中国对虾NAGase基因SNP标记的筛选及与耐高pH性状的关联分析

Correlation between the SNP of NAGase gene and susceptibility to high pH stress in Fenneropenaeus chinensis

中文关键词: 中国对虾  NAGase基因  耐高pH性状  SNP位点

英文关键词: Fenneropenaeus chinensis  NAGase gene  High pH resistance  SNP site

中文摘要:

      为探究N-乙酰-β-D-氨基葡萄糖苷酶基因(NAGase)对中国对虾(Fenneropenaeus chinensis)耐高pH性状的影响，本研究采用直接测序法及实时荧光定量PCR法检测FcNAGase的多态性，并分析其与中国对虾耐高pH性状的相关性。通过直接测序，在FcNAGase上共检测到29个潜在的SNP位点，分布频率为1.13/100 bp，其中，内含子的突变频率为0.35/100 bp，外显子的突变发生频率为0.78/100 bp。通过荧光定量PCR方法在中国对虾耐高pH性状分离群体中共检测到14个具有多态性的SNP位点，其中，观测杂合度(Ho)为0.2581~0.9390，期望杂合度(He)为0.0595~0.4490，多态信息含量(PIC)为0.0574~0.3731；利用卡方检验进行关联性分析，发现2个与中国对虾耐高pH性状显著相关的位点(P<0.05)，分别为P12 (P=0.036)和P27 (P=0.018)，本研究结果可为中国对虾分子标记辅助育种研究提供基础数据和方法。

英文摘要:

      To explore the effect of the beta-N-acetylglucosaminidase (NAGase) gene on the high pH tolerance of Fenneropenaeus chinensis, the single nucleotide polymorphism (SNP) of FcNAGase and its correlation with the high pH tolerance of F. chinensis were determined by direct sequencing and real-time fluorescent quantitative PCR. The results showed that 29 potential SNP loci were detected on the FcNAGase gene by direct sequencing, the distribution frequency was 1.13/100 bp, in which the mutation frequency of introns was 0.35/100 bp, and that of exons was 0.78/100 bp. Fourteen SNP loci were detected in the high pH tolerance population of F. chinensis using fluorescence quantitative PCR. The observed heterozygosity (Ho) ranged from 0.2581 to 0.9390, the expected heterozygosity (He) ranged from 0.0595 to 0.4490, and the polymorphism information content (PIC) ranged from 0.0574 to 0.3731. The chi-square test result revealed that two SNP loci were related to high pH tolerance traits (P<0.05), namely P12 (P=0.036) and P27 (P=0.018). The results of this experiment provided basic data and methods for molecular marker assisted breeding of F. chinensis.