太平洋牡蛎HSP70基因SNPs开发及其与温度相关性分析

Genic SNPs Development of HSP70 Genes in the Pacific Oyster and Correlation Analysis with Temperature

利用高通量二代测序联合先进的高分辨率熔解曲线技术,在牡蛎HSP70基因中开发并验证SNP标记,获43个能够分型的SNP标记,其中33个转换类型SNP,10个颠换类型SNP标记。随机挑选8个SNP位点在大连及厦门共计116个样本中进行分型分析,8个标记位点均符合H-W平衡,最小等位基因MAF在0.037 3~0.495 7之间。有2个标记基因型在不同地域群体间具有显著差异,O_H70-20标记的T等位基因在南方群体中的频率更高,表现出更高温度耐受性(P=0.025,OR=1.809,95%CI:1.074~3.044)。O_H70-41位点T等位基因频率在大连群体中明显高于厦门群体(P=0.003,OR=0.394,95%CI:0.210~0.733),该位点的T等位基因携带个体则表现出低温耐受性。NGS-HRM策略为牡蛎SNP标记开发提供简洁、高效的方法。

It has been certified that the HSP70gene were extremely expanded in Crassostrea gigas,which may have crucial roles in protecting cells against stresses.In this study,next generation sequencing(NGS)and high resolution melting(HRM)were employed to scan and develop single nucleotide polymorphisms(SNPs)of HSP70gene of pacific oyster C.gigas.There were43SNPmarkers developed,among which,33were transition and10were transversion.Eight SNPs were selected randomly for validation with116samples.All the selected markers were selected with H-W balance,while the minimum allele frequency(MAF)were between0.0373～0.4957.Significant differences of genotype/allele were found in two SNPs.Higher frequency of allele T was found in Dalian population compared to Xiamen population(P=0.025,OR=1.809,95%CI:1.074-3.044)in O_H70-20.However,in O_H70-41,more allele T was found in Xiamen population compared to Dalian population(P=0.003,OR=0.394,95%CI:0.210-0.733).NGS-HRM strategy provides simple and efficient method for SNP developing in C.gigas.The SNPs developed in this study would provide clues for further studies on stress resistance of C.gigas,and that could also be used in molecular assistant breeding of oyster.