尼罗罗非鱼ghrelin基因的多态性及其与生长性状相关SNP位点的筛选

THE POLYMORPHISM OF GHRELIN GENE OF OREOCHROMIS NILOTICUS AND IDENTIFICATION OF ITS SNP LOCI ASSOCIATED WITH THE GROWTH TRAITS

  • 摘要: 为了研究尼罗罗非鱼(Oreochromis niloticus)生长激素促分泌素基因(ghrelin)的多态性及其与生长的相关性, 研究以两个尼罗罗非鱼群体(快长群体和基础群体)的DNA样本各40份为模板, 通过PCR扩增和测序获得ghrelin基因序列。通过Dnasp v5和MEGA 5.0分析序列多态性、筛选有效SNP 位点; 采用Snapshot法对两个群体子代ghrelin基因中SNP位点进行基因分型, 然后分析SNP位点基因型与生长性状的相关性。结果表明, 快长群体ghrelin基因中的单核苷酸变异位点数(S)比基础群体要少, 而核苷酸多态性(Pi)和平均核苷酸差异数(K)要略高于基础群体。共筛得3个有效SNP 位点(S1、S2和S3), 均分布于第1个内含子中。遗传结构分析表明, 3个SNP 位点在两个群体的子代中均为低度多态性位点(PIC0.25), 但处于Hardy-Weinberg平衡(P0.05);快长群体子代中3个SNP 位点的观测杂合度、期望杂合度和多态信息含量等遗传多样性参数均小于基础群体子代的相应值, 3个SNP 位点的遗传多样性参数、基因型和基因频率在同一群体中高度一致, SNP 位点之间完全连锁。两个群体子代中3个SNP 位点处的优势基因型相同, 但快长群体子代中优势基因型频率要明显大于基础群体子代中相应基因型频率。对两个群体子代的生长性状与SNP基因型进行关联性分析的结果表明,尼罗罗非鱼个体的多项生长指标(体重、体长、体高、头长和尾柄高等)在不同基因型中存在显著差异(S1:GG AG, S2:TT AT, S3:AA AT)(P0.05)。D1双倍型(S1:GG, S2:TT, S3:AA)所对应的尼罗罗非鱼个体的多项生长指标(体重、体长、体高、头长和尾柄高等)显著高于D2双倍型(S1:AG, S2:AT, S3:AT)。以上结果表明, 尼罗罗非鱼ghrelin基因3个SNP 位点完全连锁, D1双倍型与快长性状密切相关, 可作为尼罗罗非鱼分子标记辅助育种的候选标记。

     

    Abstract: To better understand the polymorphism of ghrelin gene and its correlation with the growth performance of Nile tilapia (Oreochromis niloticus), we obtained the sequences of ghrelin gene by using PCR and sequencing, from DNA samples of two Nile tilapia populations. The two groups are the fast-growing population and the base population, and we used 40 DNA samples from each population. Dnasp v5 and MEGA 5.0 software were employed for the polymorphism analysis and the effective screening of SNP loci. We applied the snapshot method for SNP genotyping of the offspring of the two populations, and then analyzed the correlation between the growth index and the genotypes of SNP. We found that the base population had more SNP than the fast-growing population, but the average numbers of nucleotide differences (K) and nucleotide diversity (Pi) of the latter were higher than those of the former. There were three valid SNP loci in ghrelin gene and they were all located in the first intron. The genetic structure analysis demonstrated that the three SNP loci in the offspring of the two populations were low polymorphism loci (PIC0.25), but in hardy-weinberg equilibrium (P0.05). The genetic diversity parameters (observed heterozygosity, expected heterozygosity and polymorphism information content, etc) of the three SNP in the offspring of the fast-growing population were lower than those of the base population. The three SNP loci were the same in the genetic diversity parameters, the frequencies of alleles and genotypes, thus had complete genetic linkage. The offspring of the two populations had the same dominant genotypes of the three SNP loci. The offspring of the fast-growing population exhibited significantly higher frequencies of dominant genotypes than those of the base population. Moreover, we analyzed the correlation between the genotypes of SNP loci and the growth traits of the offspring of the two populations. There were significant differences between different genotypes (S1:GG AG, S2:TT AT, S3:AA AT; P0.05) in the growth traits including the body weight, the body length, the body height, the head length and the caudal peduncle depth. The growth traits (body weight, body length, body height, head and tail handle height) of D1 diplotype (S1: GG, S2: TT, S3:AA) were significantly higher than those of D2 diplotype (S1:AG, S2:AT, S3:AT). These results suggested that D1 diplotype of ghrelin gene was closely related to the growth performance of Nile tilapia, therefore it could be used as a potential molecular marker for better breeding.

     

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