三疣梭子蟹低盐耐受性相关甲基化标记的开发及验证  

Development and validation of methylation markers related to low salinity tolerance in Portunus trituberculatus

作　　者：张伟仁 卢少坤[1,2] 李荣华[1,2,3] 母昌考[1,2,3] 宋微微[1,2,3] 王春琳[1,2,3] ZHANG Weiren;LU Shaokun;LI Ronghua;MU Changkao;SONG Weiwei;WANG Chunlin(Key Laboratory of Aquacultral Biotechnology,Ministry of Education,Ningbo University,Ningbo 315211,China;Collaborative Innovation Center for Zhejiang Marine High-efficiency and Healthy Aquaculture,Ningbo University,Ningbo 315211,China;Key Laboratory of Green Mariculture(Co-construction by Ministry and Province),Ministry of Agriculture and Rural Affairs,Ningbo University,Ningbo 315211,China)

机构地区：[1]宁波大学,水产生物技术教育部重点实验室,浙江宁波315211 [2]宁波大学,浙江海洋高效健康养殖协同创新中心,浙江宁波315211 [3]宁波大学,农业农村部绿色海水养殖重点实验室,浙江宁波315211

出　　处：《水产学报》2023年第8期79-86,共8页Journal of Fisheries of China

基　　金：国家重点研发计划(2022YFD2400104);国家现代农业产业技术体系专项(CARS-48);宁波大学王宽诚幸福基金。

摘　　要：为开发三疣梭子蟹低盐耐受性相关甲基化标记,采用甲基化敏感性高分辨率熔解曲线法(methylation sensitive high resolution melting curve,MS-HRM)进行了三疣梭子蟹低盐耐受性相关甲基化位点的筛选和应用。结果显示,从三疣梭子蟹转录组数据库中共开发了8个甲基化标记,其中6个标记与低盐度耐受性显著相关,在低盐度条件下表现出显著的去甲基化或甲基化,这些标记分别位于V-ATP酶(V-type proton ATPase)、CLC型氯离子通道、琥珀酸脱氢酶、NAD(P)转氢酶NAD(P)、磷酸乙醇酸性磷酸酶、NADH还原酶基因,为三疣梭子蟹耐低盐新品种的分子标记辅助选育提供候选工具。研究表明,“宁象1号”耐低盐能力显著高于野生群体,且选育群体中位点Pt-M1、Pt-M2的去甲基化率达到100%,可能有利于低盐条件下保持体内Na^(+)和Cl^(-)的平衡。In order to develop methylation markers related to low salinity tolerance in Portunus trituberculatus,in this study,methylation sensitive high resolution melting curve method(MS-HRM)was applied to screen and validate methylation loci related to low salinity tolerance.A total of 8 methylation markers were developed from the transcriptome database of P.trituberculatus,six of which were significantly related to low salinity tolerance,exhibiting a significant demethylation or methylation pattern in the low salinity condition.They were located in Vtype proton ATPase,H^(+)/Cl^(-)exchange transporter,succinate dehydrogenase,NAD(P)transhydrogenase,phosphoglycolate phosphatase,NADH dehydrogenase genes,and these markers provided useful tools for markerassisted breeding of low-salinity-tolerant strain of P.trituberculatus.The low salinity tolerance test and genotyping results of these 6 loci in wild populations and“Ningxiang-1”selective strain showed that the survival rate of“Ningxiang-1”was significantly higher than that of wild populations under low salinity conditions,and the demethylation of loci Pt-M1 and Pt-M2 in the selective breeding population was significantly increased to 100%,which may be beneficial to maintaining the balance of Na^(+)and Cl^(-)under low salinity conditions.This study provides basic data and favorable tools for genetic improvement of low salinity resistance in P.trituberculatus.

关 键 词：三疣梭子蟹 低盐 甲基化 标记 

分 类 号：S968.252[农业科学—水产养殖]