郏县红牛遗传育种研究进展

为了研究郏县红牛遗传育种研究进展,本文梳理了夏洛来牛、南德温牛、利木赞牛、红安格斯牛、丹麦红牛改良郏县红牛的研究进展,对郏县红牛基因多态性与生长性状的关系和基因遗传变异与生长性状的关系进行了梳理,结果表明：红安格斯牛可以提高郏县红牛的肉用性能和优质牛肉肉块重量,夏洛来牛改良郏县红牛可以提高牛肉品质;ZAG基因、IGF2基因、DGAT2基因、CLPG基因、AQP7基因可以作为生长性状辅助选择的基因,POU1F1基因第6外显子被Hinf I酶切后表现多态性,BB基因型个体可以作为生长性状辅助选择的基因,SREBP-1c基因内含子7区域84 bp缺失位点可以作为生长性状辅助选择的位点。     

Association of single nucleotide polymorphisms in the growth hormone and growth hormone receptor genes with blood serum insulin-like growth factor I concentration and growth traits in Angus cattle

This study was conducted to identify polymorphisms in the promoter and coding regions of the bovine growth hormone and growth hormone receptor genes and to study association of polymorphisms identified in these genes with growth traits and serum insulin-like growth factor-I (IGF-I) concentration. The denaturing gradient gel electrophoresis method and sequencing were utilized to identify three new single nucleotide polymorphisms in the promoter region of the growth hormone gene in Angus cattle. Polymerase chain reaction-based restriction fragment length polymorphism procedures were developed for rapid determination of the single nucleotide polymorphism genotypes in the growth hormone and the growth hormone receptor genes among Angus calves from lines divergently selected for high or low blood serum IGF-I concentration. The IGF-I concentration and growth traits were analyzed using animal models. The single nucleotide polymorphism in the promoter region of the growth hormone receptor gene was associated with serum IGF-I concentration on d 42 of the postweaning test and with mean IGF-I concentration. The associated effects of the markers need to be verified in other populations.     

Association between IGF-I, IGF-IR and GHRH gene polymorphisms and growth and carcass traits in beef cattle

Molecular biology techniques are of help in genetic improvement since they permit the identification, mapping and analysis of polymorphisms of genes encoding proteins that act on metabolic pathways involved in economically interesting traits. The somatotrophic axis, which essentially consists of growth hormone releasing hormone (GHRH), growth hormone (GH), insulin-like growth factors I and II (IGF-I and IGF-II), and their associated binding proteins and receptors (GHRHR, GHR, IGF-IR and IGF-IIR), plays a key role in the metabolism and physiology of mammalian growth. The objectives of the present study were to estimate the allele and genotype frequencies of the IGF-I/SnaBI, IGF-IR/TaqI and GHRH/HaeIII gene polymorphisms in different genetic groups of beef cattle and to determine associations between these polymorphisms and growth and carcass traits. For this purpose, genotyping was performed on 79 Nellore animals, 30 Canchim (5/8 Charolais+3/8 Zebu) animals and 275 crossbred cattle originating from the crosses of Simmental (n=30) and Angus (n=245) sires with Nellore females. In the association studies, traits of interest were analyzed using the GLM procedure of SAS and least square means of the genotypes were compared by the Tukey test. Associations of IGF-I/SnaBI genotypes with body weight and subcutaneous backfat were significant (p<0.05), and nearly significant for longissimus dorsi area (p=0.06), with the BB genotype being favorable compared to the AB genotype. No significant associations were observed between this polymorphism and weight gain or carcass yield (P>0.05). The IGF-IR/TaqI and GHRH/HaeIII polymorphisms showed no association with production traits.     

深县猪mtDNA CytB基因的遗传多样性与母系起源研究

 研究过氧化物酶体增殖激活受体α (PPARα) 基因第7外显子SNP位点与牛的部分胴体、肉质性状的相关性。以6个牛品种(秦川牛、南阳牛、郏县红牛、鲁西牛、安格斯、夏南牛) 共计717个个体为研究对象,采用PCR SSCP结合DNA测序方法对PPARα基因第7外显子进行SNP检测,并该SNP位点与108头秦川牛的部分胴体、肉质性状的相关性进行分析。结果发现在PPARα基因第7外显子的184位检测到C→T突变,在南阳牛、秦川牛、郏县红牛、安格斯、鲁西牛、夏南牛这6个群体中等位基因E/F的频率分别是0.225/0.775,0.151/0.849,0.125/0.875,0.123/0.877,0.070/0.930, 0.157/0.783;遗传学指标结果显示：秦川牛、郏县红牛、安格斯、鲁西牛和夏南牛这5个群体属于低度多态(PIC<0.25),南阳牛为中度多态（PIC=0.288）;相关分析结果显示：该位点与秦川牛的背膘厚和胴体长两个性状显著相关,表现为FF基因型个体在背膘厚性状方面显著高于EE和EF基因型个体(P<0.05),FF基因型个体的胴体长显著高于EE基因型个体(P<0.05)。这一位点可能是影响牛背膘厚和胴体长的主效QTN或与之紧密连锁,可做为肉牛选育的候选分子标记。     

PLIN基因多态性与秦川牛胴体、肉质性状的相关性研究

旨在研究PLIN基因第3、4外显子多态性与秦川牛部分产肉性状的关系。以相同饲养条件下18~24月龄的405头秦川牛为研究对象,利用PCR-SSCP方法和测序技术,对PLIN基因第3、4外显子进行SNPs位点检测,并将发现的SNPs位点与部分产肉性状进行关联性分析。结果,在第3外显子的156 bp处发生T→C突变,第4外显子的14 bp处发生C→T突变。方差分析表明:AB基因型个体的宰前活体质量、胴体质量、背膘厚和眼肌面积显著高于AA和BB基因型个体(P0.05),而且相对于AA基因型个体达到差异极显著水平(P0.01);DE基因型个体的胴体长、胴体胸深和背膘厚显著高于DD和EE基因型个体(P0.05),DD基因型个体的肌肉嫩度极显著高于EE和DE基因型个体(P0.01)。研究显示,PLIN基因单核苷酸多态性对秦川牛脂肪沉积有一定的影响,可作为影响胴体和肉质的候选基因,指导肉牛的育种工作。     

牛DGAT2基因第6内含子MspI-RFLPs和TaqI-RFLPs及其与牛经济性状相关性研究

利用PCR-RFLPs方法分析牛DGAT2基因第5、6和7内含子内部的MspI和TaqI等11种酶切多态性,并在第6内含子检测到MspI-RFLPs和TaqI-RFLPs 2个多态位点。分析表明,该MspI-RFLPs普遍存在于鲁西牛、晋南牛、秦川牛、利木赞×鲁西牛、西门塔尔×延边牛、夏洛来×延边牛、安格斯×延边牛和三河牛、中国荷斯坦牛等9个试验群体中,TaqI-RFLPs只在鲁西牛、晋南牛和秦川牛3个地方品种和利木赞与鲁西牛的杂交群体中检测到,而且等位基因B(T碱基)在除鲁西牛外的群体中分布频率很低。统计分析表明,MspI-RFLPs与肉牛的体脂没有相关,但显著影响屠宰率,AA型个体的屠宰率显著高于AB和BB型(P<0.05)。DGAT2基因第6内含子MspI酶切多态性与三河牛群体平均乳脂率和平均干物质含量有极显著或显著的相关性(P<0.01和P<0.05),AA型个体的平均乳脂率和平均干物质含量均高于BB型个体的均值,但在荷斯坦牛群体中却没有得到相同的结果。研究分析了鲁西牛中TaqI-RFLPs的AA型与AB型个体之间屠宰性能的差异,结果表明这2种基因型与鲁西牛的屠宰性能没有显著的相关(P>0.05),有待在更大的鲁西牛群体中检验TaqI-RFLPs是否影响屠宰性状。     

Relationship of the bovine growth hormone gene to carcass traits in Japanese black cattle.

The bovine growth hormone gene (bGH) possesses three haplotypes, A, B and C, that differ by amino acid mutations at positions 127 and 172 in the fifth exon: (leucine 127, threonine 172), (valine 127, threonine 172) and (valine 127, methionine 172) respectively. The correlation between meat quality or carcass weight and these haplotypes was investigated in Japanese black cattle. Altogether, 940 bGH haplotypes were compared with respect to six carcass traits: carcass weight, longissimus muscle area, rib thickness, subcutaneous fat thickness, beef marbling score and beef colour. The frequency of the B haplotype was higher (0.421) than that of A (0.269) and C (0.311). High carcass weight and low beef marbling were associated with haplotype A (p < 0.05 and p < 0.01 respectively), whereas beef marbling was increased by haplotype C (p < 0.05). Estimated regression coefficient of the A haplotype substitution effect for carcass weight and beef marbling score were 5.55 (13.1% of the phenotypic SD) and -0.31 (17.0%) respectively. That of the C haplotype for beef marbling score was 0.20 (11.0%). The other traits showed no relationship to the haplotypes examined. The results of this investigation suggest that information pertaining to bGH polymorphisms in Japanese black cattle could be used to improve the selection of meat traits.     

六盘山肉牛杂交改良群体Pit-1基因多态性分析

本试验旨在对宁夏六盘山区肉牛杂交改良群体垂体特异性转录因子1(Pit-1)基因的多态性进行研究,为其杂交改良提供理论基础。采用聚合酶链式反应—限制性片段长度多态性方法（PCR-RFLP）,对该群体101个个体Pit-1基因多态性进行检测。结果表明,扩增出的451 bp片段为目的基因片段,其PCR产物经限制性内切酶HinfⅠ消化后表现出多态性,共检测到AA、BB和AB 3种基因型。其中AA基因型频率6.93%（7个）;AB基因型频率34.65%（35个）;BB型基因型频率58.42%（59个）,等位基因A频率24.26%;等位基因B频率75.74%。因此,宁夏六盘山地区的肉牛杂交改良群体Pit-1基因具有多态性。     

Study on Genetic Polymorphism and Ultrasound Measurements for Major Meat Quality Traits in Beef Cattle

In this study,we used PCR-SCCP connect with ultrasonic vivo assay techniques, analysis of correlation between Leptin gene exon Ⅱ E2JW polymorphism and part meat quality traits of backfat thickness, eye muscle area, fat content, plot eye height in Xinjiang Brown cattle at ante-mortem, post-mortem. The results showed that Leptin gene exon Ⅱ E2JW site PIC was moderate polymorphisms (0.25 < PIC < 0.5), there were three genotypes of AA, AB and BB, AB genotype were the predominant genotype, B was the predominant allele. In addition, the forecast value of AB and AA gennotypes individual muscle fat content were inconsistent with actual value.Leptin gene exon Ⅱ E2JW was identified the theroy of a candidate gene loci of beef quality traits in Xinjiang Brown cattle.Genetic polymorphism and ultrasound measurements could not only fastly and accurately measure portion meat traits, but also as a means of marker-assisted selection.     

基因多态性与超声波活体测定相结合对肉牛主要肉质性状的研究

本试验采用PCR-SSCP与超声波活体测定相结合的方法,对Leptin基因外显子Ⅱ E2JW的SNP位点与宰前/宰后新疆褐牛背膘厚度、眼肌面积、脂肪含量、眼肌高度的部分肉质性状进行关联性分析。结果表明,外显子Ⅱ E2JW位点的PIC处于中度多态（0.25 < PIC < 0.5）,E2JW多态位点存在3个基因型：AA、AB和BB,其中AB为优势基因型,B为优势等位基因。相关性结果表明,除预测值中AB、AA基因型个体的肌肉脂肪含量与实测值的分析结果不一致外,其他均表现出新疆褐牛Leptin基因外显子Ⅱ E2JW多态性与预测肉质性状和实测肉质性状关系的一致性。因此,Leptin基因外显子Ⅱ E2JW是影响新疆褐牛肉质性状的候选基因位点,基因多态性结合超声波活体测定技术不仅可以快速、准确地测量部分肉质性状,而且可作为分子标记辅助选择的一种手段。     

UCP1基因变异与绵羊生长及胴体性状的关联分析

试验旨在探讨解耦联蛋白1（UCP1）基因核苷酸变异对绵羊生长性状的性别差异和胴体性状的影响，以期能够筛选出可以提升绵羊生长及胴体性状的核苷酸变异，为提高绵羊相关重要经济性状的分子遗传标记提供材料。以9个绵羊品种为研究对象，用PCR-SSCP方法检测不同性别绵羊中UCP1基因内含子5区和外显子6区变异。利用Minitab 16.0软件中一般线性模型分析内含子5区等位基因与不同性别绵羊生长性状、公羔胴体性状的关联性。结果显示，绵羊UCP1基因内含子5区和外显子6区共检测到8个核苷酸变异，其中位点c.910 G/A突变导致p.Ala304Thr氨基酸变异。生长性状关联分析结果表明，内含子5区等位基因对绵羊生长性状的影响存在性别特异性，母羔中携带等位基因A₁的群体较缺失群体具有较低的初生重（P<0.05），公羔中携带等位基因C₁的群体较缺失群体具有较高的断尾重（P<0.05），未发现其他等位基因与羔羊的生长性状存在性别特异性。胴体性状关联分析结果表明，携带等位基因A₁的群体具有较低后腿瘦肉量、腰部瘦肉量和较高的肩部瘦肉比例（P<0.05），携带等位基因C₁的群体具有较低的后腿瘦肉比例（P<0.05），其他胴体性状均没有发现与等位基因存在显著相关，基因型分析结果与等位基因分析结果一致。结果提示，UCP1基因对绵羊的生长性状影响具有性别特异性，且携带等位基因A₁的公羔群体具有较低的胴体生产性状，为提高公羔胴体生产性能提供了理论依据。     

草原红牛胰岛素样生长因子结合蛋白6基因外显子2的遗传多态性及遗传效应分析

本试验旨在研究草原红牛不同基因型与屠宰肉用性状的关联性。选用草原红牛作为试验群体,采用PCR-SSCP方法检测胰岛素样生长因子结合蛋白6(insulin-like growth factors binding proteins 6,IGFBP6)基因外显子2的多态性,采用最小二乘法拟合线性模型将突变位点的不同基因型与牛屠宰肉用性状进行关联分析。结果表明,在IGFBP6基因外显子2上发现1个多态性位点A180G,具有3种基因型:AA、AB和BB。SPSS 13.0统计分析结果表明,IGFBP6基因外显子2 AB和BB基因型净肉重极显著高于AA基因型(P<0.01);眼肌面积和骨重显著高于AA基因型(P<0.05),而在宰前活重、胴体重、肾脂重和肋脂重方面3种基因型个体之间差异不显著(P>0.05)。结果显示,IGFBP6基因的多态性与屠宰肉用性状具有一定的关联性,可为今后草原红牛的育种保种工作提供理论依据,并对草原红牛肉用性状的改善提供参考。     

肉牛经济性状关联的微卫星标记研究进展

微卫星序列是以1～6个短核苷酸为基本重复单位的重复序列,呈共显性、重复性好、多态性丰富和操作简单,在基因定位与QTL分析、亲权分析、分子标记辅助选择等方面应用广泛.本文综述了与肉牛部分经济性状(生长发育性状、胴体性状、肉质性状、多胎性状和出生重)相关的微卫星标记研究进展.     

中国不同牛种DGAT1基因遗传多样性及经济性状关联分析研究进展

乙酰辅酯A∶二酰甘油酰基转移酶(DGAT1)是甘油三脂合成过程中唯一的关键酶,对动物机体脂肪代谢、沉积起重要作用,已成为研究奶牛泌乳性状和肉牛主要经济性状的重要候选基因之一。近年来,随着相关研究的不断深入,研究者普遍认为DGAT1基因对奶牛的产奶量、乳脂率、乳蛋白率、乳糖率、乳脂肪酸及空怀天数、输精次数、初配年龄等繁殖性状均有显著影响,且与肉牛体脂也存在相关性。作者主要介绍了DGAT1基因的遗传多态性及其与经济性状关联分析在中国各地区不同品种奶牛、肉牛、水牛和牦牛上的研究进展,指出该基因对奶牛的产奶性能及肉牛的肉质性状、生长性状具有重要的调控作用。但由于环境条件、品种及选育背景等因素影响,DGAT1基因遗传多态性对不同地区不同牛群体的经济性状影响不尽相同。因此,要将DGAT1基因实际应用在品种选育工作中还需明确该基因对研究群体的具体遗传效应,并结合其他候选基因进行综合分析。     

Identification and characterization of polymorphisms within the 5' flanking region, first exon and part of first intron of bovine GH gene

The aim of the present study was to identify and characterize polymorphisms within the 5′ flanking region, first exon and part of first intron of the bovine growth hormone gene among different beef cattle breeds: Nelore (n = 25), Simmental (n = 39), Simbrasil (n = 24), Simmental × Nelore (n = 30), Canchim × Nelore (n = 30) and Angus × Nelore (n = 30). Two DNA fragments (GH1, 464 bp and GH2, 453 bp) were amplified by polymerase chain reaction and then used for polymorphism identification by SSCP. Within the GH1 fragment, five polymorphisms were identified, corresponding to three different alleles: GH1.1, GH1.2 and GH1.3 (GenBank: AY662648 , AY662649 and AY662650 , respectively). These allele sequences were aligned and compared with bovine GH gene nucleotide sequence (GenBank: M57764 and AF118837 ), resulting in the identification of five insertion/deletions (INDELs) and five single nucleotide polymorphisms (SNPs). In the GH2 fragment two alleles were identified, GH2.1 and GH2.2 (GenBank: AY662651 and AY662652 , respectively). The allele sequences were compared with GenBank sequences ( M57764 , AF007750 and AH009106 ) and three INDELs and four SNPs were identified. In conclusion, we were able to identify six new polymorphisms of the bovine GH gene (one INDEL and five SNPs), which can be used as molecular markers in genetic studies.     

AQP7基因内含子4多态性与郏县红牛生长性状的关联分析

本试验旨在研究郏县红牛AQP7基因内含子4的遗传变异,分析其遗传多态性对生长性状的影响。采用PCR-SSCP、DNA序列分析及生物信息学技术探讨郏县红牛AQP7基因内含子4的多态性及其与生长性状的关联情况。结果发现,郏县红牛AQP7基因内含子4存在A、B、C 3个等位基因,其基因频率依次为0.838、0.076、0.086;郏县红牛AQP7基因4种不同基因型与其体重和胸围有显著相关(P< 0.05),与其他指标不存在相关性(P> 0.05),且BC基因型个体的体重和胸围均显著大于其他3种基因型(P< 0.05)。暗示BC基因型是提高郏县红牛生长性状的有利基因型,AQP7基因内含子4的遗传多态性与郏县红牛的生长性状存在相关,因此可作为该品种生长性状分子标记辅助选择的候选基因。     

Association Analysis on Polymorphisms of AQP7 Gene Intron 4 with Growth Traits in Jiaxian Cattle

The objectives were to research the genetic variations in intron 4 of AQP7 gene in Jiaxian cattle, and analyze the influence of genetic polymorphisms on growth traits.PCR-SSCP technique, DNA sequence analysis and bioinformatics were used to analyze the association between the polymorphisms of the intron 4 of AQP7 gene and growth traits in Jiaxian cattle.The results showed that there were three alleles A, B, C in this locus with the corresponding frequencies of 0.838, 0.076 and 0.086;The polymorphisms of the intron 4 of AQP7 gene had significant association with body weight and chest circumference (P< 0.05), while had no significant association with other growth traits (P> 0.05).The individuals with BC genotype was significantly higher than those of individuals with other genotypes in body weight and chest circumference (P< 0.05).It suggested that BC genotype had positive effect on growth traits, and the correlation was detected between the polymorphisms of the intron 4 of AQP7 gene and growth traits in Jiaxian cattle.So AQP7 gene could be used as a candidate gene for molecular marker assisted selection of growth traits in Jiaxian cattle.     

Association analysis of novel polymorphisms in 2′, 5′‐oligoadenylate synthetase gene with reproductive traits in indigenous and cross‐bred cattle of Indian Origin

2′, 5′‐Oligoadenylate synthetases (OAS) are important components of an interferon‐mediated antiviral pathway. No polymorphisms in exonic regions of bovine OAS1 gene have been identified and associated with reproduction traits. The objective of the study was to detect and evaluate the effects of mutations in exonic region of bovine OAS1 gene with reproduction traits in cattle. DNA samples collected from 250 individual cows of two Indian dairy breeds (Sahiwal and Frieswal) of cattle were used in the study. The genetic variants of the OAS1 gene were identified with polymerase chain reaction–single‐strand conformation polymorphism (PCR‐SSCP) and sequence analysis using seven set of primer pairs. The PCR‐SSCP analysis revealed polymorphism in the fragments comprising of exon 2, exon 5 and first fragment of exon 6 while the fragments of exons 1, 3, 4 and second fragment of exon 6 were monomorphic in Sahiwal and Frieswal cattle. The mutations in the amplified region comprising of exon 2 were found to have significant association with age at first breeding and calving, service period, dry period and pregnancy rate. Significant associations were found between SNPs in the exon 5 and service and dry periods of the animal, whereas the genetic variants in the first fragment of the exon 6 showed significant association with age at first breeding and calving. To our knowledge, this study demonstrated for the first time that the polymorphisms in OAS1 gene were associated with reproductive traits and it can be chosen as a candidate gene for improvement of reproductive performance of cattle.     

Comparative Study on the Beef Traits Between Jiaxian Red Cattle and Crossbred Beef Cattle

In order to study the beef traits and beef quality of Jiaxian Red bull with crossbred beef cattle, 5 purebred Jiaxian Red bull, 5 Red Angus cattle×Jiaxian Red bull F1 generation, 5 Simmental cattle×Jiaxian Red bull F1 generation and 5 Charolais cattle×Jiaxian Red bull F1 generation were chosen and assigned into groups A,B,C and D for continual finishing for 3 months of age under the same feeding and management conditions and slaughtered at the end of the trial. The results showed that there were no significant difference in body height, body length, chest girth and cannon circumference between Red Angus cattle×Jiaxian Red bull F1 generation and purebred Jiaxian Red bull, the same as in Charolais cattle×Jiaxian Red bull F1 generation (P>0.05). The weight of head, skin, fore hooves and tail, the skin thick, esophagus, trachea, liver, spleen, lung, kidney, and carcass meat percentage, loin eye area and the meat bone ratio in crossbred beef cattle groups were not significant differences compared with purebred Jiaxian Red bull (P>0.05), but its hind legs wide in carcass was significantly higher than that of purebred Jiaxian Red bull (P<0.05). The high-grade meat weight and high quality meat weight in Red Angus cattle×Jiaxian Red bull F1 generation were 70.22 and 105.22 kg, respectively, and was the highest in four groups,but the high-grade beef percentage among four groups did not show significant differences (P>0.05). The high quality meat percentage in Red Angus cattle×Jiaxian Red bull F1 reached 34.43%, was significantly higher than that of Jiaxian Red bull and Simmental cattle×Jiaxian Red bull F1 generation (P<0.05), and was higher than that of Charolais cattle×Jiaxian Red bull F1 generation (P>0.05). In terms of meat quality, the content of inosinic acid that related with the meat flavor in Red Angus cattle×Jiaxian Red bull F1 was the highest in four groups, while the content of unsaturated fatty acid and the flavor related amino acid, such as aspartic acid, glutamic acid, glycine and alanine, in Charolais cattle×Jiaxian Red bull F1 were the highest in four groups,and their values were 53.85% and 8.04%, respectively. To sum up, the results suggested that there was no significant difference in body development and beef traits using Red Angus cattle, Simmental cattle and Charolais cattle to improve Jiaxian Red bull, while Red Angus cattle crossbred with Jiaxian Red bull could moderately improve the weight of high-grade beef and high-quality beef, and Charolais cattle could moderately improve the beef quality of Jiaxian Red bull.