卷羽与半卷羽遗传特性分析及KRT75基因SNP位点检测

doi:10.16431/j.cnki.1671-7236.2016.11.029

摘要/Abstract

摘要：

本试验通过对卷羽性状遗传特性和卷羽、半卷羽差异及对不同羽色卷羽、半卷羽鸡的候选基因KRT75序列SNP分析，为进一步对卷羽性状基因的定位和卷羽、半卷羽鸡的开发利用提供科学依据。以纯系黄色卷羽麒麟鸡和片羽怀乡鸡为素材，正反交构建F2代资源群体，对24周龄卷羽、半卷羽和片羽鸡背羽、颈羽、翅膀主翼羽和翼肩羽观察并绘制背羽羽轴弯曲程度散点图。随机抽取25周龄黄羽、白羽麒麟鸡和贵妃鸡各10只，F1代黄羽半卷羽鸡25只，以及白羽麒麟鸡×贵妃鸡杂交F1代黑羽半卷羽鸡15只的血样提取DNA，设计引物，PCR扩增纯化后测序，Chromas 2.22和DNAMAN软件用于分析测序峰形图和序列比对。结果显示，杂交F1代正交组127只，反交139只，不论公、母表型较一致，无外显率不同，均呈现轻度卷羽（半卷羽），F1代间交配产生的F2代资源群体：卷羽鸡55只，半卷羽鸡106只，片羽鸡68只，公、母鸡均有卷羽，经过卡方适合性检验符合孟德尔遗传分离比1∶2∶1（P>0.05），因此初步验证了控制卷羽基因F呈常染色体不完全显性遗传；卷羽、半卷羽鸡羽毛均背离皮肤向上弯曲，卷羽趋势线直线斜率是半卷羽的3.5倍；白羽和黄羽麒麟鸡外显子5处均不存在69 bp缺失，此69 bp发现3个SNPs：T71C、T83C、C95T，其中片羽鸡无突变，黄羽和白羽麒麟鸡均为纯合子，黄羽半卷羽鸡2个位点为杂合子，黑羽半卷羽鸡3个位点均为杂合子；外显子6处检测到了2个SNPs分别为T662C和T770C，仅在黑羽半卷羽鸡中为杂合子，其他均为纯合子，单倍型分析从60个个体中检测到了9种单倍型，hap1/hap1纯合型是黄羽和白羽麒麟鸡特异性的基因型，黄羽和黑羽半卷羽鸡的单倍型存在明显差异，hap4单倍型是黄羽半卷羽鸡特有的单倍型，hap6、hap7、hap8、hap9为黑羽半卷羽鸡特有的单倍型。卷羽性状基因F为常染色体不完全显性遗传，卷羽和半卷羽存在明显差异，KRT75基因序列外显子5和6处的5个SNPs可能作为区别卷羽和半卷羽鸡的分子标记参考。

关键词: 卷羽; 半卷羽; 遗传特性; KRT75基因; SNP

Abstract:

The study was mainly focused on the genetic characteristics of frizzled feather trait,the phenotype difference between frizzled feather（FF）and incomplete frizzled feather（IFF),and SNP analysis of the candidate gene KRT75 in the frizzle and incomplete frizzle chickens of different feather colors,which could provide scientific gist for the frizzle gene mapping,development and utilization of the two kinds of chicken.The resource population（homozygous Yellow feather Kirin chicken×Huaixiang chicken）of hybrid F2 was established.The back,neck,wing primary and wing-bow feather were observed and the scatter diagram of rachis bending degree was drawn.The blood of 10 white frizzle feather Kirin（WFF）chicken,10 Yellow frizzle feather Kirin（YFF）chicken,10 Royal,25 Yellow feather incomplete frizzle（YFIF）chicken and 15 Black feather incomplete frizzle（BFIF）chicken from homozygous WFF chicken×Royal chicken of hybrid F1 were extracted,then the DNA of those samples were extracted.The primer of KRT75 was designed,the PCR products were also sequenced after purification,Chromas 2.22 and DNAMAN software were used to analysis of sequence peaks photos and sequence alignment,respectively.The amount of orthogonal and reciprocals cross in hybrid F1 was 127 and 139, respectively, which all were the slight（incomplete）frizzled feather phenotype,with no different penetrance.The resource population of hybrid F2 were consisted of 55 frizzle chicks including some hens and cocks,106 incomplete frizzled and 68 contour feather chicken,the proportion of which was coincided with Mendelian segregation ratio of 1:2:1 by χ²-test（P>0.05),therefore it preliminary verified that the frizzle gene F was autosomal incomplete dominant inheritance.FF and IFF curved upward deviating from the skin,and the line slope of trend line of FF was 3.5 times than that of IFF.It found that there was no 69 bp deletion mutation in exon 5 region of KRT75 gene in WFF and YFF chicken,but 3 SNPs（T71C,T83C,C95T）were deleted in the 69 bp,which were homologous CC in WFF and YFF chicken,of two were heterozygous in YFIF chicken,while all were heterozygous in BFIF chicken.2 SNPs（T662C and T770C）were also deleted in exon 6,heterozygous of which were only deleted in BFIF chicken.The haplotype analysis indicated that 9 haplotypes were detected in 60 individuals,hap1/hap1 was specific genotype of WFF and YFF chicken.The haplotypes of two incomplete frizzle feather chicken were apparently different,hap4 was specific haplotype of YFIF chicken,while hap6,hap7,hap8 and hap9 were specific haplotypes of BFIF chicken.The frizzle gene F was autosomal incomplete dominant inheritance,there were obvious differences between FF and IFF traits,5 SNPs in exon 5 and 6 of KRT75 gene probably were the reference of molecular markers as distinguishing between frizzle and incomplete frizzle chicken.

Key words: frizzled feather; incomplete frizzled feather; genetic characteristics; KRT75 gene; SNP

中图分类号:

S813.3

董晶, 张权, 张丽, 李艳青, 李珊珊, 王章, 杜炳旺. 卷羽与半卷羽遗传特性分析及KRT75基因SNP位点检测[J]. 《中国畜牧兽医》, 2016, 43(11): 3003-3010.

DONG Jing, ZHANG Quan, ZHANG Li, LI Yan-qing, LI Shan-shan, WANG Zhang, DU Bing-wang. Genetic Characteristics of Frizzled and Incomplete Frizzled Feather and SNP Detection of KRT75 Gene[J]. , 2016, 43(11): 3003-3010.

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