兰州鲇Dmrta1基因克隆及其组织时空表达规律  

Cloning of Dmrta1 Gene and Its Temporal and Spatial Expression in Tissue of Silurus lanzhouensis

作　　者：柳婷 李敏敏 赖章龙 刘彦斌 刘凯 肖伟 赛清云 田永华 杨立强 阮超岭 杨瑞兰 刘哲[1] 连总强[1,2,3,4,5] LIU Ting;LI Minmin;LAI Zhanglong;LIU Yanbin;LIU Kai;XIAO Wei;SAI Qingyun;TIAN Yonghua;YANG Liqiang;RUAN Chaoling;YANG Ruilan;LIU Zhe;LIAN Zongqiang(College of Animal Science and Technology,Gansu Agricultural University,Lanzhou 730070,China;Ningxia Fisheries Research Institute,Yinchuan 750001,China;Ningxia Fishery Engineering Technology Research Center,Yinchuan 750001,China;Academician Work Station of Ningxia Fishery Science and Technology,Yinchuan 750001,China;The College of Life and Geographic Sciences,Kashi University,Kashi 844000,China)

机构地区：[1]甘肃农业大学动物科学技术学院,甘肃兰州730070 [2]宁夏回族自治区水产研究所,宁夏银川750001 [3]宁夏渔业工程技术研究中心,宁夏银川750001 [4]宁夏渔业科技院士工作站,宁夏银川750001 [5]喀什大学生命与地理科学学院,新疆喀什844000

出　　处：《华北农学报》2023年第6期228-238,共11页Acta Agriculturae Boreali-Sinica

基　　金：国家重点研发计划项目(2018YFD0901202);宁夏重点研发计划项目(2017BN06)。

摘　　要：为了解Dmrta1基因的生物学特性及其在兰州鲇生长发育中的生理作用,以及为加快实现全雄兰州鲇群体生产提供理论依据和技术支撑。采用同源克隆和cDNA末端快速克隆(RACE)技术获得兰州鲇Dmrta1全长cDNA序列,通过qRT-PCR技术检测该基因在兰州鲇胚胎发育、仔稚幼鱼以及3龄雌雄异体和3龄雌雄同体不同组织中的表达特征。结果表明,兰州鲇Dmrta1基因cDNA全长1415 bp,ORF全长1158 bp,共编码385个氨基酸,包含20种氨基酸;经SMART预测显示,该基因属于典型的Dmrta亚家族蛋白,具有保守的DM和DMA结构域。同源性比对和系统进化树分析结果表明,兰州鲇与南方鲇的相似性最高(95.47%),且亲缘关系最近。组织表达结果表明,Dmrta1 mRNA在未受精卵粒及胚胎发育各个时期均有表达,而在未受精卵粒中表达最高(P<0.05);在兰州鲇孵化后40 d内不同发育时期中,Dmrta1 mRNA在同期XX个体组织中表达均显著高于XY个体,且在35 d迅速达到峰值(P<0.05);Dmrta1 mRNA在3龄雌雄异体兰州鲇雌性卵巢、鳃和雄性鳃组织中表达显著高于其他组织(P<0.05),且卵巢表达显著高于雌性和雄性鳃,除卵巢表达是精巢的47.07倍外,其他组织中不存在雌雄性别差异;3龄雌雄同体和3龄雌雄异体表达特征一致,都是卵巢表达显著高于鳃和精巢(P<0.05)。以上结果说明,Dmrta1基因属于母源基因,该基因可能在兰州鲇性别分化、卵子形成和鳃弓形成过程中发挥重要作用,在雌雄同体和雌雄异体兰州鲇性腺中的作用一致。In order to understand the biological characteristics of Dmrta1 gene and its physiological role in the growth and development of Silurus lanzhouensis,and to provide theoretical basis and technical support for the realization of full male S.lanzhouensis population production.Homologous cloning and cDNA terminal rapid cloning(RACE)techniques were used to obtain the full-length cDNA sequence of S.lanzhouensis Dmrta1.qRT-PCR was used to detect the expression characteristics of this gene in different tissues of S.lanzhouensis such as embryo development,larvae and juveniles,third instar hermaphroditic and third instar hermaphroditic.The results showed that the cDNA length of Dmrta1 gene was 1415 bp and that of ORF was 1158 bp,encoding 385 amino acids.SMART prediction showed that the gene belonged to a typical Dmrta subfamily protein with conserved DM and DMA domains.The sequence alignment and phylogenetic tree analysis showed that S.lanzhouensis and S.meridionalis had the highest similarity(95.47%)and were closely related.The results of tissue expression showed that Dmrta1 mRNA was expressed in all stages of unfertilized and embryonic development,and the highest expression was in unfertilized egg(P<0.05).In different developmental stages of S.lanzhouensis within 40 days after incubation,the expression of Dmrta1 mRNA in XX individuals was significantly higher than that in XY individuals,and rapidly reached the peak value on 35 days(P<0.05).The expression of Dmrta1 mRNA in the ovary and gills of female catfish of the third age was significantly higher than that in other tissues(P<0.05),and the expression in the ovary was significantly higher than that in the gills of male and female catfish.Except that the expression in the ovary was 47.07 times that in the sperms,there was no difference between male and female in other tissues.The expression characteristics of the third instar hermaphrodite and the third instar hermaphrodite were the same,and the egg expression was significantly higher than that of the gill and germ nes

关 键 词：兰州鲇 Dmrta1基因 基因克隆 早期生活史 雌雄同体 MRNA表达 

分 类 号：S917.4[农业科学—水产科学]