三疣梭子蟹(Portunus trituberculatus)雌雄性腺的差异转录组分析
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摘要
三疣梭子蟹(Portunustrituberculatus)是我国东海海域具有重要经济价值的蟹类。为探讨雌雄梭子蟹性腺基因表达差异,挖掘与性腺发育相关基因,利用高通量测序技术对三疣梭子蟹雌雄性腺进行测序和生物信息学分析,为进一步研究其性腺发育及性别分化机制提供基础。测序结果经denovo组装共获得192848个转录本,130054个unigene。差异表达分析发现,其中101742个unigene上调,37588个unigene下调。在差异表达数据库中筛选出EcR、RXR、VTG等6个可能参与三疣梭子蟹的性腺分化发育的基因,并利用荧光定量PCR验证其在雌雄性腺中的表达情况,验证结果与转录组测序结果基本一致,为深入研究三疣梭子蟹性腺分化发育调控分子机理和相关功能基因提供了重要的基因数据资源。
Portunus trituberculatus is of great economic value.To investigate the mechanism of the gonadal development and sex differentiation,the different expression of genes in male and female gonads of the swimming crab were explored using transcriptome sequencing technique and bioinformatics.In the present study,192848 transcripts and 13054 unigenes were identified through de novo assembly.Among the differential expressed genes(DEGs),101742 were up-regulated and 37588 were down-regulated.The results of sequencing were further verified by quantitative real-time PCR(qRT-PCR).The expression of six DEGs including EcR,RXR,and VTG et al,which may be involved in gonadal development and sex determination of P.trituberculatus,were analyzed.Our results have provided clues to understand the mechanisms of gonadal development and sex differentiation in the swimming crab.
Portunus trituberculatus is of great economic value.To investigate the mechanism of the gonadal development and sex differentiation,the different expression of genes in male and female gonads of the swimming crab were explored using transcriptome sequencing technique and bioinformatics.In the present study,192848 transcripts and 13054 unigenes were identified through de novo assembly.Among the differential expressed genes(DEGs),101742 were up-regulated and 37588 were down-regulated.The results of sequencing were further verified by quantitative real-time PCR(qRT-PCR).The expression of six DEGs including EcR,RXR,and VTG et al,which may be involved in gonadal development and sex determination of P.trituberculatus,were analyzed.Our results have provided clues to understand the mechanisms of gonadal development and sex differentiation in the swimming crab.
引文
水燕,史永红,徐增洪等,2013.眼柄切除快速诱导甲壳动物生长发育研究进展.广东农业科学,40(21):124-126,135
孙颖民,宋志乐,严瑞深等,1984.三疣梭子蟹生长的初步研究.生态学报,4(1):59-64
朱韬,朱冬发,邱锡尔等,2016.三疣梭子蟹fps克隆及其在卵巢发育中的表达分析.生物学杂志,33(3):10-14,19
吴旭干,汪倩,楼宝等,2014.育肥时间对三疣梭子蟹卵巢发育和营养品质的影响.水产学报,38(2):170-182
陈石林,吴旭干,成永旭等,2007.三疣梭子蟹胚胎发育过程中主要生化组成的变化及其能量来源.中国水产科学,14(2):229-235
贾复龙,孟宪亮,刘萍等,2016.三疣梭子蟹细胞Cdk7基因克隆及其在卵巢发育中的表达.中国水产科学,23(5):1032-1040
高保全,刘萍,李健等,2008.三疣梭子蟹(portunus trituberculatus)不同地理种群内自繁和种群间杂交子一代生长性状的比较.海洋与湖沼,39(3):291-296
谢熙,朱冬发,崔晓雨等,2013.三疣梭子蟹FAMeT基因克隆及其在蜕皮周期中的表达水平.水产学报,37(7):994-1001
韩坤煌,张子平,王艺磊等,2010.Cyclin-CDK-CKI及UPP参与生殖调控及在甲壳动物性腺发育中的研究进展.生物技术通报,(7):48-54
管卫兵,宣富君,戴小杰等,2009.东海三疣梭子蟹精子发生及精荚形成.中国水产科学,16(2):173-182
Cui J,Wu L T,Chan S M et al,2013.cDNA cloning and mRNAexpression of retinoid-X-receptor in the ovary of the shrimp Metapenaeus ensis.Molecular Biology Reports,40(11):6233-6244
Ding L Y,Jin M,Sun P et al,2017.Cloning,tissue expression of the fatty acid-binding protein(Pt-FABP1)gene,and effects of dietary phospholipid levels on fabp and vitellogenin gene expression in the female swimming crab Portunus trituberculatus.Aquaculture,474:57-65
Durica D S,Wu X H,Anilkumar G et al,2002.Characterization of crab EcR and RXR homologs and expression during limb regeneration and oocyte maturation.Molecular and Cellular Endocrinology,189(1-2):59-76
Fanjul-Moles M L,2006.Biochemical and functional aspects of crustacean hyperglycemic hormone in decapod crustaceans:Review and update.Comparative Biochemistry and Physiology Part C:Toxicology&Pharmacology,142(3-4):390-400
Girish B P,Swetha C,Reddy P S,2015.Expression of RXR,EcR,E75 and VtG mRNA levels in the hepatopancreas and ovary of the freshwater edible crab,Oziothelphusa senex senex(Fabricius,1798)during different vitellogenic stages.The Science of Nature,102(3-4):20
Gong J,Huang C C,Shu L et al,2016.The retinoid X receptor from mud crab:new insights into its roles in ovarian development and related signaling pathway.Scientific Reports,6:23654
Hopkins P M,Durica D,Washington T,2008.RXR isoforms and endogenous retinoids in the fiddler crab,Uca pugilator.Comparative Biochemistry and Physiology Part A:Molecular&Integrative Physiology,151(4):602-614
Jia X W,Chen Y D,Zou Z H et al,2013.Characterization and expression profile of Vitellogenin gene from Scylla paramamosain.Gene,520(2):119-130
Jiang H X,Li X L,Sun Y H et al,2018.Molecular and functional characterization of nucleoside diphosphate kinase(nm23)gene in oriental river prawn Macrobrachium nipponense during ovarian development.Aquaculture Research,49(3):1219-1231
Jin X K,Li W W,He L et al,2011.Molecular cloning,characterization and expression analysis of two apoptosis genes,caspase and nm23,involved in the antibacterial response in Chinese mitten crab,Eriocheir sinensis.Fish&Shellfish Immunology,30(1):263-272
Li K,Chen L Q,Zhou Z L et al,2006.The site of vitellogenin synthesis in Chinese mitten-handed crab Eriocheir sinensis.Comparative Biochemistry and Physiology Part B:Biochemistry and Molecular Biology,143(4):453-458
Li L,Li X J,Wu Y M et al,2017.Vitellogenin regulates antimicrobial responses in Chinese mitten crab,Eriocheir sinensis.Fish&Shellfish Immunology,69:6-14
Lv J J,Liu P,Gao B Q et al,2014.Transcriptome analysis of the Portunus trituberculatus:de novo assembly,growth-related gene identification and marker discovery.PLoS One,9(4):e94055
Meusy J J,Payen G G,1988.Female reproduction in Malacostracan Crustacea.Zoological Science,5(2):217-265
Ozsolak F,Milos P M,2011.RNA sequencing:advances,challenges and opportunities.Nature Reviews Genetics,12(2):87-98
Sarika S N,Gopinathan A,2014.DNA binding domain of retinoid receptor gene(RXR)from a field crab inhabiting the Indian peninsula.Indian Journal of Biotechnology,13(1):52-56
Tang J,Zhu D F,Cui X Y et al,2014.Molecular cloning,characterization and expression analysis of the retinoid Xreceptor in the swimming crab,Portunus trituberculatus(Miers,1876)(Decapoda,Portunidae).Crustaceana,87(3):312-327
Wang W,Wu X G,Liu Z J et al,2014.Insights into hepatopancreatic functions for nutrition metabolism and ovarian development in the crab Portunus trituberculatus:gene discovery in the comparative transcriptome of different hepatopancreas stages.PLoS One,9(1):e84921
孙颖民,宋志乐,严瑞深等,1984.三疣梭子蟹生长的初步研究.生态学报,4(1):59-64
朱韬,朱冬发,邱锡尔等,2016.三疣梭子蟹fps克隆及其在卵巢发育中的表达分析.生物学杂志,33(3):10-14,19
吴旭干,汪倩,楼宝等,2014.育肥时间对三疣梭子蟹卵巢发育和营养品质的影响.水产学报,38(2):170-182
陈石林,吴旭干,成永旭等,2007.三疣梭子蟹胚胎发育过程中主要生化组成的变化及其能量来源.中国水产科学,14(2):229-235
贾复龙,孟宪亮,刘萍等,2016.三疣梭子蟹细胞Cdk7基因克隆及其在卵巢发育中的表达.中国水产科学,23(5):1032-1040
高保全,刘萍,李健等,2008.三疣梭子蟹(portunus trituberculatus)不同地理种群内自繁和种群间杂交子一代生长性状的比较.海洋与湖沼,39(3):291-296
谢熙,朱冬发,崔晓雨等,2013.三疣梭子蟹FAMeT基因克隆及其在蜕皮周期中的表达水平.水产学报,37(7):994-1001
韩坤煌,张子平,王艺磊等,2010.Cyclin-CDK-CKI及UPP参与生殖调控及在甲壳动物性腺发育中的研究进展.生物技术通报,(7):48-54
管卫兵,宣富君,戴小杰等,2009.东海三疣梭子蟹精子发生及精荚形成.中国水产科学,16(2):173-182
Cui J,Wu L T,Chan S M et al,2013.cDNA cloning and mRNAexpression of retinoid-X-receptor in the ovary of the shrimp Metapenaeus ensis.Molecular Biology Reports,40(11):6233-6244
Ding L Y,Jin M,Sun P et al,2017.Cloning,tissue expression of the fatty acid-binding protein(Pt-FABP1)gene,and effects of dietary phospholipid levels on fabp and vitellogenin gene expression in the female swimming crab Portunus trituberculatus.Aquaculture,474:57-65
Durica D S,Wu X H,Anilkumar G et al,2002.Characterization of crab EcR and RXR homologs and expression during limb regeneration and oocyte maturation.Molecular and Cellular Endocrinology,189(1-2):59-76
Fanjul-Moles M L,2006.Biochemical and functional aspects of crustacean hyperglycemic hormone in decapod crustaceans:Review and update.Comparative Biochemistry and Physiology Part C:Toxicology&Pharmacology,142(3-4):390-400
Girish B P,Swetha C,Reddy P S,2015.Expression of RXR,EcR,E75 and VtG mRNA levels in the hepatopancreas and ovary of the freshwater edible crab,Oziothelphusa senex senex(Fabricius,1798)during different vitellogenic stages.The Science of Nature,102(3-4):20
Gong J,Huang C C,Shu L et al,2016.The retinoid X receptor from mud crab:new insights into its roles in ovarian development and related signaling pathway.Scientific Reports,6:23654
Hopkins P M,Durica D,Washington T,2008.RXR isoforms and endogenous retinoids in the fiddler crab,Uca pugilator.Comparative Biochemistry and Physiology Part A:Molecular&Integrative Physiology,151(4):602-614
Jia X W,Chen Y D,Zou Z H et al,2013.Characterization and expression profile of Vitellogenin gene from Scylla paramamosain.Gene,520(2):119-130
Jiang H X,Li X L,Sun Y H et al,2018.Molecular and functional characterization of nucleoside diphosphate kinase(nm23)gene in oriental river prawn Macrobrachium nipponense during ovarian development.Aquaculture Research,49(3):1219-1231
Jin X K,Li W W,He L et al,2011.Molecular cloning,characterization and expression analysis of two apoptosis genes,caspase and nm23,involved in the antibacterial response in Chinese mitten crab,Eriocheir sinensis.Fish&Shellfish Immunology,30(1):263-272
Li K,Chen L Q,Zhou Z L et al,2006.The site of vitellogenin synthesis in Chinese mitten-handed crab Eriocheir sinensis.Comparative Biochemistry and Physiology Part B:Biochemistry and Molecular Biology,143(4):453-458
Li L,Li X J,Wu Y M et al,2017.Vitellogenin regulates antimicrobial responses in Chinese mitten crab,Eriocheir sinensis.Fish&Shellfish Immunology,69:6-14
Lv J J,Liu P,Gao B Q et al,2014.Transcriptome analysis of the Portunus trituberculatus:de novo assembly,growth-related gene identification and marker discovery.PLoS One,9(4):e94055
Meusy J J,Payen G G,1988.Female reproduction in Malacostracan Crustacea.Zoological Science,5(2):217-265
Ozsolak F,Milos P M,2011.RNA sequencing:advances,challenges and opportunities.Nature Reviews Genetics,12(2):87-98
Sarika S N,Gopinathan A,2014.DNA binding domain of retinoid receptor gene(RXR)from a field crab inhabiting the Indian peninsula.Indian Journal of Biotechnology,13(1):52-56
Tang J,Zhu D F,Cui X Y et al,2014.Molecular cloning,characterization and expression analysis of the retinoid Xreceptor in the swimming crab,Portunus trituberculatus(Miers,1876)(Decapoda,Portunidae).Crustaceana,87(3):312-327
Wang W,Wu X G,Liu Z J et al,2014.Insights into hepatopancreatic functions for nutrition metabolism and ovarian development in the crab Portunus trituberculatus:gene discovery in the comparative transcriptome of different hepatopancreas stages.PLoS One,9(1):e84921