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

文献类型： 外文期刊

作者： Jing, Hao 1 ; Zhou, Liqing 2 ; Gong, Miao 2 ; Tu, Kang 5 ; Liu, Zhihong 2 ; Wu, Biao 2 ; Sun, Xiujun 2 ;

作者机构： 1.Shanghai Ocean Univ, Coll Fisheries & Life Sci, Shanghai 201306, Peoples R China

2.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, State Key Lab Mariculture Biobreeding & Sustainabl, Qingdao 266071, Peoples R China

3.Laoshan Lab, Lab Marine Fisheries Sci & Food Prod Proc, Qingdao 266237, Peoples R China

4.Jiangsu Ocean Univ, Sch Marine Sci & Fisheries, Lianyungang 222005, Peoples R China

5.Putian Inst Aquaculture Sci Fujian Prov, Putian 351100, Peoples R China

关键词： clam; reference gene; hypoxia; quantitative real-time PCR

期刊名称：JOURNAL OF OCEAN UNIVERSITY OF CHINA （ 影响因子：1.6； 五年影响因子：1.5 ）

ISSN： 1672-5182

年卷期： 2023 年 22 卷 4 期

页码：

收录情况： SCI

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 results. To 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 0.5 mg L-1 and DO 2.0 mg L-1) and normal condition (DO 7.5 mg L-1). Seven candidate reference genes were selected to evaluate the stability of their expression levels. The reference genes were evaluated by Delta Ct, BestKeeper, NormFinder and geNorm, and then screened by RefFinder calculation. Under hypoxic stress of 0.5 mg L-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, respectively. For hypoxic stress of 2.0 mg L-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, respectively. At 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 muscle. The 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.

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