黄芪种子小蜂气味结合蛋白基因的鉴定及表达分析  

Identification and expression analysis of odorant-binding protein gene in adult Bruchophagus huonchili(Hymenoptera:Eurytomidae)

作　　者：康婕 薛赵卓 师文丽 郑海霞[1] 牛颜冰[2] 张仙红[1] KANG Jie;XUE Zhaozhuo;SHI Wenli;ZHENG Haixia;NIU Yanbing;ZHANG Xianhong(College of Plant Protection,Shanxi Agricultural University,Taigu 030801,China;College of Life Sciences,Shanxi Agricultural University,Taigu 030801,China)

机构地区：[1]山西农业大学植物保护学院,太谷030801 [2]山西农业大学生命科学学院,太谷030801

出　　处：《植物保护》2025年第1期55-68,共14页Plant Protection

基　　金：财政部和农业农村部:国家现代农业产业技术体系建设专项(CARS-21)。

摘　　要：气味结合蛋白(odorant-binding proteins,OBPs)在黄芪种子小蜂Bruchophagus huonchili Liao et Fan识别并为害中药材黄芪种子的过程中发挥着重要作用。为探明黄芪种子小蜂的嗅觉识别机理,本研究建立了黄芪种子小蜂成虫触角转录组数据库,挖掘和鉴定嗅觉识别相关的气味结合蛋白基因。采用Illumina HiSeq平台对黄芪种子小蜂成虫触角转录组进行分析,在COG、GO、KEGG、KOG、Pfam、Swissprot、TrEMBL、eggNOG、Nr等公共数据库进行基因功能注释,根据注释信息筛选黄芪种子小蜂OBP基因;利用DNAMAN和MEGA 11.0软件分析OBP基因结构特点及进化关系;通过RT-qPCR技术检测黄芪种子小蜂雌、雄成虫触角中OBP基因的表达量及被不同挥发物诱导后的表达量。结果显示,从黄芪种子小蜂成虫触角转录组中共获得98655条unigenes,其中54670条unigenes在公共数据库被注释到,从注释结果中筛选到25个BhuoOBPs,序列比对结果显示,除BhuoOBP14、BhuoOBP19外,其余23个BhuoOBPs均包含6个保守的半胱氨酸残基,属于classic OBPs。系统进化树分析表明,大部分BhuoOBPs与丽蝇蛹集金小蜂Nasonia vitripennis、白蛾周氏啮小蜂Chouioia cunea、丽蚜小蜂Encarsia formosa、松毛虫赤眼蜂Trichogramma dendrolimi等小蜂总科昆虫的OBPs氨基酸序列聚到一起,表明它们之间亲缘关系较近。RT-qPCR结果显示,在10μg/μL的引诱剂罗勒烯诱导下,雌虫触角中BhuoOBP18和雄虫触角中BhuoOBP21表达上调最显著,分别为对照的147.4倍和615.6倍;在100μg/μL的驱避剂4-萜烯醇诱导下,雌虫触角中BhuoOBP22和雄虫触角中BhuoOBP10上调最显著,分别为对照的742.4倍和1684.4倍。本研究建立了黄芪种子小蜂成虫触角转录组数据库,鉴定并分析了BhuoOBPs基因在不同挥发物诱导下触角中的表达量,为进一步研究黄芪种子小蜂嗅觉识别机制提供了理论基础。Odorant-binding proteins(OBPs)play a crucial role in the recognition and infestation of Bruchophagus huonchili Liao et Fan to Astragalus membranaceus.To investigate the olfactory recognition mechanism,an adult antennal transcriptome database of B.huonchili was established,and OBP genes were mined and identified.Transcriptome analysis was performed using the Illumina HiSeq platform,and gene annotations were performed across multiple public databases,including COG,GO,KEGG,KOG,Pfam,Swissprot,TrEMBL,eggNOG,and Nr.OBP gene structural characteristics and evolutionary relationships were analyzed using DNAMAN and MEGA 11.0 software,while RT-qPCR was used to assess the expression of OBP genes in the antennae of female and male adult wasps and its response to different volatile inductions.A total of 98655 unigenes were obtained from the transcriptome,with 54670 unigenes annotated in the public databases.Among the 25 identified BhuoOBPs,all except BhuoOBP14 and BhuoOBP19 contained six conserved cysteine residues,belong to classic OBPs.Phylogenetic analysis indicated that most BhuoOBPs clustered with OBPs from other Chalcidoidea insects,such as Nasonia vitripennis,Chouioia cunea,Encarsia formosa,and Trichogramma dendrolimi,indicating that they are closely related to each other.RT-qPCR results showed significant upregulation of BhuoOBP18 in female and BhuoOBP21 in male antennae by 147.4-fold and 615.6-fold,respectively,under 10μg/μL ocimene induction.Under 100μg/μL terpinen-4-ol induction,BhuoOBP22 in female and BhuoOBP10 in male antennae were most significantly up-regulated,reaching 742.4-fold and 1684.4-fold,respectively.This study provides a theoretical basis for further exploration of the olfactory recognition mechanism in B.huonchili.

关 键 词：黄芪种子小蜂 转录组测序 气味结合蛋白 触角 RT-QPCR 

分 类 号：S433[农业科学—农业昆虫与害虫防治]