【作者】 张平；

【导师】 鲁绍雄； 王莉兴；

【作者基本信息】 云南农业大学 ， 农业硕士（专业学位）， 2023， 硕士

【摘要】 为揭示宣和猪新品种钙蛋白酶抑制蛋白（calpastatin,CAST）和磷酸化酶激酶催化亚基γ1（phosphorylase kinase gamma1,PHKG1）基因多态性及其与胴体和肉质性状的关联,本研究采用PCR产物直接测序法检测了111头宣和猪CAST和PHKG1基因外显子区域的SNP变异位点,利用最小二乘模型分析了2个基因各SNPs位点基因型及单倍型组合对宣和猪测定屠宰率、胴体直长、平均背膘厚、眼肌面积、后腿比例、瘦肉率6个胴体性状,以及肉色、大理石纹、p H₁、失水率、熟肉率、滴水损失、肌内脂肪含量7个肉质性状的影响。主要结果如下:（1）系统检测了宣和猪CAST和PHKG1基因外显子区域的SNPs。宣和猪CAST基因外显子区域共检出10个SNPs位点,其中第14、17、28和29外显子各1个,第1、15和19外显子各2个,包括5个错义突变位点:第1外显子EX1-T315G（Leu/Arg）、第15外显子EX15-A47G（Lys/Arg）、第17外显子EX17-C22G（Leu/Val）、第28外显子EX28-C11T（Thr/Met）和第29外显子EX29-A54C（Arg/Ser）。PHKG1基因外显子区域共检出6个SNPs位点,分别位于第3外显子（2个）和第10外显子（4个）,其中第10外显子的EX10-C224T（Ser/Leu）为错义突变位点。（2）从CAST和PHKG1基因多态性角度揭示了宣和猪的遗传多样性。宣和猪CAST基因10个SNPs位点的平均多态信息含量为0.2980（0.2211～0.3710）、平均杂合度为0.3727（0.2531～0.4920）,PHKG1基因6个SNPs位点的平均多态信息含量为0.1982（0.1248～0.3740）、平均杂合度为0.2335（0.1338～0.4980）,2个基因16个SNPs位点的平均多态信息含量和杂合度分别为0.2606和0.3205,猪群遗传多样性较为丰富。（3）揭示了宣和猪CAST基因多态性对胴体和肉质性状的影响。宣和猪CAST基因第28外显子区域的EX28-C11T位点与眼肌面积和后腿比例显著关联（P<0.05）;EX1-T315G、EX1-C316T与大理石纹评分和失水率,EX14-T27C、EX15-A12G、EX15-A47G位点与肉色评分和滴水损失,EX17-C22G位点与失水率,EX28-C11T、EX29-A54C位点与滴水损失显著关联（P<0.05或P<0.01）;各位点不同单倍型组合对所分析的6个胴体性状和大理石纹评分、p H₁、肌内脂肪含量3个肉质性状有显著影响（P<0.05或P<0.01）。CAST基因各SNP位点基因型及单倍型组合的具体效应会因性状的不同而有所差异。（4）揭示了宣和猪PHKG1基因多态性对胴体和肉质性状的影响。宣和猪PHKG1基因EX3-A43G位点与屠宰率、后腿比例,EX3-C55T、EX10-C132T、EX10-C213T位点与屠宰率,EX10-C224T位点与瘦肉率显著关联;EX10-T27C位点与熟肉率,EX3-C55T、EX10-C132T、EX10-C213T位点与肌内脂肪含量显著关联（P<0.05或P<0.01）;不同单倍型组合对屠宰率、平均背膘厚、眼肌面积、后腿比例和瘦肉率5个胴体性状以及肉色评分、p H₁、失水率和肌内脂肪含量4个肉质性状有显著影响（P<0.05或P<0.01）。各性状的增效基因型或单倍型组合会因性状的不同而有所差异。更多还原

【Abstract】 In order to reveal the the polymorphisms of calpastatin（CAST）and phosphorylase kinase gamma1（PHKG1）gene and their associations with carcass and meat quality traits in the new developed breed Xuanhe pigs,the single nucleotide polymorphisms（SNP）of CAST and PHKG1 PCR detected by PCR product directly sequencing in 111 Xuanhe pigs.The effects of different genotypes and haplotype combinations of SNPs of the two genes on 6carcass traits,dressing percentage（DP）,carcass straight length（CSL）,average backfat thickness（ABF）,loin eye area（LEA）,ham percentage（HP）,lean meat percentage（LMP）,and 7 meat quality traits,meat color（MC）,marbling score（MS）,p H₁,water loss percentage（WLP）,cooked rate（CR）,drip loss percentage（DLP）and intramuscular fat content（IMF）were analyzed by least square model.The main results were as follows:（1）The SNPs in the exon regions of CAST and PHKG1 genes in Xuanhe pigs was revealed.A total of 10 SNPs were detected in the exon region of CAST gene in Xuanhe pigs.There is one in each of the exon 14,17,28 and 29;there were two in each of exon 1,15 and19,respectively.Among the 10 SNPs,EX1-T315G（Leu/Arg）,EX15-A47G（Lys/Arg）,EX17-C22G（Leu/Val）,EX28-C11T（Thr/Met）and EX29-A54C（Arg/Ser）were missense mutations.A total of 6 SNPs were detected in the exon region of PHKG1 gene,which distributed in the exon 3（2 SNPs）and exon 10（4 SNPs）,and the SNP EX10-C224T（Ser/Leu）was a missense mutation.（2）The genetic diversity of Xuanhe pigs was revealed from the polymorphisms of CAST and PHKG1 gene.The average polymorphic information content（PIC）and heterosygosity（H）of the ten SNPs of CAST gene in Xuanhe pigs were 0.2980（0.2211-0.3710）and 0.3727（0.2531-0.4920）;and the average PIC and H of the six SNPs of PHKG1gene in Xuanhe pigs were 0.1982（0.1248-0.3740）and 0.2335（0.1338-0.4980）,and the average PIC and H of the sixteen SNPs of the two genes were 0.2606 and 0.3205,respectively.It indicated that the genetic diversity of Xuanhe pigs was relative abundant.（3）The association CAST gene polymorphism with carcass and meat quality traits in Xuanhe pigs was revealed.The SNP EX28-C11T in the exon 28 of CAST gene of Xuanhe pigs was significantly associated with LEA and HP（P<0.05）;significant associations were showed that EX1-T315G and EX1-C316T with MS and WLP,EX14-T27C,EX15-A12G and EX15-A47G with MC and DLP,EX17-C22G with WLP,EX28-C11T and EX29-A54C with DLP（P<0.05 or P<0.01）.It showed significant effects of different haplotype combinations of the SNPs on all the six carcass traits and three meat quality traits,MS,p H₁and IMF（P<0.05 or P<0.01）.However,the specific effects of the genotypes and haplotype combinations of CAST gene might be various for different traits.（4）The association of PHKG1 gene polymorphism and carcass and meat quality traits in Xuanhe pigs was revealed.Sinificant associations of SNPs of PHKG1 gene in Xuanhe pigs were identified that EX3-A43G with DR and HP,EX3-C55T,EX10-C132T and EX10-C213T with DR,EX10-C224T with LMP,EX10-T27C with CR,EX3-C55T,EX10-C132T and EX10-C213T with IMF（P<0.05 or P<0.01）.It showed significant effects of different haplotype combinations of the SNPs on five carcass traits,DR,ABF,LEA,HP and LMP,and four meat quality traits,MC,p H₁,WLP and IMF（P<0.05 or P<0.01）.The effects of the genotypes and haplotype combinations of PHKG1 gene on the carcass and meat quality traits were also various for different and trats.更多还原