不同繁殖状态的梨小食心虫触角转录组分析及嗅觉相关基因鉴定  

Analysis of adult antennal transcriptome and identification of olfaction-related genes in oriental fruit moth Grapholita molesta with different reproductive status

作　　者：曹炜晔 王萌 李俊峰 许子昕 王怡 马瑞燕[1] 李捷 孔维娜[1] Cao Weiye;Wang Meng;Li Junfeng;Xu Zixin;Wang Yi;Ma Ruiyan;Li Jie;Kong Weina(College of Plant Protection,Shanxi Agricultural University,Taiyuan 030031,Shanxi Province,China;College of Horticulture,Shanxi Agricultural University,Taiyuan 030031,Shanxi Province,China)

机构地区：[1]山西农业大学植物保护学院,太原030031 [2]山西农业大学园艺学院,太原030031

出　　处：《植物保护学报》2024年第4期882-898,共17页Journal of Plant Protection

基　　金：中央引导引导地方科技发展资金资助项目(YDZJSX2022A034);国家现代农业(梨)产业技术体系资助项目(CARS-28);山西省科技创新人才团队专项(202304051001006);山西省重点研发计划(202302140601011)。

摘　　要：为明确不同繁殖状态对雌、雄成虫触角上嗅觉基因的影响,以梨小食心虫Grapholita molesta为研究对象,利用高通量测序RNA-seq技术分析未交配、受孕、产卵的雌成虫和未交配、交配1次、交配2次的雄成虫的差异表达基因,并进行实时荧光定量PCR(quantitative real-time PCR,qRTPCR)验证。结果显示:从梨小食心虫雌、雄成虫触角转录组中共获得19581条unigene序列,其中16946条unigene序列在至少1个数据库中获得功能注释,从中筛选出了161个嗅觉相关基因。其中,气味结合蛋白(odorant binding protein,OBP)和化学感受蛋白(chemosensory protein,CSP)的表达量在3种状态的雄成虫间存在差异;交配2次雄成虫的信息素结合蛋白(pheromone binding protein,PBP)的表达量高于未交配雄成虫;与未交配或受孕状态相比,产卵雌成虫的气味受体(odorant receptor,OR)和OBP表达量上调;与受孕或产卵状态相比,未交配雌成虫中CSP高表达;产卵雌成虫PBP和离子型受体(ionotropic receptor,IR)的表达量均高于未交配雌成虫。对差异表达OR,OBP和PBP基因的qRT-PCR验证结果与转录组测序结果一致,表明本研究转录组数据可靠。To determine the effects of different reproductive states on the olfactory genes in the antennae of adult oriental fruit moth Grapholita molesta,differentially expressed genes(DEGs)in unmated,gravid,and ovipositional females,as well as unmated,once-mated,and twice-mated males,were analyzed using high-throughput RNA-seq technology.The DEGs were verified by quantitative real-time PCR(qRT-PCR).A total of 19581 unigenes were obtained,with 16946 sequences functionally annotated in at least one database,from which 161 olfactory-related genes were identified.Odorant binding proteins(OBPs)and chemosensory proteins(CSPs)showed differential expressions among the three male reproductive states.Pheromone binding protein(PBP)expression was higher in twice-mated males compared to unmated males.In females,compared to unmated and gravid states,odorant receptor(OR)and OBP expressions were upregulated inovipositional females.CSP expression was higher in unmated females than in gravid or ovipositional females.Additionally,PBP and ionotropic receptor(IR)expression levels were higher in ovipositional females compared to unmated females.The qRT-PCR results for differentially expressed OR,OBP,and PBP genes were consistent with the transcriptome sequencing results,indicating the reliability of the transcriptome data in this study.

关 键 词：梨小食心虫 成虫繁殖状态 触角转录组 嗅觉差异表达基因 

分 类 号：S433[农业科学—农业昆虫与害虫防治]