Evaluation of Reference Genes for Quantitative Real-Time PCR Analysis in Manila Clam Ruditapes philippinarum Under Hypoxic Stress

作     者：JING Hao ZHOU Liqing GONG Miao TU Kang LIU Zhihong WU Biao SUN Xiujun JING Hao;ZHOU Liqing;GONG Miao;TU Kang;LIU Zhihong;WU Biao;SUN Xiujun

作者机构：College of Fisheries and Life ScienceShanghai Ocean UniversityShanghai 201306China State Key Laboratory of Mariculture Biobreeding and Sustainable GoodsYellow Sea Fisheries Research InstituteChinese Academy of Fishery SciencesQingdao 266071China Laboratory for Marine Fisheries Science and Food Production ProcessesLaoshan LaboratoryQingdao 266237China School of Marine Science and FisheriesJiangsu Ocean UniversityLianyungang 222005China Putian Institute of Aquaculture Science of Fujian ProvincePutian 351100China 

出 版 物：《Journal of Ocean University of China》 (中国海洋大学学报（英文版）)

年 卷 期：2023年第22卷第4期

页      面：1059-1067页

核心收录：

学科分类：0908[农学-水产] 09[农学] 

基　　金：supported by research grants from the Science and Technology Innovation Program of the Laoshan Laboratory(No.LSKJ202203803) the National Natural Science Foundation of China(No.32273107) supported by the Central Public-Interest Scientific Institution Basal Research Fund,Yellow Sea Fisheries Research Institute,CAFS(No.20603022022001) the project of Putian Science and Technology Department(No.2021NJJ002) the Shinan District Science and Technology Plan Project(No.2022-2-026-ZH) 

主　　题：clam reference gene hypoxia quantitative real-time PCR 

摘      要：Quantitative real-time PCR(qRT-PCR)has been widely used for gene expression analysis,and selection of reference genes is a key point to obtain accurate *** find out optimal reference genes for qRT-PCR in Manila clam Ruditapes philippinarum in response to hypoxia,different tissues were used and compared to evaluate the stability of candidate reference genes under low oxygen stress(DO 0.5mgL^(−1) and DO 2.0mgL^(−1))and normal condition(DO 7.5mgL^(−1)).Seven candidate reference genes were selected to evaluate the stability of their expression *** reference genes were evaluated by Delta Ct,BestKeeper,NormFinder and geNorm,and then screened by RefFinder *** hypoxic stress of 0.5mgL^(−1),the most suitable reference gene for gill and hepatopancreas was RPL31,and the optimal reference genes for axe foot and adductor muscle were TUB and HIS,*** hypoxic stress of 2.0mgL^(−1),the most stable reference gene for gill and hepatopancreas was RPL31,and the optimal reference genes for axe foot and adductor muscle were RPS23 and EF1A,*** the normal condition,HIS and EF1A were identified as the optimal internal reference genes in gill and hepatopancreas respectively,and GFRP2 was the best internal reference gene for axe foot and adductor *** present findings will provide important basis for the selection of reference genes for qRT-PCR analysis of gene expression level in bivalves under hypoxic stress,which might be helpful for the analysis of other molluscs too.