乌鳢三种性腺组织的转录组比较分析  

Comparative Transcriptome Analysis of Three Types of Gonads in Northern Snakehead(Channa argus)

作　　者：黄苏静 吴玉霞 夏威威 张啸天 赵建[1,2] 李凯彬[1] 罗青[1] 刘海洋 费树站 陈昆慈[1,2] 欧密 HUANG Su-Jing;WU Yu-Xia;XIAWei-Wei;ZHANG Xiao-Tian;ZHAO Jian;LI Kai-Bin;LUO Qing;LIU Hai-Yang;FEI Shu-Zhan;CHEN Kun-Ci;OU Mi(Pearl River Fisheries Research Institute,Chinese Academy of Fishery Sciences/Key Laboratory of Utilization and Aquaculture of Tropical and Subtropical Aquatic Resources,Ministry of Agriculture and Rural Affairs,Guangzhou 510380,China;College of Fisheries and Life Sciences,Shanghai Ocean University,Shanghai 201306,China)

机构地区：[1]中国水产科学研究院珠江水产研究所/农业农村部热带亚热带水产种质资源利用与养殖重点实验室,广州510380 [2]上海海洋大学水产与生命学院,上海201306

出　　处：《农业生物技术学报》2024年第2期379-395,共17页Journal of Agricultural Biotechnology

基　　金：财政部和农业农村部:国家现代农业产业技术体系(CARS-46);广东省基础与应用基础研究基金(2021A1515012075,2021A1515010875);广州市科技计划项目(202201010120,202102021070);中国水产科学研究院基本科研业务费专项(2023XT0202,2023TD37);广东省淡水水产种质资源库运行项目(2022-SBH-00-001)。

摘　　要：乌鳢(Channa argus)是我国重要的淡水经济鱼类,具有显著的性别二态性。为了研究乌鳢性别分化和性腺发育的分子机制,挖掘精、卵巢差异表达的功能基因和信号通路,本研究采用Illumina HiSeq 2000测序平台对6月龄乌鳢XX雌鱼(XX-F)、XY雄鱼(XY-M)和YY超雄鱼(YY-M)的性腺组织进行了转录组测序(RNA-seq)分析。性腺组织两两样品的表达量相关性分析显示,XY-M精巢和YY-M精巢具有相似的表达模式,与XX-F卵巢表达模式相差甚远,与性腺的组织学结果一致。三种性腺组织的基因差异表达分析显示,XY-M vs XX-F中有12317个差异表达基因(differentially expressed genes,DEGs),其中5427个上调、6890个下调;XY-M vs YY-M中有212个DEGs,其中59个上调、153个下调;YY-M vs XX-F中有11282个DEGs,其中5038个上调、6244个下调。利用qRT-PCR对性别分化和性腺发育相关的DEGs进行验证,发现抗缪勒管激素2型受体(anti-müllerian hormone receptor type 2,Amhr2)、抗缪勒管激素(anti-müllerian hormone,Amh)、double sex和mab3相关转录因子1(double sex and mab3-related transcription factor1,Dmrt1)、性腺体细胞衍生因子(gonadal somatic cell-derived factor,Gsdf)等10个基因在XY-M和YY-M精巢中高表达,骨形态发生蛋白15(bone morphogenetic protein 15,Bmp15)、性腺型芳香化酶基因Cyp19a1a(cytochrome P450 family 19 subfamily A member 1a)、叉头框转录因子2(forkhead transcription factor gene 2,Foxl2)等9个基因在XX-F卵巢中高表达。qRT-PCR验证结果与转录组分析结果一致,说明Amhr2、Amh、Dmrt1和Gsdf等基因与乌鳢精巢发育调控相关,Bmp15、Cyp19a1a和Foxl2等基因与乌鳢卵巢发育调控相关。另外,对DEGs进行KEGG代谢通路富集分析,发现较多的DEGs富集在性别分化和代谢相关通路,如丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)、无翅型MMTV整合位点家族成员(wingless-type MMTV integration site family,Wnt)以及转化生长因子-β(transforming growth Northern snakehead(Channa argus)is an important freshwater fish in China with significantly sexual dimorphism.In order to illuminate the molecular mechanism of sex differentiation and gonadal development of C.argus,and explore the functional genes and signaling pathways that differentially express in testis and ovary,RNA-seq was performed on gonads of 6-month-old normal XX females(XX-F),normal XY males(XY-M),and YY super-males(YY-M)by Illumina HiSeq 2000.Correlation analysis of pairwise samples of gonadal tissues indicated that similar expression patterns were found in XY-M and YY-M testis,which were different from that in XX-F ovary,and the result was consistent with the histological observation of gonads.A tremendous amounts of differentially expressed genes(DEGs)were found in the pairwise comparison among XX-F,XY-M,and YY-M gonads.12317 DEGs were obtained in XY-M testis compared with XX-F ovary,in which 5427 were up-regulated and 6890 were down-regulated.212 DEGs were obtained in XY-M testis compared with YY-M testis,in which 59 were up-regulated and 153 were downregulated.11282 DEGs were obtained in YY-M testis compared with XX-F ovary,in which 5038 were upregulated and 6244 were down-regulated.qRT-PCR was used to verify DEGs that were related to sex differentiation and gonadal development with significant differences among testis and ovary.It displayed that 10 genes had relatively high expression in testis of XY-M and YY-M,such as anti-müllerian hormone receptor type 2(Amhr2),anti-müllerian hormone(Amh),doublesex-mab3-related transcription factor1(Dmrt1)and gonadal somatic cell-derived factor(Gsdf),9 genes had relatively high expression in ovary of XX-F,including bone morphogenetic protein 15(Bmp15),cytochrome P450 family 19 subfamily A member 1a(Cyp19a1a)and forkhead transcription factor gene 2(Foxl2).The results of qRT-PCR validation were consistent with the results of the RNA-seq,illustrating that Amhr2,Amh,Dmrt1 and Gsdf genes were related to the regulation of testicular development,and Bmp15,Cyp19a1a

关 键 词：乌鳢 转录组 差异表达基因(DEG) 性别分化 性腺发育 

分 类 号：S917.4[农业科学—水产科学]