柯乐猪PRLR基因多态性与繁殖性状的关联性

doi:10.3969/j.issn.1004-1524.2023.03.08

浙江农业学报 ›› 2023, Vol. 35 ›› Issue (3): 556-564.DOI: 10.3969/j.issn.1004-1524.2023.03.08

柯乐猪PRLR基因多态性与繁殖性状的关联性

杨酸¹(), 郭小江², 杨红文², 熊力², 李晨², 谭元成¹, 王春源¹, 张依裕¹^,^*()

1.贵州大学动物科学学院,高原山地动物遗传育种与繁殖教育部重点实验室,贵州贵阳 550025
2.贵州省农业农村厅,贵州贵阳 550001

收稿日期:2022-05-07 出版日期:2023-03-25 发布日期:2023-04-07
通讯作者: ^*张依裕,E-mail: zyy8yyc@163.com
作者简介:杨酸(1998—),女,贵州龙里人,硕士研究生,研究方向为动物遗传育种与繁殖。E-mail: 1535390590@qq.com
基金资助:
贵州省科技支撑计划(黔科合支撑〔2021〕一般147号);贵州省优秀青年科技人才培养计划(黔科合平台人才〔2021〕5630号);贵州省生猪产业发展项目(2021)

Correlation of PRLR gene polymorphisms and reproductive traits in Kele pig

YANG Suan¹(), GUO Xiaojiang², YANG Hongwen², XIONG Li², LI Chen², TAN Yuancheng¹, WANG Chunyuan¹, ZHANG Yiyu¹^,^*()

1. Key Laboratory of Plateau Animal Genetics, Breeding and Reproduction, Ministry of Education, College of Animal Science, Guizhou University, Guiyang 550025, China
2. Department of Agriculture and Rural Affairs of Guizhou Province, Guiyang 550001, China

Received:2022-05-07 Online:2023-03-25 Published:2023-04-07

摘要/Abstract

摘要：

本研究旨在探究催乳素受体(prolactin receptor,PRLR)基因的单核苷酸突变位点(single nucleotide polymorphism,SNP)及其对柯乐猪繁殖性状的遗传效应,以期为提高柯乐猪的繁殖性能研究提供基因标记。采用PCR产物直接测序法对204头柯乐猪PRLR基因外显子区的SNP位点进行筛查鉴定,同时利用SPSS软件中单因子方差法分析各SNPs不同基因型和双倍型与7个繁殖性状的相关性,并通过在线软件分析碱基变异对mRNA二级结构的影响。结果显示,在柯乐猪PRLR基因第8外显子检测到g.20655220 C>T突变,其导致丝氨酸变成脯氨酸,为错义突变;在第4内含子检测到2个同义突变位点g.20648859 C>T和g.20648931 C>T,3个SNPs均存在3种基因型,g.20648931 C>T属于低度多态,其余2个位点属于中度多态;经卡方χ²检验,g.20655220 C>T位点的基因型分布符合Hardy-Weinberg平衡(P>0.05),g.20648859 C>T和g.20648931 C>T 位点的基因型分布极显著(P<0.01)偏离Hardy-Weinberg平衡;g.20655220 C>T位点导致PRLR基因mRNA序列二级结构发生改变,致使mRNA二级结构稳定性降低;3个SNPs共组成5种单倍型和9种双倍型,优势单倍型H1(CCC)频率为0.610,优势双倍型H1H1(CCCCCC)频率为0.382。关联性分析结果显示,g.20655220 C>T位点CC基因型母猪的初生个体重、断奶窝重和断奶个体重均显著(P<0.05)高于CT基因型,g.20648931 C>T位点CT基因型母猪的初生个体重、初生窝重和断奶窝重均显著(P<0.05)高于CC基因型和TT基因型,且该位点在初生个体重上,CC基因型显著(P<0.05)高于TT基因型;双倍型H1H5(CTCTCT)在产仔数和活仔数上为优势双倍型,H2H3(CCTTCT)在断奶窝重和断奶个体重上为较优双倍型。综上,PRLR基因可作为候选基因参与柯乐猪繁殖性状的分子标记辅助选择。

关键词: PRLR基因, 柯乐猪, 多态性, 繁殖性状

Abstract:

The aim of this study was to investigate the polymorphisms of the prolactin receptor gene (PRLR) and their effects on reproductive performance in Kele pigs, with a view to finding molecular markers for improving reproductive performance in Kele pigs. In this study, SNPs in the exon region of the PRLR gene were screened in 204 Kele pigs and identified by direct PCR product sequencing. At the same time, the correlation between different genotypes and diplotypes of SNPs and seven reproductive traits was analysed by single-factor variance method in SPSS software, and the influence of base variation on the secondary structure of mRNA was analysed by online software. The results showed that g.20655220C>T mutation was detected in the 8th exon of PRLR gene, which caused serine to proline change, which was a missense mutation. Two synonymous mutations g.20648859C>T and g.20648931C>T were detected in the fourth intron. There were three genotypes in all three SNPs, g.20648931C>T was low polymorphism and the other two loci were medium polymorphism. Chi-squared test showed that the genotype distribution of g.20655220C>T loci was in Hardy-Weinberg equilibrium (P>0.05). The genotypes of g.20648859C>T and g.20648931C>T deviated significantly(P<0.01) from Hardy-Weinberg equilibrium; g.20655220C>T locus led to changes in the secondary structure of the PRLR gene mRNA sequence, resulting in decreased stability of the secondary structure of the mRNA; the three SNPs consisted of 5 haplotypes and 9 diplotypes. The dominant haplotype H1 (CCC) frequency was 0.610 and the dominant diplotype H1H1 (CCCCCC) frequency was 0.382. Correlation analysis showed that the average birth weight, weaning litter weight and weaning average weight of CC genotype were significantly(P<0.05) higher than those of CT genotype. CT genotype of g.20648931c>T locus had significantly (P<0.05) higher birth weight, birth litter weight and weaning litter weight than those of CC genotype and TT genotype, and CC genotype had significantly (P<0.05) higher birth weight than that of TT genotype. H1H5 (CTCTCT) was the dominant doublet type in terms of litter size and live litter size, whereas H2H3 (CCTTCT) was the superior doublet type in terms of litter weight and average weaning weight. In conclusion, PRLR gene could be used as a candidate gene for molecular marker-assisted selection of breeding traits in Kele pig.

Key words: PRLR gene, KeLe pig, polymorphism, reproductive traits

中图分类号:

S828

杨酸, 郭小江, 杨红文, 熊力, 李晨, 谭元成, 王春源, 张依裕. 柯乐猪PRLR基因多态性与繁殖性状的关联性[J]. 浙江农业学报, 2023, 35(3): 556-564.

YANG Suan, GUO Xiaojiang, YANG Hongwen, XIONG Li, LI Chen, TAN Yuancheng, WANG Chunyuan, ZHANG Yiyu. Correlation of PRLR gene polymorphisms and reproductive traits in Kele pig[J]. Acta Agriculturae Zhejiangensis, 2023, 35(3): 556-564.

图/表 9

参考文献 30

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引物名称 Primer name	正向引物 Forward primer (5'→3')	反向引物 Reverse primer (5'→3')	基因位置 Location in gene	产物长度 Product length/bp	退火温度 Tm/℃
P1	ACAGTCACCTCCGGGAAAAC	TTGTTGGGAGACACGCGAAT	g.20640695~g.20640852	158	59
P2	CTGGATCCCGACAGCGATTC	CTTGGATCCATCCGCGTGAA	g.20655030~g.20655245	216	60
P3	CCCTAGGCCACTCACTTGCT	AATTAGTCCTGGGAGGGCTGT	g.20648455~g.20649007	553	58
P4	ATGATCTGGGCAGTGGCTTT	TGCTGGCACTTACCTCCAAC	g.20652417~g.20653041	625	60

引物名称 Primer name	正向引物 Forward primer (5'→3')	反向引物 Reverse primer (5'→3')	基因位置 Location in gene	产物长度 Product length/bp	退火温度 Tm/℃
P1	ACAGTCACCTCCGGGAAAAC	TTGTTGGGAGACACGCGAAT	g.20640695~g.20640852	158	59
P2	CTGGATCCCGACAGCGATTC	CTTGGATCCATCCGCGTGAA	g.20655030~g.20655245	216	60
P3	CCCTAGGCCACTCACTTGCT	AATTAGTCCTGGGAGGGCTGT	g.20648455~g.20649007	553	58
P4	ATGATCTGGGCAGTGGCTTT	TGCTGGCACTTACCTCCAAC	g.20652417~g.20653041	625	60

SNP位点 SNP loci	基因型(频率,样本数) Genotype(frequency, sample number))			等位基因(频率) Allele(frequency)		He	Ne	PIC	χ²
g.20655220 C>T	CC (0.583,119)	CT (0.358,73)	TT (0.059,12)	C(0.762)	T(0.238)	0.362	1.569	0.297	0.033
g.20648859 C>T	CC (0.657,134)	CT (0.255,52)	TT (0.088,18)	C(0.784)	T(0.216)	0.338	1.511	0.281	12.405
g.20648931 C>T	CC (0.755,154)	CT (0.196,40)	TT (0.049,10)	C(0.853)	T(0.147)	0.251	1.335	0.219	9.730

SNP位点 SNP loci	基因型(频率,样本数) Genotype(frequency, sample number))			等位基因(频率) Allele(frequency)		He	Ne	PIC	χ²
g.20655220 C>T	CC (0.583,119)	CT (0.358,73)	TT (0.059,12)	C(0.762)	T(0.238)	0.362	1.569	0.297	0.033
g.20648859 C>T	CC (0.657,134)	CT (0.255,52)	TT (0.088,18)	C(0.784)	T(0.216)	0.338	1.511	0.281	12.405
g.20648931 C>T	CC (0.755,154)	CT (0.196,40)	TT (0.049,10)	C(0.853)	T(0.147)	0.251	1.335	0.219	9.730

倍型 Ploidy	SNP位点(样本数) SNP locus(Sample number)	g.20655220 C>T	g.20648859 C>T	g.20648931 C>T	频率 Frequency/%
单倍型	H1(243)	C	C	C	0.610
Haplotypes	H2(28)	C	T	C	0.069
	H3(40)	C	T	T	0.098
	H4(77)	T	C	C	0.189
	H5(20)	T	T	T	0.049
双倍型	H1H1(78)	C C	C C	C C	0.382
Diplotypes	H1H2(11)	C C	C T	C C	0.054
	H1H3(22)	C C	C T	C T	0.108
	H1H4(44)	C T	C C	C C	0.216
	H1H5(10)	C T	T T	T T	0.049
	H2H3(8)	T T	T T	T T	0.039
	H2H4(9)	C T	C T	C C	0.044
	H3H5(10)	C T	T T	T T	0.049
	H4H4(12)	T T	C C	C C	0.059

柯乐猪PRLR基因多态性与繁殖性状的关联性

Correlation of PRLR gene polymorphisms and reproductive traits in Kele pig

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SNP位点 SNP loci	g.20655220 C>T	g.20648859 C>T	g.20648931 C>T
g.20655220 C>T		0.111	0.022
g.20648859 C>T	0.001		1.000
g.20648931 C>T	0	0.627

位点 Locus	基因型 Genotype	每窝总产仔数 Total number born	每窝产活仔数 Number born alive	初生窝重 Newborn litter weight/kg	初生个体重 Individual birth weight/kg	每窝断奶仔猪数 Number of weaned piglets	断奶窝重 Weight of weaning litter/kg	断奶个体重 Weaning individual weight/kg
g.20655220	CC(119)	8.765±2.547	8.050±2.587	8.785±3.018	1.109±0.229 a	6.967±2.278	39.551±13.000 a	5.776±1.277 a
C>T	CT(73)	8.671±2.582	7.726±2.864	7.950±3.090	1.044±0.205 b	6.616±2.542	35.423±13.395 b	5.417±0.817 b
	TT(12)	9.500±2.468	8.417±3.089	9.283±4.229	1.086±0.209 ab	6.833±2.362	36.600±10.525 ab	5.443±0.589 ab
g.20648859	CC(134)	8.716±2.530	7.948±2.640	8.407±2.993	1.073±0.214	6.828±2.306	37.490±12.621	5.567±0.961
C>T	CT(52)	8.769±2.478	8.019±2.797	8.927±3.417	1.125±0.233	6.846±2.321	38.747±13.351	5.724±1.295
	TT(18)	9.222±2.981	7.833±3.130	8.131±3.416	1.053±0.231	6.833±2.975	38.508±16.323	5.809±1.545
g.20648931	CC(154)	8.617±2.487	7.857±2.631	8.327±2.987 b	1.076±0.221 b	6.747±2.288	37.144±12.757 b	5.570±1.010
C>T	CT(40)	9.225±2.636	8.425±2.782	9.684±3.440 a	1.158±0.207 a	7.300±2.3345	42.326±13.089 a	5.930±1.466
	TT(10)	9.400±3.098	7.600±3.658	6.740±2.970 b	0.916±0.145 c	6.300±3.401	31.845±15.107 b	5.311±0.781

双倍型 Diplotype	每窝总产仔数 Total number born	每窝产活仔数 Number born alive	初生窝重 Newborn litter weight/kg	初生个体重 Individual birth weight/kg	每窝断奶仔猪数 Number of weaned piglets	断奶窝重 Weight of weaning litter/kg	断奶个体重 Weaning individual weight/kg
H1H1(78)	8.731±2.567 ab	8.051±2.613	8.586±2.980 abc	1.084±0.223 abc	6.936±2.275	38.482±12.672 abc	5.635±1.095 ab
H1H2(11)	8.364±2.248 ab	7.364±2.942	7.809±3.263 abc	1.091±0.310 abc	6.091±2.300	35.455±14.903 abc	5.836±1.527 ab
H1H3(22)	9.000±2.582 ab	8.364±2.441	9.587±2.859 a	1.165±0.204 a	7.318±2.255	42.739±11.863 ab	6.011±1.397 ab
H1H4(44)	8.477±2.492 ab	7.636±2.589	7.853±2.576 bc	1.050±0.201 bc	6.636±2.441	35.973±13.129 bc	5.480±0.775 b
H1H5(10)	9.900±2.601 a	8.800±3.795	9.750±4.875 ab	1.091±0.217 abc	7.100±2.726	37.810±14.445 abc	5.351±0.934 b
H2H3(8)	9.000±3.024 ab	8.125±2.532	9.869±3.284 ab	1.224±0.206 a	7.500±2.390	46.838±14.531 a	6.433±2.054 a
H2H4(9)	7.444±1.944 b	7.111±2.147	7.767±2.763 abc	1.108±0.238 abc	6.333±2.062	34.056±13.041 b	5.302±1.029 b
H3H5(10)	9.400±3.098 ab	7.600±3.658	6.740±2.969 b	0.916±0.145 b	6.300±3.41	31.845±15.103 bc	5.311±0.781 b
H4H4(12)	9.500±2.468 ab	8.417±3.088	9.283±4.229 abc	1.086±0.210 abc	6.833±2.125	36.600±10.525 bc	5.443±0.589 ab

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