| 摘要: |
| 为分析谷胱甘肽硫-转移酶M 基因(glutathione S-transferases M, GSTM)与鱼类低温耐受性的相关性, 本实验运用PCR-SSCP 技术研究了130 尾斑马鱼(Danio rerio)GSTM 基因5'UTR、3'UTR 和第一内含子序列的单核苷酸多态性(Single nucleotide polymorphisms, SNPs), 同时分析了筛选到的基因型与其低温耐受性状的关联性。结果显示, 在其5'UTR 区域检测到AB、BC、AC 3 种基因型个体, 共A、B、C 3个等位基因, 其观测杂合度和期望杂合度分别是1.000 和0.570, 多态信息含量为0.472, 所检测群体在该座位偏离了Hardy-Weinberg 平衡; 第1 内含子中检测到DD、DE 和EE 3 种基因型个体, 共D、E 两个等位基因, 其观测杂合度和期望杂合度分别为0.408 和0.477, 多态信息含量为0.362, 群体在该座位符合Hardy-Weinberg 平衡。3'UTR 区域中没有发现多态性。上述2 个SNP 座位与斑马鱼低温耐受性状的关联分析结果表明, 5'UTR 区 3 种基因型与低温耐受性状均没有显著相关性(χ2 = 4.029, P>0.05)。而第1 内含子3 种基因型与低温耐受性状显著相关(χ2 = 8.498, P< 0.05): DD 基因型在耐低温群体中占优势(50.00%), 并表现为对受低温胁迫斑马鱼的保护性因素(OR = 0.520, 95% CI = 0.255–1.061),而DE 基因型在不耐低温群体中占优势(51.31%), 表现为低温胁迫下斑马鱼的危险因素(OR = 3.012,95% CI = 1.413–6.419)。研究结果为GSTM基因SNPs 位点与斑马鱼低温耐受性能关联分析提供了依据,也将为海水经济鱼类抗寒标记筛选育种提供参考。 |
| 关键词: 斑马鱼(Danio rerio) GSTM PCR-SSCP SNPs 低温耐受相关 |
| DOI：10.11759/hykx20140304002 |
| 分类号: |
| 基金项目:国家973 计划资助项目(2010CB126304); 国家863 计划资助项目(2012AA10A408, 2012AA092203); 国家科技基础条件平台建设运行项目-水产种质资源平台运行服务(2006DKA30470017) |
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| Correlation analysis between polymorphisms of GSTM gene and cold-tolerance trait in zebrafish, Danio rerio |
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| Abstract: |
| In order to explore the correlation between Glutathione S-transferases M (GSTM) gene and cold tolerance traits in fish, the single nucleotide polymorphisms (SNPs) of GSTM gene in 130 zebrafish were investigated using PCR-SSCP method, and the association between the polymorphisms and cold tolerance traits was also analyzed. In the 5'UTR region, AB, BC and AC genotypes were detected, and there were A, B and C alleles. The observed heterozygosity and expected heterozygosity of this locus were 1.000 and 0.570, respectively. The polymorphism information content was 0.472 and the locus in this zebra stock was significantly deviated from Hardy-Weinberg equilibrium. In intron 1 of the GSTM gene, three genotypes (denoted DD, DE and EE) were found, and there were D and E alleles. The observed heterozygosity and expected heterozygosity of this locus were 0.408 and 0.477, respectively. The polymorphism information content was 0.362 and the genotypes distribution fitted to the Hardy-Weinberg equilibrium. And no polymorphism site was identified in 3'UTR region. The correlation between different genotypes and cold-tolerance traits of zebrafish was analyzed by Pearson’s chi-square test. The results indicated that there was no significant correlation between the genotypes of 5'UTR and cold-tolerance traits (χ2 = 4.029, P > 0.05). While the genotypes of intron 1 were significantly associated with the cold-tolerance traits (χ2 = 8.498, P < 0.05). Among them, the DD genotype was dominant in the cold-tolerance group (50.00%) and performed as a protective factor for zebrafish under cold stress (OR = 0.520, 95% CI = 0.255–1.061). The DE genotype was dominant in the cold-sensitive group (51.31%) and acted as a risk factor for zebrafish enduring cold stress (OR = 3.012, 95% CI = 1.413–6.419). These results could provide a basis for a further research on GSTM gene in association with cold tolerance in zebrafish, and a reference for the Marker Assistant Selection Breeding in marine commercial fish culture. |
| Key words: Danio rerio GSTM PCR-SSCP SNPs association to cold tolerance |
