The effect of adipocyte and heart fatty acid-binding protein genes on intramuscular fat and backfat content in Meishan crossbred pigs

Effects of genetic variation in porcine adipocyte and heart fatty acid-binding protein genes, A-FABP and H-FABP, respectively, on intramuscular fat (IMF) content and backfat thickness (BFT) were examined in F2 crossbreds of Meishan and Western pigs. The involvement of each FABP gene in IMF accretion was studied to confirm previous results for Duroc pigs. The F2 crossbred pigs were genotyped for various markers including microsatellite sequences situated within both FABP genes. Linkage analysis assigned the A-FABP and H-FABP genes to marker intervals S0001-S0217 (20 cM) on SSC4 and Sw316-S0003 (16.6 cM) on SSC6, respectively, refining previous chromosomal assignments. Next, the role of both chromosome regions/genes on genetic variation in IMF content and BFT was studied by 1) screening SSC4 and SSC6 for QTL affecting both traits by performing a line-cross analysis and 2) estimation of the effect of individual A-FABP and H-FABP alleles on both traits. In the first analysis, suggestive and chromosome-wise significant evidence for a QTL affecting IMF was detected on SSC6. The H-FABP gene is a candidate gene for this effect because it resides within the large region containing this putative QTL. The second analysis showed a considerable but nonsignificant effect of H-FABP microsatellite alleles on IMF content. Suggestive evidence for a QTL affecting BFT was found on SSC6, but H-FABP was excluded as a candidate gene. In conclusion, present and previous results support involvement of H-FABP gene polymorphisms in IMF accretion independently from BFT in pigs. Therefore, implementation of these polymorphisms in marker-assisted selection to control IMF content independently from BFT may be considered. In contrast to previous findings for Duroc pigs, no evidence was found for an effect of the A-FABP gene on IMF or BFT in this population.     

Evaluation of FABP2 as candidate gene for a fatty acid composition QTL in porcine chromosome 8

The objective of this work was to analyse the porcine Fatty acid binding protein 2, intestinal ( FABP2 ) gene as a candidate gene for a fatty acid composition quantitative trait loci (QTL) previously described on porcine chromosome 8 in an Iberian by Landrace F₂ cross (IBMAP). Re-sequencing of the porcine FABP2 gene in three Iberian and eight Landrace parental animals resulted in the identification of three single-nucleotide polymorphisms, all of them localized in intron 1. The polymorphism FABP2 :g.412T>C, localized in intron 1, and two additional microsatellites were genotyped in the IBMAP population in order to perform an association test of the FABP2 gene and to better define the QTL position previously described. Association analyses of the FABP2 :g.412T>C with the fatty acid composition traits were not significant in simple association and marker-assisted association tests, suggesting that the FABP2 region sequenced is not responsible for the QTL. However, the addition of three new markers to the pedigree allowed us to define the S0144–SW61 marker interval as the most likely QTL position, facilitating the future study of other candidate genes for this QTL.     

New alleles in calpastatin gene are associated with meat quality traits in pigs

Suggestive QTL affecting raw firmness scores and average Instron force, tenderness, juiciness, and chewiness on cooked meat were mapped to pig chromosome 2 using a three-generation intercross between Berkshire and Yorkshire pigs. Based on its function and location, the calpastatin (CAST) gene was considered to be a good candidate for the observed effects. Several missense and silent mutations were identified in CAST and haplotypes covering most of the coding region were constructed and used for association analyses with meat quality traits. Results demonstrated that one CAST haplotype was significantly associated with lower Instron force and cooking loss and higher juiciness and, therefore, this haplotype is associated with higher eating quality. Some of the sequence variation identified may be associated with differences in phosphorylation of CAST by adenosine cyclic 3', 5'-monophosphate-dependent protein kinase and may in turn explain the meat quality phenotypic differences. The beneficial haplotype was present in all the commercial breeds tested and may provide significant improvements for the pig industry and consumers because it can be used in marker-assisted selection to produce naturally tender and juicy pork without additional processing steps.     

Single nucleotide polymorphism association study for backfat and intramuscular fat content in the region between SW2098 and SW1881 on pig chromosome 6

This study was carried out to identify SNP associated with fatness traits on pig chromosome 6. In total, 11,067 putative genomic variations were detected in 125 complete bacterial artificial chromosome sequences corresponding to the region between SW2098 and SW1881, which harbors multiple QTL affecting intramuscular fat content (IMF) and backfat thickness (BFT). Among 173 putative SNP validated by MassArray, 120 SNP were used in an association study on 541 offspring produced by a cross of Korean native pig and Landrace breeds. The significance level of each SNP was determined using single marker regression analysis. Further, significant threshold values were determined using a false discovery rate. Nine out of 120 SNP showed significant effects on BFT or IMF or both. Of the 9 significant SNP, 4 were significantly associated with IMF, 7 were significantly related to BFT, and 2 SNP (Kps8172 and Kps6413) showed significant effects on both traits. Moreover, multiple regression analysis considering all significant SNP was used to correct spurious false positives due to linkage disequilibrium. Consequently, only 1 SNP (Kps6413) was significant for IMF, whereas 4 SNP including Kps6413 showed significant effects on BFT. The significant SNP had generally additive effects and on average explained 1.72% of the genetic variation for IMF and 3.92% for BFT, respectively. These markers can potentially be applied in pig breeding programs for improving IMF and BFT traits after validation in other populations.     

Expression analyses of candidate genes related to meat quality traits in squabs from two breeds of meat‐type pigeon

In this study, meat quality traits were compared between squabs from two pigeon breeds: one Chinese indigenous breed, the Shiqi (SQ) meat‐type pigeon, and an imported breed, the white king (WK) meat‐type pigeon. Breed differences were detected in the content of intramuscular fat (IMF) in the breast muscle. SQ squabs had significantly higher IMF content than the WK birds. The shear force value (an objective measure of meat tenderness) of SQ birds was also relatively lower than that of the WK squabs. Further analysis of fatty acids profile revealed that SQ squabs exhibited significant advantage in the synthesis of polyunsaturated fatty acids, while WK squabs were significantly higher in the sum of monounsaturated fatty acids. Breast muscle in the SQ squabs was also significantly higher in the ratio of polyunsaturated fatty acids to saturated fatty acids, as well as the sum of omega 6 fatty acids. Variability of expression levels of functional genes in relation to fat accumulation and meat tenderness was analysed by qRT‐PCR. Gene expression analyses showed that the hepatic expression of LPL (lipoprotein lipase), FABP4 (fatty acid‐binding protein 4), and CAPN2 (calpain‐2) were significantly higher in the SQ squabs. In the breast muscle tissue, the FABP3 (fatty acid‐binding protein 3) and CAPN2mRNA abundance was significantly higher in SQ squabs. Our results suggested that these differentially expressed genes might be candidate genes used in the programmes of targeted selection for squabs with higher IMF content, tender meat, and more favourable fatty acids composition.     

Validation of the QTL on SSC4 for meat and carcass quality traits in a commercial crossbred pig population

Porcine chromosome 4 harbours many quantitative trait loci (QTL) affecting meat quality, fatness and carcass composition traits, detected in resource pig populations previously. However, prior to selection in commercial breeds, QTL identified in an intercross between divergent breeds require confirmation, so that they can be segregated. Consequently, the objective of this study was to validate several QTL on porcine chromosome 4 responsible for meat and carcass quality traits. The experimental population consisted of 14 crossbred paternal half-sib families. The region of investigation was the q arm of SSC4 flanked by the markers S0073 and S0813. Regression analysis resulted in the validation of three QTL within the interval: Minolta a * loin, back fat thickness and the weight of trimmed ham. The results were additionally confirmed by factor analysis. Candidate genes were proposed for meat colour, which was the most evident QTL validated in this study.     

影响猪肌内脂肪含量的QTL研究进展

肌内脂肪含量是评定猪肉品质的重要指标之一。分子生物学技术的飞速发展及其应用，为人们在DNA水平上研究猪重要经济性状的遗传机制奠定了坚实的基础。本文从可能的主效基因、可以考虑的候选基因以及其他影响猪肌内脂肪含量的QTL等方面概述了猪肌内脂肪含量的QTL研究进展，并对在育种实践中利用DNA标记辅助选择来改良猪肌内脂肪含量的应用前景、研究方向和存在的问题进行初步的探讨。     

Exclusion of the swine leukocyte antigens as candidate region and reduction of the position interval for the Sus scrofa chromosome 7 QTL affecting growth and fatness

Pig chromosome 7 (SSC 7) has been shown to be rich in QTL affecting performance and quality traits. Most studies mapped the QTL close to the swine leukocyte antigens (SLA), which has a large effect on adaptability and natural selection. Previous comparative mapping studies suggested that the 15-cM region limited by markers LRA1 (mapped at 55 cM) and S0102 (mapped at 70 cM) contains hundreds of genes. To decrease the number of candidate genes, we improved the mapping resolution with a genetic chromosome dissection through a backcross recombinant progeny test program between Meishan (MS) and European (EU; i.e., Large White or Landrace) breeds. Three first-generation backcross--(EU x MS) x EU--and two second-generation backcross--([EU x MS] x EU) x EU--sires carrying a recombination in the QTL mapping interval were progeny-tested (i.e., measured for a total of 44 growth, fatness, carcass and meat quality traits). Progeny family size varied from 29 to 119 pigs. Animals were genotyped for markers covering the region of interest. Progeny-test results allowed the QTL interval to be decreased from 15 to 20 cM down to 10 cM, and even less than 6 cM if we assumed that the EU pigs used in this study share only one QTL allele. Except for a putative QTL affecting some carcass composition traits, the SLA is excluded as a candidate region, suggesting that it might be possible to apply a marker-assisted selection strategy for this QTL, while controlling SLA allele diversity. The strong QTL effects remaining in animals with only 12.5% (issued from first-generation backcross boars) and 6.25% (issued from second-generation back-cross boars) Meishan genetic background shows that epistatic interactions are likely to be limited. Finally, the QTL does not have strong effects on meat quality traits.     

Genome‐wide study on intramuscular fat in Italian Large White pig breed using the PorcineSNP60 BeadChip

Genome‐wide association study results are presented for intramuscular fat in Italian Large White pig breed. A total of 886 individuals were genotyped with PorcineSNP60 BeadChip. After quality control performed with plink software and in R environment, 49 208 markers remained for the association analysis. The genome‐wide association studies was conducted using linear mixed model implemented in GenABEL. We detected seven new SNPs of genes till now not found associated to intramuscular fat (IMF). Three markers map in a wide intergenic region rich of QTL linked to fat traits, one map 388 kb upstream the gene SDK1, one map inside PPP3CA gene, one inside SCPEP1 gene and the last is not mapped in the porcine genome yet. Associations here presented indicate a moderate effect of these genes on IMF. In particular, PPP3CA, that is involved in the oxidative metabolism of skeletal muscle, could be considerated as an interesting candidate gene for IMF content in pigs. However, further studies are needed to clarify the role of these genes on the physiological processes involved in IMF regulation. These results may be useful to control this trait that is important in terms of nutritional, technological and organoleptic characteristics of fresh meat and processed products.     

Impact of MYOD family genes on pork traits in Large White and Landrace pigs

Summary Porcine myogenic differentiation genes ( MYOD ) family play a key role in growth and muscle development and are therefore considered as candidate genes for meat production traits. The objective of the study was to investigate the polymorphisms at four loci belonging to the MYOD genes family and analyse their associations with variation in meat production traits in Czech pig breeds. To verify the associations between the polymorphisms and the selected meat traits, altogether 254 pigs, including full- and half-sibs, of Large White and Landrace breeds were tested. The studied meat characteristics were weight of neck, loin, shoulder and ham, lean meat content (LMC), backfat thickness, intramuscular fat (IMF), remission, dry matter content and test daily gain. Statistically significant associations were observed between MYOG gene and fat and neck weight, and between MYF5 gene and IMF and LMC. High significant differences were observed between genotypes AA and AB of MYOD1 in IMF and between genotypes AB and BB of MYF5 in loin weight.     

拷贝数变异全基因组关联分析及数量性状基因座定位联合鉴定猪体高性状候选基因

旨在利用全基因组拷贝数变异区域（copy number variation regions,CNVRs）关联分析以及全基因组数量性状基因座（quantitative trait locus,QTLs）定位联合筛选出影响猪体高性状的候选基因。本研究利用快速检测基因组拷贝数变异软件CNVcaller对本实验室构建的大白×民猪F2代资源群体的重测序数据进行拷贝数变异检测。利用混合线性模型（mixed-linear model,MLM）将性别和胎次作为固定效应对体高性状进行拷贝数变异全基因组关联分析（CNVR-GWAS）。采用软件R/qtl进行QTL分析,并使用置换检验（permutation test,PT）进行检验。将CNVR-GWAS与QTL结果进行联合注释,结合GO富集和KEGG通路分析,对影响猪体高的位点和基因进行挖掘。利用实时荧光定量PCR（qPCR）方法验证候选基因。结果表明,本群体在全基因组范围内共有3 099个CNVRs,其中有两个CNVRs与体高性状在全基因组范围内显著相关,分别位于7号染色体的25 358 001~26 696 400 bp处（CNVR1）和54 087 201~54 090 000 bp处（CNVR2）。在混合线性模型分析的结果中发现,CNVR1拷贝数增加（P<0.01）和CNVR2拷贝数缺失（P<0.01）对猪的体高性状具有显著影响。基因组显著水平可找到2个显著影响猪体高的QTLs,分别为BH-1和BH-2,其中BH-2对体高性状的影响较大。CNVR1和BH-2重叠区存在1个嗅觉受体基因OR12D3和18个未被注释的基因。qPCR验证OR12D3的拷贝数变异与利用混合线性模型统计推断出的结果一致。初步推测,OR12D3基因的拷贝数变异可能与猪体高性状相关。     

Identification of quantitative trait loci for carcass composition and pork quality traits in a commercial finishing cross

A QTL study for carcass composition and meat quality traits was conducted on finisher pigs of a cross between a synthetic Piétrain/Large White boar line and a commercial sow cross. The mapping population comprised 715 individuals evaluated for a total of 30 traits related to growth and fatness (4 traits), carcass composition (11 traits), and meat quality (15 traits). Offspring of 8 sires (n = 715) were used for linkage analysis and genotyped for 73 microsatellite markers covering 14 chromosomal regions representing approximately 50% of the pig genome. The regions examined were selected based on previous studies suggesting the presence of QTL affecting carcass composition or meat quality traits. Thirty-two QTL exceeding the 5% chromosome-wise significance level were identified. Among these, 5 QTL affecting 5 different traits were significant at the 1% chromosome-wise level. The greatest significance levels were found for a QTL affecting loin weight on SSC11 and a QTL with an effect on the Japanese color scale score of the loin on SSC4. About one-third of the identified QTL were in agreement with QTL previously reported. Results showed that QTL affecting carcass composition and meat quality traits segregated within commercial lines. Use of these results for marker-assisted selection offers opportunities for improving pork quality by within-line selection.     

Meat quality traits were unaffected by a quantitative trait locus affecting leg composition traits in Texel sheep

A QTL affecting leg muscle and fat traits has been identified within the New Zealand Texel population. The QTL maps to a region on OAR 2 with a two-marker haplotype test established at markers BULGE20 and BM81124. These markers encompass the likely position of Growth Differentiation Factor 8 (GDF8). The pleiotropic effects of this QTL on meat quality traits are tested. Objective measures of meat quality including pH, color (L*, a*, and b*), and tenderness (as assessed by Warner-Bratzler shear force measurements) were assessed on longissimus and semi-membranosus muscles of 540 progeny from six Texel sires. Four of these sires were subsequently identified as segregating for leg muscle and fat traits. For these segregating sires, comparison of progeny that had inherited the favorable haplotype from their sire with those that had received the alternate haplotype revealed no significant differences in the meat quality traits assessed. This finding suggests that the muscling QTL does not have pleiotropic effects on meat quality. A general scan for meat quality QTL was carried out using genotype data for eight markers from FCB128 to RM356 flanking 122cM of OAR 2 using Haley-Knott regression. This analysis revealed two QTL for a single sire. A QTL detected in the region of Marker INRA40 for color L* mapped to a site close to the muscling QTL, but there was evidence to suggest it is at a distinct locus. The QTL in the region of Marker RM356 might map distal to Marker RM356, as no peak was observed. This QTL, which seems to affect pH, color a*, color b*, and Warner-Bratzler shear measurements, requires further characterization.     

Genetic mapping of the QTL affecting body weight in chickens using a F2 family

1. To identify the quantitative trait loci (QTL) affecting growth in chickens, we carried out QTL analysis on chicken growth traits using a population of 227 F2 crosses between a Satsumadori (slow-growing, light-weight Japanese native breed used as a meat chicken) male and a White Plymouth Rock (early-maturing, heavy weight broiler). 2. We chose 78 microsatellite loci from 331 publicly available on 14 linkage groups, with respect to their utility and location. 3. Two QTLs affecting body weight at 13 and 16 weeks were mapped at 220 cM on chromosome 1 (LOD scores, 2.8 and 4.5, respectively, at 13 and 16 weeks), and at 60 cM on chromosome 2 (LOD scores, 6.2 and 8.1, respectively, at 13 and 16 weeks). 4. The closest loci to the QTLs were LEI71 on chromosome 1 and LMU13 and MCW184 on chromosome 2. 5. The sites of the QTLs agreed closely with those already reported. Therefore, it seems likely that QTLs affecting growth of chickens are located at these sites.     

Mapping genes for fatness and growth on pig chromosome 13: a search in the region close to the pig PIT1 gene

Previous research has shown that PIT1 polymorphisms in several resource populations and the chromosomal region near PIT1 in some populations have been significantly associated with fatness and growth QTLs on pig chromosome 13. To confirm these previous results and to clarify the role of the PIT1 gene in the putative QTL region, this research project was enlarged to include two microsatellite markers flanking each side of the PIT1 gene ( Swr1008 , S0068 , Sw398 and Sw1056 ). The ISU Chinese × US resource families were used and the traits analysed were birth weight, 21 day weight, 42 day weight, longissimus muscle area, back-fat thickness at several locations, meat colour, marbling and firmness on the carcass, and growth rate for selected time periods. The total number of F₂ pigs used ranged from 241 to 330. The data were analysed using interval mapping for each breed-cross separately as well as with the pooled data, and single marker least squares analyses for the pooled data. Significant evidence of a QTL for first rib back-fat thickness was detected approximately 20 cM from the PIT1 gene by using both single marker (p < 0.01) and interval mapping analyses in the pooled data (p < 0.0001) as well as in one family (p < 0.01). Evidence of a QTL for birth weight was detected at the estimated PIT1 position in the interval mapping analysis by using the pooled data (p < 0.014) and verified by the single marker analyses. These results confirmed the previously published QTL work on pig chromosome 13 for the birth weight QTL but suggest that other genes in the region may be partly responsible for the earlier results on the back-fat thickness QTL in our resource families.     

Quantitative trait locus mapping for meat quality traits in an Iberian x Landrace F2 pig population

An experimental F2 cross between Iberian and Landrace pig strains was performed to map quantitative trait loci (QTL) for diverse productive traits. Here we report results for meat quality traits from 369 F2 animals with records for pH 24 h postmortem (pH 24 h), muscle color Minolta measurements L* (lightness), a* (redness), and b* (yellowness), H* (hue angle), C* (chroma), intramuscular fat (IMF) and haematin pigment content measured in the longissimus thoracis. Pigs were genotyped for 92 markers covering the 18 porcine autosomes (SSC). Results of the genome scan show evidence for QTL for IMF (SSC6; F = 27.16), pH 24 h (SSC3; F = 7.73), haematin pigments (SSC4 and SSC7; F = 8.68 and 9.47 respectively) and Minolta color measurements L* (SSC4 and SSC7; F =16.42 and 7.17 respectively), and a* (SSC4 and SSC8; F = 8.05 and 7.36 respectively). No QTL were observed for the color measurements b*, H*, and C*. Alternative models fitting epistasis between QTL were also tested, but detected epistatic interactions were not significant at a genome-wise level. In this work we identify genomic regions related with meat quality traits. Improvement by traditional selection methods is complicated, and finer mapping would be required for their application in introgression programs.     

Mapping QTL for porcine muscle fibre traits in a White Duroc × Erhualian F2 resource population

Muscle fibre traits are related with meat quality in meat animals. In this study, a whole‐genome scan with 183 microsatellite markers covering the pig genome was performed to identify quantitative trait loci (QTL) for cross‐sectional area, numerical percentage and relative area of type I, IIA and IIB myofibres, fibre number per square centimetre and total fibre number in the longissimus muscle by using 120 F₂ animals in a White Duroc × Erhualian intercross. In total, 20 QTL were mapped on pig chromosomes (SSC) 1, 2, 7, 8, 9, 11, 15, 16 and X, of which eight reached genome‐wide significance levels and explained large proportions (6.53–34.63%) of phenotypic variance. Five QTL detected in this study confirmed the previous QTL reports and the others were detected for the first time. Chinese Erhualian alleles are generally associated with muscle fibre traits favourable for meat quality.     

An evaluation of four candidate genes for use in selection programmes aimed at increased intramuscular fat in Duroc swine

Summary A sufficient level of intramuscular fat (IMF) is needed to enhance consumer acceptance of pork products, and is currently receiving greater attention within swine genetic improvement programmes. An examination of previously described and novel genetic variants within candidate genes for IMF deposition was performed to evaluate potential use of genetic markers in marker-assisted selection (MAS). Biological candidate genes implicated to play a role in adipogenesis were investigated within two different lines of purebred Duroc pigs. These included MC4R , FABP3 , DLK1 , and TCF7L2 . Significant variation in IMF within the control line was described by the MC4R genotype and a novel Bsr fI single nucleotide polymorphism within the FABP3 gene. Genetic markers for DLK1 and TCF7L2 evaluated in this population are not currently recommended for selection in Duroc swine. Existence of MC4R and FABP3 mutations may be useful markers in MAS aimed at IMF improvement, provided that gene effects are segregating and the presence of an association is detected within the population. However, additional work to confirm the use of the investigated genetic markers in selection programmes is needed.