低盐胁迫下三疣梭子蟹Aldh基因的表达及DNA甲基化分析  

Expression and DNA methylation analysis of Aldh gene in swimming crab(Portunus trituberculatus)under low salinity stress

作　　者：郭俊阳 吕建建[2,3] 孙东方 周现法 刘萍 高保全[2,3] GUO Junyang;L Jianjian;SUN Dongfang;ZHOU Xianfa;LIU Ping;GAO Baoquan(Key Laboratory of Marine Biological Resources and Environment of Jiangsu Province,College of Marine Science and Fisheries,Jiangsu Ocean University,Lianyungang 222005,China;National Key Laboratory of Mariculture Biobreeding and Sustainable Goods,Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao 266071,China;Function Laboratory for Marine Fisheries Science and Food Production Processes,Qingdao National Laboratory for Marine Science and Technology,Qingdao 266235,China)

机构地区：[1]江苏海洋大学海洋科学与水产学院,江苏省海洋生物资源与环境重点实验室,江苏省海洋生物技术重点实验室,江苏连云港222005 [2]中国水产科学院黄海水产研究所,海水养殖生物育种与可持续产出全国重点实验室,山东青岛266071 [3]青岛海洋科学与技术国家实验室,海洋渔业科学与食物产出过程功能实验室,山东青岛266235

出　　处：《大连海洋大学学报》2024年第5期773-780,共8页Journal of Dalian Ocean University

基　　金：国家重点研发计划(2022YFD2400104);国家自然科学基金(42076116,41876187);财政部和农业农村部:国家现代农业产业技术体系资助(CARS-48);中国水产科学研究院基本科研业务费(2023TD50)。

摘　　要：为探究三疣梭子蟹(Portunus trituberculatus)耐低盐性状的表观遗传调控机制,对三疣梭子蟹(体质量为35.5 g±2.8 g)醛脱氢酶(Aldh)的基因结构、相似性、系统进化、组织表达,以及低盐胁迫下的基因表达、丙二醛(MDA)含量和DNA甲基化模式进行了研究。结果表明:三疣梭子蟹Aldh基因的保守结构域与中华绒螯蟹的保守结构域相似性最高;实时荧光定量PCR分析显示,Aldh基因在三疣梭子蟹各个组织中均有表达,其中在鳃中的表达水平最高,低盐胁迫下该基因的表达水平显著降低(P<0.05),MDA含量显著增加(P<0.05);亚硫酸氢盐测序法(BS-PCR)分析显示,低盐胁迫72 h后该基因的甲基化水平显著增加,与基因表达呈负相关性(P<0.05)。研究表明,低盐环境下DNA甲基化可能通过抑制Aldh基因表达,导致体内MDA浓度升高从而造成组织损伤。本研究结果为三疣梭子蟹耐低盐性状的表观遗传调控机制解析提供了重要参考依据。In order to explore the epigenetic regulatory mechanism of low-sainity tolerance traits in swimming crab(Portunus trituberculatus),structure,similarity,phylogeny,and tissue expression of aldehyde dehydrogenase(Aldh)gene were investigated in swimming crab with body weight of(35.5±2.8)g and gene expression,malondialdehyde(MDA)content,and DNA methylation patterns were probed in the swimming crab exposed to salinity of 11 mg/L(LC50 in 72 h from the pretest).The results showed that swimming crab had the maximal similarity in conserved structural domains in Aldh in swimming crab to those found in Chinese mitten handed crab Eriocheir sinensis.Real time PCR analysis revealed that the Aldh gene was expressed in all the test tissues,with the maximal expression level in the gills.Moreover,significant decrease in the expression level of the Aldh gene in the swimming crab exposed to low salinity stress(P<0.05),coupled with a noticeable increase in MDA content.The BS-PCR results indicated a significant increase in the methylation level of the Aldh gene in swimming crab exposed to low salinity stress following for 72 hours with negative correlation with gene expression(P<0.05).It was found that DNA methylation of the Aldh gene was likely involved in regulation of the response to low salinity stress in the gill tissue of swimming crab,primarily by suppressing of the expression of the Aldh gene.The finding offers valuable reference for understanding of the epigenetic regulatory mechanisms underlying the low salinity tolerance trait in swimming crab.

关 键 词：三疣梭子蟹 低盐胁迫 DNA甲基化 基因表达 

分 类 号：S917.4[农业科学—水产科学]