中华蜜蜂幼虫肠道的高表达基因分析
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摘要
中华蜜蜂(中蜂)是中国特有的本土蜜蜂资源,也是养蜂生产中的常用蜂种之一,具有重要的生态和经济价值。本研究利用RNA-seq技术对中蜂5和6日龄幼虫肠道进行深度测序,得到的有效读段(clean reads)数分别为29290630与26636038,Q20分别为98.45%和98.38%。GO富集分析结果显示,Ac5中的高表达基因(HEGs)富集在44个GO term,其中基因富集数最多的为代谢进程(753 unigenes),其次是催化活性(679 unigenes)与结合(657 unigenes);Ac6中的HEGs富集在44个GO term,其中基因富集数最多的为代谢进程(792 unigenes),其次是结合(760 unigenes)和催化活性(744 unigenes)。KEGG代谢通路(pathway)富集分析结果显示,Ac5的HEGs富集在129个pathway,其中基因富集数最多的是核糖体(103 unigenes)、内质网蛋白质加工(68 unigenes)和碳代谢(67 unigenes);Ac6的HEGs富集在129个代谢通路,其中基因富集数最多的是核糖体(103 unigenes)、氧化磷酸化(88 unigenes)及内质网蛋白质加工(68 unigenes)。进一步对HEGs进行Venn分析,结果显示,二者共有的HEGs为2 015个,特有的HEGs分别为219和491个。研究结果不仅可提供中蜂幼虫肠道发育过程中的基因表达谱数据,也为分子水平上深入研究中蜂幼虫肠道发育提供重要信息。
Apis cerana cerana is not only a specific honeybee species resource in China,but also a frequently-used honeybee species in apiculture. In the present study,the 5-( Ac5) and 6-day-old( Ac6) larval guts of A. c. cerana were sequenced using RNA-seq. After filtration,29290630 and 26636038 clean reads with a Q20 of 98. 45% and98. 38% were obtained in Ac5 and Ac6. GO enrichment analysis results showed that the highly expressed genes( HEGs) in Ac5 were enriched in 44 GO terms,among them metabolic process( 753 unigenes),catalytic activity( 679 unigenes) and binding( 657 unigenes) were the largest groups; the HEGs in Ac6 were enriched in 44 GO terms,and the mostly enriched one was metabolic process( 792 unigenes),followed by binding( 760 unigenes) andcatalytic activity( 744 unigenes). KEGG enrichment analysis results suggested that the HEGs of Ac5 were enriched in 129 pathways,among them ribosome( 103 unigenes) was the largest group,followed by protein processing in endoplasmic reticulum( 68 unigenes) and carbon metabolism( 67 unigenes); the HEGs of Ac6 were enriched in 129 pathways,the mostly enriched ones were ribosome( 103 unigenes),oxidative phosphorylation( 88 unigenes) as well as protein processing in endoplasmic reticulum( 68 unigenes). Our findings can not only offer the gene expression profiles during the developmental process of the larval gut of A. c. cerana at transcriptome level,but also provide key information for further investigation of the larval gut's development.
Apis cerana cerana is not only a specific honeybee species resource in China,but also a frequently-used honeybee species in apiculture. In the present study,the 5-( Ac5) and 6-day-old( Ac6) larval guts of A. c. cerana were sequenced using RNA-seq. After filtration,29290630 and 26636038 clean reads with a Q20 of 98. 45% and98. 38% were obtained in Ac5 and Ac6. GO enrichment analysis results showed that the highly expressed genes( HEGs) in Ac5 were enriched in 44 GO terms,among them metabolic process( 753 unigenes),catalytic activity( 679 unigenes) and binding( 657 unigenes) were the largest groups; the HEGs in Ac6 were enriched in 44 GO terms,and the mostly enriched one was metabolic process( 792 unigenes),followed by binding( 760 unigenes) andcatalytic activity( 744 unigenes). KEGG enrichment analysis results suggested that the HEGs of Ac5 were enriched in 129 pathways,among them ribosome( 103 unigenes) was the largest group,followed by protein processing in endoplasmic reticulum( 68 unigenes) and carbon metabolism( 67 unigenes); the HEGs of Ac6 were enriched in 129 pathways,the mostly enriched ones were ribosome( 103 unigenes),oxidative phosphorylation( 88 unigenes) as well as protein processing in endoplasmic reticulum( 68 unigenes). Our findings can not only offer the gene expression profiles during the developmental process of the larval gut of A. c. cerana at transcriptome level,but also provide key information for further investigation of the larval gut's development.
引文
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[11]CORNMAN R S,LOPEZ D,EVANS J D.Transcriptional response of honey bee larvae infected with the bacterial pathogen Paenibacillus larvae[J].PLo S One,2013,8(6):e65424.
[12]JULIE A,BARBARA MISME-AUCOUTURIER,BERNARD V,et al.Transcriptome analyses of the honeybee response to Nosema ceranae and insecticides[J].Plo S One,2014,9(3):e91686.
[13]CRISTINO A S,BARCHUUK A R,FREITAS F C,et al.Neuroligin-associated micro RNA-932 targets actin and regulates memory in the honeybee[J].Nature Communications,2014,5:5529.
[14]GLI N/SKI Z,JAROSZ J.Infection and immunity in the honey bee Apis mellifera[J].Apiacta,2001,36:12-24.
[15]HONEY BEE GENOME SEQUENCING CONSORTIUM.Insights into social insects form the genome of the honeybee Apis mellifera[J].Nature,2006,443(7114):931-949.
[16]PARK D,JUNE J W,CHOI B S,et al.Uncovering the novel characteristics of Asian honey bee,Apis cerana,by whole genome sequencing[J].BMC Genomics,2015,16:1.
[2]杨冠煌.中华蜜蜂[M].北京:中国农业科技出版社,2001.
[3]张复兴.现代养蜂生产[M].北京:中国农业大学出版社,1998.
[4]GILLIAM M.Identification and roles of non-pathogenic microflora associated with honey bees[J].FEMS Microbiology Letters,1997,155(1):1-10.
[5]JEYAPRAKASH A,HOY M A,ALLSOPP MH.Bacterial diversity in worker adults of Apis mellifera capensis and Apis mellifera scutellata(Insecta:Hymenoptera)assessed using 16S r RNA sequences[J].Journal of Invertebrate Pathology,2003,84(2):96-103.
[6]LI JL,QIN HR,WU J,et al.The prevalence of parasites and pathogens in Asian honeybees Apis cerana in China[J].PLo S One,2012,7(11):e47955.
[7]王倩,孙亮先,肖培新,等.室内人工培育中华蜜蜂幼虫技术研究[J].山东农业科学,2009(11):113-116.WANG Q,SUN L X,XIAO P X,et al.Study on technology for indoor artificial feeding of Apis cerana cerana larvae[J].Shandong Agricultural Sciences,2009(11):113-116.(in Chinese with English abstract)
[8]张曌楠,熊翠玲,徐细建,等.蜜蜂球囊菌的参考转录组de novo组装及SSR分子标记开发[J].昆虫学报,2017,60(1):34-44.ZHANG Z N,XIONG C L,XU X J,et al.De novo assembly of a reference transcriptome and development of SSR markers for Ascosphaera apis[J].Acta Entomologica Sinica,2017,60(1):34-44.(in Chinese with English abstract)
[9]MORTAZAVI A,WILLIAMS BA,MCCUE K,et al.Mapping and quantifying mammalian transcriptomes by RNA-Seq[J].Nature Methods,2008,5(7):621-628.
[10]丁桂玲.中华蜜蜂(Apis cerana)群体多样性的研究[D].北京:中国农业科学院,2006.DING G L.Study on the population diversity of Apis cerana[D].Beijing:Chinese Academy of Agricultural Sciences,2006.(in Chinese with English abstract)
[11]CORNMAN R S,LOPEZ D,EVANS J D.Transcriptional response of honey bee larvae infected with the bacterial pathogen Paenibacillus larvae[J].PLo S One,2013,8(6):e65424.
[12]JULIE A,BARBARA MISME-AUCOUTURIER,BERNARD V,et al.Transcriptome analyses of the honeybee response to Nosema ceranae and insecticides[J].Plo S One,2014,9(3):e91686.
[13]CRISTINO A S,BARCHUUK A R,FREITAS F C,et al.Neuroligin-associated micro RNA-932 targets actin and regulates memory in the honeybee[J].Nature Communications,2014,5:5529.
[14]GLI N/SKI Z,JAROSZ J.Infection and immunity in the honey bee Apis mellifera[J].Apiacta,2001,36:12-24.
[15]HONEY BEE GENOME SEQUENCING CONSORTIUM.Insights into social insects form the genome of the honeybee Apis mellifera[J].Nature,2006,443(7114):931-949.
[16]PARK D,JUNE J W,CHOI B S,et al.Uncovering the novel characteristics of Asian honey bee,Apis cerana,by whole genome sequencing[J].BMC Genomics,2015,16:1.