PLPP3基因多态性对北京黑猪背部脂肪厚度和脂肪酸含量的影响

doi:10.16431/j.cnki.1671-7236.2025.01.025

摘要/Abstract

摘要： 【目的】探究磷脂磷酸酶3(PLPP3)基因多态位点与北京黑猪背部脂肪厚度及其脂肪酸含量的相关性,为北京黑猪背部脂肪厚度和脂肪酸含量的遗传改良提供参考。【方法】本研究采集239头北京黑猪耳组织样,获得基因组重测序数据。测定北京黑猪不同部位(肩部最厚处、第6和第7胸椎结合处、胸腰结合处、腰荐结合处)的背部脂肪厚度及左侧胴体第6和7胸椎背部脂肪中5种脂肪酸(棕榈酸、硬脂酸、油酸、亚油酸、α-亚麻酸)的含量。利用测序结果研究PLPP3基因的单核苷酸多态性(SNP)位点的群体遗传特性、单倍型连锁情况,将SNP位点与各部位背部脂肪厚度及脂肪酸含量进行关联分析。进行启动子区SNP转录因子(transcription factor,TF)预测,并运用实时荧光定量PCR技术检测PLPP3功能位点不同基因型个体背部脂肪中基因的表达差异。【结果】该北京黑猪群体中PLPP3基因包含8个SNPs,共5个SNPs显示中度多态性(0.25＜PIC＜0.5),3个SNPs为低度多态性(PIC＜0.25),g.155894136 C＞T位点呈现哈代-温伯格平衡状态(P＞0.05)。单倍型连锁分析显示,g.155800228 G＞A和g.155800259 A＞C位点之间存在较强的连锁(D’=91%);g.155891807 G＞A和g.155891715 A＞G位点之间存在一定的连锁性(D’＞80%)。基因多态位点与背部脂肪厚度性状的关联分析结果表明,g.155800222 C＞A、g.155891715 A＞G、g.155891807 G＞A、g.155894136 C＞T位点不同基因型个体的背部脂肪厚度存在显著差异(P＜0.05)。基因多态位点与脂肪酸含量的关联分析显示,g.155800222 C＞A、g.155800228 G＞A位点不同基因型个体背部脂肪中的棕榈酸含量和亚油酸/α-亚麻酸存在显著差异(P＜0.05)。启动子区SNPs TF预测发现,g.155800228 G＞A位点的变异,导致与其结合的TF由SPI1变为MYB。进一步分析g.155800228 G＞A位点不同基因型个体背部脂肪组织PLPP3基因的表达情况,发现AA基因型个体PLPP3基因mRNA水平显著高于GG基因型(P＜0.05)。【结论】 PLPP3基因的多态位点不同基因型个体的北京黑猪不同部位背部脂肪厚度及部分脂肪酸含量存在显著差异。这些多态位点可作为北京黑猪背部脂肪厚度和脂肪酸含量的潜在分子标记,为北京黑猪的选育提供理论支撑。

关键词: 北京黑猪; PLPP3基因; 背部脂肪厚度; 脂肪酸

Abstract: 【Objective】 The purpose of this study was to explore the polymorphisms of phospholipid phosphatase 3 (PLPP3) gene and its correlation with back fat thickness and fatty acid content of Beijing Black pigs,in order to provide reference for genetic improvement of back fat thickness and fatty acid content of Beijing Black pigs. 【Method】 In this study,the ear tissue samples of 239 Beijing Black pigs were collected to obtain genomic resequencing data.The thickness of back fat in different parts of 239 Beijing Black pigs (the thickest of the shoulder,junction of the 6-7 thoracic vertebrae,thoracolumbar junction,the joint of the lumbar sacral) and the contents of 5 fatty acids (palmitic acid,stearic acid,oleic acid,linoleic acid,α-linolenic acid) in the back fat of junction of the 6-7 thoracic vertebrae of the left carcass were determined.The sequencing results were utilized to study the population genetic characteristics and haplotype linkage of single nucleotide polymorphism (SNP) sites in the PLPP3 gene,and conduct association analysis between SNPs and back fat thickness and fatty acid content in various parts.The transcription factor of SNP in promoter region were predicted,and the differential gene expression in back fat of individuals with different genotypes of PLPP3 functional SNPs were detected by Real-time quantitative PCR. 【Result】 A total of 8 SNPs of PLPP3 gene were detected in the Beijing Black pig population,of which 5 were moderate polymorphisms (0.25＜PIC＜0.5) and 3 were low polymorphisms (PIC＜0.25),g.155894136 C＞T was in Hardy-Weinberg equilibrium (P＞0.05).Haplotype linkage analysis showed that there was a strong linkage between g.155800228 G＞A and g.155800259 A＞C (D’=91%).There was a certain linkage between g.155891807 G＞A and g.155891715 A＞G(D’＞80%).The association analysis between gene polymorphism and backfat thickness showed that there were significant differences in back fat thickness among individuals with different genotypes,including g.155800222 C＞A,g.155891715 A＞G,g.155891807 G＞A,g.155894136 C＞T (P＜0.05).The association analysis between gene polymorphism and fatty acid content showed that there were significant differences in the content of palmitic acid and the ratio of linoleic acid/α-linolenic acid in back fat among individuals with different genotypes,including g.155800222 C＞A,g.155800228 G＞A (P＜0.05).TFs prediction of SNPs in promoter region found that the mutation of g.155800228 G＞A caused the binding transcription factor to change from SPI1 to MYB.Further analysis of PLPP3 gene expression differences among different genotypes at g.155800228 G＞A showed that PLPP3 gene mRNA level in AA individuals was significantly higher than that in GG individuals (P＜0.05). 【Conclusion】 In summary,the different genotypes of PLPP3 gene were significantly differences with the thickness of back fat and the content of some fatty acids in different parts of Beijing Black pigs.It could be used as a potential molecular marker of back fat thickness and fatty acids content of Beijing Black pigs,and provided theoretical support for breeding of Beijing Black pigs.

Key words: Beijing Black pigs; PLPP3 gene; back fat thickness; fatty acids

中图分类号:

S813.1

祝雪丽, 马嘉宁, 许可, 孙超, 刘欣, 蒲蕾. PLPP3基因多态性对北京黑猪背部脂肪厚度和脂肪酸含量的影响[J]. 中国畜牧兽医, 2025, 52(1): 278-288.

ZHU Xueli, MA Jianing, XU Ke, SUN Chao, LIU Xin, PU Lei. Association Analysis of PLPP3 Gene Polymorphisms with Back Fat Thickness and Fatty Acids Content in Beijing Black Pigs[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(1): 278-288.

参考文献

[1] ZHANG Y,ZHANG J,GONG H,et al.Genetic correlation of fatty acid composition with growth,carcass,fat deposition and meat quality traits based on gwas data in six pig populations[J].Meat Science,2019,150:47-55.
[2] 田威龙,兰干球,张龙超,等.北京黑猪DKK3和CCR1基因多态性检测及其与背膘厚的关联分析[J].畜牧兽医学报,2022,53(7):2083-2093. TIAN W L,LAN G Q,ZHANG L C,et al.Detection of DKK3 and CCR1 genes polymorphisms and their association with backfat thickness in Beijing Black pigs[J].Chinese Journal of Animal and Veterinary Sciences,2022,53(7):2083-2093.(in Chinese)
[3] WANG Q,WANG X.The effects of a low linoleic acid/alpha-linolenic acid ratio on lipid metabolism and endogenous fatty acid distribution in obese mice[J].International Journal of Molecular Sciences,2023,24(15):12117.
[4] PAN H,HUANG T,YU L,et al.Transcriptome analysis of the adipose tissue of Luchuan and Duroc pigs[J].Animals,2022,12(17):2258.
[5] HUMTSOE J O,LIU M,MALIK A B,et al.Lipid phosphate phosphatase 3 stabilization of beta-catenin induces endothelial cell migration and formation of branching point structures[J].Molecular and Cellular Biology,2010,30(7):1593-1606.
[6] GAUDET P,LIVSTONE M S,LEWIS S E,et al.Phylogenetic-based propagation of functional annotations within the gene ontology consortium[J].Briefings in Bioinformatics,2011,12(5):449-462.
[7] DUFLOT T,TU L,LEUILLIER M,et al.Preventing the increase in lysophosphatidic acids:A new therapeutic target in pulmonary hypertension?[J].Metabolites,2021,11(11):784.
[8] LI B,XI W,BAI Y,et al.Fto-dependent M⁶A modification of PLPP3 in circscmh1-regulated vascular repair and functional recovery following stroke[J].Nature Communications,2023,14(1):489.
[9] DAVIDSON J,ROTONDO D.The potential role of the lipid phosphate phosphatase 3 (PLPP3) gene in cardiovascular disease[J].Current Opinion in Lipidology,2020,31(4):258-259.
[10] 田威龙.基于组学技术探究影响北京黑猪背部脂肪品质的重要候选基因及变异[D].南宁:广西大学,2022. TIAN W L.Exploration of candidate genes and variations affecting backfat quality of Beijing Black pigs based on omics technology[D].Nanning:Guangxi University,2022.(in Chinese)
[11] 倪黎纲,周春宝,徐盼,等.苏姜猪不同生长阶段背膘组织差异表达基因及其调控通路分析[J].中国畜牧兽医,2023,50(7):2595-2605. NI L G,ZHOU C B,XU P,et al.Analysis of differentially expressed genes and regulation pathway of backfat tissue in SuJiang pigs at different growth stages[J].China Animal Husbandry & Veterinary Medicine,2023,50(7):2595-2605.(in Chinese)
[12] 吴姿仪,陈健梅,程博文,等.利用全基因组关联分析鉴定猪生长性状相关的遗传变异及候选基因[J].中国畜牧杂志,2022,58(8):193-200. WU Z Y,CHEN J M,CHENG B W,et al.Genome-wide association analysis was used to identify genetic variants and candidate genes associated with pig growth traits[J].Chinese Journal of Animal Science,2022,58(8):193-200.(in Chinese)
[13] CHANG W,FA H,XIAO D,et al.microRNA-184 alleviates insulin resistance in cardiac myocytes and high fat diet-induced cardiac dysfunction in mice through the LPP3/DAG pathway[J].Molecular and Cellular Endocrinology,2020,508:110793.
[14] BRINDLEY D N,PILQUIL C.Lipid phosphate phosphatases and signaling[J].Journal of Lipid Research,2009,50 (Suppl):S225-S230.
[15] HOOKS S B,RAGAN S P,LYNCH K R.Identification of a novel human phosphatidic acid phosphatase type 2 isoform[J].FEBS Letters,1998,427(2):188-192.
[16] 胡卫芹.水稻脂磷酸磷酸酶在脂质代谢和生长中的作用[D].武汉:华中农业大学,2016. HU W Q.Role of lipid phosphate phosphatase in lipid metabolism and growth in rice[D].Wuhan:Huazhong Agricultural University,2016.(in Chinese)
[17] DESTAILLATS F,OLIVEIRA M,BASTIC S V,et al.Comparison of the incorporation of dha in circulatory and neural tissue when provided as triacylglycerol (TAG),monoacylglycerol (MAG) or phospholipids (PL) provides new insight into fatty acid bioavailability[J].Nutrients,2018,10(5):620.
[18] KIM K H,LOPEZ-CASILLAS F,BAI D H,et al.Role of reversible phosphorylation of acetyl-CoA carboxylase in long-chain fatty acid synthesis[J].FASEB Journal,1989,3(11):2250-2256.
[19] 裴梦甜,王鑫潇,余庆,等.稻瘟病菌MYB转录因子MoIsw2调控基因的鉴定和功能分析[J].福建农林大学学报(自然科学版),2023 53(5):598-609. PEI M T,WANG X X,YU Q,et al.Identification and functional analysis of MYB transcription factor Molsw2 regulatory gene madnaoorthe orvzae[J].Journal of Fujian Agriculture and Forestry University(Natural Science Edition),2023,53(5):598-609.(in Chinese)
[20] WEIDEMULLER P,KHOLMATOV M,PETSALAKI E,et al.Transcription factors:Bridge between cell signaling and gene regulation[J].Proteomics,2021,21(23-24):e2000034.
[21] OKSUZ O,HENNINGER J E,WARNEFORD-THOMSON R,et al.Transcription factors interact with RNA to regulate genes[J].Molecular Cell,2023,83(14):2449-2463.
[22] LANE J M,DOYLE J R,FORTIN J P,et al.Development of an OP9 derived cell line as a robust model to rapidly study adipocyte differentiation[J].PLoS One,2014,9(11):e112123.
[23] KOGELMAN L J,CIRERA S,ZHERNAKOVA D V,et al.Identification of co-expression gene networks,regulatory genes and pathways for obesity based on adipose tissue rna sequencing in a porcine model[J].BMC Medical Genomics,2014,7:57.
[24] SHI T,XIE J,XIONG Y,et al.Human HS1BP3 induces cell apoptosis and activates ap-1[J].BMB Reports,2011,44(6):381-386.
[25] WEIDEMULLER P,KHOLMATOV M,PETSALAKI E,et al.Transcription factors:Bridge between cell signaling and gene regulation[J].Proteomics,2021,21(23-24):e2000034.
[26] MILLARD P S,KRAGELUND B B,BUROW M.R2R3 MYB transcription factors-Functions outside the DNA-binding domain[J].Trends in Plant Science,2019,24(10):934-946.
[27] TAKAO S,FORBES L,UNI M,et al.Convergent organization of aberrant MYB complex controls oncogenic gene expression in acute myeloid leukemia[J].eLife,2021,10:e65905.
[28] LI M,JIANG P,CHENG K,et al.Regulation of MYB by distal enhancer elements in human myeloid leukemia[J].Cell Death & Disease,2021,12(2):223.
[29] ZUBER J,RAPPAPORT A R,LUO W,et al.An integrated approach to dissecting oncogene addiction implicates a MYB-coordinated self-renewal program as essential for leukemia maintenance[J].Genes & Development,2011,25(15):1628-1640.
[30] SHI M,YU L,SHI J,et al.A conserved MYB transcription factor is involved in regulating lipid metabolic pathways for oil biosynthesis in green algae[J].New Phytologist,2022,235(2):576-594.