雌雄罗氏沼虾脑和胸神经节的转录组比较分析

Comparative transcriptome analysis of brain and thoracic gan⁃ glion in female and male Macrobrachium rosenbergii

  • 摘要: 【目的】基于转录组测序技术筛选出罗氏沼虾(Macrobrachium rosenbergii)雌雄样本脑和胸神经组织中的差异表达基因(DEGs),为揭示其生长发育的性别二态性及开展分子辅助育种提供理论依据。【方法】通过TRIzol提取罗氏沼虾雌雄样本脑和胸神经节组织总RNA,构建cDNA测序文库后通过Illumina HiSeq 2000平台进行转录组测序,经过滤、质量控制及拼接组装获得Unigenes,使用DESeq筛选出DEGs,并进行GO功能注释分析和KEGG信号通路富集分析。【结果】转录组数据经de novo拼接成84627条Unigenes。以P<0.05和|log2 Fold Change|>1为判断标准,发现雌雄罗氏沼虾脑组织间存在5287个DEGs,雄性组织中显著上调DEGs有2416个、显著下调DEGs有2871个;胸神经节组织间存在6211个差异表达基因,雄性组织中显著上调DEGs有3280个、显著下调DEGs有2931个。雌雄罗氏沼虾脑和胸神经节组织DEGs在GO数据库中注释到生物学过程、细胞组分和分子功能三大功能条目,分别涉及56和57个亚类,包含顶体外膜、生殖结构发育、组织形态发生、细胞过程、代谢过程、细胞组分及结合等; KEGG信号通路富集分析结果表明,DEGs主要参与RNA运输、剪接体、内质网蛋白加工、转录、翻译、折叠和降解等过程。筛选得到的TUBA1B、COX、TRY、MHC、MLC、HSP90IAGBP等10个DEGs与罗氏沼虾肌肉生长及性腺发育相关,对雌雄罗氏沼虾生长发育差异调控起重要作用。【结论】雌雄罗氏沼虾脑和胸神经节组织中的TUBA1B、COX、TRY、MHC、MLC、HSP90、TRA2、PCNA、MT-ND4IAGBP等DEGs,富集在RNA转运、内质网蛋白加工及MAPK信号通路等,参与肌肉生长和性腺发育调控,有助于罗氏沼虾生长发育性别二态性的形成。

     

    Abstract: 【Objective】 Transcriptomic sequencing technology was applied for screening differentially expressed genes (DEGs)in brain and thoracic ganglion of male and female M. rosenbergii,and to provide theoretical basis for revealing the sexual dimorphism in growth and development,and for carring out the molecular-guided breeding of M. rosenbergii.【Method】Total RNA from brain and thoracic ganglion of male and female M. rosenbergii samples were extracted using TRIzol,and cDNA sequencing libraries were constructed,and then transcriptome sequencing was performed on the Illumina HiSeq 2000 platform,and Unigenes were obtained by filtration,quality control,splicing and assembly. The DEGs were screened out by using DESeq and were analyzed for GO functional annotation and KEGG signal pathway enrichment.【Result】 The transcriptome data were spliced into 84627 Unigenes by de novo. Using P<0.05 and|log2 Fold Change|> 1 as judgment criteria,5287 DEGs were detected in the brain of male and female M. rosenbergii via comparing with that of female M. rosenbergii. Out of them,2416 DEGs showed significantly up-regulated expression,and 2871 DEGs showed significantly down-regulated expression in male tissues. A total of 6211 DEGs were detected in the thoracic ganglion of male M. rosenbergii via comparing with that of female M. rosenbergii. Out of them,3280 DEGs showed significantly up-regulated expression,and 2931 DEGs showed significantly down-regulated expression in male tissues. The DEGs that were obtained from brain and thoracic ganglion of female and male M. rosenbergii,according to GO database, were successfully annotated to three major functional entries of biological process,cellular component and molecular function,and classified into 56 and 57 subcategories respectively,including outer acrosomal membrane,reproductive structure development,tissue morphogenesis,cellular process,metabolic process,cell part and binding and so on. The DEGs,according to the results of KEGG signal pathway enrichment analysis,were mainly involved in RNA transport, spliceosome, endoplasmic reticulum protein processing,transcription,translation,protein folding,and degradation, etc. The 10 DEGs related to muscle growth and gonad development were screened out,including TUBA1B,COX, TRY, MHC,MLC,HSP90 and IAGBP,which played important roles in regulating differential growth and development of female and male M. rosenbergii.【Conclusion】DEGs such as TUBA1B,COX,TRY,MHC,MLC,HSP90,TRA2, PCNA,MT-ND4 and IAGBP in the brain and thoracic ganglion of female and male M. rosenbergii are enriched in RNA transport,endoplasmic reticulum protein processing,and MAPK signal pathway,and are involved in the regulation of muscle growth and gonad development, and contribute to the formation of sexual dimorphism in growth of M. rosenbergii.

     

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