全基因组选择信号检测方法在家畜研究中的应用进展

长期的自然和人为选择使家畜品种经济性状得到了显著改善,其相关的基因组区域也发生了特定遗传变异。随着时间的推移部分基因多态性已经下降或消失,而在群体中保留包含单一单倍型的多个基因。这种基因组上特定区域基因多态性频率的变异被称为选择信号。识别选择信号可以提供家畜驯化机制并进一步揭示表型相关的基因变异。目前,高密度SNP芯片及大规模重测序技术已成功应用于家畜选择信号鉴定研究。全基因组选择信号检测方法有等位基因频率检测法、连锁不平衡检测法和群体分化分析法。作者综述了全基因组范围内选择信号检测方法及其在家畜研究中的应用进展,为从事家畜育种及生物进化研究人员提供参考。     

利用基因组信息提高畜禽生产性能研究进展

为了满足人们对畜产品需求的快速增长，必须在加快畜禽产业发展的同时把对环境的影响降到最低，提高畜禽遗传特性有望促进这一问题的解决。进入21世纪以来，以基因组选择为核心的分子育种技术迎来了发展机遇，利用该技术可实现早期准确选择，从而大幅度缩短世代间隔，加快群体遗传进展，并显著降低育种成本。虽然在某些畜种中（如奶牛），基因组选择取得了成功，群体也获得较大遗传进展，但仍无法满足快速增长的需求。因此，亟需寻找能够进一步加快遗传进展的方法。研究表明，在SNP标记数据中加入目标性状的已知功能基因信息，可以提高基因组育种值预测的准确性，进而加快遗传进展。而挖掘更多基因组信息的同时，开发更优化的分析方法可以更有助于目标的实现。文章总结了主要畜禽物种的可用基因组数据，包括牛、绵羊、山羊、猪和鸡以及这些数据是如何有助于鉴定影响重要性状的遗传标记和基因，从而进一步提高基因组选择的准确性。     

基于选择信号分析揭示猪终端父本群体间性状趋同的关键基因

【目的】试验旨在揭示终端父本皮特兰猪与杜洛克猪在人工选择作用下重要经济性状呈现出表型趋同的基因组变化特征。【方法】利用376头皮特兰猪、451头杜洛克猪品系Ⅰ、841头杜洛克猪品系Ⅱ和497头杜洛克猪品系Ⅲ群体的50K SNP芯片数据,以100 kb窗口、50 kb步长计算综合单倍型评分(iHS)和等位基因频率差(△AF),分别取前5%作为猪群体内、群体间的基因组选择信号候选区域;利用bedtools分别对iHS、△AF按照左右200 kb进行合并,每2个群体间合并后的iHS、△AF统计量的重叠区域定义为性状趋同区域,并挖掘该区域与猪重要经济性状相关的平行选择信号。【结果】iHS结果显示,在皮特兰猪和杜洛克猪4个群体内共检测到5 112个选择信号候选区域,总长约487.51 Mb。基于△AF方法,于皮特兰猪和杜洛克猪每2个群体间共检测到9 579个选择信号显著区域,总长约913.50 Mb。基于合并后的iHS和△AF,共检测到52个性状趋同区域,总长约4.67 Mb,注释到88个与猪的繁殖、胴体和肉质等性状相关的平行选择候选基因。【结论】皮特兰猪和杜洛克猪群体间存在性状趋同的基因组选择区域有52个,平行选择信号主要涉及猪的繁殖、胴体及肉质等重要经济性状,这与瘦肉型猪种相同的育种方向相关,这些关键基因的发现可为后续商业猪品种遗传改良提供参考。     

Production of transgenic livestock: promise fulfilled

The introduction of specific genes into the genome of farm animals and its stable incorporation into the germ line has been a major technological advance in agriculture. Transgenic technology provides a method to rapidly introduce "new" genes into cattle, swine, sheep, and goats without crossbreeding. It is a more extreme methodology, but in essence, not really different from crossbreeding or genetic selection in its result. Methods to produce transgenic animals have been available for more than 20 yr, yet recently lines of transgenic livestock have been developed that have the potential to improve animal agriculture and benefit producers and/or consumers. There are a number of methods that can be used to produce transgenic animals. However, the primary method to date has been the microinjection of genes into the pronuclei of zygotes. This method is one of an array of rapidly developing transgenic methodologies. Another method that has enjoyed recent success is that of nuclear transfer or "cloning." The use of this technique to produce transgenic livestock will profoundly affect the use of transgenic technology in livestock production. Cell-based, nuclear transfer or cloning strategies have several distinct advantages for use in the production of transgenic livestock that cannot be attained using pronuclear injection of DNA. Practical applications of transgenesis in livestock production include enhanced prolificacy and reproductive performance, increased feed utilization and growth rate, improved carcass composition, improved milk production and/or composition, and increased disease resistance. One practical application of transgenics in swine production is to improve milk production and/or composition. To address the problem of low milk production, transgenic swine over-expressing the milk protein bovine alpha-lactalbumin were developed and characterized. The outcomes assessed were milk composition, milk yield, and piglet growth. Our results indicate that transgenic overexpression of milk proteins may provide a means to improve swine lactation performance.     

Genome-wide detection of signatures of selection in three Valdostana cattle populations

The Valdostana is a local dual purpose cattle breed developed in Italy. Three populations are recognized within this breed, based on coat colour, production level, morphology and temperament: Valdostana Red Pied (VPR), Valdostana Black Pied (VPN) and Valdostana Chestnut (VCA). Here, we investigated putative genomic regions under selection among these three populations using the Bovine 50K SNP array by combining three different statistical methods based either on allele frequencies (F_ST) or extended haplotype homozygosity (iHS and Rsb). In total, 8, 5 and 8 chromosomes harbouring 13, 13 and 16 genomic regions potentially under selection were identified by at least two approaches in VPR, VPN and VCA, respectively. Most of these candidate regions were population-specific but we found one common genomic region spanning 2.38 Mb on BTA06 which either overlaps or is located close to runs of homozygosity islands detected in the three populations. This region included inter alia two well-known genes: KDR, a well-established coat colour gene, and CLOCK, which plays a central role in positive regulation of inflammatory response and in the regulation of the mammalian circadian rhythm. The other candidate regions identified harboured genes associated mainly with milk and meat traits as well as genes involved in immune response/inflammation or associated with behavioural traits. This last category of genes was mainly identified in VCA, which is selected for fighting ability. Overall, our results provide, for the first time, a glimpse into regions of the genome targeted by selection in Valdostana cattle. Finally, this study illustrates the relevance of using multiple complementary approaches to identify genomic regions putatively under selection in livestock.     

德国肉用美利奴羊全基因组选择信号检测

旨在研究德国肉用美利奴羊基因组选择信号。利用绵羊高密度SNPs芯片,运用iHS方法检测德国肉用美利奴羊基因组选择信号,并通过生物信息学分析揭示其潜在受选择基因。选择信号检测结果表明,在全基因组范围内共检测到220个具有选择信号的基因组区段,这些区段内与895个候选基因紧密相关。基因富集结果表明,这些基因主要与蛋白翻译、骨骼发育、生物合成调控、肌肉器官发育、视黄酸受体活性、转录因子结合、核糖体组分等相关。研究结果为充分利用德国肉用美利奴羊的种用价值以及高效开展该品种羊的群体改良工作提供了参考依据。     

Detection of selection signatures for agonistic behaviour in cattle

The identification of genomic regions including signatures of selection produced by domestication and its subsequent artificial selection processes allows the understanding of the evolution of bovine breeds. Although several studies describe the genomic variability among meat or milk production cattle breeds, there are limited studies orientated towards bovine behavioural features. This study is focused on mapping genomic signatures of selection which may provide insights of differentiation between neutral and selected polymorphisms. Their effects are studied in the Lidia cattle traditionally selected for agonistic behaviour compared with Spanish breeds showing tamed behaviour. Two different approaches, BayeScan and SelEstim, were applied using genotypic 50K SNP BeadChip data. Both procedures detected two genomic regions bearing genes previously related to behavioural traits. The frequencies of the selected allele in these two regions in Lidia breed were opposite to those found in the tamed breeds. In these genomic regions, several putative genes associated with enriched metabolic pathways related to the behavioural development were identified, as neurochondrin gene (NCDN) or glutamate ionotropic receptor kainate type subunit 3 (GRIK3) both located at BTA3 or leucine‐rich repeat and Ig domain containing 2 (LINGO2) and phospholipase A2‐activating protein (PLAA) at BTA8.     

全基因组测序在畜禽中应用的研究进展

在基因组研究方面,目前全基因组测序已由第一代测序技术发展到第三代测序技术,全基因组测序与传统方法相比具有更加全面、精准、高效等优势。随着测序技术的发展和费用的降低,全基因组测序（whole genome sequencing,WGS）技术逐渐成为基因组研究应用最广泛的技术。全基因组测序已经在畜禽起源进化、重要经济性状基因挖掘、分子育种等方面取得了诸多成果。通过全基因组重测序,能够发现拷贝数变异（copy number variation,CNV）及单核苷酸多态性（single nucleotide polymorphism,SNP）变异,丰富现有的CNV和SNP数据库,为抗病、生长、食欲、代谢调节、表型、环境适应机制及重要经济性状基因的分析提供重要数据。作者针对全基因组测序技术在主要畜禽上的研究进展,综述了全基因组测序在畜禽的品种遗传多样性、群体演变机制、功能基因挖掘等研究中的应用,并探讨了全基因组测序存在的问题,旨在为畜禽种质资源保护和分子育种实践提供参考。     

畜禽基因组选择方法研究进展

畜禽的选种选育在生产中至关重要,育种值估计是选种选育的核心。基因组选择（genomic selection,GS）是利用全基因组范围内的高密度标记估计个体基因组育种值的一种新型分子育种方法,目前已在牛、猪、鸡等畜禽育种中得到应用并取得了良好的效果。该方法可实现畜禽育种早期选择,降低测定费用,缩短世代间隔,提高育种值估计准确性,加快遗传进展。基因组选择主要是通过参考群体中每个个体的表型性状信息和单核苷酸多态性（single nucleotide polymorphism,SNP）基因型估计出每个SNP的效应值,然后测定候选群体中每个个体的SNP基因型,计算候选个体的基因组育种值,根据基因组育种值的高低对候选群体进行合理的选择。随着基因分型技术快速发展和检测成本不断降低,以及基因组选择方法不断优化,基因组选择已成为畜禽选种选育的重要手段。作者对一些常用的基因组选择方法进行了综述,比较了不同方法之间的差异,分析了基因组选择存在的问题与挑战,并展望了其在畜禽育种中的应用前景。     

整合生物学先验信息的全基因组选择方法及其在家畜育种中的应用进展

相较于传统的育种方法,全基因组选择（genomic selection,GS）通过对拟留种的个体进行早期选择和增加选择的准确性进而加快育种的遗传进展。通过改进GS方法无法再缩短育种的世代间隔,因而如何提高GS的准确性以获得额外的遗传进展一直是GS研究的核心问题。当前,各种组学技术不断成熟,从公开的资料或前期的研究积累获取生物学先验信息已比较容易。因而,如何在GS模型中整合已知的先验信息进而提高GS的准确性以获得额外的遗传进展成为当前育种研究的热点问题。本文对生物学先验信息的类型以及整合先验信息的GS方法进行综述,探讨了这些方法在家畜育种中的应用和前景,以期为家畜育种中开展整合生物学先验信息的GS研究提供借鉴与参考。     

Research Progress on Genomic Selection Methods in Livestock and Poultry

Selection and breeding are very important in production of livestock and poultry,and breeding value estimation is the core of selection and breeding.Genomic selection (GS) is a novel molecular breeding method to estimate genomic breeding value using high-density markers across the whole genome.At present,GS has been successfully applied in cattle,pig,chicken and so on,and made significant progress.This method can achieve early selection,decrease the testing costs,shorten generation interval,improve the accuracy of breeding value estimation and accelerate genomic progress.GS estimates the effect of SNP by phenotype information and SNP genotype of each individual in the reference population,and measures the SNP genotype to calculate the genomic estimated breeding value in the candidate population,then selects the best individuals according to the genomic estimated breeding value.With the rapid development of genotyping technology and the decrease of detection cost,and the continuous optimization and high efficiency of genomic selection methods,genomic selection has become an important research method in the selection and breeding of livestock and poultry.The authors reviewed some of the widely used genomic selection methods,compared the differences between different methods,analyzed the problems and challenges of genomic selection,and looked forward to its application prospects in breeding.     

芯片技术在畜禽育种中的应用研究进展

中国畜禽品种资源丰富,且有许多优良性状基因,但这些优良性状基因并没有被充分利用,因此,在基因水平上开展遗传资源的开发和利用是畜禽经济性状改良的重要方向。目前,虽然传统系谱选择方法在育种工作中发挥了重要作用,但存在准确率低、育种周期长等缺点。随着分子生物学技术的快速发展,近年来先进的基因组测序和基因分型技术大大促进了畜禽育种方法的革新。从低通量、耗时的限制性片段多态标记（RFLP）到如今高通量、高密度的单核苷酸多态性（SNP）标记,基因检测效率有了大幅度提高。基因芯片技术在分子标记辅助选择和全基因组选择育种研究中逐渐得到广泛应用,成为畜禽育种的新技术手段和新热点。主要介绍了高、低密度SNP芯片技术在畜禽育种中的研究及应用,并简述了其技术优势、存在问题及挑战、应用展望,旨在表明基因芯片技术必将会成为畜禽分子育种工作中一项重要的基础技术,在畜禽种业快速发展过程中起到重要的推动作用,以期为基因芯片技术在畜禽育种中得到进一步应用提供理论参考,推进中国畜禽育种遗传进展,提升中国畜禽种业的科技竞争力。     

基因组选择在家畜改良上的研究进展

基因组选择（GS）是全基因组范围内的分子标记辅助选择,目前被证明是利用DNA标记信息改善复杂性状最有效的方法。本文简要概述了基因辅助选择以及标记辅助选择;重点介绍了GS,包括GS的实施策略与育种值估计方法,GS的准确性获得以及对GS方法的比较,总结了当前家畜上利用GS加速遗传改良的应用进展;并对家畜在GS上的应用前景进行展望。     

Genetic aspects of piglet survival and related traits: a review

In livestock, mortality in general, and mortality of the young, is societal worries and is economically relevant for farm efficiency. Genetic change is cumulative; if it exists for survival of the young and genetic merit can be estimated with sufficient accuracy, it can help alleviate the pressure of mortality. Lack of survival is a moving target; livestock production is in continuous change and labor shortage is a given. There is now ample evidence of clear genetic variance and of models able to provide genomic predictions with enough accuracy for selection response. Underlying traits such as birth weight, uniformity in birth weight, gestation length, number of teats, and farrowing duration all show genetic variation and support selection for survival or, alternatively, be selected for on their own merit.     

CRISPR/Cas9技术在畜禽育种中的研究进展

利用传统方法在畜禽特定基因座上进行基因组修饰时,只能通过在体细胞中进行同源重组再经细胞核移植实现。传统同源重组方法的困难性和低效性阻碍了基因修饰在畜禽遗传育种中的广泛应用。近年来,位点特异性核酸内切酶的发现为靶向基因修饰提供了一条更直接的途径,主要由于这些酶能直接在DNA序列上进行一步式的基因编辑。成簇规律间隔短回文重复序列/Cas关联蛋白9(clustered regularly interspersed short palindromic repeat/CRISPR associated protein 9,CRISPR/Cas9)是一种RNA导向的DNA内切酶,精准定位于特定的靶位点,高效完成RNA导向的DNA识别及编辑。CRISPR/Cas9技术作为精准而强大的第3代基因组编辑工具,已经成功应用于猪、牛、山羊、绵羊和鸡上,这些CRISPR/Cas9基因编辑畜禽可作为研究人或畜禽生理和病理的生物模型、生产功能性蛋白质的生物反应器或器官移植的供体。特别是在畜禽生产方面,CRISPR/Cas9基因编辑可用于改善生产遗传特性及畜产品质量,提高畜禽对疾病的抵抗力。作者对当前畜禽中特定位点基因组修饰的CRISPR/Cas9技术的原理及基因组编辑在畜禽育种中应用的最新进展进行了综述,以期为推进CRISPR/Cas9技术在畜禽育种中的研究提供参考。     

Physiological criteria and genetic selectionDes critères physiologiques dans la sélectionPhysiologische kriterien und genetische selektion

The application of genetic techniques to the improvement of farm livestock is dependent upon the rate of change which can be offered. One factor affecting the rate of response to genetic selection is the ability to recognise individuals which carry favourable alleles, and for reproduction and lactation this ability is severely restricted by the limitation of the expression of the traits to mature females. Males however carry the genes for these traits. If they were expressed in males, and if their expression could be measured conveniently, this restriction might be alleviated.With many loci involved it is argued that biochemical measurement of the direct products of gene action is unlikely to be a useful way to recognise the presence of favourable alleles. The possibility that their expression may be integrated into measurable physiological systems has therefore been studied. Hormone levels and testis growth in young males have been shown to be related to female reproduction, and there are signs that hormone and metabolite responses to changes in energy balance may be related to dairy merit.It is concluded that traits of males could be used to improve the efficiency of genetic selection and that current results provide a basis for the development of acceptable criteria. The more general implications of physiological study of genetic variation are also discussed.     

家畜全基因组关联分析研究进展

全基因组关联分析（genome-wide association study,GWAS）是一种研究经济性状候选基因的分析方法。近年来,随着家畜全基因组测序的完成,大量的单核苷酸多态性（single nucleotide polymorphisms,SNPs）被标识,GWAS也越来越多地应用于家畜重要性状的研究领域中,在动物遗传育种中,通过对家畜基因组进行GWAS分析研究,找到控制家畜主要经济性状的重要SNPs,从而挖掘重要经济性状的候选基因。作者详细综述了GWAS的分析方法及其在重要家畜育种中的研究进展。GWAS分析方法包括基因组控制法（genommic control）、分层分析法（stratification analysis）、主成分分析法（principal components analysis,PAC）和混合线性模型分析法（mixed-linear-model association,MLMA）,通路分析方法包括非核算法（基因功能富集分析（gene set enrichment analysis,GSEA）和分层贝叶斯优取（hierarchical Bayes prioritization,HBP））和核算法。依据不同的目标性状选择合理的分析方法,提高GWAS分析结果的准确性,为进一步利用GWAS分析各种性状的遗传基础提供合理的借鉴。     

P-glycoprotein selection in strains of Haemonchus contortus resistant to benzimidazoles

Anthelmintic resistance in parasitic nematodes of livestock is a chronic problem in many parts of the world. Benzimidazoles are effective, broad-spectrum anthelmintics that bind to and selectively depolymerise microtubules. Resistance to the benzimidazoles, however, developed quickly and is caused by genetic changes in genes encoding beta-tubulins, subunits of microtubules. In Haemonchus contortus, resistance to avermectins has been correlated with genetic changes at a gene encoding a P-glycoprotein, a cell membrane transport protein that has a very high affinity for ivermectin. The substrate specificity of P-glycoprotein is very broad, and resistance to benzimidazoles can be modulated by lectins specific for P-glycoprotein. We investigated the possibility that genetic changes in P-glycoprotein might be correlated with benzimidazole resistance in nematodes. An analysis of restriction fragment length polymorphisms of a P-glycoprotein gene from a sensitive and a cambendazole-selected strain of H. contortus, derived from the sensitive strain, showed a significant difference in allele frequencies between strains. The frequency of one allele in particular increased substantially. The same allele was also found at a high frequency in an independently derived thiabendazole-selected field isolate. We present genetic evidence of selection at a P-glycoprotein locus during selection for benzimidzole resistance in H. contortus.     

拷贝数变异在畜禽中的研究进展

拷贝数变异（copy number variation,CNV）具有多种形式的变异结构,在品种多样性、生物进化和疾病相关性等研究中起着重要作用,并具有片段长度大、覆盖范围广等特点。随着分子生物学的发展及DNA测序技术的日渐成熟,人们对遗传变异的研究不断向DNA分子水平深入,多态性标记在畜禽育种中已逐渐成为动物育种研究的趋势和主流。由于CNV对基因的调控和表达所造成的影响更为显著,因此,CNV在重要畜禽中的研究越来越多。目前,已检测出大量有关畜禽重要经济性状的基因序列变异,并有许多研究均表明CNV与动物的重要经济特征及疾病的发生有关。笔者主要通过参考国内外相关的研究报道,简述了CNV的相关研究背景、概念、突变机制,归纳总结了CNV对牛、羊、猪、鸡的经济性状、繁殖性状和疾病调控的影响,以期通过对这些重要畜禽的基因组学研究揭示其适应性遗传机理和表型性状差异的遗传基础,开发相应的分子遗传标记,为畜禽的标记辅助选育提供理论基础。     

全基因组SNP分型技术在畜禽遗传育种研究中的应用

随着畜禽资源分子鉴定、物种进化、全基因组育种等热点领域的逐渐兴起,准确的全基因组SNP分型成为了畜禽基因组研究的关键。基因芯片、重测序、简化基因组测序及靶向捕获测序等全基因组SNP分型技术已广泛应用于畜禽基因组研究中。本文概述了全基因组SNP分型技术的原理及其在全基因组关联分析、选择信号分析和畜禽遗传资源背景分析等方面的应用,以期为畜禽基因组研究和育种应用提供借鉴和参考。