2 sensory neuron membrane protein (SNMP) genes, were
identified. After comparison of the transcriptome of male and female antennae,
114 differentially expressed genes (DEGs) including 5 olfaction-related genes (OBP8, OBP17, CSP3, CSP34 and OR15) were screened. Of these DEGs, 66 genes were highly expressed
in the antennae of females and 48 in the antennae of males. [Conclusion] This study acquired the antennal
transcriptome data of G. mellonella and identified olfaction-related genes. The results provide a foundation for
the further study of gene functions and the molecular mechanisms of olfactory
reception in G. mellonella.