山猪肉质性状主要相关基因的遗传特性及其对肉质的作用和影响
摘要
中国是猪肉消费大国,随着人们生活水平的不断提高和养猪业的进一步发展,人们对猪肉品质的要求日益提高,因此,改善猪的肉质已成为目前生猪生产上的最大挑战。猪肉品质是一个复杂的经济性状,其表现会受到众多因素的影响,而微效多基因调控机制是决定猪胴体、肉质等数量性状的分子基础。本研究采用候选基因法对肉质相关候选基因进行分子遗传分析,并利用荧光定量PCR技术对候选基因在不同杂交猪品种中的表达差异进行研究,以期了解肉质相关候选基因的多态性及基因表达对胴体、肉质性状的影响。
本研究对17个候选基因进行了SNPs的检测,结果在10个基因中检测到19个SNPs，对其中9个肉质相关基因进行多态性分析,研究这些基因的基因型频率和等位基因频率的分布规律。结果表明,在不同猪品种中,除了MYF5基因,其它基因的基因型频率的差异均达到显著(p<0.05)或极显著(p<0.01)水平,在不同性别的山猪杂交猪之间的基因型分布没有显著差异(p>0.05),在不同山猪血统含量的杂交猪(山猪二元猪和山猪三元猪)之间,MYOD1的基因型分布存在显著差异(p<0.05),LPL基因的基因型分布显示出极显著差异(p<0.01)。在山猪中,占优势的等位基因分别是MC4R的A等位基因,LEPR的A等位基因,HSL的A等位基因,MYF5的A等位基因,MYOD1的C等位基因,H-FABP的T等位基因以及ADRB3基因的A等位基因。在不同山猪杂交猪中研究了上述基因的多态性位点与猪胴体、肉质性状间的相关性,结果表明,除MYOD1基因外,其余分子标记均与猪中某些胴体、肉质性状存在显著或极显著相关。研究发现,MC4R的AA基因型更有利于体重以及脂肪沉积；LEPR的AA基因型系水能力更强,且更有利于优良肉色性状的表达;HSL的GG基因型有利于提高瘦肉率,而AA基因型有利于提高肌内脂肪含量；LPL的CC基因型具有较薄的背膘厚、较大的眼肌面积和较低的失水率,TT基因型则具有较高的肉色参数值和肌内脂肪含量,指示选择CC基因型猪将有利于地方猪品种的遗传改良；MYF5的AA基因型的肉嫩度更高,肉质系水力更强,具有更好的食用口感；ADRB3的G等位基因更有利于增加肌内脂肪含量,选择G等位基因的引入将有利于国内猪种的遗传改良；H-FABP的T等位基因有利于提高猪肉嫩度和脂肪含量；PPARγ的GG基因型系水能力更强。
采用荧光定量PCR方法检测11个候选基因在山猪杂交猪背最长肌中的表达情况,结果表明,在二元杂交猪、三元杂交猪之间,LEPR、PPARα、ADRB3、 AdPLA、SCD基因的表达差异达到显著水平,LPL基因的表达差异达到极显著水平；在不同性别的杂交猪之间,PPARα、ADRB3基因的表达差异达到显著水平。而在各个基因的不同基因型之间,LEPR、LPL、MYF5、PPARγ、ADRB3基因的表达差异达到显著水平。LEPR的AT基因型、PPARγ的AG基因型更有利于基因表达；ADRB3的AG基因型则会减弱基因表达；而LPL的T等位基因、MYOD1的A等位基因更有利于基因表达。在不同山猪杂交猪中研究了候选基因的表达差异与猪胴体、肉质性状间的相关性,结果表明,除了PEPCK基因,其余基因的表达差异均与猪中某些胴体、肉质性状存在显著或极显著相关,其中,MC4R、LEPR、LPL、PPARα基因表达的增强有利于体脂含量、肌内脂肪含量的增加,而不利于胴体性状；MYF5基因表达的增强有利于增加脂肪沉积,但会使猪肉的瘦肉率和嫩度降低；MYOD1基因表达的增强有利于pH24值提高,影响猪肉的食用品质；ADRB3表达量的提高有利于胴体性状,而不利于脂肪性状;PPARγ表达量的提高也不利于胴体性状,但同时会使嫩度降低；AdPLA表达量的提高有助于脂肪的积累,但不利于蛋白质含量的增加；SCD基因表达量的提高有助于肌内脂肪的沉积,增强肉质食用口感。
本研究鉴定出一批对猪胴体、肉质性状具有影响的等位基因或基因型,并建立这些等位基因或基因型的检测方法：另外分析各基因的遗传变异类型、表达水平等对肉质性状的作用和影响,为猪肉质性状的遗传控制理论研究和主效基因的评价与利用提供新的研究途径。
Pork is one of the most important meats in our country. With the development of swine industry and improvement of people's life, the requirments of meat quality rise year by year, so the improvement of meat quality has become the largest challenge in pig production. Meat quality is a complex economical trait which affected by a lot of factors, but the regulatory mechanism of micro-effect polygenes determines porcine meat quality, carcass characteristics and other relative quantitative traits, In this study, we selected candidate genes associated with meat quality by candidate gene approach to detect molecular markers in pigs, and used qRT-PCR to examine the expression differences of candidate genes associated with carcass and meat quality in M.longissimus dor si muscle of crossbred pigs. The objective of this study was to investigate the effects of polymorphisms in ten candidate genes and their expression as well as genders and bloodlines on carcass and meat quality traits in pigs.
We tested the polymorphism of17candidate genes, and investigated19single nucleotide polymorphisms (SNPs) among10genes. The genotype frequency and allele frequency for differences of genotype distributions were calculated for each polymorphism of nine meat quality related genes in different pig breeds. Except for MYF5, the genotypic and allelic frequencies of the remaining genes exhibited high rates of polymorphisms, and a x2test revealed that differences in the genotypic frequencies were very significant (p<0.Ol) or significant (p<0.05) in different pig breeds. Genotype distributions didn't show significant (p>0.05) differences in two genders. Genotype distributions of MYOD1and LPL showed significant (p<0.05) differences in two bloodlines. The analysis revealed that MC4R C.1426A allele, LEPR c.232T allele, HSL c.442A allele, MYF5c.1205A allele, MYOD1c.510C, H-FABP c.1322T and ADRB3c.l192A were dominant in shanzhu pigs. An association analysis revealed that all polymorphic markers tested, with the exception of MYOD1gene, showed very significant (p<0.01) or significant (p<0.05) associations with aspects of carcass and meat quality traits in crossbred pigs. Each allele's main effects were as follows:MC4R c.1426A allele increased body weight and fat deposition. LEPR c.232A allele increased water-holding capacity. HSL c.442G allele increased rib eye area and decreased intramuscular fat (IMF). LPL c.114T allele increased IMF and yellowness and LPL c.114C allele increased rib eye area and water-holding capacity. MYF5c.1205A allele improved tenderness and water-holding capacity. ADRB3 c.1192G allele increased IMF. H-FABP c.1322T allele improved tenderness and IMF. PPARγ c.175G allele increased water-holding capacity.
We used qRT-PCR to examine the expression differences of11candidate genes in M.longissimus dorsi muscle of crossbred pigs. The expression of LEPR, PPARa, ADRB3, AdPLA, SCD and LPL showed significant (p<0.05) differences between two crossbreds respectively. PPARa and ADRB3showed significant (p<0.05) differences in two genders respectively. Variance analysis (ANOVA) revealed that expression differences among different genotypes of LEPR, LPL, MYF5, PPARy and ADRB3genes were significant (p<0.05). LEPR AT genotype was better for gene expression than AA or TT genotype. PPARy AG genotype was better for gene expression than AA or GG genotype. ADRB3AG genotype decreased gene expression. LPL c.114T allele increased gene expression. MYOD1c.510A increased gene expression. An association analysis revealed that expression differences of candidate genes, with the exception of PEPCK gene, showed very significant (p<0.01) or significant (p<0.05) associations with aspects of carcass and meat quality traits in M.longissimus dorsi muscle of crossbred pigs. MC4R, LEPR, LPL and PPARa expression increased body fat content and intramuscular fat content. MYF5expression increased lipid deposition but decreased lean meat percentage and tenderness. MYOD1expression increased pH24value. ADRB3expression improved carcass traits. PPARy expression decreased carcass quality and tenderness. AdPLA expression increased intramuscular fat content but decreased protein content. SCD expression increased intramuscular fat content and tenderness.
These results highlight some alleles or genotypes for porcine carcass and meat quality traits, develop rapid and simple detective methods for these alleles or genotypes, analysis the effects of these genes' SNPs and expression on carcass and meat quality traits, and provide some data for the evaluation and utilization of meat quality related gene in the effort to improve pigs'meat quality.
本研究对17个候选基因进行了SNPs的检测,结果在10个基因中检测到19个SNPs，对其中9个肉质相关基因进行多态性分析,研究这些基因的基因型频率和等位基因频率的分布规律。结果表明,在不同猪品种中,除了MYF5基因,其它基因的基因型频率的差异均达到显著(p<0.05)或极显著(p<0.01)水平,在不同性别的山猪杂交猪之间的基因型分布没有显著差异(p>0.05),在不同山猪血统含量的杂交猪(山猪二元猪和山猪三元猪)之间,MYOD1的基因型分布存在显著差异(p<0.05),LPL基因的基因型分布显示出极显著差异(p<0.01)。在山猪中,占优势的等位基因分别是MC4R的A等位基因,LEPR的A等位基因,HSL的A等位基因,MYF5的A等位基因,MYOD1的C等位基因,H-FABP的T等位基因以及ADRB3基因的A等位基因。在不同山猪杂交猪中研究了上述基因的多态性位点与猪胴体、肉质性状间的相关性,结果表明,除MYOD1基因外,其余分子标记均与猪中某些胴体、肉质性状存在显著或极显著相关。研究发现,MC4R的AA基因型更有利于体重以及脂肪沉积；LEPR的AA基因型系水能力更强,且更有利于优良肉色性状的表达;HSL的GG基因型有利于提高瘦肉率,而AA基因型有利于提高肌内脂肪含量；LPL的CC基因型具有较薄的背膘厚、较大的眼肌面积和较低的失水率,TT基因型则具有较高的肉色参数值和肌内脂肪含量,指示选择CC基因型猪将有利于地方猪品种的遗传改良；MYF5的AA基因型的肉嫩度更高,肉质系水力更强,具有更好的食用口感；ADRB3的G等位基因更有利于增加肌内脂肪含量,选择G等位基因的引入将有利于国内猪种的遗传改良；H-FABP的T等位基因有利于提高猪肉嫩度和脂肪含量；PPARγ的GG基因型系水能力更强。
采用荧光定量PCR方法检测11个候选基因在山猪杂交猪背最长肌中的表达情况,结果表明,在二元杂交猪、三元杂交猪之间,LEPR、PPARα、ADRB3、 AdPLA、SCD基因的表达差异达到显著水平,LPL基因的表达差异达到极显著水平；在不同性别的杂交猪之间,PPARα、ADRB3基因的表达差异达到显著水平。而在各个基因的不同基因型之间,LEPR、LPL、MYF5、PPARγ、ADRB3基因的表达差异达到显著水平。LEPR的AT基因型、PPARγ的AG基因型更有利于基因表达；ADRB3的AG基因型则会减弱基因表达；而LPL的T等位基因、MYOD1的A等位基因更有利于基因表达。在不同山猪杂交猪中研究了候选基因的表达差异与猪胴体、肉质性状间的相关性,结果表明,除了PEPCK基因,其余基因的表达差异均与猪中某些胴体、肉质性状存在显著或极显著相关,其中,MC4R、LEPR、LPL、PPARα基因表达的增强有利于体脂含量、肌内脂肪含量的增加,而不利于胴体性状；MYF5基因表达的增强有利于增加脂肪沉积,但会使猪肉的瘦肉率和嫩度降低；MYOD1基因表达的增强有利于pH24值提高,影响猪肉的食用品质；ADRB3表达量的提高有利于胴体性状,而不利于脂肪性状;PPARγ表达量的提高也不利于胴体性状,但同时会使嫩度降低；AdPLA表达量的提高有助于脂肪的积累,但不利于蛋白质含量的增加；SCD基因表达量的提高有助于肌内脂肪的沉积,增强肉质食用口感。
本研究鉴定出一批对猪胴体、肉质性状具有影响的等位基因或基因型,并建立这些等位基因或基因型的检测方法：另外分析各基因的遗传变异类型、表达水平等对肉质性状的作用和影响,为猪肉质性状的遗传控制理论研究和主效基因的评价与利用提供新的研究途径。
Pork is one of the most important meats in our country. With the development of swine industry and improvement of people's life, the requirments of meat quality rise year by year, so the improvement of meat quality has become the largest challenge in pig production. Meat quality is a complex economical trait which affected by a lot of factors, but the regulatory mechanism of micro-effect polygenes determines porcine meat quality, carcass characteristics and other relative quantitative traits, In this study, we selected candidate genes associated with meat quality by candidate gene approach to detect molecular markers in pigs, and used qRT-PCR to examine the expression differences of candidate genes associated with carcass and meat quality in M.longissimus dor si muscle of crossbred pigs. The objective of this study was to investigate the effects of polymorphisms in ten candidate genes and their expression as well as genders and bloodlines on carcass and meat quality traits in pigs.
We tested the polymorphism of17candidate genes, and investigated19single nucleotide polymorphisms (SNPs) among10genes. The genotype frequency and allele frequency for differences of genotype distributions were calculated for each polymorphism of nine meat quality related genes in different pig breeds. Except for MYF5, the genotypic and allelic frequencies of the remaining genes exhibited high rates of polymorphisms, and a x2test revealed that differences in the genotypic frequencies were very significant (p<0.Ol) or significant (p<0.05) in different pig breeds. Genotype distributions didn't show significant (p>0.05) differences in two genders. Genotype distributions of MYOD1and LPL showed significant (p<0.05) differences in two bloodlines. The analysis revealed that MC4R C.1426A allele, LEPR c.232T allele, HSL c.442A allele, MYF5c.1205A allele, MYOD1c.510C, H-FABP c.1322T and ADRB3c.l192A were dominant in shanzhu pigs. An association analysis revealed that all polymorphic markers tested, with the exception of MYOD1gene, showed very significant (p<0.01) or significant (p<0.05) associations with aspects of carcass and meat quality traits in crossbred pigs. Each allele's main effects were as follows:MC4R c.1426A allele increased body weight and fat deposition. LEPR c.232A allele increased water-holding capacity. HSL c.442G allele increased rib eye area and decreased intramuscular fat (IMF). LPL c.114T allele increased IMF and yellowness and LPL c.114C allele increased rib eye area and water-holding capacity. MYF5c.1205A allele improved tenderness and water-holding capacity. ADRB3 c.1192G allele increased IMF. H-FABP c.1322T allele improved tenderness and IMF. PPARγ c.175G allele increased water-holding capacity.
We used qRT-PCR to examine the expression differences of11candidate genes in M.longissimus dorsi muscle of crossbred pigs. The expression of LEPR, PPARa, ADRB3, AdPLA, SCD and LPL showed significant (p<0.05) differences between two crossbreds respectively. PPARa and ADRB3showed significant (p<0.05) differences in two genders respectively. Variance analysis (ANOVA) revealed that expression differences among different genotypes of LEPR, LPL, MYF5, PPARy and ADRB3genes were significant (p<0.05). LEPR AT genotype was better for gene expression than AA or TT genotype. PPARy AG genotype was better for gene expression than AA or GG genotype. ADRB3AG genotype decreased gene expression. LPL c.114T allele increased gene expression. MYOD1c.510A increased gene expression. An association analysis revealed that expression differences of candidate genes, with the exception of PEPCK gene, showed very significant (p<0.01) or significant (p<0.05) associations with aspects of carcass and meat quality traits in M.longissimus dorsi muscle of crossbred pigs. MC4R, LEPR, LPL and PPARa expression increased body fat content and intramuscular fat content. MYF5expression increased lipid deposition but decreased lean meat percentage and tenderness. MYOD1expression increased pH24value. ADRB3expression improved carcass traits. PPARy expression decreased carcass quality and tenderness. AdPLA expression increased intramuscular fat content but decreased protein content. SCD expression increased intramuscular fat content and tenderness.
These results highlight some alleles or genotypes for porcine carcass and meat quality traits, develop rapid and simple detective methods for these alleles or genotypes, analysis the effects of these genes' SNPs and expression on carcass and meat quality traits, and provide some data for the evaluation and utilization of meat quality related gene in the effort to improve pigs'meat quality.
引文
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