肌肉嫩度仪相关论文：江口萝卜猪UCP3-基因变异及其表达量与肉质性状关联性分析.pdf

基金项目：贵州省科技支撑计划项目(黔科合20172590号)收稿日期：2018-08-08接受日期：2018-09-04江口萝卜猪UCP3基因变异及其表达量与肉质性状关联性分析张继1吴雪1邱淦远1刘鹏程1杨茂林2刘若余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，贵阳 550025；2贵州省江口县科技局，铜仁554400*通讯作者，摘要解耦联蛋白3(uncoupling protein 3,UCP3)作为重要调控蛋白，主要参与机体能量代谢调节，在脂质代谢，脂肪酸氧化和脂肪沉积中起着重要作用，对氧化磷酸化反应途径是否产生ATP起决定性作用。为研究UCP3基因与猪肉质性状之间的关系，本研究测定了165头江口萝卜猪(Sus scrofa)11项肉质性能指标，利用混合DNA池扩增UCP3基因全部外显子，测序筛选SNPs位点，用单个样本DNA扩增SNPs位点对应外显子并测序，以验证位点和分型，分析各位点不同基因型江口萝卜猪肉质性能差异。采用qRT-PCR检测了UCP3基因在背最长肌中的表达量，进行基因型、肉质性能以及基因表达量的相关性研究。研究结果发现了 6 个 SNPs 位点，g.3127AG、g.3205TC、g.10439CT、g.10441CA、g.10466GA、g.10479AG/T。群体遗传参数分析发现除g.10479AG/T位点外的其他位点杂合度高，说明群体内等位基因分布比较均匀；有效等位基因数高，表明群体内变异程度大；多态信息含量均大于0.25，为中度多态，表明位点可以提供一定的遗传信息。2检验显示6个位点均极显著偏离Hardy-Weinberg平衡。连锁不平衡分析发现，g.3127AG 和 g.3205TC 位点、g.10439CT 和 g.10466GA 位点为完全连锁不平衡，g.10439CT、g.10441CA、g.10466GA、g.10479AG/T之间为强连锁不平衡。SNPs位点与肉质的关联性分析发现g.10479AG/T位点TT型和GG型个体大理石纹评分显著高于AA型和AG型个体，剪切力则反之；基因型与背最长肌中UCP3基因表达量的关联性分析发现，g.10439CT、g.10441CA、g.10466GA以及g.10479AG/T 4个位点上均为突变型个体表达量极显著高于另外两种基因型；UCP3基因在背最长肌中的表达量与两项肉质指标存在极显著相关性，其中与剪切力负相关，与肌内脂肪含量正相关。本研究结果表明，g.10439CT、g.10441CA、g.10466GA以及g.10479AG/T位点为影响江口萝卜猪肉质性能的重要SNPs位点，且UCP3基因表达量提高有利于肌内脂肪的沉积，此研究结果为筛选有助于江口萝卜猪肌内脂肪沉积的分子标记提供了参考依据。关键词江口萝卜猪；解耦联蛋白3(UCP3)；SNP；肉质性状；相关性分析中图分类号S828文献标识码ACorrelation Analysis of UCP3 Gene Variation,Expression and MeatQuality Traits in Jiangkou Luobo Pig(Sus scrofa)ZHANG Ji1WU Xue1QIU Gan-Yuan1LIU Peng-Cheng1YANG Mao-Lin2LIU Ruo-Yu1*1 College of Animal Sciences,Guizhou University/Key Laboratory of Animal Genetics,Breeding and Reproduction in the Plateau MountainousRegion,Ministry of Education,Guiyang 550025,China;2 Science and Technology Bureau of Jiangkou County,Tongren 554400,China*Corresponding author,AbstractUncoupling protein 3(UCP3),as an important regulatory protein,is mainly involved in theregulation of the bodys energy metabolism and plays an important role in lipid metabolism,fatty acidOnline system:http:/www.jabiotech.org农 业 生 物 技 术 学 报Journal of Agricultural Biotechnology2020,28(03):490500DOI:10.3969/j.issn.1674-7968.2020.03.012解耦联蛋白(uncoupling protein,UCPs)是线粒体上的质子载体，以质子漏方式把H+从外膜传递到内膜(Ricquier,Frdric,2000)。在氧化磷酸化过程中，可以降低膜内外的电化学梯度，在电子传递过程中实现质子回流，从而起到让呼吸链与ATP合成过程解耦联的作用，使能量以热能的形式散发，氧化磷酸化合成 ATP 的效率下降(张雷等,2010)。UCP3基因是UCPs家族成员之一，与UCP2基因紧密连锁并定位于猪的 9 号染色体(Stone et al.,2015)。当受到脂肪酸刺激时，UCP3基因表达量上调，且UCP3蛋白参与脂肪酸和其他阴离子跨膜转运(Lapice et al.,2017)。UCP3 活性对脂肪酸氧化和氨基酸代谢及其相关的生化途径具有重要调节作用(Aguer et al.,2016)。所以，很多畜禽研究者将其作为一个影响肉品质的重要功能候选基因展开研究(李秋玲等,2006;伍革民等,2007;胡倩,2008)。方美英等(2002)在对猪(Sus scrofa)UCP3基因多态性的研究中，在内含子3发现了T272G变异位点。李艳华等(2006)分析了长白猪、杜洛克等5个国内外猪种UCP3基因的多态位点，发现位于启动子区的SNPs位点会造成转录因子结合位点的改变，影响对UCP3基因的转录和调控，从而影响猪的生产性能。陈哲(2011)在UCP3基因cDNA上检测到A448G突变位点，并发现该位点等位基因A对于改善猪脂肪性状具有积极作用。王阳等(2011)研究发现UCP3参与猪骨骼肌细胞脂肪代谢的调节，从而影响地方猪种的肉质和其他性状。但是关于UCP3基因表达量与肉质性能之间的相关性尚未见研究报道。本研究以贵州优良地方品种江口萝卜猪为实验oxidation and fat deposition.It plays a decisive role in the production of ATP in oxidative phosphorylation.Inorder to study the relationship betweenUCP3gene and pork quality traits,11 meat quality indexes of 165Jiangkou Luobo pigs(Sus scrofa)were determined.SNPs loci were screened by sequencing all exons ofUCP3gene amplified by mixed DNA pool.Single sample DNA was amplified and sequenced for validation andtyping.At the same time,the difference of meat quality among individuals with different genotypes wasanalyzed.The expression ofUCP3gene of individuals with different genotypes was detected to study therelationship between meat quality andUCP3gene expression.This study identified 6 SNPs,they wereg.3127AG,g.3205TC,g.10439CT,g.10441CA,g.10466GA and g.10479AG/T.Population geneticparameters analysis showed that the heterozygosity except g.10479AG/T locus was high,which indicatedthat the distribution of alleles in the population was relatively uniform;the number of effective alleles washigh,which indicated that the degree of variation in the population was large;and the content of polymorphicinformation was more than 0.25,which indicated that the loci could provide some genetic information.2testshowed that the genotype distribution of g.3127AG and g.3205TC loci was in Hardy-Weinberg equilibrium,while the genotype distribution of g.10439CT,g.10441CA and g.10466GA loci deviated significantlyfrom Hardy-Weinberg equilibrium.Linkage disequilibrium analysis showed that g.3127AG and g.3205TCloci,g.10439CT and g.10466GA loci were completely linkage disequilibrium,g.10441CA,g.10466GAand g.10479AG/T were strongly linkage disequilibrium.The correlation analysis between SNPs loci andmeat quality showed that marbling scores of TT and GG individuals at g.10479AG/T loci were significantlyhigher than those of AA and AG individuals,whereas shear stress was the opposite.Genotype-expressioncorrelation analysis showed that g.10439CT,g.10441CA,g.10466GA and g.10479AG/T loci weresignificantly higher than those of others.UCP3gene expression was negatively correlated with shear stressand positively correlated with intramuscular fat content.The results showed that g.10439CT,g.10441CA,g.10466GA and g.10479AG/T sites were important SNPs sites affecting the meat quality of Jiangkou Luobopigs,and the increase ofUCP3gene expression was beneficial to the deposition of intramuscular fat.Theresults provide a reference for the screening of molecular markers that contribute to intramuscular fatdeposition in Jiangkou Luobo pig.KeywordsJiangkou Luobo pig;Uncoupling protein 3(UCP3);SNP;Meat quality;Correlation analysis江口萝卜猪UCP3基因变异及其表达量与肉质性状关联性分析CorrelationAnalysis of UCP3 Gene Variation,Expression and Meat Quality Traits in Jiangkou Luobo Pig(Sus scrofa)491农业生物技术学报Journal ofAgricultural Biotechnology材料，屠宰后测定其11项主要肉质性能指标，并提取背最长肌中的基因组DNA建立混合DNA池，扩增UCP3基因全部外显子并测序，初步筛选多态位点，然后以单个样本DNA为模板，扩增筛选到的位点对应外显子，用以测序验证并分型。对应肉质指标，对不同基因型猪进行肉质性能差异分析。提取背最长肌总RNA，检测不同基因型江口萝卜猪背最长肌中UCP3基因的平均表达量做差异显著性分析，分析基因型、表达量与及肉质性能三者之间的关联性，以期探寻UCP3基因多态性对江口萝卜猪肉质性能的影响，为科学的选种选育提供基础材料。1材料与方法1.1实验动物实验动物为贵州省江口萝卜猪(Sus scrofa)繁育中心提供的1012月龄去势雄性江口萝卜猪，共165头无亲缘关系，体重为7080 kg。屠宰后取腰大肌和背最长肌测定肉质性能，取部分组织置于75%酒精中保存，用于基因组DNA提取，另外在背最长肌中取部分组织用于RNA提取，用预装冻存液的冻存管置于-80低温冰箱保存。1.2主要试剂与仪器2TaqMasterMix(康为世纪,北京)、Ezup柱式动物基因组DNA抽提试剂盒(上海生工)、琼脂糖(上海生工)、TRIzol(RNAiso Plus,TaKaRa,日本)、iTaqUniversal SYBR Green Supermix(Bio-rad,美国)、RT-PCR Kit(K1622,赛默飞,美国)。荧光定量 PCR 仪(CFX Connect,Bio-Rad,美国)、胴体PH测定仪(PH-STAR,德国,Matthaus)、肌肉嫩度仪(RTA-meat,上海腾拔)、胴体肉色测定仪(OPTO-STAR,Matthaus,德国)、肉质品质分析仪(series3000,NI,澳大利亚)。1.3肉质性能测定大理石纹、剪切力、熟肉率、滴水损失等肉质性能指标参照 猪肌肉品质测定技术规范(NY/T821-2004)测定。使用胴体PH测定仪测定pH值，使用胴体肉色测定仪测定肉色，肉色以Opto值(比色值)表示，Opto值参考标准：Opto53为差：肉质有缺陷的 PSE 猪肉(pale soft exudative meat,PSE肉)；53Opto63为良：较满意；Opto63为优：肉质好的猪肉(龚建军,2005)。使用肉品质分析仪测定背最长肌粗蛋白含量、水分含量和肌内脂肪含量，测定前需用索氏抽提法测定5个肉样，然后将数据输入肉品质分析仪中校正仪器，其余肉样即可直接测定。1.4DNA池构建与cDNA合成利用动物基因组DNA抽提试剂盒提取基因组DNA，提取的DNA用超微量紫外分光光度计检测纯度和浓度。将所有样本DNA浓度调整至50 ng/L，再各取5L构建混合DNA池，于-20冰箱保存备用。背最长肌的总 RNA 采用 TRIzol法提取，并检测浓度和纯度，用逆转录试剂盒进行cD-NA合成，总RNA用量为2g。1.5基因引物设计参 考 NCBI 上 登 录 号 为 NC_010451.4 的 猪UCP3基因 DNA 参考序列，利用软件 Primer Pre-mier 5针对UCP3基因的7个外显子序列设计特异性引物，并根据 mRNA 参考序列设计荧光定量PCR引物，内参基因选用-actin。由生工生物技术(上海)有限公司合成，基因引物信息见表1。1.6PCR扩增反应体系(20L)：2TaqPCR Master Mix 10L、10 pmol/L 上下游引物各 2L、DNA 模板 2L、ddH2O 4L。PCR扩增结束后取4L扩增产物经琼脂糖凝胶电泳检查扩增效果。PCR反应程序：95预变性 2 min；95变性 30 s、退火 30 s、72延伸 30 s、总共 35 个循环；72终止延伸 2min。qRT-PCR反应体系(10L)：2iTaqUniversalSYBR Green Supermix 5L、cDNA 模 板 1L，ddH2O 3L，10 pmol/L 上、下游引物各 0.5L。反应程序：95预变性3 min；95变性5 s，60.5退火30 s，72延伸30 s，共40个循环，并绘制扩增曲线。40 个循环完成后从 60至 95以每 5 s0.5的速率上升，并收集荧光信号绘制熔解曲线。每个样品做3个重复。1.7统计分析将以DNA池为模板的PCR扩增产物送由生物公司测序，用SeqMan查看峰图筛选套峰位点，再与参考序列比对，初步确定多态位点，然后以单个DNA为模板扩增相应序列并测序，用于对每个位点的分型。统计各个位点不同基因型个体数，计算492包括基因纯合度(homozygosity,Ho)、基因杂合度(heterozygosity,He)、有效等位基因数(effect num-ber of allele,Ne)、多态信息含量(polymorphism in-formation content,PIC)在内的各项群体遗传参数，并用2检验Hardy-Weinberg平衡。各位点之间的连锁关系用在线软件SHEsis(http:/analysis.bio- 结束后各取 4L产物进行琼脂糖凝胶电泳检测，可以看到电泳条带清晰明亮，无杂带，无拖尾，说明引物特异性好，扩增片段可用于直接测序，且对照DNA Marker可以看出扩增片段大小与实验设计引物对应片段大小一致，与实验设计相符。2.2PCR测序分型扩增得到的江口萝卜猪UCP3基因所有外显子序列送由生工生物工程（上海）股份有限公司双向测序，在所有外显子序列中共筛选到6个多态位点，将猪UCP3基因参考序列第1外显子的第1位碱基定义为+1，则这6个位点分别命名为：g.3127AG、g.3205TC、g.10439CT、g.10441CA、g.10466GA和g.10479AG/T。g.3127AG和g.3205TC位点位于 5UTR，其余 4 个位点均位于 3UTR，其中g.10479AG/T位点存在AT型和AG型两种突变类型(图2)。测序验证多态位点和分型见图3。将不同于参考序列的纯合型个体定义为突变型，杂合个体定义为杂合型，与参考序列相同的纯合型个体定义为野生型，其中g.10479AG/T位点只检测到4种基因型，分别为突变型GG和TT，野生型AA，以及杂合型AG，其余位点均检测到3种基因型。2.3群体遗传参数分析如表2所示，UCP3基因上的6个SNPs位点中，g.10479AG/T位点A为优势等位基因，等位基因频率为0.560 6，其中T等位基因频率较低，为0.121 2，由于该位点存在3个等位基因，但只检测到4种基因型，造成此位点等位基因分布不均匀，有效等位基因数低，但杂合度为0.569 8，多态信息含量接近于高度多态，说明此位点变异程度大遗传信息较丰富。位于5UTR的2个位点和位于3UTR的3个位点遗传参数趋于一致，推测是因为位点之间距离较近，导致遗传上存在连锁。但是可以看到几个位点杂合度、有效等位基因数均较高，多态信息含量显示为中度多态，表明群体内变异程度大，可以提供一定的遗传信息。2检验显示所有位点基因型分布均极显著偏离Hardy-Weinberg平衡，这可能是由于长期的人工选择淘汰，导致一些基因型个体数量较少所致。引物名称Primer nameUCP3E1UCP3E2UCP3E3-4UCP3E5-6UCP3E7UCP3-actin片段大小/bpProduct length4725278921214932194165引物序列(53)Primer sequencesF:CTCCAGTCAAGTGATCGGGC;R:TGGGGCAAGGGCTTTATGF:GCTGGGATGCAGGGTCA;R:TTGGGCGATGGTGATGGF:CGCCACGAACCTGTCATT;R:TGGTTAGGGTGTTGGTTTAGAATF:TCTCTAAGACAGACACGGATAAT;R:GTTGCTGATGGCACTTTGACF:CCTCTGCCCGAGTTCCCT;R:GCTTTGTGCTCCCATTTTACGF:CCAACATCACGAGGAATGCC;R:CGAGTTCATGTACCGGGTCTF:CTTCTTGGGCATGGAGTC;R:TAGAGGTCCTTCCTGATGTTm/62.860.057.050.562.861.061.0用途UsagePCRPCRPCRPCRPCRqRT-PCRqRT-PCR表1UCP3基因引物信息Table 1Primer information of UCP3 geneUCP3：解耦联蛋白3；-actin：-肌动蛋白；下同UCP3:Uncoupling protein 3;The same below江口萝卜猪UCP3基因变异及其表达量与肉质性状关联性分析CorrelationAnalysis of UCP3 Gene Variation,Expression and Meat Quality Traits in Jiangkou Luobo Pig(Sus scrofa)493农业生物技术学报Journal ofAgricultural Biotechnology2.4连锁不平衡分析运用SHEsis在线软件对江口萝卜猪UCP3基因6个SNPs位点进行连锁不平衡分析，结果如表3和图4。其D值和r2值为位点之间连锁程度的判断依据，其中D值为1时位点之间完全连锁，r2=1为完美连锁，D=0或r2=0为无连锁或连锁平衡，r20.33提示为强连锁。UCP3基因g.3127AG和g.3205TC位点、g.10439CT和g.10466GA 位点，D=1，r2=1 为完全连锁不平衡，g.10439CT、g.10441CA、g.10466GA、g.10479AG/T 4个位点之间D1，r20.33，为强连锁不平衡。此结果表明g.3127AG和g.3205TC 位点以及 g.10439CT 和 g.10466GA位点是紧密连锁作为一个整体遗传，而其余位点之间趋向于相互独立遗传。2.5基因型与肉质性状的关联性分析UCP3基因上共检测到6个SNPs位点，表4可以看出在大理石纹评分和剪切力两项指标上，g.10479AG/T位点野生型和杂合型个体大理石纹评分与两种突变型个体相比较低，差异显著而剪切力则反之。其余位点各项肉质指标和肌肉组成成分含量差异均不显著，但是也可以看到，g.10439CT、g.10441CA以及g.10466GA位点上突变型个体大理石纹评分和肌内脂肪含量与野生型以及杂合型相比明显较高，剪切力为突变型个体明显低于野生型和杂合型。总体而言，这几个SNPs位点不同基因型的江口萝卜猪主要在大理石纹、剪切力和肌内脂肪含量3项指标上存在明显差异，且突变型均为大理石纹评分和肌内脂肪含量较高，剪切力较低，肉质性能表现较好，这提示UCP3基因3UTR的SNPs位点变异可能促进江口萝卜猪肌内脂肪的沉积。2.6不同基因型个体UCP3基因表达量差异分析本实验每个位点每种基因型选择10个RNA样本，随机选择13号肉猪样本cDNA荧光定量PCR扩增Ct值为对照，计算UCP3基因各多态位点不同基因型江口萝卜猪背最长肌中UCP3基因相对表达量平均值(图5)。可以看到，UCP3基因5UTR上的g.3127AG和g.3205TC位点各基因型表达量差异不大，说明2个位点对UCP3基因的表达量影响较 小。3UTR 上 的 g.10439CT、g.10441CA、图1UCP3基因的PCR扩增Figure 1PCR amplification ofUCP3geneM：DL2000 DNAMarker；15：外显子扩增片段；6：UCP3基因荧光定量PCR扩增片段；7：内参基因荧光定量PCR扩增片段15:Exon amplification fragment;6:Real time quantitative PCR amplification fragment ofUCP3gene;7:Real time quantitativePCR amplification fragment of internal reference geneM1bpbp20001000750500250100472bp20001000750500250100M2bp527bp20001000750500250100M3bp892bp20001000750500250100bp1214M4bp20001000750500250100M5bp932bp20001000750500250100M6bp194bp20001000750500250100M7bp165494g.3205TC(TT)g.3205TC(TC)g.3205TC(CC)g.10439CT(CC)g.10439CT(CT)g.10439CT(TT)g.10441CA(CC)g.10441CA(CA)g.10441CA(AA)g.10466GA(GG)g.10466GA(GA)g.10466GA(AA)g.10479AG/T(AA)g.10479AG/T(GG)g.10479AG/T(AG)g.10479AG/T(TT)g.3127AG(AA)g.3127AG(AG)g.3127AG(GG)g.10466GA以及g.10479AG/T位点基因表达量差异显著，为突变型高、野生型和杂合型低，说明这4个位点基因的变异对UCP3基因在背最长肌中的表达有积极影响。2.7UCP3基因表达量与肉质性能相关性分析利用SPSS18.0中的双变量相关性分析，研究图3SNPs位点不同基因型测序图谱Figure 3SNPs loci sequencing map of different genotypes图2SNPs位点混池测序图谱Figure 2SNPs locus pooled sequencing mapg.3127AGg.3205TCg.10439CTg.10441CAg.10466GAg.10479AG/T江口萝卜猪UCP3基因变异及其表达量与肉质性状关联性分析CorrelationAnalysis of UCP3 Gene Variation,Expression and Meat Quality Traits in Jiangkou Luobo Pig(Sus scrofa)495农业生物技术学报Journal ofAgricultural BiotechnologyUCP3基因在背最长肌中的表达量与肉质性状之间的相关性(表5)。可以看到，UCP3基因在背最长肌中的表达量与肉色、水份含量等指标为正相关，与pH、粗蛋白含量等指标为负相关。其中，肌内脂肪含量呈极显著正相关，与剪切力呈极显著负相关。可见，UCP3基因的表达量与肌内脂肪的沉积密切相关，UCP3基因在背最长肌中的表达量越高，越有利于肌内脂肪的沉积，肉质性能就越好。位点Locusg.3127AGg.3205TCg.10439CTg.10441CAg.10466GAg.10479AG/T样本数Samplenumber1010550101055080256080256080256080254020基因型GenotypeAAAGGGTTTCCCCCCTTTCCCAAAGGGAAAAAAGGGTT基因型频率Frequency ofgenotype0.06060.63640.30300.06060.63640.30300.48480.15150.36360.48480.15150.36360.48480.15150.36360.48480.15150.24240.1212等位基因AlleleAGTCCTCAGAAGT等位基因频率Allele frequency0.37880.62120.37880.62120.56060.43940.56060.43940.56060.43940.56060.31820.1212Ho0.52940.52940.50730.50730.50730.4302He0.47060.47060.49270.49270.49270.5698Ne1.88901.88901.97101.97101.97102.3244PIC0.35990.35990.37130.37130.37130.4939HWE（2）17.777817.777879.115679.115679.1156229.8186表2UCP3基因群体遗传参数Table 2Genetic parameters of UCP3 gene populationHo：纯合度；He：杂合度；Ne：有效等位基因数；PIC：多态信息含量；PIC5：高度多态，0.25PIC0.5：中度多态，PIC0.25：低度多态；HWE(2)：基因型哈代温伯格平衡，20.01(2)=9.21Ho:Homozygosity;He:Heterozygosity;Ne:Number of effective alleles;PIC:Information content of gene polymorphism;PIC5:High diversity,0.25PIC0.5:Moderate diversity,PIC0.25:Low diversity;HWE(2):Genotype Hardy-Weinberg equilib-rium,20.01(2)=9.21位点Locusg.3127AGg.3205TCg.10439CTg.10441CAg.10466GAg.10479AG/Tg.3127AG1.0000.1650.1780.1650.139g.3205TC1.0000.1650.1780.1650.139g.10439CT0.5950.5950.9411.0000.771g.10466GA0.5950.5951.0001.0000.771g.10441CA0.5990.5991.0000.9410.726g.10479AG/T0.6020.6021.0001.0001.000表3江口萝卜猪UCP3基因SNPs位点连锁不平衡分析Table 3Linkage disequilibrium analysis of UCP3 gene SNPs in Jiangkou Luobo pigs表对角线上方为D值，对角线下方为r2值Above the diagonal of the table is the value of D,below the diagonal is the value of r2496位点Locusg.3127AGg.3205TCg.10439CTg.10441CAg.10466GAg.10479AG/T数量No.1010550101055080256080256080256080254020基因型Geno-typeAAAGGGTTTCCCCCCTTTCCCAAAGGGAAAAAAGGGTTpH16.330.026.290.266.360.306.330.026.290.266.360.306.320.266.290.336.380.296.320.266.290.336.380.296.320.266.290.336.380.296.390.286.430.336.340.266.370.22熟肉率Cookedmeatpercentage64.611.4065.061.3764.011.0264.611.4065.061.3764.011.0264.951.4463.771.2264.251.2464.951.4463.771.2264.251.2464.951.4463.771.2264.251.2464.730.8963.981.2163.561.4364.001.44PH245.720.185.810.225.860.255.720.185.810.225.860.255.840.235.910.245.880.365.840.235.910.245.880.365.840.235.910.245.880.365.820.255.800.245.840.235.740.10肉色Meatcolor(Opto)62.011.7964.281.9962.882.3762.011.7964.281.9962.882.3763.751.9463.432.0164.652.6763.751.9463.432.0164.652.6763.751.9463.432.0164.652.6763.772.5162.393.0163.662.5562.143.47大理石纹Marblingscores3.490.573.550.623.560.773.490.573.550.623.560.773.430.563.450.613.830.773.430.563.450.613.830.773.430.563.450.613.830.773.200.53a3.210.44a4.070.66b4.020.59b失水率Rate ofwater loss19.210.9919.950.9718.880.9119.210.9919.950.9718.880.9119.000.8618.710.6918.661.0119.000.8618.710.6918.661.0119.000.8618.710.6918.661.0118.561.1118.920.8119.040.9919.120.77滴水损失Drip loss1.850.911.820.841.900.651.850.911.820.841.900.651.990.811.910.841.960.761.990.811.910.841.960.761.990.811.910.841.960.762.050.832.040.791.890.611.970.59剪切力Shearingforce(N)42.884.4532.784.5631.295.0942.884.4532.784.5631.295.0935.443.3433.884.8227.123.6335.443.3433.884.8227.123.6335.443.3433.884.8227.123.6342.134.54a40.883.25a23.323.98b22.333.09b水份/%Moisture67.991.5368.712.0568.092.3167.991.5368.712.0568.092.3168.332.7167.871.9767.942.0168.332.7167.871.9767.942.0168.332.7167.871.9767.942.0167.772.3268.441.7067.912.0168.070.85肌内脂肪/%Intramuscu-la