鸡Wnt3a基因组织表达差异与SNPs对鸡毛囊密度性状的遗传效应

doi:10.16431/j.cnki.1671-7236.2021.05.018

摘要/Abstract

摘要： Wnt家族成员3a （Wnt family member 3a，Wnt3a）基因是Wnt信号通路的重要成员，Wnt信号通路在动物皮肤毛囊的发生发育中具有重要作用。试验以Wnt3a为候选基因，旨在探讨鸡Wnt3a基因的组织表达差异与单核苷酸多态性（single nucleotide polymorphisms，SNPs）位点对鸡皮肤毛囊密度性状的遗传效应。用实时荧光定量PCR检测Wnt3a基因在鸡不同组织中的表达差异；用PCR扩增直接测序法对Wnt3a基因的SNPs位点进行筛查，验证和分析不同SNPs位点基因型背部毛囊密度的差异；用实时荧光定量PCR技术检测不同品种鸡皮肤组织中Wnt3a基因mRNA表达差异。结果表明，Wnt3a mRNA在皮肤组织中表达量最高，显著高于其他组织（P<0.05），其次是肝脏、睾丸、卵巢和下丘脑，心脏、胸肌、垂体和脾脏等组织表达量较低。在Wnt3a基因第2和3外显子区各筛选到1个SNP位点：g.2587569 G>A和g.2555812 T>C；在第3内含子区筛选到1个SNP位点：g.2555377 T>C。卡方检验结果显示，3个位点均极显著偏离Hardy-Weinberg平衡。群体遗传参数分析发现，g.2555812 T>C位点为中度多态，有效等位基因数为1.7，遗传变异程度较g.2587569 G>A和g.2555377 T>C位点高。SNP位点分型与毛囊密度的关联分析结果表明，g.2555377 T>C位点CC基因型个体的毛囊密度显著高于TT和CT基因型个体（P<0.05）。在毛囊密度具有显著差异的不同品种鸡的背部皮肤组织中，Wnt3a mRNA表达差异显著（P<0.05）。该研究结果为进一步深入解析Wnt3a在鸡皮肤毛囊生长发育中的作用、筛选与毛囊密度性状相关的辅助育种标记提供了基础资料。

关键词: 鸡; Wnt3a基因; 皮肤毛囊; SNP; mRNA表达

Abstract: Wnt family member 3a (Wnt3a) gene is one of the important members in the Wnt signaling pathway,and the Wnt signaling pathway plays important roles in the development of animal skin and feather follicles.In this study,Wnt3a was selected as a candidate gene to detect the mRNA expression pattern for different tissues and the genetic effect of single nucleotide polymorphisms sites (SNPs) on feather follicle density in chicken.Real-time quantitative PCR was used to detect the mRNA expression profiles among different tissues.PCR amplification and direct sequencing technologies were used to identify the SNPs of the Wnt3a gene and analyze the difference of feather follicle density among different SNP genotypes.Furthermore,the Wnt3a mRNA expression difference of skin tissue between different chicken breeds was detected by Real-time-quantitative PCR.The results showed that the mRNA expression of skin was significantly higher than that of other tissues (P<0.05),the following tissues were liver,testis,ovary and hypothalamus,and heart,chest muscle,pituitary gland and spleen were the lowest.This study identified 3 SNPs,they were g.2587569 G>A,g.2555812 T>C and g.2555377 T>C,located in exons 2 and 3,and intron 3,respectively.Chi-square test showed that the genotype distribution of these SNPs had deviated significantly from Hardy-Weinberg equilibrium.Population genetic parameters analysis showed that g.2555812 T>C was moderately polymorphic,the number of effective alleles was 1.7,which indicated that the degree of variation in the population was higher than other SNPs.The correlation analysis between SNPs and feather follicle density showed that CC genotype at g.2555377 T>C were significantly higher than that of TT and CT genotypes (P<0.05).The mRNA expression level of the Wnt3a gene in skin tissues was significantly different between varieties,which showed a significant difference in feather density (P<0.05).The results of the study would provide a reference for further analyzing the role of the Wnt3a gene in chicken skin and feather follicles development and screening of molecular markers that contribute to feather follicle density traits.

Key words: chicken; Wnt3a gene; skin feather follicle; SNP; mRNA expression

中图分类号:

S834⁺.8

束婧婷, 姬改革, 屠云洁, 章明, 巨晓军, 单艳菊, 刘一帆, 邹剑敏. 鸡Wnt3a基因组织表达差异与SNPs对鸡毛囊密度性状的遗传效应[J]. 中国畜牧兽医, 2021, 48(5): 1672-1680.

SHU Jingting, JI Gaige, TU Yunjie, ZHANG Ming, JU Xiaojun, SHAN Yanju, LIU Yifan, ZOU Jianmin. Tissue Expression Analysis of Wnt3a Gene and Genetic Effects of SNPs on Feather Follicle Density in Chicken[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(5): 1672-1680.

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