香港牡蛎糖原合酶与糖原磷酸化酶基因多态性研究及糖原含量性状相关分子标记挖掘

Polymorphisms of glycogen synthase and glycogen phosphorylase genes and identification of glycogen content-associated molecular markers in Crassostrea hongkongensis

香港牡蛎是我国南方沿海地区的主要经济贝类,糖原含量是衡量其品质高低的重要指标之一。本研究在转录组测序基础上,以三墩群体(SD,60只个体)和大风江群体(DFJ,90只个体)为研究对象,采用ELISA方法检测每个香港牡蛎糖原含量,并利用片段长度差异等位基因特异性PCR(Fragment length discrepant allele specific PCR,FLDAS-PCR)技术开展糖原合酶和糖原磷酸化酶基因多态性研究,开发单核苷酸多态性(Single nucleotide polymorphism,SNP)标记。结果共开发25个SNPs,其中4个SNPs(Ch_GS680,Ch_GS1248,Ch_GP992和Ch_GP1962)在实验群体中分型成功,SPSS关联分析显示SNP位点Ch_GS680与糖原含量显著关联(P<0.05),暗示其可作为香港牡蛎糖原含量性状的潜在分子标记,研究结果为高糖原含量香港牡蛎新品种的分子标记辅助选育奠定了基础。

Crassostrea hongkongensis is a primary economic shellfish in the coastal areas of southern China.Glycogen content is one of the crucial indicators used to measure its quality.Based on transcriptome sequencing,the glycogen content of each individual of C.hongkongensis collected from Sandun and Dafengjiang was assessed by enzyme linked immunosorbent assay—sandwich technique.The gene polymorphisms and single nucleotide polymorphisms(SNPs)of glycogen synthase and glycogen phosphorylase of C.hongkongensis were analyzed using the fragment length discrepant allele specific PCR technology.The results showed that a total of 25 SNPs were validated,of which 4 SNPs(Ch_gs680,Ch_gs1248,Ch_gp992,and Ch_gp1962)were successfully genotyped in the experimental population.However,only SNP locus Ch_GS680 was significantly associated with glycogen content(P<0.05),as shown by association analysis with SPSS software,indicating that SNP Ch_GS680 may be a potential molecular marker for the glycogen content feature of C.hongkongensis.These findings provide a foundation for the selection of new C.hongkongensis varieties with high glycogen content using molecular markers.