缢蛏糖原磷酸化酶基因多态性与糖原含量的关联分析  

The association of single nucleotide polymorphisms of the glycogen phosphorylase gene with glycogen content in the razor clam Sinonovacula constricta

作　　者：陈燕园 林志华 刘圣[3] 姚韩韩 董迎辉 Chen Yanyuan;Lin Zhihua;Liu Sheng;Yao Hanhan;Dong Yinghui(Zhejiang Key Laboratory of Aquatic Germplasm Resources,College of Biological&Environmental Sciences,Zhejiang Wanli University,Ningbo 315100,China;National Demonstration Center for Experimental Fisheries Science Education,Shanghai Ocean University,Shanghai 201306,China;Ninghai Marine Biological Seed Industry Research Institute,Zhejiang Wanli University,Ninghai 315604,China)

机构地区：[1]浙江万里学院生物与环境学院浙江省水产种质资源高效利用技术研究重点实验室,浙江宁波315100 [2]上海海洋大学水产科学国家级实验教学示范中心,上海201306 [3]浙江万里学院宁海海洋生物种业研究院,浙江宁海315604

出　　处：《海洋学报》2022年第2期77-83,共7页

基　　金：国家重点研发计划“蓝色粮仓”科技创新课题(2018YFD0901405);浙江省农业新品种选育重大科技专项(2021C02069-7);国家现代贝类产业技术体系项目(CARS-49);国家海洋水产种质资源库项目;宁波市“科技创新2025”重大专项(2019B10005)。

摘　　要：为探究缢蛏糖原磷酸化酶基因(Sc-GPH)与糖原含量的关系,本研究克隆获得Sc-GPH cDNA全长,检测其在不同组织、不同月份的表达模式,并在Sc-GPH基因编码区筛选与糖原含量关联的SNP位点。结果表明,Sc-GPH cDNA全长为3963 bp,开放阅读框为2541 bp,编码846个氨基酸;氨基酸序列多重比对和系统发育树显示,缢蛏与欧洲大扇贝、虾夷扇贝、长牡蛎等贝类亲缘关系较近,而与哺乳类、甲壳类、昆虫类亲缘关系较远。实时荧光定量PCR结果显示,Sc-GPH在8个组织中均有表达,其中在外套膜和足中表达量最高(p<0.01),推测与其糖原存储能力有关;不同月份的缢蛏外套膜和足中,Sc-GPH在8月表达量最高,而此时糖原含量较低,说明Sc-GPH的表达可能受缢蛏生殖周期的影响。以缢蛏"甬乐1号"群体为实验材料,在Sc-GPH基因编码区筛选到4个与糖原含量相关SNP位点,其中c.930T>C位点在台州野生群体中得到进一步验证,该位点为高糖原缢蛏分子标记辅助选育提供了候选标记。To clarify the relationship between the glycogen phosphorylase gene(Sc-GPH) and glycogen content in Sinonovacula constricta, the full-length cDNA sequence of Sc-GPH is obtained using RACE technology, its expression pattern in different tissues, different months are analyzed, and the SNP loci of Sc-GPH associated with glycogen content are explored. The full-length cDNA of Sc-GPH gene is 3 963 bp and the ORF is 2 541 bp, encoding 846 amino acids. The multiple alignment of amino acid sequences and phylogenetic tree show that S. constricta are closely related to shellfish such as Pecten maximus, Mizuhopecten yessoens, Crassostrea gigas, but farther to mammals, arthropodas and insects. The results of real-time quantitative PCR show that Sc-GPH gene is expressed in eight tissues, with the highest expression in mantle and foot, suggesting they are related to glycogen storage capacity. In different months, the expression levels of Sc-GPH in the mantle and foot is the highest in August, at which time the glycogen content decreased, indicating that the expression of Sc-GPH seems to be driven by reproductive cycle of S. constricta. In “YL1” population, four SNPs related to glycogen content are screened in the coding region of Sc-GPH gene, and c.930 T>C is further verified in Taizhou wild population, which provides a candidate marker for molecular marker assisted breeding of high glycogen S. constricta varieties.

关 键 词：缢蛏 糖原磷酸化酶 糖原含量 SNP 关联分析 

分 类 号：Q786[生物学—分子生物学] S917.4[农业科学—水产科学]