摘要
旨在基于基因多态性分析Wnt家族成员3a(wnt family member 3a,Wnt3a)基因遗传变异位点与崇仁麻鸡皮肤毛囊密度的相关性。本研究随机选取600只在相同环境和相同管理水平下饲养至上市日龄(120 d),健康且状况良好的崇仁麻鸡,其中原种母鸡200只,公鸡100只和B系母鸡200只,公鸡100只。屠宰后选取背部、胸部和大腿部皮肤相同的位置测量毛囊密度,通过皮肤组织切片HE染色的方法观察毛囊的形态结构并测量毛囊直径和皮肤厚度。采血提DNA用PCR扩增后直接测序的方法筛选Wnt3a基因遗传变异位点,并与鸡皮肤毛囊密度进行相关性分析。通过实时荧光定量PCR技术检测Wnt3a基因在不同皮肤部位的表达量。结果表明:1)原种和B系同性别同皮肤部位相比较,原种母鸡背部毛囊密度极显著高于B系(P<0.01),大腿部毛囊密度显著高于B系(P<0.05),背部和大腿部毛囊直径显著低于B系(P<0.05)。原种公鸡背部皮肤毛囊密度极显著高于B系(P<0.01),胸部毛囊密度极显著低于B系(P<0.01),背部和大腿部毛囊直径显著低于B系(P<0.05)。B系公鸡胸部皮肤厚度显著高于原种(P<0.05),B系公、母鸡大腿皮肤厚度均显著高于原种(P<0.05)。2)Wnt3a基因筛选出的3个SNPs位点与各部位皮肤毛囊密度进行相关性分析后发现,g.2555812 T>C和g.2555377 T>C位点的CC基因型个体背部皮肤毛囊密度显著高于其他基因型个体背部(P<0.05)。3)实时荧光定量PCR结果显示,Wnt3a基因在原种和B系鸡的3个皮肤部位的毛囊组织上都有比较高的表达量,且在原种背部的表达量显著高于在B系背部的表达量(P<0.05)。综上所述,崇仁麻鸡原种皮肤毛孔相较于B系更加细密,Wnt3a基因g.2555812 T>C位点的CC基因型和g.2555377 T>C位点的CC基因型是崇仁麻鸡皮肤毛囊的有利基因型,可选作鸡皮肤毛囊密度性状的分子标记。本研究探讨了作为毛囊生长发育的候选基因Wnt3a的遗传效应及SNPs与皮肤毛囊性状的相关性,以期为优质鸡胴体包装性状的分子标记筛选与标记辅助选择提供参考依据。
The aim of this study was to analyze the correlation between genetic variant loci of Wnt family member 3a(Wnt 3a)gene and skin hair follicle density in Chongren partridge chicken based on gene polymorphism.Six hundred Chongren partridge chickens,200 protospecies hens and 100 roosters and 200 B strain hens and 100 roosters,reared under the same environment and the same management level until date of listing(120 d),were randomly selected in good health and condition.After slaughter,hair follicle density was measured at the same locations on the skin of the back,chest and thighs,the morphological structure of hair follicles was observed by HE staining of skin tissue sections,and the diameter of hair follicles and skin thickness were measured.The genetic variant loci of Wnt 3a gene were screened by PCR amplification followed by direct sequencing,and their correlation with the hair follicle density of chicken skin was analyzed.The expression of Wnt 3a gene in different skin sites was detected by real-time fluorescence quantitative PCR technique.The results showed that:1)Comparing the same sex and skin area of protospecies and B strain,the hair follicle density on the back of protospecies hens was extremely significantly higher than that of B strain(P<0.01),the hair follicle density on the thighs was significantly higher than that of B strain(P<0.05),and the hair follicle diameter on the back and thighs was significantly lower than that of B strain(P<0.05).The density of skin follicles on the back of the protospecies cock was highly significant higher than that of B strain(P<0.01),the density of hair follicles on the chest was highly significant lower than that of B strain(P<0.01),and the diameter of hair follicles on the back and thighs was significantly lower than that of B strain(P<0.05).The thickness of the skin on the chest of males in B strain was signifi-cantly higher than that of protospecies(P<0.05),and the thickness of the skin on the thighs of both males and females in B strain was significantly higher than that of protospecies(P<0.05).2)Three SNPs loci were screened for the Wnt 3a gene and their correlation with the skin follicle density at each site was analyzed.g.2555812 T>C and g.2555377 T>C loci were found to have significantly higher skin follicle density on the back of individuals with CC genotype than that of individuals with other genotypes(P<0.05).3)Real-time fluorescence quantitative PCR results showed that the Wnt 3a gene in hair follicle tissues of all 3 skin sites of the protospecies and the B strain had relatively high expression,and the expression in the dorsal part of the protospecies was significantly higher than that in the dorsal part of the B strain(P<0.05).In conclusion,the skin pores of protospecies of Chongren partridge chicken were finer compared to the B strain,and the CC genotype of the Wnt 3a gene at the g.2555812 T>C locus and the CC genotype at the g.2555377 T>C locus were favorable genotypes for the skin follicles of Chongren partridge chicken and could be used as molecular markers for the chicken skin follicle density trait.In this study,we investigated the genetic effects of Wnt 3a as a candidate gene for hair follicle growth and develop-ment and the correlation between SNPs and skin hair follicle traits,in order to provide a reference basis for molecular marker screening and marker-assisted selection of quality chicken carcass packaging traits.
作者
陈春
康昭风
魏岳
黎观红
武艳平
谢金防
CHEN Chun;KANG Zhaofeng;WEI Yue;LI Guanhong;WU Yanping;XIE Jinfang(Institute of Animal Husbandry and Veterinary Medicine,Jiangxi Academy of Agricultural Sciences,Nanchang 330200,China;College of Animal Science and Technology of Jiangxi Agricultural University,Nanchang 330095,China)
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2023年第7期2810-2823,共14页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
江西省重点研发计划项目(2021BBF61011)
江西省家禽产业技术体系(JXARS-09)
江西现代农业科研协同创新专项(JXXTCX202107)
国家肉鸡产业技术体系(nycytx-42-Z09)。
关键词
崇仁麻鸡
毛囊性状
Wnt3a基因
SNP位点
荧光定量
Chongren partridge chicken
hair follicle traits
Wnt 3a gene
SNP loci
fluorescence quantification