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。     

贵州荷斯坦奶牛甘油二酯酰基转移酶基因多态性及其与产奶性状的关联分析

甘油二酯酰基转移酶（diacylglycerol acyl-transferase 1,DGAT1）是控制甘油三脂合成的关键酶。近年来DGAT1基因被鉴定出来,被认为是奶牛乳脂率的一个重要功能候选基因。该研究以贵州荷斯坦奶牛为试验动物,利用奶牛DGAT1基因序列设计引物,以RCR-单链构象多态性（PCR-SSCP）方法检测了奶牛DGAT1基因第8外显子的碱基突变。结果共检测到AA、AB和BB 3种基因型,基因型频率分别为0.5588、0.3824和0.0588,等位基因A和B的频率分别为0.7549和0.2451;该碱基突变对305 d校正产奶量和乳脂率的影响均达到显著水平（P<0.05）,乳蛋白率影响不显著（P>0.05）;多重比较结果表明,AA和AB型对305 d校正产奶量和乳脂率均显著高于BB型（P<0.05）。结果显示,DGAT1基因突变对贵州荷斯坦奶牛泌乳性状有较大的遗传效应,可用于其泌乳性状的分子标记辅助选择。     

奶牛产奶性状候选基因研究进展

产奶性状是奶牛的主要经济性状，主要包括产奶量、乳脂量、乳蛋白量、乳脂率、乳蛋白率5个方面。产奶性状改良是当今遗传育种界的研究热点之一。利用候选基因策略寻找影响产奶性状的主效基因，从而应用于奶牛产奶性状的改良，从分子遗传学角度来说是可行的。本文综述了与奶牛产奶性状有关的基因：乳蛋白基因、Weaver基因、催乳素基因、生长素和生长素受体基因、淀粉酶基因、核外基因和其他基因（DGAT1和DGAT2基因、垂体特异性转录因子）以及主效QTL的研究进展。     

家畜DGAT1基因的生物学功能及其在遗传育种中的应用研究

DGAT1基因属于酰基辅酶A胆固醇酰基转移酶(acyl CoA:cholesterol acyltransferase,ACAT)基因家族,DGAT1基因催化反应的底物为二酰甘油和乙酰辅酶A(fatty acyl CoAs),该基因一方面参与脂肪的合成,在细胞中起到重要的中心代谢作用,另一方面作为影响家畜泌乳性状和脂肪沉积的重要候选基因,被认为对乳脂合成、脂肪含量和食用价值均有显著影响。本文通过对DGAT1基因的生物学功能、分子结构以及该基因多态性与各类家畜经济性状的关联性进行综述,阐明该基因在家畜发育中的遗传表达规律,并对DGAT1基因在家畜育种方面的研究前景做出展望。     

影响奶牛产奶性状候选基因研究进展

奶牛的产奶性状是一种重要的数量经济性状，由微效多基因决定。奶牛产奶性状的更新以及改良是当今遗传育种研究的热点之一。利用候选基因的研究策略去寻找影响奶牛产奶性状的关键基因，进而从根本上改良奶牛产奶性状，从分子遗传学的角度来说具有实际意义。本文综述了与奶牛产奶性状相关基因ABCG2、BLG、CSN3、DGAT1、OPN、POU1F1、PPARGC1A、SCD1和SYP19等的研究进展，以期为提高奶牛产奶性能的研究提供帮助。 [关键词] 奶牛|产奶性状|候选基因     

奶牛产奶性状候选基因的研究进展

产奶性状主要包括产奶量、乳蛋白率、乳脂率、乳糖率和干物质率等方面,是奶牛的重要数量经济性状,由微效多基因所决定。通过改良产奶性状来提高奶牛产奶性能,具有重要的经济应用价值。本文综述了与奶牛产奶性状有关的候选基因:DGAT1、SCD1、OPN、PPARGC1A、OLR1和ABCG2基因,以期为今后提高奶牛生产性能的研究提供帮助。     

科技

肉牛胴体性状的遗传机制被揭示近日,牛遗传育种创新团队基于全基因组重测序数据,整合选择信号、单性状关联分析以及多性状关联分析,发现了与胴体性状相关的选择信号及候选基因,揭示了肉牛胴体性状的遗传机制,为肉牛胴体性状的选育奠定了理论基础。相关成果发表在《Genomics(基因组学)》。胴体性状作为肉牛重要经济性状之一,直接决定了牛肉的产量和质量,是肉牛育种中的关键目标。解析肉牛胴体性状的遗传机制与机理并挖掘相关的候选标记与基因,不仅可以为肉牛胴体性状的选育提供相关理论基础,还可以推动肉牛育种进展,切实地提高我国牛肉产量。目前,国内外已大量开展了对肉牛胴体性状遗传基础解析工作,捕获到NCAPG、LCORL等重要候选基因,但利用全基因组重测序数据对胴体性状遗传机制解析的分析方法仍需创新。     

德宏奶水牛DGAT1基因多态性及其与产奶性状的相关性分析

探讨德宏奶水牛（摩拉水牛×德宏水牛）DGAT1基因多态性与产奶性状的关系,以期为德宏奶水牛产奶性状标记辅助选择侯选基因的选择提供理论依据。以81头泌乳的德宏奶水牛为研究对象,利用PCR—SSCP方法结合候选基因直接测序法检测DGAT1基因第8外显子及第8内含子区的多态性,并采用最小二乘法分析DGAT1基因多态位点对产奶量、乳脂率及乳蛋白率的影响。结果在德宏奶水牛群体中共检测到CTGG,CCGG和CCGT三种基因型,测序结果显示,在检测的群体中未发现K232A位点的突变,而在DGAT1基因第8内含子的第14位检测到C→T突变,第35位检测到G→T突变。多重比较表明,CCGT基因型对德宏奶水牛的乳脂率有显著影响（P〈0．05）。     

畜禽经济性状核外遗传效应的研究进展

真核生物存在两个遗传系统,一个在细胞核内,一个在细胞质内,形成了细胞核和细胞质对性状遗传的相互作用。核外遗传也称细胞质遗传,是指细胞核染色体以外的遗传物质的结构、功能及遗传信息传递的体系。长期以来,畜禽经济性状主要集中在核基因变异及其遗传效应,很多与生长发育、繁殖、产奶、屠宰、肉质、产蛋、产毛、产绒等功能相关的候选基因被鉴定,而关于核外遗传效应的研究则较少。文章简要介绍了主要畜禽经济性状核外遗传效应,综述了奶牛、肉牛、羊、猪、鸡等动物线粒体基因多态性与重要经济性状的研究进展,总结了目前动物核外遗传效应研究存在的问题,并提出了应开展的研究方向。     

河北地区中国荷斯坦牛DGAT1基因多态性与泌乳性状遗传效应分析

本研究以河北地区870头中国荷斯坦牛为试验材料,采用PCR-RFLP法对DGAT1基因K232A位点进行了遗传多态性分析,并利用混合动物模型分析了DGAT1基因K232A位点突变对乳脂率、乳脂量、乳蛋白率、乳蛋白量、305d产奶量5个泌乳性状的影响。结果共检测到KK、KA和AA 3种基因型,基因频率分别为0.0402、0.9276和0.0321;等位基因K和A的频率分别为0.5040和0.4960;该碱基突变对乳脂率的影响达到极显著水平(P<0.01),对30d产奶量的影响达到显著水平(P<0.05),对乳脂量、乳蛋白率和乳蛋白量的影响不显著(P>0.05);多重比较结果表明,KK基因型的乳脂率极显著高于KA和AA基因型(P<0.01);KA和AA基因型的305d产奶量显著高于KK基因型(P<0.05);AA和KA基因型的泌乳性状无显著差异(P>0.05)。结果显示,DGAT1基因突变对河北荷斯坦奶牛泌乳性状有较大的遗传效应,可用于泌乳性状的分子标记辅助选择。     

奶牛乳质性状相关基因及其单核苷酸多态性的研究进展

乳质性状为奶牛重要的经济性状,主要包括乳蛋白量、乳蛋白率、乳脂量和乳脂率等。乳质性状改良是当今遗传育种界的研究热点之一,利用候选基因策略寻找影响乳质性状的功能基因或功能性单核苷酸多态,并将其应用于奶牛乳质性状的分子育种实践已经成为后基因组时代的发展趋势。目前针对乳质性状的功能基因或功能性单核苷酸多态已经开展了大量的研究工作,作者就参与泌乳调控的乳质相关基因、脂肪合成与次生物质代谢通路,以及其他一些与乳质性状相关基因及功能性单核苷酸的研究现状进行简要综述。     

采用四引物ARMS-PCR方法检测奶牛乳腺组织DGAT1基因单核苷酸多态性

试验旨在快速有效地检测泌乳奶牛的二酰甘油转酰基酶1（diacylgycerol acyltransferase 1,DGAT1）乳脂量性状的优势等位基因K232A单核苷酸多态性,确定DGAT1基因型进而确定泌乳性状,为中国荷斯坦奶牛分子标记辅助选择提供技术支持。选取6头泌乳初期（3头为高乳产量牛,3头为低乳产量牛）中国北方荷斯坦奶牛为研究对象,提取乳腺组织基因组DNA,分别设计1对外引物和1对内引物,建立一种四引物ARMS-PCR体系快速检测奶牛乳腺组织DGAT1基因单核苷酸多态性。结果发现,外引物扩增片段长度为512 bp,为PCR反应的阳性对照,基因型为232K扩增片段长度为369 bp,基因型为232A扩增片段长度为181 bp。PCR结果显示,6头牛的乳腺组织样本均由外部引物扩增出长度为512 bp的片段,泌乳期高乳产量奶牛和泌乳期低乳产量奶牛乳腺组织的特异性扩增片段长度均为181 bp。表明本研究选取的6头奶牛样本DGAT1基因K232A多态性均为232A型。提示该PCR鉴定方法能够快速有效地鉴定奶牛DGAT1基因型,可用于中国荷斯坦奶牛分子标记辅助选择。     

Detection of Dairy Cow Mammary Tissue DGAT1 Gene Single Nucleotide Polymorphisms Using Tetra-primers ARMS-PCR Method

This study was aimed to detect the single nucleotide polymorphisms of diacylgycerol acyltransferase 1 (DGAT1) K232A, and determine the DGAT1 genotype and milk traits of dairy cow, which would provide a new technique for marker-assisted selection in China Holstein dairy cows. In the present study, six Northern China Holstein dairy cows (three were lactating cows with high quatity milk and three were lactating cows with low quatity milk) were used to detect mammary tissue DGAT1 gene K232A polymorphisms. Genome DNA was extracted from each cow, a pair of external primers and a pair of internal primers were designed to amplify DGAT1 gene. The results showed that PCR-amplified fragments were 512 bp (external band), 369 bp (232K allele) and 181 bp (232A allele), respectively. The exterenal band functions as the internal PCR-positive control. The tetra-primer ARMS-PCR amplifications yielded a 512 bp fragment and a 181 bp fragment, indicating that the six dairy cows were all homozygous 232A. The results indicated that the tetra-primers ARMS-PCR was a quick and convenient method to identify dairy cow DGAT1 gene K232A polymorphisms, which was suitable for marker-assisted selection in China Holstein dairy cows.     

Effect of DGAT1 and TG gene polymorphisms on intramuscular fat and on milk production traits in different cattle breeds in Hungary

The objective of this study was to estimate the effect of the thyroglobulin (TG) locus on beef quality traits in some beef cattle breeds and to investigate the effect of the DGAT1 locus on milk production traits in the Hungarian Holstein Friesian population. TG and DGAT1 genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. At the TG locus TT bulls showed the highest fat percentage values in the longissimus dorsi muscle (m. longissimus dorsi); the difference between CC and TT genotypes was significant. DGAT1 GC/GC cows had the highest milk, fat and protein yield values. Due to the relatively small number of GC/GC cows the difference proved to be significant only between AA/AA and AA/GC genotypes.     

Mitochondrial respiratory metabolism and performance of cattle

Two groups of cows were sampled to study variation of mitochondrial metabolism. The first group included 10 lactating and five gestating Holstein cows, representing available phenotypic extremes for milk yield. The second group included 13 Angus, 13 Brangus and 13 Hereford cows, representing available extremes within breeds for growth breeding value ratios. Consistently poor mitochondrial samples were obtained from gestating cows; these were excluded from analyses. Linear correlations of Holstein mitochondrial respiratory activities with sire-predicted difference for milk and milk fat ranged from -.35 to .15. Correlations of dam and cow indices and cow yields for milk with acceptor-stimulated mitochondrial respiration, oxidative phosphorylation efficiency (adenosine diphosphate:oxygen ratio [ADP:O ratio]) and adenosine triphosphate (ATP) synthesis rate were .25 to .48, consistently higher than for milk fat, -.06 to .17. No differences among beef breeds in mitochondrial respiration rates, coupling of respiration to phosphorylation, ADP:O ratio or ATP synthesis rate were detected by repeated measures analyses of variance. No patterns were evident among correlations of the mitochondrial characteristics and growth and milk traits in beef cattle. These initial findings indicate that variability in the mitochondrial respiratory activities measured has less relation to weaning and yearling growth traits of beef cattle than to milk yield of Holstein cattle.     

中国荷斯坦奶牛RPL23A、ACACB基因3个SNPs检测及其与产奶性状的关联分析

旨在开展奶牛群体高乳成分功能基因的验证与筛选,利用微流控芯片自主选育技术分析北京地区母牛群体高乳蛋白、高乳脂基因频率,同时分析基因多态性及其与产奶性状的相关性。本研究对北京地区8个大型奶牛场1 596头中国荷斯坦奶牛RPL23A、ACACB基因的多态性进行了检测,所有个体均为3胎以内的泌乳牛,收集每头牛所有测定日的产奶量、乳脂率、乳蛋白率等数据,同时对3个多态位点不同基因型与产奶性状进行了关联分析。RPL23A基因的SNP位点g.20146771G>A,在第一泌乳期,与产奶量、乳脂量、乳脂率和乳蛋白量均达到极显著关联（P<0.01）,第二泌乳期,g.20146771G>A与5个产奶性状均呈极显著关联（P<0.01）。ACACB基因的SNP位点g.63962768C>T,在第一泌乳期,与产奶量、乳脂量、乳脂率和乳蛋白量均呈极显著关联（P<0.01）,在第二泌乳期,与产奶量、乳脂量、乳脂率和乳蛋白率呈极显著关联（P<0.01）。结果表明,RPL23A、ACACB基因可以作为影响中国荷斯坦奶牛产奶性状的候选基因用于标记辅助选择,以上基因位点可能通过直接或间接的途径影响奶牛的乳脂或乳蛋白性状,对产奶性状起到重要调控作用。本研究为荷斯坦奶牛后续的标记辅助选择奠定了良好基础。     

Effects of DGAT1 variants on milk production traits in German cattle breeds

Various QTL mapping experiments led to the detection of a QTL in the centromeric region of cattle chromosome 14 that had a major effect on the fat content of milk. Recently, the gene encoding diacylglycerol O-acyltransferase (DGAT1) was proposed to be a positional and functional candidate for this trait. This study investigated the effects of a nonconservative lysine to alanine (K232A) substitution in DGAT1, which very likely represents the causal mutation, on milk production traits. Existing granddaughter designs for Fleckvieh and German Holstein, the two major dairy/dual-purpose breeds in Germany, were used to estimate allele frequencies and gene substitution effects for milk, fat, and protein yield, as well as fat and protein content. A restriction fragment length polymorphism assay was applied to diagnose the K232A substitution in DGAT1. Estimates of the allele frequencies for the lysine-encoding variant were based on maternally inherited alleles in sons and amounted to 0.072 for Fleckvieh and 0.548 for German Holstein. Effects of DGAT1 variants on content traits were pronounced; estimates of the gene substitution effect for the lysine-encoding variant were 0.35 and 0.28% for fat content and 0.10 and 0.06% for protein content in Fleckvieh and German Holstein, respectively. Conversely, negative effects of the lysine variant of -242 to -180 kg for Fleckvieh and -260 to -320 kg for German Holstein were revealed for milk yield from first to third lactation, resulting in enhanced fat yield of 7.5 to 14.8 kg in Fleckvieh and 7.6 to 10.7 kg in German Holstein. For protein yield, however, mainly negative effects of -3.6 to 0.2 kg in Fleckvieh and -4.8 to -5.2 kg in German Holstein were observed. Pearson correlations between residuals of milk yield and content traits were decreased when omitting DGAT1 effects in the analysis, thereby indicating that DGAT1 contributes to negative correlations between these traits. Molecular tests allow for the direct selection among variants; however, the benefits of the alternative alleles depend on economic weights given to the different milk production traits in the breeding goal.     

Evaluation in beef cattle of six deoxyribonucleic acid markers developed for dairy traits reveals an osteopontin polymorphism associated with postweaning growth

The objectives of this study were to 1) estimate the allelic frequencies in US beef cattle of 6 DNA markers reported to be associated with variation in dairy production traits; and 2) evaluate the association of these markers with beef production traits. Several genetic markers have been associated with milk yield or composition, including polymorphisms in secreted phosphoprotein 1 (SPP1; also called osteopontin), growth hormone receptor (GHR), casein S1 (CSN1S1), diacylglycerol O-acyltransferase 1 (DGAT1), peroxisome proliferator-activated receptor gamma co-activator-1alpha (PPARGC1A), and ATP-binding cassette subfamily G (white) member 2 (ABCG2). Allelic frequencies for these 6 markers, and their association with 21 phenotypes, were evaluated in 2 crossbred beef cattle populations that sample influential industry sires. Five of 6 markers were segregating in beef cattle populations; the exception was ABCG2. The SPP1 marker was associated with yearling weight (P = 0.025), live weight at slaughter (P = 0.016), postweaning ADG (P = 0.007), and HCW (P = 0.007) in a large, multisire population representing the 7 most populous beef breeds in the United States. Postweaning growth trait associations were confirmed in an independent population of similar construction, including sires from tropically adapted breeds. The SPP1 marker was associated with yearling weight (P = 0.034), live weight at slaughter (P = 0.011), and postweaning ADG (P = 0.015) and showed a trend toward association with HCW (P = 0.083) in this population. Whereas DGAT1, GHR, and CSN1S1 polymorphisms showed association with some traits in individual populations, the lack of consistent predictive merit between populations indicates they may not be suited for beef cattle selection. No significant associations were observed for the PPARGC1A marker and any of 21 recorded traits, indicating this marker had no apparent value in selection for the beef cattle traits tested in these populations. The SPP1 marker had consistent associations and effect sizes (10.5 to 11.5 kg of live weight at slaughter) in both populations, providing strong evidence for utility of the SPP1 marker for postweaning growth in beef cattle.