猪雌激素受体（ESR）基因对产仔数性状的影响

猪产仔数是重要的经济性状,产仔数的提高将会大大地增加商品肉猪的产量,给现代养猪生产带来巨大的经济效益。ＥＳＲ基因是影响猪产仔数的主效基因,而且与猪的生长发育性状及酮体性状之间不存在负的基因多效性影响。采用ＰＣＲ－ＲＦＬＰｓ的方法,通过对５个不同品种的、特别是我国地方品种的２６２头母猪进行了基因多态性分析,研究表明ＥＳＲ基因在这种种群对产仔数均有极显著影响,ＥＳＲ位点ＢＢ基因型母猪平均比ＡＡ基因型母     

Association with litter size of new polymorphisms on ESR1 and ESR2 genes in a Chinese-European pig line

The objective of this study was to search for polymorphisms in the coding region of the estrogen receptors 1 and 2 (ESR1 and ESR2 )and to analyze the effects of these variants and the well known intronic ESR1 Pvu II polymorphism on litter size in a Chinese-European pig line. We identified five silent single nucleotide polymorphisms (SNP) in the ESR1 cDNA: c.669T > C (exon 3), c.1227C > T (exon 5), c.1452C > T (exon 7), c.1665T > C and c.1755A > G (exon 8). One pair of these SNP (c.1665T > C and c.1755A > G) co-segregated in the analyzed line, and the SNP c.669T > C showed the same segregation pattern as the Pvu II polymorphism. These polymorphisms were tested in this study, although the c.1452C > T SNP within exon 7 was not analyzed due to its low informativeness. In the ESR2 cDNA, one missense SNP was found within exon 5, which caused an amino acid substitution in the coded protein: "c.949G > A (p.Val317Met)" and was tested on sow litter size. Information on 1622 litter records from 408 genotyped sows was analyzed to determine whether these SNP influenced the total number of piglets born (TNB) or the number of born alive (NBA). The polymorphisms ESR1: [Pvu II; c.669T > C], ESR1: [c.1665T > C; c.1755A > G] and ESR2: c.949G > A showed no statistically significant association with litter size. However, the ESR1: c.1227T allele was significantly associated with TNB. The additive substitution effect was estimated to be 0.40 piglets born per litter (P < 0.03), and no dominance effects were observed. This SNP could be useful in assisted selection for litter size in some pig lines, as a new genetic marker in linkage disequilibrium with the causative mutation.     

绵羊ESR2基因组织表达及其多态性与产羔数关联分析

为了探究雌激素受体2 (Estrogen receptor 2, ESR2)基因在绵羊各组织的表达及其多态性与产羔数之间的关系,本研究利用半定量PCR和实时荧光定量PCR技术检测ESR2基因在不同繁殖力小尾寒羊群体组织中的相对表达量,同时采用Sequenom MassARRAY誖SNP技术对多羔品种绵羊(小尾寒羊,湖羊,策勒黑羊)和单羔品种绵羊(苏尼特羊,草原型藏羊,滩羊) ESR2基因g.73324006C>T位点进行检测,并与小尾寒羊产羔数进行关联分析。半定量PCR表明,ESR2基因在单、多羔小尾寒羊子宫中高表达,在其它组织中等或低丰度表达;单羔群体、多羔群体间荧光定量PCR表明,ESR2基因在单羔小尾寒羊垂体表达量显著高于多羔小尾寒羊(p<0.05);群体遗传学分析表明,g.73324006C>T在小尾寒羊群体中表现为低度多态(PIC<0.25),在滩羊群体中处于中度多态(0.25T在小尾寒羊群体处于哈代温伯格平衡状态(p>0.05);关联分析表明,g.73324006C>T位点多态性与小尾寒羊第一胎、第二胎、第三胎产羔数及平均产羔数均显著关联(p<0.05),CC型各胎产羔数均高于TC型。与FecB (A746G)基因组合后发现,GG-CC和AG-CC基因型母羊产羔数显著高于AA-TC、AA-CC、AG-TC基因型组合(p<0.05)。综上,ESR2与小尾寒羊产羔数密切相关,g.73324006C>T可作为绵羊产羔性状选育的潜在分子标记。     

Comparison of genetic models for analysing the effects of a PvuII polymorphism in the oestrogen receptor 1 (ESR1) gene on prolificacy in an Iberian x Meishan pig population

The effect of a previously reported PvuII polymorphism in oestrogen receptor 1 (ESR1) was analysed in an F(2) population of Iberian x Meishan pigs. We tested three hypotheses: (1) that a causal mutation was fixed in the parental populations, (2) that a causal mutation existed that was in complete linkage disequilibrium with the alleles of the PvuII polymorphism and (3) that a causal mutation existed in linkage disequilibrium within each parental population. The third model was the most plausible based on the available data. ESR1 alleles displayed different patterns of linkage disequilibrium with the causal mutation in each of the parental populations and the PvuII polymorphism was clearly not the causal mutation. As a consequence, the use of the ESR1 mutation for selection must be evaluated for a particular pig population before it is applied.     

Genotyping of the porcine ryanodine receptor 1 (RYR1) and estrogen receptor 1 (ESR1) genes by high resolution melting (HRM) approach

During the last decade, DNA mutations in the porcine ryanodine receptor 1 gene (RYR1, C1843T) and the estrogen receptor 1 gene (ESR1, T1665G), have been widely used in marker-assisted selection (MAS) for the pig industry. These 2 well-characterized SNPs in RYR1 and ESR1 are responsible for porcine stress syndrome (PSS) and litter size, respectively. Here, we describe a reliable, high-efficiency method for the genotyping of these 2 genes using the high-resolution melting (HRM) method. The HRM approach exhibited high-accuracy and repeatability, comparable with the classic PCR-restriction fragment length polymorphism (PCR-RFLP) approach, and is potentially suitable for large-scale genotyping in commercial pig farms.     

Impact of the ESR gene on litter size and production traits in Czech Large White pigs

To evaluate the effect of the PvuII polymorphism of the oestrogen receptor gene on litter size and production traits in Czech Large White swine, data from 1250 sows and 3600 litters were analysed with two four-trait animal models. The traits in the first model were number of piglets born alive in a sow's first litter, number of piglets born alive in second and subsequent litters, lifetime daily gain and lean meat percentage. The second model included number of piglets born, number of piglets born alive, number of piglets weaned and litter weight at weaning from first and subsequent litters. The oestrogen receptor (ESR) locus significantly affected prolicacy in the first parity and averaged over all parities (P < 0.05), with allele A superior to allele B. In the first parity, AA sows produced approximately 0.5 more live piglets per litter than BB sows. Averaged over all parities, this difference was c. 0.25 piglets. Results for total number of piglets born and number of piglets weaned were similar to results for numbers born alive. No significant dominance effect was found for prolificacy traits. For litter weight at weaning, no significant additive effect was observed at the ESR locus, but a significant negative dominance effect (-1.5 kg) was estimated averaged across parities (litters of AB sows were similar to litters of BB sows for this trait). No pleiotropic effect of the ESR polymorphism on average daily gain or lean meat percentage was found.     

No detectable association of the ESR PvuII mutation with sow productivity in a Meishan x Large White F2 population

The polymorphism at the PvuII recognition site in the ESR gene showed no statistically significant association with sow productivity traits in a Meishan x Large White F2 population. Estimates of the effect on litter size were, however, in the opposite direction and statistically different from previously published estimates. Taken together with results from other publications, results here indicate that this PvuII polymorphism displays different degrees of linkage disequilibrium with a gene or genes controlling litter size in different populations.     

撒坝猪血清酯酶多态性与繁殖性能关系的研究

采用垂直板聚丙烯酰胺凝胶电泳法(PAGE), 对115头撒坝猪的血清酯酶(ES)多态性进行了检测,计算了该位点的基因型频率、基因频率和位点多态杂合度(h),并用二因素有互作的最小二乘模型对血清酯酶多态性与繁殖性能的关系进行了分析。结果表明,撒坝猪血清酯酶3种基因型AA、AB和BB的频率分别为0.2696、0.5826和0.1478。两个等位基因A和B的基因频率分别为0.5609和0.4391。该位点的杂合度为0.4926。在3种基因型中,不同基因型母猪的繁殖性能在产仔数、仔猪初生窝重、20日龄窝重、断奶仔猪数和断奶窝重等性状上存在着显著差异(P<0.05);公、母猪不同基因型交配组合在产仔数、断奶仔猪数、仔猪断奶体重和断奶窝重等性状上亦有显著差异(P<0.05)。显示出猪的血清酯酶多态性可望作为繁殖性能选种的遗传标记。 Abstract：　Serum esterase polymorphisms of 115 Saba pigs were investigated by using the method of vertical polyacrylamide gel electrophoresis. The genotype frequency, gene frequency and heterozygosity of this locus were calculated. The relationship between the serum esterase polymorphism and reproductive performance was analyzed by the least square analysis of two factors (ES genotype of boar and sow) with interaction. The results demonstrated that the genotype frequency of AA, ABandBBwas 0.2696, 0.5826 and 0.1478 respectively, the gene frequency of the alleleAandBwas 0.5609 and 0.4391 respectively. The heterozygosity of this locus was 0.4926. There are significant differences (P<0.05) on litter size, litter weight at birth, litter weight at 20 days, litter size at weaning and litter weight at weaning of different genotypic sows. The significant differences (P<0.05) were showed on litter size, litter size at weaning, weaning weight and litter weight at weaning of different genotypic mating combinations. It indicated that the serum esterase polymorphism was expected to be the genetic marker of pig reproductive performance.     

Non-additive effects of RBP4, ESR1 and IGF2 polymorphisms on litter size at different parities in a Chinese-European porcine line

Background

The aim of this work was to study the effects on litter size of variants of the porcine genes RBP4, ESR1 and IGF2, currently used in genetic tests for different purposes. Moreover, we investigated a possible effect of the interaction between RBP4-MspI and ESR1-PvuII polymorphisms. The IGF2-intron3-G3072A polymorphism is actually used to select lean growth, but other possible effects of this polymorphism on reproductive traits need to be evaluated.

Methods

Detection of polymorphisms in the genomic and cDNA sequences of RBP4 gene was carried out. RBP4-MspI and IGF2-intron3-G3072A were genotyped in a hyperprolific Chinese-European line (Tai-Zumu) and three new RBP4 polymorphisms were genotyped in different pig breeds. A bivariate animal model was implemented in association analyses considering the number of piglets born alive at early (NBA₁₂) and later parities (NBA₃₊ ) as different traits. A joint analysis of RBP4-MspI and ESR1-PvuII was performed to test their possible interaction. In the IGF2 analysis, paternal or maternal imprinting effects were also considered.

Results

Four different RBP4 haplotypes were detected (TGAC, GGAG, GAAG and GATG) in different pig breeds and wild boars. A significant interaction effect between RBP4-MspI and ESR1-PvuII polymorphisms of 0.61 ± 0.29 piglets was detected on NBA₃₊. The IGF2 analysis revealed a significant increase on NBA₃₊ of 0.74 ± 0.37 piglets for the paternally inherited allele A.

Conclusions

All the analyzed pig and wild boar populations shared one of the four detected RBP4 haplotypes. This suggests an ancestral origin of the quoted haplotype. The joint use of RBP4-MspI and ESR1-PvuII polymorphisms could be implemented to select for higher prolificacy in the Tai-Zumu line. In this population, the paternal allele IGF2-intron3-3072A increased litter size from the third parity. The non-additive effects on litter size reported here should be tested before implementation in other pig breeding schemes.     

FSHβ subunit gene is associated with major gene controlling litter size in commercial pig breeds

An insertion fragment in porcine FSHβ subunit gene was cloned by PCR. Sequencing data show that the insertion is a retroposon of 292 bp siting in intronⅠ at the site between +809 and +810 base. Based on these results, a PCR programme was created to genotype animal individuals in different pig breeds at FSHβ locus and polymorphism of FSHβ gene was analyzed. With the combination of genotype and litter size of sows, it was demonstrated that FSHβ locus is closely associated with major gene controlling litter size in commercial pig breeds, such as Yorkshire, Landrace, Durco. Averagely the AA sows give more 1.5 piglets than BB sows do per litter.     

FSHB subunit gene is associated with major gene controlling litter size in commercial pig breeds

An insertion fragment in porcine FSHβ subunit gene was cloned by PCR. Sequencing data show that the insertion is a retroposon of 292 bp siting in intron I at the site between + 809 and +810 base. Based on these results, a PCR programme was created to genotype animal individuals in different pig breeds at FSW locus and polymorphism of FSHP gene was analyzed. With the combination of genotype and litter size of sows, it was demonstrated that FSHβ locus is closely associated with major gene controlling litter size in commercial pig breeds, such as Yorkshire, Landrace, Durco. Averagely the AA sows give more 1.5 piglets than BB sows do per litter.     

Sow level risk factors for early piglet mortality and crushing in organic outdoor production

Piglet mortality is a major problem in organic pig production affecting both farm economy and animal welfare. Knowledge is scarce on the risk factors of piglet mortality in Danish commercial organic pig production. The objectives of this study were to evaluate season, litter size, parity, sow body condition and stillborn littermates as risk factors for early piglet mortality and crushing of liveborn piglets from parturition until castration at day 3 to 5 postpartum (pp). The study was conducted over a 1-year period in nine commercial Danish organic pig herds practicing outdoor farrowing all year round. Data included recordings on 3393 farrowings with 50 284 liveborn piglets of which 14.8% died before castration. A subset of the dead piglets were collected and necropsied to identify crushed piglets. The average number of liveborn piglets per litter was 14.8 (SD=3.7) and the average time from parturition until castration was 4.1 (SD=1.7) days. A negative binomial regression analysis was used to model the effect of the predictive variables on the early piglet mortality accounting for different time periods from parturition to castration. An increase in maternal body condition score (BCS) and parity significantly increased the risk of dying between parturition and castration. Early mortality was found to be lowest during spring (March to May) and highest during summer (June to August). Being born into a litter with one or more stillborn littermates increased the risk of early mortality. The risk factors for crushing of piglets were evaluated using a logistic analysis. A significant effect of parity and litter size was found where the odds of at least one piglet in a litter with mortality was diagnosed as crushed increased with increasing parity and litter size. In conclusion, being born during summer (June to August), high parity and maternal BCS and stillborn littermates were found to be risk factors for piglet mortality between parturition and castration. In addition, parity and increasing litter size were found to be risk factors for crushing of piglets in litters with mortality.     

The effect of estrogen receptor genotype on litter size and placental traits at term in F2 crossbred gilts

The effect of estrogen receptor (ESR) genotype (two alleles, A and B) on litter size of 275 Large White x Meishan F2 crossbred gilts (73 AA, 126 AB and 76 BB gilts) was tested. In addition, for 63 of these gilts (18 AA, 24 AB, and 21 BB) the effect of ESR genotype on average placental traits at term was tested, since individual placental information was available for 88% of the 628 liveborn piglets. Without affecting average birth weight of the piglets, ESR genotype significantly affected litter size, i.e. AB gilts had larger litters than BB gilts (P < 0.05). Total number born was 11.38+/-0.38, 11.88+/-0.28, and 10.68+/-0.35, while number born alive was 10.45+/-0.39, 11.07+/-0.29, and 9.85+/-0.36 for AA, AB and BB gilts, respectively. Since the B allele in previous research was associated with largest litters, the hypothesis that ESR is a marker rather than the major gene itself is discussed. Average placental length, surface area, and weight including and excluding amnion were not affected by ESR genotype. However, placentae of AB gilts had a significantly lower number of areolae per placenta than BB gilts and had a lower number of areolae/cm2 placenta than AA and BB gilts. Number of areolae was 8945+/-663, 7240+/-619, and 9694+/-633, for AA, AB and BB gilts, respectively. Although the reason for the low number of areolae on placentae in AB gilts is not yet known, the results suggest that the ESR linked major gene for litter size might be involved in the development and activity of endometrial glands.     

应用多重PCR和基因扫描技术对五指山猪13个家系32个微卫星基因座的遗传分析

五指山猪是我国著名的珍稀畜种,体型小、性成熟早、遗传稳定、近交程度高,可用于医学、药学和生物材料等方面。作者利用多重PCR和基因扫描技术对五指山猪13个家系的32个微卫星基因座进行遗传检测。统计各家系的等位基因组成,计算各家系的平均杂合度和多态信息含量(PIC);结果,各家系32基因座的平均等位基因数为13.66个,平均PIC为0.731,平均杂合度为0.559。表明海南五指山猪具有丰富的遗传多样性。这些结果对于海南五指山猪的保种、定向选育、开发利用都具有重要的指导意义。     

Differential gene expression in ovaries of pregnant pigs with high and low prolificacy levels and identification of candidate genes for litter size

Previous results from a genome scan in an F(2) Iberian × Meishan pig intercross showed several chromosome regions associated with litter size traits in this species. In order to identify candidate genes underlying these quantitative trait loci (QTL), we performed an ovary gene expression analysis during the sow's pregnancy. F(2) sows were ranked by their estimated breeding values for prolificacy: six sows with the highest estimated breeding value (EBV) (i.e., high prolificacy) and six sows with the lowest EBV (low prolificacy) were selected. Samples were hybridized using an Affymetrix GeneChip porcine genome array. Statistical analysis with a mixed model approach identified 221 differentially expressed probes, representing 189 genes. These genes were functionally annotated in order to identify genetic pathways overrepresented in this list. Among the functional groups most represented was, in first position, immune system response activation against external stimulus. The second group consisted of integrated genes that regulate maternal homeostasis by complement and coagulation cascades. A third group was involved in lipid and fatty acid enzymes of metabolic processes, which participate in the steroidogenesis pathway. In order to identify powerful candidate genes for prolificacy, the second approach of this study was to merge microarray data with the QTL positional information affecting litter size, previously detected in the same experimental cross. As a result, we have identified 27 differentially expressed genes colocalizing with QTL for litter size traits, which fulfill the biological, positional, and functional criteria.     

母猪生殖器官大小和产仔数的分子遗传基础

采用单核苷酸多态分析技术（SNP）分析母猪3个生殖激素受体基因（ESR、PRLR和FSHR）的变异,屠宰103头母猪并测定其生殖器官大小;统计母猪的产仔数;利用SAS或SPSS分析软件分析基因变异与母猪的生殖器官大小,产仔数多少的连锁关系,以探讨母猪生殖器官大小和产仔数多少的分子遗传基础,结果表明,如母猪携带的基因型为位点,ESR的BB型,位点FSHRB的BB型,位点ESRB的AA型,位点PRLR的AA型,则母猪的生殖器官较大,产仔数也较多;在位点ESRB或位点PRLR中,带有AA基因型母猪不仅产仔数显著地高于AB、BB型,而且生殖器官也显著大于AB、BB型;在位点ESR和FSHRB中,带有BB基因型母猪的生殖器官,产仔数显著高于带有AB或AA型母猪。     

A genome‐wide linkage analysis for reproductive traits in F2 Large White × Meishan cross gilts

Female reproductive performance traits in pigs have low heritabilities thus limiting improvement through traditional selective breeding programmes. However, there is substantial genetic variation found between pig breeds with the Chinese Meishan being one of the most prolific pig breeds known. In this study, three cohorts of Large White × Meishan F₂ cross‐bred pigs were analysed to identify quantitative trait loci (QTL) with effects on reproductive traits, including ovulation rate, teat number, litter size, total born alive and prenatal survival. A total of 307 individuals were genotyped for 174 genetic markers across the genome. The genome‐wide analysis of the trait‐recorded F₂ gilts in their first parity/litter revealed one QTL for teat number significant at the genome level and a total of 12 QTL, which are significant at the chromosome‐wide level, for: litter size (three QTL), total born alive (two QTL), ovulation rate (four QTL), prenatal survival (one QTL) and teat number (two QTL). Further support for eight of these QTL is provided by results from other studies. Four of these 12 QTL were mapped for the first time in this study: on SSC15 for ovulation rate and on SSC18 for teat number, ovulation rate and litter size.     

Genome-wide association analysis reveals genomic regions on Chromosome 13 affecting litter size and candidate genes for uterine horn length in Erhualian pigs

Litter size has a great impact on the profit of swine producers. Uterine development is an important determinant of reproduction efficiency and could hence affect litter size. Chinese Erhualian pig is one of the most prolific breeds in the world, even though large phenotypic variation in litter size was observed within Erhualian sows. To dissect the genetic basis of the phenotypic variation, we herein conducted genome-wide association studies for total number born and number born alive (NBA) of Erhualian sows. In total, one significant single nucleotide polymorphism (SNP) (P<1.78e−06) and 11 suggestive SNPs (P<3.57e−05) were identified on 10 chromosomes, confirming seven previously reported quantitative trait loci (QTL) and uncovering six QTL for litter size or uterus length. One locus on Sus scrofa chromosome (SSC) 13 (79.28 to 90.43 Mb) harbored a cluster of suggestive SNPs associated with multiparous NBA. The SNP (rs81447100) within this region was confirmed to be significantly (P<0.05) associated with litter size in Erhualian (n=313), Sutai (n=173) and Yorkshire (n=488) populations. Retinol binding protein 2 and retinol binding protein 1 functionally related to the development of uterus were located in a region of 2 Mb around rs81447100. Moreover, four genes related to embryo implantation and development were also detected around other significant SNPs. Taken together, our findings provide a potential marker (rs81447100) for the genetic improvement of litter size not only in Chinese Erhualian pigs but also in European commercial pig breeds like Yorkshire, and would facilitate the final identification of causative variant(s) underlying the effect of SSC13 QTL on litter size.     

猪产仔数分子标记及其效应分析

初步确定了2个新的猪产仔数分子标记,雌激素受体基因ESR的第8外显子处的ESRB位点、催乳素受体基因的第7外显子的FSHRB位点。通过比较和分析多个产仔数的效应,初步确定了4个有利于产仔数提高的分子标记基因型,基因位点ESR、FSHRB的基因型BB的产仔数显著地高于AB、AA型;位点ESRB、PRLR的基因型AA的产仔数显著地高于AB、BB型;4个基因位点多态性与仔猪生长性能、母猪乳头数不存在显著的影响。     

影响二花脸猪高产仔性能的生理及遗传机制研究进展

二花脸猪是我国太湖流域以产仔数高而闻名于世界的优质猪种。已有的研究发现,二花脸猪排卵数高、胚胎死亡率低和子宫容积大是其高产性能的重要生理特征,通过候选基因法鉴别到其高产的重要候选基因——促卵泡素β亚基(follicular-stimulating hormone beta, FSH?),采用QTL定位方法在猪第2、6、7、8、12、13和15号染色体上定位到影响其产仔性状的QTL。但是,目前影响二花脸猪产仔数的主效基因尚不清楚。本文主要介绍了二花脸猪的高排卵数、低胚胎死亡率和高子宫容积特性对高产仔性能形成的生理作用;概括了基于传统方法鉴别出的影响二花脸猪高产仔性能的候选基因和QTL,以及基于组学方法鉴别出的二花脸猪高产候选基因;探讨了如何利用基因组、转录组、蛋白质组和功能组等多组学手段深入研究二花脸猪高产性能的成因,以期为深入解析二花脸猪高产性能的分子遗传机制提供参考。