Variation at the insulin-like growth factor 1 gene and its association with body weight traits in the chicken

The insulin-like growth factor 1 (IGF1) is essential for normal embryonic and postnatal growth in mammals. In this study, a total of 342 F(2) individuals, derived from Broiler crossing to Baier layer (Northeast Agricultural University Resource Population, NEAURP), were used to investigate the associations of haplotypes in the chicken IGF1 (cIGF1) gene with body weight traits. Primers for the 5'-flanking, exon 3 and 3'-flanking regions of cIGF1 were designed according to chicken genome database. Single nucleotide polymorphisms (SNPs) between parental lines were detected by sequencing, and PCR restriction fragment length polymorphism (PCR-RFLP) and PCR single-stranded-conformation polymorphism (PCR-SSCP) methods were used to genotype the SNPs in the population. Haplotypes were constructed with the three SNPs detected. The association analysis showed that haplotypes based on three cIGF1 polymorphisms (c.-366A>C, c.528G>A and c.*1024C>T) were associated with body weight traits, suggesting that cIGF1 or a tightly linked gene had effects on body weight in the chicken.     

Three indel variants in chicken LPIN1 exon 6/flanking region are associated with performance and carcass traits

1. LPIN1 is a Mg²⁺-dependent phosphatidic acid phosphatase. Variation in chicken LPIN1 exon 6 and its flanking regions were identified and three indel variants in 6 breeds and their associations with performance traits were studied.

2. Seven variants were detected from 6 breeds, which contained a synonymous tri-allelic variant (c.924A/T/C) and three indels. The exon 6 variants detected from chicken breeds were conserved among bird species. The indel variation frequency presented clear differences among breeds.

3. Two coding indels (c.1014-1018del3 and c.1125-1138del12) were multiples of three nucleotides and maintained the open reading frames of LPIN1 proteins. However, they were predicted to result in the clear change of the RNA secondary structure of chicken LPIN1 exon 6 and LPIN1 protein conformation.

4. The association analysis showed that c.871–15-22del6 variation had a significant effect on body weight at hatch (BW0) and 2 weeks (BW2); c. 1014-1018del3 variation had a significant effect on BW4, BW6, caecum length and gizzard weight (GW) traits; c.1125-1138del12 variation had a significant effect on BW12, shank length at 4 weeks (SL4), carcass weight, lactate dehydrogenase traits (LDH), glucose (GLU) and albumin (ALB) traits. The genotype combination for c.1014-1018del3 and c.1125-1138del12 also presented significant effects on SL4, SL8, GW, leg muscle weight, ALB, GLU and LDH.

5. The study demonstrated that chicken LPIN1 has an important effect on body, carcass and organ weight, serum LDH, GLU and ALB level.

     

Genomewide association study of immune traits in chicken F2 resource population

Immune traits play pivotal roles in animal immune capacity development and disease resistance. Single nucleotide polymorphisms (SNPs) are common forms of genetic variations among individuals, which are thought to account for the majority of inherited phenotypic variations. In this study, we performed genomewide association, using the Illumina 60K SNP BeadChip studies to detect molecular markers and candidate genes associated with immune traits in an F2 population. Sixteen immune traits were measured. We identified 85 significant SNPs (p < 2.98 × 10^?6) with 5% as the genomewide significance threshold, 380 SNPs of suggestive significance (p < 5.96 × 10^?5) from simple model (general linear model, GLM) and 15 SNPs of suggestive significance (p < 5.96 × 10^?5) from the compressed mixed linear model (MLM), which were also found in GLM (six significant SNPs and seven suggestive SNPs). Three significant SNPs (GGaluGA151406, Gga_rs14554319 and Gga_rs13593979) and candidate genes (LYRM4 and KTN1) were found to be associated with avian influenza antibody titres, and the first two SNPs are from the results of two‐model analysis. For the immune organs, through the analysis of GLM, 19 SNPs were found to be significantly associated with the thymus weight, 61 SNPs were significantly associated with the bursa of Fabricius weight, six of which were located within a 34‐Mb region (125 846 474–159 649 698 bp) on chicken chromosome 1 (GGA1). A candidate region relevant to haematological traits from GLM was found in GGA4 and 9 loci were located on it. Three loci (GGaluGA348521, Gga_rs16098446 and GGaluGA348518) within 179 kb (16 286 868–16 466 134 bp) on GGAZ from GLM provided evidence that this genomic segment may be relevant to red blood cell volume distribution width (RDW). Our study provides a list of significant SNPs and candidate genes that will be valuable information for unveiling the underlying molecular mechanism of immune regulation.     

峨眉黑鸡屠宰性能及肉质性状研究

选取36周龄峨眉黑鸡公、母各6只,对其体尺、屠宰性能、肉品质及化学成分进行了测定分析。结果显示:峨眉黑鸡多项指标达到了优质肉鸡标准,同时,多项体尺、屠宰性能、肉品质和化学成分指标存在显著的性别间差异。研究表明,峨眉黑鸡的公鸡、母鸡均具有较好的产肉性能和优良的肌肉品质。     

Identification of a single nucleotide polymorphism of the insulin-like growth factor binding protein 2 gene and its association with growth and body composition traits in the chicken

Insulin-like growth factor binding protein 2 regulates a broad spectrum of biological activities involved in growth, development, and differentiation. The current study was designed to investigate the associations of IGFBP2 gene polymorphisms with chicken growth and body composition traits. The Northeast Agricultural University Resource Population (NEAURP) was established by crossing broiler sires, derived from lines at Northeast Agricultural University, that were divergently selected for abdominal fat, with Baier layer dams, a Chinese local breed. The F1 birds were inter-crossed to produce an F2 population. Body weight and body composition traits were measured in the NEAURP. The PCR primers for the intron 2 region of IGFBP2 were designed based on chicken genomic sequence. Nucleotide polymorphisms between parental lines were detected by DNA sequencing. A C/T SNP in intron 2 was detected, and PCR-RFLP methods were then developed to genotype the F2 individuals. The results showed that the IGFBP2 SNP was associated with multiple traits, including BW, metatarsus length, shank length, femur length, shank weight, femur weight, metatarsus claw weight, and abdominal fat weight in the 1,028 NEAURP F2 individuals. This research suggests that IGFBP2 or a tightly linked gene has broad effects on growth and development in the chicken.     

鸡早期产蛋性能与ESR1基因内含子1多态性相关分析

为探寻鸡ESR1基因内含子1多态性与早期产蛋性能存在的关联性,采用DNA测序和PCR-SSCP技术对124只绿壳蛋鸡的基因型与性状进行相关性研究。结果表明,鸡ESR1基因In1片段中发现5种基因型:T1T1、T1T2、T2T2、T1T3和T3T3;试验群体处于高度多态,群体遗传偏离了Hardy-Weinberg平衡(P<0.05)。方差分析显示,各基因型个体间的早期产蛋性状指标达到了显著(P<0.05)或极显著(P<0.01)水平,其中T1T3基因型个体性能处于优势,对该群体进行人为选择可提高产蛋性能。     

Polymorphism and expression of insulin-like growth factor 1 (IGF1) gene and its association with growth traits in chicken

1. The objectives of the study were to detect polymorphism in the coding region of the IGF1 gene, explore the expression profile and estimate association with growth traits in indigenous and exotic chickens.

2. A total of 12 haplotypes were found in Cornish, control layer and Aseel breeds of chicken in which the h1 haplotype was most frequent.

3. Nucleotide substitutions among haplotypes were found at 21 positions in the IGF1 gene in which 4 substitutions resulted in non-synonymous mutations in the receptor binding domain of the IGF1 protein.

4. The haplogroup showed a significant effect on body weight at 24 and 42 d of age in the control layer line, body weight at 42 d and daily weight gain between 29 and 42 d in the control broiler line, daily weight gain between 29 and 42 d in Cornish, and body weights at 42 d as well as daily weight gain between 29 and 42 d in Aseel birds.

5. IGF1 expression varied among the breeds during embryonic and post-hatch periods. The expression among the haplogroups varied in different chicken tissues. The effect of haplogroup on myofibre number in pectoral muscle was non-significant, although there was significant variation in numbers between d 1 and d 42, and between broiler and layer lines.

6. It was concluded that the coding region of the IGF1 gene was polymorphic, expressed differentially during the pre-hatch and post-hatch periods, and haplogroups showed significant association with growth traits in chicken.     

Study on association of single nucleotide polymorphism of CAPN1 gene with muscle fibre and carcass traits in quality chicken populations

This study was aimed at investigating the effect of the calpain 1 (CAPN1) gene on carcass and meat quality traits in eight meat-type chicken populations, including five pure lines (developed from Chinese local breeds) and three cross-breeds. Primer pairs for the Coding Sequence (CDS) region in CAPN1 were designed from the chicken genomic sequence database. Polymorphisms were detected by polymerase chain reaction (PCR)-Single Strand Conformation Polymorphism (SSCP) and DNA sequencing. Three single nucleotide polymorphisms (SNP; C2546T, G3535A and C7198A) were detected among individuals in each population. The associations of their haplotypes (H1 = CGA, H2 = CGC, H3 = CAA, H4 = CAC, H5 = TGA and H7 = TAA) with chicken breast muscle fibre and carcass traits were analysed. Results showed that the haplotypes were associated with live weight (LW), carcass weight (CW), breast muscle weight (BMW) and leg muscle weight (LMW) (p < 0.05), and were also related to eviscerated percentage (%EP) and breast muscle fibre density (p < 0.01). H1H3 haplotype was dominant for LW, CW and BMW; H1H5 haplotype was dominant for EP; H3H4 haplotype was dominant for LMW and H1H1 haplotype was dominant for BFD. It was concluded that the CAPN1 gene may be a major gene affecting meat quality traits of chicken or it is linked with the major gene. H1H3, H1H5 and H3H4 were the most advantageous haplotypes for carcass traits whereas H1H1 was the positive haplotype for breast muscle fibre trait.     

zyxin基因外显子7(exon 7)SNP位点与京海黄鸡屠宰及肉品质性能的相关性

以京海黄鸡为试验材料,采用PCR-SSCP技术检测斑联蛋白(zyxin)基因第7外显子SNP,探讨zyxin基因该位点多态性与京海黄鸡屠宰性能以及肉品质之间的关系.结果发现该位点存在1个SNP突变位点,表现为3种基因型,分别为AA、AB和BB.统计分析表明公鸡的肝质量、屠宰率,BB基因型显著高于AA型(P＜0.05);腹脂质量、胸肌率、腿肌率,AA型显著高于BB型(P＜0.05).母鸡群体中,只有爪质量AA型显著高于BB型(P＜0.05).3种基因型对内品质的影响均不显著(P＞0.05).     

A novel 31-bp indel in the paired box 7 (PAX7) gene is associated with chicken performance traits

1. A novel 31-bp indel polymorphism in intron 3 of the chicken paired box 7 (PAX7) gene was identified and genotyped in an F₂ resource population of Gushi chicken crossed with Anka broiler to analyse its associations with chicken growth, carcass and meat quality traits.

2. Results showed that the 31-bp indel was significantly associated with body weight at 4, 6, 8, 10, 12 weeks of age and body size indices including shank length, shank girth, body slanting length at 8 and 12 weeks of age. Significant associations were found for carcass weight, semi-evisceration weight, evisceration weight, breast muscle fibre diameter, leg muscle fibre diameter, breast muscle fibre density, while no significant association with leg muscle fibre density was observed.

3. It was concluded that the 31-bp indel in intron 3 of the PAX7 gene was associated with chicken growth, carcass and meat quality traits where the 31-bp deletion had a negative effect on chicken growth and carcass traits and positive effect on meat quality traits.     

DRD4基因多态性与鸡繁殖性状的相关性分析

旨在分析多巴胺受体4基因(dopamine receptor 4,DRD4)多态性与鸡繁殖性状的关系。对DRD4基因共设计3对引物,筛选出7个多态位点,对瑶山鸡母系60个个体进行了与繁殖性状的关联分析。结果表明:DRD4基因5365(C→T)、5747(C→T)对开产日龄存在显著影响(P0.05);5546(G→A)对首次就巢日龄、首次就巢持续时间和300日龄就巢次数存在显著影响(P0.05)。结果提示:DRD4基因5365(C→T)、5747(C→T)可作为开产日龄辅助选择标记;DRD4基因5546(G→A)可作为首次就巢日龄、首次就巢持续时间和300日龄就巢次数辅助选择标记。     

鸡钙蛋白酶抑制蛋白基因多态性及其与肉质性状的相关性

为探讨钙蛋白酶抑制蛋白(Calpastatin,CAST)基因作为影响鸡肉质性状候选基因的可能性,本研究以8个优质鸡新品系为试验材料,采用PCR-SSCP方法对CAST基因的部分编码区进行单核苷酸多态性(SNPs)检测。结果发现了CAST基因1个多态性位点(G→T),使试验鸡群表现出AA、BB和AB这3种基因型。利用SAS(8.01)软件统计分析了3种基因型与优质鸡肉质性状的相关性,结果表明,CAST基因的多态性对鸡肌内脂肪含量、肌纤维密度和直径有显著的影响(P0.05),对其他肉质性状无显著影响(P0.05);推测CAST基因可能对鸡肉质性状具有很大的影响或与控制肉质性状的主基因连锁。     

Polymorphisms of the PNPLA3 gene and their associations with chicken growth and carcass traits

1. An F₂ resource population of Gushi chickens crossed with Anka broilers was used to investigate the genetic effects of the chicken PNPLA3 gene on growth and adipose accumulation.

2. Associations between three SNPs (g.40006G?>?T, g.42344T?>?C and g.42404A?>?T) and broiler traits were determined using linkage disequilibrium, haplotype construction and association analysis.

3. The g.40006G?>?T mutation was associated with body weights at 4, 6, 8, 10 and 12 weeks of age, carcass weight, evisceration weight and semi-evisceration weight (P? 4. Haplotypes of the g.42344T?>?C and g.42404A?>?T mutations were associated with body weight at 12 weeks, carcass weight, evisceration weight, and semi-evisceration weight (P? 5. The results suggest that the PNPLA3 gene may be in linkage with the causative mutation or a QTL controlling growth traits in chickens. In contrast to human studies, the polymorphisms were not associated with fat related traits.     

草原红牛二酞甘油酞基转移酶2（DGAT2）基因的多态性及与泌乳性状关联性分析

为研究草原红牛二酞甘油酞基转移酶2（DGAT2）的多态性及与泌乳性状相关性,采用PCR-RFLP方法检测DGAT2基因的遗传多态性。结果检测到AA、AB和BB 3种基因型,统计分析表明该多态位点与草原红牛乳中脂肪和干物质含量呈显著相关,脂肪：AA型极显著高于AB型和BB型（P〈0.01）,AB型和BB型之间差异不显著（P〉0.05）;干物质：AA型显著高于AB型（P〈0.05）,极显著高于BB型（P〈0.01）,而AB型又显著高于BB型（P〈0.05）;其他泌乳性状的基因型间差异不显著（P〉0.05）。此结果提示,DGAT2基因对草原红牛乳中脂肪和干物质含量具有较大的遗传效应,可以初步推断DGAT2是控制这些性状的众多基因之一,可能是影响草原红牛乳中脂肪和干物质含量的一个主效基因或与主效基因相连锁,可作为选育草原红牛低脂及高干物质奶牛的分子标记,用于标记辅助选择意义重大。     

Polymorphisms in GnRHI and GnRHII genes and their association with egg production and egg quality traits in chicken

1. Two candidate genes, namely, Gonadotropin releasing hormone I (GnRHI) and Gonadotropin releasing hormone II (GnRHII) play pivotal roles in ovulation and egg production in chicken. The objective of this study was to explore polymorphism in these genes and to estimate the effects of polymorphism of these two genes on egg production and egg quality traits in White Leghorn laying hens.

2. Single strand conformation polymorphism followed by sequencing was performed to detect polymorphism in these genes.

3. The coding regions of the GnRHI and GnRHII genes were found to be polymorphic. In the GnRH1 gene, 12 haplotypes were determined, of which the h1 haplotype was predominant and the h5, h9 and h11 haplotypes were the least frequent ones. In the GnRHII gene, eight haplotypes were found, of which the h1 haplotype was the most frequent and the h6 was the least frequent haplotype in the White Leghorn population.

4. The haplogroups of GnRHI had a significant effect on body weight and egg production up to 64 weeks of age, yolk content, Haugh units and egg shell parameters. The h1h2 haplogroup of the GnRHI gene showed the highest egg production, with 211.0 ± 24.3 eggs up to 64 weeks of age, while the highest yolk content and Haugh unit was found in h3h10 haplogrouped birds. The haplogroups of GnRHII had a significant effect on age at sexual maturity (ASM) where the shortest ASM was found in the h1h4 birds (147.3 ± 5.9 d) and the longest ASM was observed in the h1h3 birds (160.6 ± 23.4 d).

5. It was concluded that GnRHI and GnRHII genes are polymorphic and have a significant effect on body weight, egg production and egg quality traits in White Leghorn laying hens.     

肌联蛋白基因SNPs位点多态性与延边黄牛生长性状间的关联分析

为了研究延边黄牛肌联蛋白(Titin,TTN)基因的多态性对延边黄牛生长性状的影响,选取208头健康的32月龄延边黄牛,利用PCRRFLP和PCR-SSCP方法进行TTN基因遗传变异检测。结果表明:延边黄牛TTN基因存在2个突变位点,分别为A47751G突变,导致氨基酸Ile→Val的错义突变,C73301A同义突变;χ2检验显示,2个突变位点在所检测延边黄牛群体中处于Hard-Weinberg平衡状态(P005);关联分析显示,延边黄牛TTN基因2个突变位点的不同基因型与体高、体斜长、胸围和尻长上差异显著(P005)。将2个突变位点合并基因型分析结果表明,组合AACC基因型个体在体高和体斜长显著高于AATT型个体(P005),初步认为TTN基因对延边黄牛生长性状的有显著影响,其中组合AACC可能是影响延边黄牛生长性状的最佳基因型组合。     

Polymorphism in GHRH gene and its association with growth traits in Chinese native cattle

Growth hormone-releasing hormone (GHRH) is secreted by the hypothalamus and stimulates growth hormone (GH) released from the pituitary. Mutations detected in GHRH gene showed associations with animal production traits. The purpose of this study was to investigate the association of the GHRH gene with growth traits in Chinese native cattle. PCR-SSCP and sequencing were used to detect mutations of the GHRH gene in this study. One novel mutation 4251nt (C>T) was found and the frequencies of C allele were 0.8778 and 0.8476 for Qinchuan and Nanyang cattle, respectively. Body weight with the CT genotype was significantly higher (P<0.05 or P<0.01) than those with CC genotype for different growth periods (6, 12, 18, and 24 months old) in Nanyang cattle. Our findings suggested that polymorphism in bovine GHRH might be one of the important genetic factors to influence body weight.     

Associations between polymorphisms in the chicken VIP gene,egg production and broody traits

1.?In this study, the vasoactive intestinal peptide (VIP) gene was screened for variations by cloning and sequencing to analyse the association with chicken egg production and broodiness traits.

2.?Sequencing revealed that 69 polymorphisms were found in a 9305-bp length of the chicken VIP gene, of which 39 were located in introns, 28 in 5′ regulatory region, 1 in exon 6 (synonymous) and 1 in 3′-UTR. The nucleotide diversity corrected for sample size was 1·96 × 10^?3.

3.?Five polymorphisms, C-3134T, “AGG” indel at ?2648 to ?2650, C + 338T, G + 780T and A + 4691G, were genotyped in 644 individuals of Ningdu Sanhuang chickens to evaluate their effects on egg production and broodiness traits. Marker-trait association analyses showed that the “AGG” indel was associated with total number of eggs from 90 to 300 d of age and total number of qualified eggs from 90 to 300 d of age. In contrast, C + 338T was associated with duration of broodiness.     

奶牛催乳素基因多态性及其与产奶性状的关联分析

催乳素(PRL)基因是影响奶牛泌乳性状的重要候选基因。通过PCR-RFLP结合测序方法检测了荷斯坦奶牛PRL基因5'侧翼区调控序列的多态性,并利用最小二乘法分析其与泌乳性状的关联性。结果显示:在PRL基因5'侧翼序列的906位点处发现A/G突变。共发现AA、AG和GG三种基因型,基因型频率分别为0.239 3、0.522 8和0.177 9,A和G的等位基因频率为0.530 7和0.469 3。多态信息含量检测结果显示该位点处于中度多态,卡方检验显示该位点处于Hardy-Weinberg不平衡状态(P0.05)。关联分析结果表明,该位点多态对产奶量、乳脂率均有影响。