苏尼特母羊GDF9基因多态性及其与产羔性状的关联分析

doi:10.12160/j.issn.1672-5190.2026.03.009

摘要/Abstract

摘要： [目的] 探究苏尼特母羊生长分化因子9（GDF9）基因启动子区与第一外显子的单核苷酸多态性（SNP）及其与产羔性状的关联性,筛选与苏尼特母羊高繁殖力相关的分子标记,为苏尼特母羊繁殖性状遗传改良、分子标记辅助育种及肉羊规模化养殖效益提升提供数据支撑。[方法] 选取95只健康苏尼特母羊作为试验材料,其中产单羔个体48只、产双羔个体47只,采集血液样本并采用血液基因组DNA提取试剂盒提取总DNA;参照绵羊GDF9基因序列设计特异性PCR引物,对GDF9基因启动子区及第一外显子区域进行PCR扩增,通过Sanger测序技术完成基因分型;利用MegAlign、Chromas软件进行序列拼接、比对与基因型判定,统计各突变位点的基因型频率、等位基因频率,计算群体遗传参数（纯合度Ho、杂合度He、有效等位基因数Ne、多态信息含量PIC）,采用卡方检验分析各位点Hardy-Weinberg平衡状态,并解析GDF9基因多态性与苏尼特母羊产羔性状的关联关系。[结果] 在苏尼特母羊GDF9基因中成功检测到5个SNP多态位点,分别为chr5：41769149、chr5：41768729、chr5：41768538、chr5：41768521、chr5：41768463。群体遗传学分析结果显示,chr5：41769149、chr5：41768538、chr5：41768521、chr5：41768463位点的多态信息含量（PIC）均小于0.25,属于低度多态位点;chr5：41768729位点PIC值介于0.25~0.5,为中度多态位点。卡方平衡检验表明,chr5：41768521与chr5：41768729位点显著偏离Hardy-Weinberg平衡状态（P<0.05）,其余3个位点均处于平衡状态（P>0.05）。性状关联分析显示,chr5：41768521位点优势基因型为CC,优势等位基因为C,双羔羊该等位基因频率（0.93）显著高于单羔羊（0.84）;chr5：41768729位点优势基因型为AA,优势等位基因为A,双羔羊该等位基因频率（0.84）显著高于单羔羊（0.73）,上述2个位点多态性与苏尼特母羊产羔性状呈显著相关;其余3个位点多态性与产羔性状无显著关联。[结论] 苏尼特母羊GDF9基因的chr5：41768521和chr5：41768729位点多态性与产羔性状显著关联,可作为苏尼特母羊高繁殖力性状选育的有效分子标记。研究结果丰富了绵羊繁殖性状分子遗传研究内容,为苏尼特母羊繁殖性能提升、分子育种体系构建及肉羊产业高质量发展提供了参考。

关键词: 苏尼特母羊, GDF9 基因, 产羔性状, SNP多态性, 分子标记

Abstract: [Objective] To explore the single nucleotide polymorphisms （SNPs） in the promoter region and the first exon of the growth differentiation factor 9（GDF9） gene in Sonid ewes and its association with litter size traits, and to screen molecular markers related to high fecundity of Sonid ewes, aiming to provide data support for genetic improvement of reproductive traits of Sonid ewes, molecular marker-assisted breeding and improvement of large-scale breeding efficiency of mutton sheep. [Methods] A total of 95 healthy Sonid ewes were selected as experimental animals, including 48 single-lambing individuals and 47 twin-lambing individuals. Blood samples were collected and total DNA was extracted using a blood genomic DNA extraction kit. Specific PCR primers were designed according to the GDF9 gene sequence of sheep. The promoter region and the first exon region of GDF9 gene were amplified using PCR, and the genotyping was performed using Sanger sequencing. MegAlign and Chromas software were used for sequence assembly, alignment and genotype determination. The genotype frequency and allele frequency of each mutation site were counted, and the population genetic parameters （homozygosity Ho, heterozygosity He, effective number of alleles Ne, polymorphism information content PIC） were calculated. Chi-square test was used to analyze the Hardy-Weinberg equilibrium state of each locus, and the association between GDF9 gene polymorphisms and litter size traits of Sonid ewes was analyzed. [Results] Five SNP polymorphic loci were successfully detected in the GDF9 gene of Sonid ewes, including chr5: 41769149, chr5: 41768729, chr5: 41768538, chr5: 41768521 and chr5: 41768463, respectively. The results of population genetics analysis showed that the polymorphic information content （PIC） of chr5: 41769149, chr5: 41768538, chr5: 41768521 and chr5: 41768463 was less than 0.25, which were low polymorphic loci; the PIC value of chr5: 41768729 locus was between 0.25 and 0.5, which was a moderately polymorphic locus. Chi-square equilibrium test showed that the loci chr5: 41768521 and chr5: 41768729 significantly deviated from Hardy-Weinberg equilibrium （P<0.05）, while the remaining three loci were in equilibrium （P>0.05）. Trait association analysis showed that the dominant genotype of chr5: 41768521 was CC and the dominant allele was C, and the allele frequency of twin-lambing ewes （0.93） was significantly higher than that of single-lambing ewes （0.84）. The dominant genotype of chr5: 41768729 locus was AA and the dominant allele was A, and the allele frequency of twin-lambing ewes （0.84） was significantly higher than that of single-lambing ewes （0.73）. The polymorphisms of the above two loci were significantly associated with litter size traits of Sonid ewes and the remaining three loci were not significantly associated with litter size traits. [Conclusion] The polymorphisms of chr5: 41768521 and chr5: 41768729 loci of GDF9 gene in Sonid ewes are significantly associated with litter size traits, which can be used as effective molecular markers for the breeding of high fecundity traits in Sonid ewes. The results enrich the molecular genetic research of sheep reproductive traits, and provide a reference for the improvement of reproductive performance of Sonid ewes, the construction of molecular breeding system and the high-quality development of mutton sheep industry.

Key words: Sonid ewes, GDF9 gene, litter size traits, SNP polymorphism, molecular markers

中图分类号:

特日格勒, 刘宇, 付绍印, 何小龙. 苏尼特母羊GDF9基因多态性及其与产羔性状的关联分析[J]. 畜牧与饲料科学, 2026, 47(3): 69-75.

Terigele, LIU Yu, FU Shaoyin, HE Xiaolong. Polymorphisms of the GDF9 Gene and Its Association Analysis with Litter Size Traits in Sonid Ewes[J]. Animal Husbandry and Feed Science, 2026, 47(3): 69-75.

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