湘村黑猪sirt2基因多态性及其与肉质性状的关联分析和组织表达研究

doi:10.16431/j.cnki.1671-7236.2019.04.015

摘要/Abstract

摘要：

本研究旨在探索沉默信号调节子家族（silent information regulator 1-7，SIRT1-7）sirt2基因在湘村黑猪不同组织间的表达及其多态性与肉质性状间的关联性，以期寻找与湘村黑猪肉质性状相关的分子标记。采用PCR-RFLP方法和基因测序技术对湘村黑猪sirt2基因多态位点进行分析，采用实时荧光定量PCR技术对sirt2基因在湘村黑猪心脏、肝脏、脾脏、肺脏、肾脏、胰腺、后腿肌和背最长肌8个组织中的相对表达量进行分析，利用SAS 9.4软件对sirt2基因突变位点不同基因型与肌肉色值、pH_{45 min}、pH_{24 h}、滴水损失、失水率、肌内脂肪、嫩度和眼肌面积进行关联分析。结果显示，在湘村黑猪sirt2基因第8外显子扩增片段中的240 bp处发现1处C→T碱基突变，编码氨基酸由精氨酸（Arg）变为半胱氨酸（Cys），为错义突变，并形成CC、CT和TT 3种基因型，CC基因型为优势基因型，C为优势等位基因，经χ²检验表明该突变位点偏离哈代-温伯格平衡；该位点群体纯合度较高，有效等位基因数为1.301，多态信息含量为0.205，为低度多态（PIC<0.25）。基因多态性与肉质性状关联分析结果表明，TT基因型肉色L^*值和滴水损失显著低于CC和CT基因型（P<0.05），CC基因型失水率显著高于CT和TT基因型（P<0.05）。sirt2基因在湘村黑猪8个组织中均有表达，其中在背最长肌中相对表达量最高，与肺脏中表达量差异不显著（P>0.05），但显著高于心脏、肝脏、脾脏、肾脏、胰腺和后腿肌（P<0.05），且心脏、肝脏、脾脏、肾脏和胰腺中相对表达量差异均不显著（P>0.05）。本试验结果表明，sirt2基因对湘村黑猪肉质性状的发育有一定影响，可作为影响湘村黑猪肉质性状的候选基因进行深入研究。

关键词: 湘村黑猪; sirt2基因; 肉质性状; 组织表达; 关联分析

Abstract:

This study was aimed to investigate the expression of the silent information regulator family (SIRT1-7) sirt2geneof different tissues inXiangcunBlackpig,analyzetheassociation between sirt2 gene polymorphism and meat quality traits,and find the molecular markers associated with meat quality traits in Xiangcun Black pig.The PCR-RFLP method and gene sequencing technology were used to analyze the polymorphism of sirt2 gene in Xiangcun Black pig.Real-time PCR was used to analyze the relative expression of sirt2 gene in eight tissues including heart,liver,spleen,lung,kidney,pancreas,hind leg muscle and longissimus dorsi.At the same time,the different genotypes of sirt2 gene mutation site and muscle color value,pH_{45 min},pH_{24 h},drip loss,water loss rate,intramuscular fat,tenderness and loin eye muscle area were analyzed by SAS 9.4 software.The results showed that a mutation (C to T) was found at 240 bp in the amplified fragment of sirt2 gene exon 8 in Xiangcun Black pig,and the encoded amino acid changed from arginine (Arg) to cysteine (Cys).The mutation type was a missense mutation and formed three genotypes of CC,CT and TT,CC genotype was the dominant genotype,and C was the dominant allele.The χ² test indicated that the mutation site deviated from Hardy-Weinberg equilibrium.The homozygosity of this locus was higher,the number of effective alleles was 1.301,and the polymorphic information content was 0.205,which was low polymorphism (PIC<0.25).Correlation analysis between gene polymorphism and meat quality traits showed that the muscle L^* value and drip loss of TT genotype were significantly lower than that of CC and CT genotypes (P<0.05).The water loss rate of CC genotype was significantly higher than that of CT and TT genotypes (P<0.05).sirt2 gene was expressed in 8 tissues of Xiangcun Black pig,and the relative expression was the highest in longissimus dorsi,which was not significantly different with lung (P>0.05),but was significantly higher than heart,liver,spleen,kidney,pancreas and hind leg muscle (P<0.05),and the relative expressions in heart,liver,spleen,kidney and pancreas were not significant (P>0.05).The results showed that sirt2 gene had a certain effect on the development of meat quality traits in Xiangcun Black pig,which could be used as a candidate gene affecting the meat quality in Xiangcun Black pig.

Key words: Xiangcun Black pig; sirt2 gene; meat quality traits; tissue expression; association analysis

中图分类号:

S813.3

崔清明, 张星, 彭英林, 高峰, 陈晨, 邓缘, 任慧波, 胡雄贵, 朱吉, 罗璇, 刘莹莹, 左剑波. 湘村黑猪sirt2基因多态性及其与肉质性状的关联分析和组织表达研究[J]. 《中国畜牧兽医》, 2019, 46(4): 1086-1093.

CUI Qingming, ZHANG Xing, PENG Yinglin, GAO Feng, CHEN Chen, DENG Yuan, REN Huibo, HU Xionggui, ZHU Ji, LUO Xuan, LIU Yingying, ZUO Jianbo. Study on Tissue Expression and Association Analysis Between sirt2 Gene Polymorphism and Meat Quality Traits in Xiangcun Black Pig[J]. , 2019, 46(4): 1086-1093.

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