宁乡猪FABPs、SLC13A5和NR1H4基因多态性及其与IMF含量的关联分析

doi:10.11843/j.issn.0366-6964.2022.11.003

摘要/Abstract

摘要： 旨在研究宁乡猪FABPs、SLC13A5和NR1H4基因的多态性及其与肌内脂肪（IMF）含量的关联性。本试验采用PCR-RFLP和Sanger测序的方法对172头宁乡阉公猪FABPs、SLC13A5和NR1H4基因的多态性进行检测，并探究多态性与宁乡猪IMF含量的关联性。结果表明，宁乡猪H-FABP-Hinf Ⅰ酶切位点存在多态性，其中Hh基因型个体的IMF含量最高，与HH、hh基因型个体的IMF含量差异极显著（P<0.01）；H-FABP-Hae Ⅲ酶切只检测到DD和Dd两种基因型，Dd基因型个体的IMF含量极显著高于DD基因型个体（P<0.01）；H-FABP基因的Hinf Ⅰ和Hae Ⅲ多态性位点共形成3种单倍型和5种单倍型组合，其中单倍型组合HD/hd个体具有最佳的IMF含量。H-FABP的Msp I酶切结果及A-FABP基因测序结果显示，宁乡猪不存在多态现象；SLC13A5基因多态性位点g.50705299T/C和NR1H4基因多态性位点g.83607915G/A处不同基因型间IMF含量的差异均不显著（P>0.05）。综上所述，H-FABP基因Hinf Ⅰ和Hae Ⅲ酶切位点多态性与宁乡猪IMF含量显著关联，其形成的单倍型组合HD/hd的IMF含量最高。以上结果可为宁乡猪IMF含量性状的早期分子选育提供参考。

关键词: 宁乡猪, FABPs, SLC13A5, NR1H4, 肌内脂肪

Abstract: The study aimed to explore the polymorphisms of FABPs, SLC13A5 and NR1H4 genes in Ningxiang pigs and their association with intramuscular fat (IMF) content. In the experiment, the polymorphisms of FABPs, SLC13A5 and NR1H4 genes in 172 Ningxiang barrows were detected using PCR-RFLP and Sanger sequencing, and the association between them and IMF content was explored. The results showed that the Hinf Ⅰ digestion site of H-FABP gene was polymorphic in Ningxiang pigs, and the IMF content of individuals with Hh genotype was the highest, which was significantly different from that of individuals with HH and hh genotypes (P<0.01). Only DD and Dd genotypes were detected in H-FABP using Hae Ⅲ enzyme, and the IMF content of individuals with Dd genotype was significantly higher than that of individuals with DD genotype (P<0.01). There were 3 haplotypes and 5 haplotype combinations in the analysis of the Hinf Ⅰ and Hae Ⅲ sites of H-FABP gene, and the pigs with haplotype combination HD/hd had the best IMF content. The results of H-FABP Msp I digestion and A-FABP sequencing showed that there was no polymorphism at the detected sites in Ningxiang pigs. No significant difference was observed in IMF content among different genotypes at the polymorphism site of SLC13A5 (g.50705299T/C) and NR1H4 (g.83607915G/A) gene (P>0.05). In conclusion, the polymorphisms of H-FABP Hinf Ⅰ and Hae Ⅲ sites were significantly associated with IMF content in Ningxiang pigs, and the pigs with haplotype combination HD/hd had the highest IMF content. The above results could provide reference for the early molecular breeding of IMF content trait in Ningxiang pigs.

Key words: Ningxiang pigs, FABPs, SLC13A5, NR1H4, IMF

中图分类号:

S828.2

李艺阳, 殷诗舒, 廖印长, 徐康, 张跃博, 何俊. 宁乡猪FABPs、SLC13A5和NR1H4基因多态性及其与IMF含量的关联分析[J]. 畜牧兽医学报, 2022, 53(11): 3737-3747.

LI Yiyang, YIN Shishu, LIAO Yinchang, XU Kang, ZHANG Yuebo, HE Jun. Polymorphisms of FABPs, SLC13A5 and NR1H4 Genes in Ningxiang Pigs and Their Association with IMF Content[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(11): 3737-3747.

参考文献 42

[1]	肖瑜.宁乡猪全基因组重测序及其特异性标记筛选研究[D].长沙:湖南农业大学, 2019.XIAO Y.Whole genome resequencing of Ningxiang pig and its specific marker screening[D].Changsha:Hunan Agricultural University, 2019.(in Chinese)
[2]	任慧波.地方优良猪品种——宁乡猪[J].农村百事通, 2019(13):36.REN H B.Local excellent pig breed-Ningxiang pig[J].Nongcun Baishitong, 2019(13):36.(in Chinese)
[3]	吴婷, 敖翔, 何健.肌内脂肪与肉品质关系及其营养调控策略[J].养猪, 2021(4):50-54.WU T, AO X, HE J.Relationship between intramuscular fat and meat quality and its nutritional regulation strategy[J].Swine Production, 2021(4):50-54.(in Chinese)
[4]	宿甲子, 孙肖明, 孟宪梅.肌内脂肪对猪肉品质的影响及研究进展[J].畜牧兽医科技信息, 2020(10):19-20.SU J Z, SUN X M, MENG X M.Effects of intramuscular fat on pork quality and research progress[J].Chinese Journal of Animal Husbandry and Veterinary Medicine, 2020(10):19-20.(in Chinese)
[5]	陆鹏.走近宁乡花猪[J].湖南农业, 2021(1):7.LU P.Approaching Ningxiang pig[J].Hunan Agricuture, 2021(1):7.(in Chinese)
[6]	李志娟.猪心脏型脂肪酸结合蛋白研究进展[J].养猪, 2021(2):55-56.LI Z J.Research progress of heart fatty acid-binding protein[J].Swine Production, 2021(2):55-56.(in Chinese)
[7]	HOVENIER R, KANIS E, VERHOEVEN J A M.Repeatability of taste panel tenderness scores and their relationships to objective pig meat quality traits[J].J Anim Sci, 1993, 71(8):2018-2025.
[8]	庞卫军, 杨公社.8个猪种和野猪H-FABP基因分子标记与IMF含量的关系[J].农业生物技术学报, 2005, 13(6):733-738.PANG W J, YANG G S.Relationship between molecular marker of eight pig and one wild pig H-FABP gene and IMF content[J].Journal of Agricultural Biotechnology, 2005, 13(6):733-738.(in Chinese)
[9]	李慧.巴淮猪及亲本猪四个肉质相关基因多态性和巴淮猪胴体肉质分析[D].滁州:安徽科技学院, 2019.LI H.Four meat quality-related gene polymorphisms in Bahuai pigs and their parents and analysis of carcass meat quality in Bahuai pigs[D].Chuzhou:Anhui Science and Technology University, 2019.(in Chinese)
[10]	GERBENS F, DE KONING D J, HARDERS F L, et al.The effect of adipocyte and heart fatty acid-binding protein genes on intramuscular fat and backfat content in Meishan crossbred pigs[J].J Anim Sci, 2000, 78(3):552-559.
[11]	杨虎.沙子岭猪和大白猪背最长肌转录组与蛋白组研究[D].长沙:湖南农业大学, 2015.YANG H.Transcriptome and proteome study on longissimus dorsi muscle between Shaziling and Yorkshire pigs[D].Changsha:Hunan Agricultural University, 2015.(in Chinese)
[12]	SHANG P, ZHANG B, ZHANG J, et al.Expression and single-nucleotide polymorphisms of the H-FABP gene in pigs[J].Gene, 2019, 710:156-160.
[13]	DE KONING D J, JANSS L L G, RATTINK A P, et al.Detection of quantitative trait loci for backfat thickness and intramuscular fat content in pigs (Sus scrofa)[J].Genetics, 1999, 152(4):1679-1690.
[14]	PANG W J, BAI L, YANG G S.Relationship among H-FABP gene polymorphism, intramuscular fat content, and adipocyte lipid droplet content in main pig breeds with different genotypes in western China[J].Acta Genet Sin, 2006, 33(6):515-524.
[15]	高广雄, 李斌, 高宁, 等.杜长大商品猪肌内脂肪含量的全基因组关联分析[J].南方农业学报, 2022, 53(4):985-990.GAO G X, LI B, GAO N, et al.Genome-wide association study of intramuscular fat content in a Duroc-Landrace-Yorkshire pig population[J].Journal of Southern Agriculture, 2022, 53(4):985-990.(in Chinese)
[16]	杨帅, 史明月, 李文霞, 等.猪肌内脂肪沉积相关基因的筛选及其表达特性分析[J].中国畜牧兽医, 2022, 49(5):1651-1661.YANG S, SHI M Y, LI W X, et al.Screening and expression analysis of genes related to porcine intramuscular fat deposition[J].China Animal Husbandry & Veterinary Medicine, 2022, 49(5):1651-1661.(in Chinese)
[17]	谭娅, 李亮, 雷宇航, 等.高低肌内脂肪猪背最长肌关键基因表达差异分析[J].南方农业学报, 2022, 53(4):899-907.TAN Y, LI L, LEI Y H, et al.Expression difference analysis of hub genes between high and low intramuscular fat content in longissimus dorsi of pigs[J].Journal of Southern Agriculture, 2022, 53(4):899-907.(in Chinese)
[18]	孙菁希, 逄世龙, 孙敬春, 等.基于转录组测序鉴定猪肌内脂肪沉积相关生物学通路及候选基因[J].中国畜牧兽医, 2021, 48(12):4508-4519.SUN J X, PANG S L, SUN J C, et al.Identification of biological pathways and candidate genes related to porcine intramuscular fat deposition using transcriptome sequencing[J].China Animal Husbandry & Veterinary Medicine, 2021, 48(12):4508-4519.(in Chinese)
[19]	OCKNER R K, MANNING J A, POPPENHAUSEN R B, et al.A binding protein for fatty acids in cytosol of intestinal mucosa, liver, myocardium, and other tissues[J].Science, 1972, 177(4043):56-58.
[20]	宋波, 帅素容.脂肪酸结合蛋白A-FABP和E-FABP的研究进展[J].畜牧与兽医, 2017, 49(10):135-139.SONG B, SHUAI S R.The progress in fatty acid binding protein A-FABP and E-FABP[J].Animal Husbandry & Veterinary Medicine, 2017, 49(10):135-139.(in Chinese)
[21]	于嘉霖, 徐雅楠, 韩璐, 等.脂肪酸结合蛋白4对BCG诱导巨噬细胞自噬的调控作用[J].畜牧兽医学报, 2020, 51(9):2265-2274.YU J L, XU Y N, HAN L, et al.Role of fatty acid binding protein 4 in regulating macrophage autophagy induced by BCG infection[J].Acta Veterinaria et Zootechnica Sinica, 2020, 51(9):2265-2274.(in Chinese)
[22]	刘志成, 强巴央宗, 张浩, 等.藏猪FABPs基因PCR-RFLP多样性研究[J].中国农业大学学报, 2009, 14(6):6-10.LIU Z C, CHAMBA Y, ZHANG H, et al.Research on PCR-RFLP polymorphisms of FABPs genes in Tibet pigs[J].Journal of China Agricultural University, 2009, 14(6):6-10.(in Chinese)
[23]	张陈华, 王阳, 丁月云, 等.圩猪H-FABP基因多态性分析及其与IMF含量的相关性[J].中国农业科学, 2011, 44(5):1063-1070.ZHANG C H, WANG Y, DING Y Y, et al.Polymorphism analysis of H-FABP gene and correlation with IMF content in Wei Pig[J].Scientia Agricultura Sinica, 2011, 44(5):1063-1070.(in Chinese)
[24]	刘剑锋, 王立贤, 张贵香, 等.H-FABP基因型对中畜黑猪I系生长性能的影响[J].畜牧兽医学报, 2005, 36(6):555-558.LIU J F, WANG L X, ZHANG G X, et al.Influence of PCR-RFLP genotypes of porcine H-FABP gene on production performances in Zhongxu black line I population[J].Acta Veterinaria et Zootechnica Sinica, 2005, 36(6):555-558.(in Chinese)
[25]	王存芳.猪H-FABP和A-FABP基因的多态性及其与肌内脂肪性状关系的研究[D].泰安:山东农业大学, 2002.WANG C F.Studies on polymorphisms of H-FABP and A-FABP genes and their associations with IMF trait in pigs[D].Taian:Shandong Agricultural University, 2002.(in Chinese)
[26]	GERBENS F, VERBURG F J, VAN MOERKERK H T B, et al.Associations of heart and adipocyte fatty acid-binding protein gene expression with intramuscular fat content in pigs[J].J Anim Sci, 2001, 79(2):347-354.
[27]	柳小春, 郭松长, 施启顺.H-FABP基因对猪肉品质的影响[C]//中国的遗传学研究——中国遗传学会第七次代表大会暨学术讨论会论文摘要汇编.海口:中国遗传学会, 2003.LIU X C, GUO S C, SHI Q S.Effects of H-FABP gene on pork quality[C]//Proceedings of the Seventh Congress and Symposium of the Genetics Society of China.Haikou:The Genetics Society of China, 2003.(in Chinese)
[28]	林万华, 黄路生, 任军, 等.中外十个猪种H-FABP基因遗传变异的研究[J].遗传学报, 2002, 29(1):12-15.LIN W H, HUANG L S, REN J, et al.Research on genetic variation of heart fatty acid-binding protein gene in ten pig breeds[J].Acta Genetica Sinica, 2002, 29(1):12-15.(in Chinese)
[29]	许卫华, 康丽珠, 林秀娇, 等.槐猪H-FABP基因多态性及其与肌内脂肪含量相关性研究[J].福建师范大学学报:自然科学版, 2014, 30(2):76-80.XU W H, KANG L Z, LIN X J, et al.Polymorphism analysis of H-FABP gene and its correlation with IMF Content in Huai Pig[J].Journal of Fujian Normal University:Natural Science Edition, 2014, 30(2):76-80.(in Chinese)
[30]	张桂香, 曹红鹤, 王立贤, 等.9个猪种H-FABP基因5'-游区和第二内含子的遗传变异[J].畜牧兽医学报, 2002, 33(4):340-343.ZHANG G X, CAO H H, WANG L X, et al.Genetic variation in 5'-upstream region and the second intron of H-FABP gene in nine pig breeds[J].Acta Veterinaria et Zootechnica Sinica, 2002, 33(4):340-343.(in Chinese)
[31]	房嘉园, 王春丽, 马泽, 等.猪IGF-1R胞外域编码区两种单倍型对基因表达的影响[J].吉林农业大学学报, 2021:1-7.FANG J Y, WANG C L, MA Z, et al.Effects of two haplotypes in the extracellular coding region of porcine IGF-1R on gene expression[J].Journal of Jilin Agricultural University, 2021:1-7.(in Chinese)
[32]	黄大鹏, 黄玉兰, 陈建华.三江白猪心脏脂肪酸结合蛋白(H-FABP)基因多态性及多态片段的序列分析[J].中国兽医学报, 2008, 28(3):336-338, 342.HUANG D P, HUANG Y L, CHEN J H.Polymorphism and sequence analysis of polymorphic fragments of H-FABP gene of Sanjiang white pigs[J].Chinese Journal of Veterinary Science, 2008, 28(3):336-338, 342.(in Chinese)
[33]	黄玉兰, 黄大鹏, 李祥辉.三江白猪脂肪型脂肪酸结合蛋白基因多态性及多态性片段的序列分析[J].中国生物制品学杂志, 2011, 24(7):791-793, 801.HUANG Y L, HUANG D P, LI X H.Polymorphism of adipocyte fatty acid-binding protein gene of Sanjiang white pigs and sequencing of polymorphic fragments[J].Chinese Journal of Biologicals, 2011, 24(7):791-793, 801.(in Chinese)
[34]	李祥辉, 黄大鹏, 黄玉兰.4个不同品种猪A-FABP基因多态性及其与IMF相关性研究[J].中国畜牧杂志, 2011, 47(7):11-14.LI X H, HUANG D P, HUANG Y L.Polymorphism of A-FABP gene in four different pig breeds and its correlation with IMF[J].Chinese Journal of Animal Science, 2011, 47(7):11-14.(in Chinese)
[35]	王顺利, 杨明华, 黄英, 等.A-FABP基因多态性与猪肌内脂肪含量的相关性分析[J].黑龙江畜牧兽医, 2017(9):96-99.WANG S L, YANG M H, HUANG Y, et al.Correlation analysis between A-FABP polymorphisms and intramuscular fat contents in Pigs[J].Heilongjiang Animal Science and Veterinary Medicine, 2017(9):96-99.(in Chinese)
[36]	GERBENS F, JANSEN A, VAN ERP A J M, et al.The adipocyte fatty acid-binding protein locus:characterization and association with intramuscular fat content in pigs[J].Mamm Genome, 1998, 9(12):1022-1026.
[37]	高妍, 张永宏, 熊秋宏, 等.猪A-FABP基因5'调控区多态性及与肉质性状的相关分析[J].畜牧与兽医, 2010, 42(5):34-37.GAO Y, ZHANG Y H, XIONG Q H, et al.Polymorphism of 5' regulatory region of porcine A-FABP gene and its correlation with meat quality traits[J].Animal Husbandry & Veterinary Medicine, 2010, 42(5):34-37.(in Chinese)
[38]	DE AGUIAR VALLIM T Q, TARLING E J, EDWARDS P A.Pleiotropic roles of bile acids in metabolism[J].Cell Metab, 2013, 17(5):657-669.
[39]	GRIMAUDO S, DONGIOVANNI P, PIHLAJAMÄKI J, et al.NR1H4 rs35724 G>C variant modulates liver damage in nonalcoholic fatty liver disease[J].Liver Int, 2021, 41(11):2712-2719.
[40]	YANG H, JIANG J, XU X L, et al.Molecular characterization, tissue expression profile and SNP analysis of the porcine NR1H4 gene[J].Mol Biol Rep, 2014, 41(10):7009-7014.
[41]	LUO W Z, CHENG D X, CHEN S K, et al.Genome-wide association analysis of meat quality traits in a porcine Large White×Minzhu intercross population[J].Int J Biol Sci, 2012, 8(4):580-595.
[42]	王玲玉.猪SLC13A5和Wnt10b基因的克隆、组织表达及其与生长性状的关联分析[D].长沙:湖南农业大学, 2015.WANG L Y.Cloning, Tissue expression profile and SNP analysis of the porcine SLC13A5 and Wnt10b gene[D].Changsha:Hunan Agricultural University, 2015.(in Chinese)