摘要
【目的】化学感受蛋白(chemosensory proteins,CSPs)是一类小分子的可溶性蛋白,在昆虫中发挥多种作用。本研究旨在组装中国北方主要草原害虫之一——亚洲小车蝗Oedaleus asiaticus的触角转录组,鉴定出化学感受蛋白基因以及分析其在成虫不同组织中的表达水平。【方法】利用RNA-Seq对亚洲小车蝗成虫触角进行转录组测序和组装;通过筛选转录组数据库、克隆及测序,鉴定出化学感受蛋白基因;应用q PCR分析CSP基因在成虫不同组织(触角、去除触角和口器的头、上唇、去掉下唇须的下唇、下唇须、下颚须、胸部、跗节、翅和腹部)中的表达模式。【结果】成功构建了亚洲小车蝗成虫触角转录组,共获得61 629个unigenes,平均长度为733 nt,总长度和N50分别为45 175 449和1 130 nt。其中26 064个unigenes(42. 29%)注释到6个数据库(NR,NT,Swiss-Prot,KEGG,COG和GO)。通过Blast验证、克隆和测序鉴定出17个CSP基因; BlastP和系统发育分析表明亚洲小车蝗CSPs (OasiCSPs)与东亚飞蝗Locusta migratoria CSPs (LmigCSPs)和沙漠蝗Schistocerca gregaria CSPs(SgreCSPs)关系最密切。q PCR分析表明,8个CSP基因在成虫不同组织中的表达水平存在显着差异,特别是,Oasi CSP8在下唇须和下颚须中高表达,而OasiCSP11和Oasi CSP13在触角中表达量最高。OasiCSP15在化学感受器官(触角、上唇、去掉下唇须的下唇、下唇须、下颚须和跗节)中的表达量远高于非化学感受器官(去掉触角和口器的头部、胸部、翅和腹部)中的表达量,而OasiCSP12在几乎所有测定的组织中具有相似的表达分布。【结论】OasiCSPs在亚洲小车蝗化学感受和发育过程中可能起着多种作用,这些结果为进一步研究这些化学感受蛋白在亚洲小车蝗中的功能奠定了基础。
【Aim】Chemosensory proteins(CSPs) are soluble carrier proteins that may play multiple roles in insects. The objective of this study is to assembly the antennal transcriptomes of Oedaleus asiaticus,one of the most important grasshopper pests in northern China,to identify the chemosensory protein genes,and to analyze their tissue expression profiles. 【Methods 】 The antennal transcriptomes of the O. asiaticus adults were sequenced and assembled by RNA-Seq. The CSP genes were identified by screening the transcriptome data,and their expression levels in different adult tissues(antenna,head without antennae and mouthparts,labrum,labium without labial palps,labial palp,maxillary palp,thorax,tarsus,wing,and abdomen) were analyzed by qPCR. 【Results】The antennal transcriptomes of the O. asiaticus adults were successfully assembled. A total of61 629 unigenes with a mean length of 733 nt were obtained,and the total length and N50 were 45 175 449 nt and 1 130 nt,respectively. Among them,26 064 unigenes(42. 29%) were annotated to six databases(NR,NT,Swiss-Prot,KEGG,COG and GO). A total of 17 putative CSP genes were identified by blasting,cD NA cloning and sequencing. BlastP best hit results and phylogenetic analysis both showed that the putative OasiCSPs were most closely related to LmigCSPs from Locusta migratoria and SgreC SPs from Schistocerca gregaria. q PCR analysis showed that there were significant differences in the expression levels of eight different CSP genes in different adult tissues. Especially,OasiCSP8 was much highly expressed in the labial palps and maxillary palps,and OasiCSP11 and OasiCSP13 had the highest expression levels in the antenna. OasiCSP15 had much higher expression levels in chemosensory tissues(antenna,labrum,labium without labial palps,labial palp, maxillary palp, and tarsus) than in non-chemosensory tissues(head without antenna and mouthpart,thorax,wing,and abdomen). However,OasiCSP12 had similar expression distribution in nearly all the tested tissues. 【Conclusion 】 The results suggest that OasiCSPs might play multiple roles in the chemosensory process and development of O. asiaticus. Our findings provide a foundation for the further functional studies of these CSPs in O. asiaticus.
引文
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