摘要蛋用性状是鸭(Anas platyrhynchos)育种工作者长期关注的重要经济性状之一,筛选与蛋用性状相关的遗传标记对于鸭分子辅助育种具有重要意义。为探究视黄醇结合蛋白4 (retinol-binding protein 4, RBP4)基因多态性与鸭蛋用性状的关系及其分子标记,本研究以农华白鸭(Nonghua White duck)为研究对象,利用PCR与直接测序相结合的方法对RBP4基因外显子1~6及内含子1~3区域的SNP突变位点进行检测,并分析其与产蛋性能和蛋品质性状的关联性。结果显示：农华白鸭RBP4基因内含子1上第266个核苷酸位点发生了SNP突变(intron 1, g.266T>G),检测群体中存在TT、TG和GG 3种基因型,其中T为优势等位基因;群体遗传学分析表明：该位点杂合度(heterozygosity, He)、纯合度(homozygosity, Ho)、有效等位基因数(effective number of alleles, Ne)和多态信息含量(polymorphic information content, PIC)分别为0.408 7、0.407 9、1.697 7和0.277,呈中度多态(0.25<P<0.50);χ2检验结果显示：该位点不处于Hardy-Weinberg平衡状态(P<0.05);关联性分析表明：g.266T>G位点基因型与蛋白高度极显著相关(P<0.01),TT基因型蛋白高度极显著高于TG和GG基因型(P<0.01);且该突变位点可能对产蛋数、蛋壳强度和蛋重有一定影响,GG基因型个体产蛋数和蛋壳强度高于TG和TT基因型,但其蛋重低于TG和TT基因型。综上,RBP4基因内含子1上g.266T>G SNP突变位点对农华白鸭蛋品质性状有显著影响,可作为鸭蛋用性状选择的候选分子标记,这为农华白鸭的分子标记辅助育种提供了理论基础。
Abstract：Egg production trait is one of the economically important traits that have long attracted attentions from duck (Anas platyrhynchos) breeders, and screening genetic markers associated with egg trait is of great significance for molecular marker-assisted breeding in duck. To explore the association of retinol-binding protein 4 (RBP4) gene polymorphisms with duck egg trait and identify the potential molecular markers, in this study, Nonghua White ducks were used as the experimental animals to detect single nucleotide polymorphisms (SNPs) in exons 1~6 and introns 1~3 of the RBP4 gene using PCR amplification combined with direct sequencing technologies, and their correlations with laying performance and egg quality traits were also analyzed. The results showed that one SNP namely g.266T>G was identified at the 266th nucleotide within intron 1 of the RBP4 gene in Nonghua White duck, and the resulting 3 genotypes (TT, TG and GG) were present in this population, with 'T' being the dominant allele. Population genetics analysis showed that the heterozygosity (He), homozygosity (Ho), effective number of alleles (Ne) and polymorphic information content (PIC) at this mutation site was 0.408 7, 0.407 9, 1.697 7 and 0.277, respectively, being moderately polymorphic (0.25<P<0.50). Results from χ2 test showed that this SNP site was not in Hardy-Weinberg equilibrium (P<0.05). Correlation analysis showed that genotypes at g.266T>G site were extremely significantly correlated with egg-white height (P<0.01), as manifested by higher egg-white height observed in individuals with TT genotype compared to those with TG and GG genotypes (P<0.01). Besides, this SNP could have the potential to influence egg production number, eggshell strength and egg weight, because individuals with GG genotype had higher egg production number and eggshell strength but lower egg weight compared to those with TG and TT genotypes. In summary, the g.266T>G SNP in intron 1 of the RBP4 gene has a significant impact on egg quality traits in Nonghua White duck and can be exploited as a candidate molecular marker for selection of duck egg production trait. This study provides a theoretical basis for future molecular marker-assisted breeding in Nonghua White duck.
路遥, 胡深强, 唐彬铖, 欧阳清渊, 李雪健, 包德威, 卿恩华, 李亮, 王继文. RBP4基因多态性与农华白鸭蛋用性状的关联性分析[J]. 农业生物技术学报, 2021, 29(8): 1566-1573.
LU Yao, HU Shen-Qiang, TANG Bin-Cheng, OU-YANG Qing-Yuan, LI Xue-Jian, BAO De-Wei, QING En-Hua, LI Liang, WANG Ji-Wen. Association Analysis of RBP4 Gene Polymorphisms with Egg Traits in Nonghua White Duck (Anas platyrhynchos). 农业生物技术学报, 2021, 29(8): 1566-1573.
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