弓背青鳉两个地理群体Dmrt基因家族的鉴定与表达分析  

Identification and Expression Analysis of the Dmrt Gene Family in Two Geographical Groups of Oryzias curvinotus

作　　者：陈子阳 姚泽彬 李金蓬 邓爱萍 王中铎[1] CHEN Ziyang;YAO Zebin;LI Jinpeng;DENG Aiping;WANG Zhongduo(Key Laboratory of Aquaculture in South China Sea for Aquatic Economic Animal of Guangdong Higher Education Institutes,Fisheries College,Guangdong Ocean University,Zhanjiang,524088)

机构地区：[1]广东海洋大学水产学院南海水产经济动物增养殖重点实验室,湛江524088

出　　处：《基因组学与应用生物学》2025年第2期157-169,共13页Genomics and Applied Biology

基　　金：广东省自然科学基金面上项目(2022A1515011441)资助。

摘　　要：Dmrt(double-sex and mab-3 related transcription factor)基因家族编码的蛋白质是一类转录因子,具有保守的DM(double-sex/mab-3)结构域。在鱼类生长发育和进化中,Dmrt在多种生理过程尤其是性别决定和性腺发育方面起到至关重要的作用。弓背青鳉(Oryzias curvinotus)和日本青鳉(O.latipes)均具有XX/XY性别决定系统和雄性决定基因dmrt1-bY/dmy(DM domain gene on Y chromosome),但是Dmrt基因家族在弓背青鳉中的作用尚不清楚。本研究利用生物信息学方法对弓背青鳉Dmrt基因家族的系统进化关系、基因结构、保守基序、保守结构域以及在不同组织中基因的表达水平进行分析,用实时定量PCR(real-time quantitative PCR,RT-qPCR)方法验证转录组信息。结果显示,弓背青鳉湛江群体中鉴定出Ocu_ZJ_dmrt1、Ocu_ZJ_dmy、Ocu_ZJ_dmrt2a、Ocu_ZJ_dmrt2b.1、Ocu_ZJ_dmrt2b.2、Ocu_ZJ_dmrt3、Ocu_ZJ_dmrtA1和Ocu_ZJ_dmrtA2,弓背青鳉三亚群体中鉴定出Ocu_SY_dmrt1、Ocu_SY_dmrt2a、Ocu_SY_dmrt2b、Ocu_SY_dmrt3、Ocu_SY_dmrtA1和Ocu_SY_dmrtA2。保守结构域分析显示,14个Dmrt蛋白质序列均具有DM保守结构域。利用弓背青鳉湛江群体胚胎发育转录组以及湛江和三亚群体雌雄性腺转录组数据对Dmrt基因家族进行表达水平分析,结果显示,Dmrt基因家族在胚胎各个发育时期表达不一,在两个群体的性腺中均有表达,其中Ocu_ZJ_dmy、Ocu_ZJ_dmrt1和Ocu_SY_dmrt1在雄性高表达,Ocu_ZJ_dmrt2a和Ocu_SY_dmrt2a在雌性性腺表达更高,三亚群体未检测到dmy的表达。通过RT-qPCR验证了11个差异表达基因(differentially expressed genes,DEGs)的RNA-seq数据的可靠性。本研究为青鳉属物种Dmrt基因家族的研究提供必要的基础数据,为更好地认识Dmrt基因家族在弓背青鳉性别决定和性腺、胚胎发育中的作用提供参考。The Dmrt(double-sex and mab-3 related transcription factor)gene family is a class of transcription factors with a conserved DM(double-sex/mab-3)structural domain.In fish development and evolution,Dmrt plays a crucial role in various physiological processes,especially sex determination and gonadal development.Both Oryzias curvinotus and O.latipes have XX/XY sex determination system and male determination gene dmrt1-bY/dmy(DM domain gene on Y chromosome),but the role of Dmrt gene family in O.curvinotus is not clear.In this study,we used bioinformatics tools to analyze the phylogenetic relationship,gene structure,conserved motifs,conserved structural domains and gene expression levels in different tissues of the Dmrt gene family in O.curvinotus,and then validating transcriptome information using the real-time quantitative PCR(RT-qPCR)method.The results showed that Ocu_ZJ_dmrt1,Ocu_ZJ_dmy,Ocu_ZJ_dmrt2a,Ocu_ZJ_dmrt2b.1,Ocu_ZJ_dmrt2b.2,Ocu_ZJ_dmrt3,Ocu_ZJ_dmrtA1 and Ocu_ZJ_dmrtA2 were identified in Zhanjiang group and Ocu_SY_dmrt1,Ocu_SY_dmrt2a,Ocu_SY_dmrt2b,Ocu_SY_dmrt3,Ocu_SY_dmrtA1 and Ocu_SY_dmrtA2 in Sanya group,respectively.The conserved domain analysis showed that all 14 Dmrt protein sequences had DM conserved structural domains.The expression levels of the Dmrt gene family were analyzed using the embryonic developmental transcriptomes of the Zhanjiang group and the female and male gonads transcriptomic data of the Zhanjiang and Sanya groups of O.curvinotus,respectively,the results showed that Dmrt gene family was differentially expressed at different developmental stages of embryos and was expressed in the gonads of both groups,with Ocu_ZJ_dmy、Ocu_ZJ_dmrt1 and Ocu_SY_dmrt1 being highly expressed in males while Ocu_ZJ_dmrt2a and Ocu_SY_dmrt2a being more highly expressed in female gonads,and no dmy expression was detected in Sanya group.The reliability of the RNA-seq data of 11 differentially expressed genes(DEGs)was ve-rified by RT-qPCR.This study will provide the necessary basic data for the study of the Dmrt

关 键 词：弓背青鳉 dmy缺失群体 Dmrt基因家族 胚胎发育转录组 雌雄性腺转录组 

分 类 号：S917.4[农业科学—水产科学]