奶牛重要经济性状全基因组关联分析研究进展

产奶性状是奶牛最重要的生产性状, 随着平衡育种理念的提出和发展, 繁殖性状、体型性状、健康性状和长寿性等功能性状也逐渐被重视并纳入育种规划中。鉴定产奶性状和功能性状主效基因或遗传标记并将之应用于奶牛标记辅助选择可望加快遗传进展。随着高密度SNP标记的高通量检测技术的发展, 全基因组关联分析已成为鉴定畜禽重要经济性状基因的重要途径。文章对奶牛产奶性状和功能性状全基因组关联分析研究进展进行综述。     

Vps28基因敲除小鼠模型的构建及其对泌乳和免疫性状影响的研究

尽管基于QTL定位和关联分析研究挖掘了许多奶牛产奶性状候选功能基因,然而仅有少数几个基因经过体内外的功能验证.本课题组前期全基因组关联分析(GWAS)研究结果表明Vps28基因与奶牛产奶性状表型显著相关,并在奶牛乳腺组织中特异性高表达.为了深入了解该基因对泌乳性状的作用和功能,本研究利用CRISPR/Cas9技术构建了...     

羊重要性状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)是一种复杂性状功能基因鉴定的分析策略,已成为挖掘畜禽重要经济性状候选基因的重要手段。随着绵羊和山羊基因组完成和公布,以及不同密度的SNP (single nucleotide polymorphism)芯片的推出并进行商业化推广,不仅大大丰富了羊标记辅助选择可利用的分子标记,而且还为开展重要性状的分子机理的探索提供了重要技术支撑。本文主要针对羊角、羊毛、羊奶、生长发育、肉质、繁殖和疾病等重要性状的GWAS研究所用的群体、主要研究方法和研究结果进行了综述,并对GWAS方法研究现状进行了归纳,以期为进一步利用GWAS进行羊的各种性状的遗传基础研究提供参考。     

全基因组关联分析的进展与反思

全基因组关联分析(genomewide association study,GWAS)是应用人类基因组中数以百万计的单核苷酸多态性(single nucleotide polymorphism,SNP)为标记进行病例-对照关联分析,以期发现影响复杂性疾病发生的遗传特征的一种新策略。近年来,随着人类基因组计划和基因组单倍体图谱计划的实施,人们已通过GWAS方法发现并鉴定了大量与人类性状或复杂性疾病关联的遗传变异,为进一步了解控制人类复杂性疾病发生的遗传特征提供了重要的线索。然而,由于造成复杂性疾病/性状的因素较多,而且GWAS研究系统较为复杂,因此目前GWAS本身亦存在诸多的问题。本文将从研究方式、研究对象、遗传标记,以及统计分析等方面,探讨GWAS的研究现状以及存在的潜在问题,并展望GWAS今后的发展方向。     

关联分析及其在植物中的应用

关联分析是新近开始在植物数量性状研究和植物育种中应用的一种分析方法.它以连锁不平衡为基础鉴定某一群体内性状与遗传标记或候选基因间的关系,是对分子育种中QTL分析的补充和提高.本文在介绍连锁不平衡的定义和度量方法的基础上,讨论连锁不平衡程度和群体结构对关联分析的影响,综述了关联分析在植物方面的研究进展,并最后讨论了关联分析在植物数量性状和分子育种研究中可能的应用.     

陆地棉主要产量相关性状的SSR标记关联分析

高产优质育种是我国棉花育种的主要目标。寻找与目标性状关联的分子标记,可克服常规育种的盲目性,提高分子标记辅助选择育种的准确性。本研究对118份陆地棉种质资源的衣分、单铃重、单株铃数及子指等4个产量相关性状进行2年2点的表型鉴定,并利用覆盖全基因组的、有多态性的214对SSR标记进行标记与性状的关联分析。结果表明:118份材料的4个产量相关性状表型变异丰富,平均变异系数的变幅在6.1%~19.1%之间,且在各环境中表现较为稳定;基因型分析表明,214对标记共检测到460个等位变异,基因多样性指数平均为0.5151,PIC值平均为0.4587,表明该批标记具有较多的等位变异数和较高的基因多样性;群体结构分析表明该批材料可分为4个亚群,且各类群中材料与地理来源无对应关系;关联分析结果显示,在显著条件下(-log10P1.3,P0.05),共有39个标记位点能够在2个及2个以上的环境中同时检测到,其中有4个标记位点同时与2个以上性状相关联,进一步比较发现,有7个位点与前人研究结果一致,其余32个位点为新发现的位点。研究结果可为陆地棉产量性状遗传改良的分子标记辅助选择提供理论依据。     

玉米叶夹角形成的遗传基础与分子机制解析*

玉米产量取决于植株捕获光能和固定CO₂合成有机化合物的效率。叶夹角是株型重要性状之一,较小叶夹角有利于提高玉米植株光合作用效率和种植密度,因而有利于提高玉米产量。研究表明玉米叶夹角为多基因控制的复杂数量性状,其遗传力较高,主要受基因的加性效应调控。目前,利用数量性状位点(quantitative trait loci, QTL)定位和全基因组关联分析(genome-wide association study, GWAS)等方法已鉴定数百个玉米叶夹角相关QTL;结合突变体分析等方法,已克隆数十个调控叶夹角关键基因,这为了解玉米叶夹角遗传机制提供了重要参考。由于前人研究所采用群体、分析方法及参考基因组版本不同,各研究之间所鉴定QTL差异较大,因此无法客观揭示叶夹角性状的遗传规律。为此,通过总结前人所定位叶夹角相关QTL和单核苷酸多态性(single nucleotide polymorphism,SNP)位点并构建一致性图谱,鉴定出叶夹角性状定位热点区间,并对调控叶夹角的已知基因进行功能分类。这不仅为了解玉米叶夹角的遗传结构、推动叶夹角相关重要基因克隆提供数据支撑,也对进一步开发叶夹角相关分子标记,指导玉米分子育种和提高玉米产量提供有益指导。     

玉米雌穗性状遗传分析与形成机制*

雌穗是玉米重要的生殖器官,雌穗发育决定成熟果穗大小及单穗粒重,进而直接影响玉米产量。雌穗性状主要包括穗长、穗粗、穗行数、行粒数、穗重、单穗粒重等,均为多基因控制的数量遗传性状,且其遗传结构各不相同。解析雌穗性状的遗传基础,优化雌穗结构,是玉米增产的重要途径。前人通过数量性状位点(quantitative trait locus mapping,QTL)定位和全基因组关联分析(genome-wide association study, GWAS)等方法,已经鉴定出较多雌穗性状相关的遗传位点,但是目前已鉴定功能的基因较少,所建立的遗传位点一致性图谱并不完整,因此难以全面揭示雌穗性状遗传结构。通过综合前人雌穗性状遗传定位进展,现将已鉴定QTL位点和显著关联SNP整合至玉米B73参考基因组V4版本,并鉴定出雌穗性状定位热点区间,对深入解析雌穗性状遗传结构、指导雌穗性状基因克隆和理解雌穗发育分子机制均具有重要意义。     

水产动物重要经济性状相关功能基因的研究进展

随着基因组学技术的快速发展,已经在水产动物中鉴定出了大量的功能基因,并通过建立基因多态性与重要经济性状之间的关联,挖掘出了与经济性状连锁的优势等位基因和基因型。对基因资源的研究不仅有助于理解经济性状的分子调控机制,而且将为分子辅助育种提供理论指导和技术支持。综述了近年来水产动物重要经济性状相关基因的研究进展,以期为解析功能基因的分子调控机制和分子辅助育种提供基础理据和参考资料。     

桃基因组及全基因组关/联分析研究进展

桃(Prunus persica [L.] Batsch)是蔷薇科重要的核果类果树, 适应性强, 栽培范围广, 果实口感好, 深受消费者喜欢。提高桃果实品质及增加抗病、抗虫性一直是桃遗传育种者关注的焦点。文章对近年来桃遗传分子标记连锁图谱和物理图谱构建、分子标记开发应用、全基因组和转录组测序工作中所取得的最新成果进行综述, 同时阐述了高密度SNP芯片标记技术在桃以及其它作物上所开展的全基因组关联分析应用实例, 为桃进一步开展全基因组关联分析, 挖掘目标性状QTLs以及高效育种选择标记提供理论基础     

Genome-wide association mapping in Norwegian Red cattle identifies quantitative trait loci for fertility and milk production on BTA12

Reproductive performance is a critical trait in dairy cattle. Poor reproductive performance leads to prolonged calving intervals, higher culling rates and extra expenses related to multiple inseminations, veterinary treatments and replacements. Genetic gain for improved reproduction through traditional selection is often slow because of low heritability and negative correlations with production traits. Detection of DNA markers associated with improved reproductive performance through genome-wide association studies could lead to genetic gain that is more balanced between fertility and production. Norwegian Red cattle are well suited for such studies, as very large numbers of detailed reproduction records are available. We conducted a genome-wide association study for non-return rate, fertility treatments and retained placenta using almost 1 million records on these traits and 17 343 genome-wide single-nucleotide polymorphisms. Genotyping costs were minimized by genotyping the sires of the cows recorded and by using daughter averages as phenotypes. The genotyped sires were assigned to either a discovery or a validation population. Associations were only considered to be validated if they were significant in both groups. Strong associations were found and validated on chromosomes 1, 5, 8, 9, 11 and 12. Several of these were highly supported by findings in other studies. The most important result was an association for non-return rate in heifers in a region of BTA12 where several associations for milk production traits have previously been found. Subsequent fine-mapping verified the presence of a quantitative trait loci (QTL) having opposing effects on non-return rate and milk production at 18 Mb. The other reproduction QTL did not have pleiotropic effects on milk production, and these are therefore of considerable interest for use in marker-assisted selection.     

Genetic variants in mammary development,prolactin signalling and involution pathways explain considerable variation in bovine milk production and milk composition

Background

The maintenance of lactation in mammals is the result of a balance between competing signals from mammary development, prolactin signalling and involution pathways. Dairy cattle are an interesting case study to investigate the effect of polymorphisms that affect the function of genes in these pathways. In dairy cattle, lactation yields and milk composition (for example protein percentage and fat percentage) are routinely recorded, and these vary greatly between individuals. In this study, we test 8058 single nucleotide polymorphisms in or close to genes in these pathways for association with milk production traits and determine the proportion of variance explained by each pathway, using data on 16 812 dairy cattle, including Holstein-Friesian and Jersey bulls and cows.

Results

Single nucleotide polymorphisms close to genes in the mammary development, prolactin signalling and involution pathways were significantly associated with milk production traits. The involution pathway explained the largest proportion of genetic variation for production traits. The mammary development pathway also explained additional genetic variation for milk volume, fat percentage and protein percentage.

Conclusions

Genetic variants in the involution pathway explained considerably more genetic variation in milk production traits than expected by chance. Many of the associations for single nucleotide polymorphisms in genes in this pathway have not been detected in conventional genome-wide association studies. The pathway approach used here allowed us to identify some novel candidates for further studies that will be aimed at refining the location of associated genomic regions and identifying polymorphisms contributing to variation in lactation volume and milk composition.     

The role of bovine causal genes underlying dairy traits in Spanish Churra sheep

In dairy cattle, quantitative trait nucleotides (QTNs) underlying quantitative trait loci (QTL) for milk production traits have been identified in bovine DGAT1, GHR and ABCG2 genes. The SPP1 gene has also been proposed to be a regulator of lactation. In sheep, QTL underlying milk production traits have been reported only recently, and no proven QTN has been identified. Taking into account the close phylogenetic relationship between sheep and cattle, this study examined the possible effects of the aforementioned genes on sheep milk production traits. We first studied the genetic variability of the DGAT1, GHR, ABCG2 and SPP1 genes in 15 rams of the Spanish Churra dairy sheep breed. Second, we performed an association analysis between SNPs identified in these genes and three milk production traits recorded in a commercial population of Churra sheep. This analysis revealed only three significant associations at the nominal level (P-value <0.05) involving allelic variants of the ABCG2 gene, whereas no significant association was found for the DGAT1, GHR and SPP1 genes. When the Bonferroni correction was applied to take into account the multiple tests performed, none of the associations identified at the nominal level remained significant. Nevertheless, taking into account the high level of false-negative findings that can arise when applying the stringent Bonferroni correction, we think that our results provide a valuable primary assessment of strong candidate genes for milk traits in sheep.     

Whole-genome resequencing reveals selection signatures associated with milk production traits in African Kenana dairy zebu cattle

Milk production and composition are the most economically important traits affecting profitability in dairy cattle. In this study, we aimed at detecting signatures of positive selection in Kenana, known as one of the high milk production African indigenous zebu cattle, using next-generation sequencing data. To detect genomic signatures of positive selection, we applied three methods based on population comparison, fixation index (F_ST), cross population composite likelihood ratio (XP-CLR) and nucleotide diversity (Pi). Further analysis showed that several candidate genes such as CSN3, IGFBP-2, RORA, ABCG2, B4GALT1 and GHR are positively selected for milk production traits in Kenana cattle. The candidate genes and enriched pathways identified in this study may provide a basis for future genome-wide association studies and investigations into genomic targets of selection in dairy cattle.     

Application of Selection Mapping to Identify Genomic Regions Associated with Dairy Production in Sheep

In Europe, especially in Mediterranean areas, the sheep has been traditionally exploited as a dual purpose species, with income from both meat and milk. Modernization of husbandry methods and the establishment of breeding schemes focused on milk production have led to the development of “dairy breeds.” This study investigated selective sweeps specifically related to dairy production in sheep by searching for regions commonly identified in different European dairy breeds. With this aim, genotypes from 44,545 SNP markers covering the sheep autosomes were analysed in both European dairy and non-dairy sheep breeds using two approaches: (i) identification of genomic regions showing extreme genetic differentiation between each dairy breed and a closely related non-dairy breed, and (ii) identification of regions with reduced variation (heterozygosity) in the dairy breeds using two methods. Regions detected in at least two breeds (breed pairs) by the two approaches (genetic differentiation and at least one of the heterozygosity-based analyses) were labeled as core candidate convergence regions and further investigated for candidate genes. Following this approach six regions were detected. For some of them, strong candidate genes have been proposed (e.g. ABCG2, SPP1), whereas some other genes designated as candidates based on their association with sheep and cattle dairy traits (e.g. LALBA, DGAT1A) were not associated with a detectable sweep signal. Few of the identified regions were coincident with QTL previously reported in sheep, although many of them corresponded to orthologous regions in cattle where QTL for dairy traits have been identified. Due to the limited number of QTL studies reported in sheep compared with cattle, the results illustrate the potential value of selection mapping to identify genomic regions associated with dairy traits in sheep.     

Genome-wide association study of milk production traits in a crossbred dairy sheep population using three statistical models

Milk production is one of the most important characteristics of dairy sheep, and the identification of genes affecting milk production traits is critical to understanding the genetics and improve milk production in future generations. Three statistical techniques, namely GWAS, ridge-regression BLUP and BayesC , were used to identify SNPs in significant association with three milk production traits (milk yield, fat yield and protein yield) in a crossbred dairy sheep population. The results suggested that chromosomes 1, 3, 4, 5, 7 and 11 were likely to harbor genes important to milk production because these chromosomes had the greatest top-100-SNP variance contributions on the three milk production traits. The GWAS analysis identified between 74 and 288 genome-wide significant SNP (P < 0.05) whereas the BayesCπ model revealed between six and 63 SNPs, each with >95% posterior probability of inclusion as having a non-zero association effect on at least one of the three milk production traits. Positional candidate genes for milk production in sheep were searched, based on the sheep genomic assembly OAR version 3.1, such as those which map position coincided with or was located within 0.1 Mbp of a genome-wide suggestive or significant SNP. These identified SNPs and candidate genes supported some previous findings and also added new information about genetic markers for genetic improvement of lactation in dairy sheep, but keeping in mind that the majority of these positional candidate genes are not necessarily true causative loci for these traits and future validations are thus necessary.     

Genes of the RNASE5 pathway contain SNP associated with milk production traits in dairy cattle

Background

Identification of the processes and mutations responsible for the large genetic variation in milk production among dairy cattle has proved challenging. One approach is to identify a biological process potentially involved in milk production and to determine the genetic influence of all the genes included in the process or pathway. Angiogenin encoded by angiogenin, ribonuclease, RNase A family 5 (RNASE5) is relatively abundant in milk, and has been shown to regulate protein synthesis and act as a growth factor in epithelial cells in vitro. However, little is known about the role of angiogenin in the mammary gland or if the polymorphisms present in the bovine RNASE5 gene are associated with lactation and milk production traits in dairy cattle. Given the high economic value of increased protein in milk, we have tested the hypothesis that RNASE5 or genes in the RNASE5 pathway are associated with milk production traits. First, we constructed a “RNASE5 pathway” based on upstream and downstream interacting genes reported in the literature. We then tested SNP in close proximity to the genes of this pathway for association with milk production traits in a large dairy cattle dataset.

Results

The constructed RNASE5 pathway consisted of 11 genes. Association analysis between SNP in 1 Mb regions surrounding these genes and milk production traits revealed that more SNP than expected by chance were associated with milk protein percent (P < 0.05 significance). There was no significant association with other traits such as milk fat content or fertility.

Conclusions

These results support a role for the RNASE5 pathway in milk production, specifically milk protein percent, and indicate that polymorphisms in or near these genes explain a proportion of the variation for this trait. This method provides a novel way of understanding the underlying biology of lactation with implications for milk production and can be applied to any pathway or gene set to test whether they are responsible for the variation of complex traits.     

Database of cattle candidate genes and genetic markers for milk production and mastitis

A cattle database of candidate genes and genetic markers for milk production and mastitis has been developed to provide an integrated research tool incorporating different types of information supporting a genomic approach to study lactation, udder development and health. The database contains 943 genes and genetic markers involved in mammary gland development and function, representing candidates for further functional studies. The candidate loci were drawn on a genetic map to reveal positional overlaps. For identification of candidate loci, data from seven different research approaches were exploited: (i) gene knockouts or transgenes in mice that result in specific phenotypes associated with mammary gland (143 loci); (ii) cattle QTL for milk production (344) and mastitis related traits (71); (iii) loci with sequence variations that show specific allele-phenotype interactions associated with milk production (24) or mastitis (10) in cattle; (iv) genes with expression profiles associated with milk production (207) or mastitis (107) in cattle or mouse; (v) cattle milk protein genes that exist in different genetic variants (9); (vi) miRNAs expressed in bovine mammary gland (32) and (vii) epigenetically regulated cattle genes associated with mammary gland function (1). Fourty-four genes found by multiple independent analyses were suggested as the most promising candidates and were further in silico analysed for expression levels in lactating mammary gland, genetic variability and top biological functions in functional networks. A miRNA target search for mammary gland expressed miRNAs identified 359 putative binding sites in 3'UTRs of candidate genes.     

A Post-GWAS Replication Study Confirming the PTK2 Gene Associated with Milk Production Traits in Chinese Holstein

Our initial genome-wide association study (GWAS) demonstrated that two SNPs (ARS-BFGL-NGS-33248, UA-IFASA-9288) within the protein tyrosine kinase 2 (PTK2) gene were significantly associated with milk production traits in Chinese Holstein dairy cattle. To further validate if the statistical evidence provided in GWAS were true-positive findings, a replication study was performed herein through genotype-phenotype associations. The two tested SNPs were found to show significant associations with milk production traits, which confirmed the associations observed in the original study. Specifically, SNPs lying in the PTK2 gene were also detected by sequencing 14 unrelated sires in Chinese Holsteins and a total of thirty-three novel SNPs were identified. Thirteen out of these identified SNPs were genotyped and tested for association with milk production traits in an independent resource population. After Bonferroni correction for multiple testing, twelve SNPs were statistically significant for more than two milk production traits. Analyses of pairwise D’ measures of linkage disequilibrium (LD) between all SNPs were also explored. Two haplotype blocks were inferred and the association study at haplotype level revealed similar effects on milk production traits. In addition, the RNA expression analyses revealed that a non-synonymous coding SNP (g.4061098T>G) was involved in the regulation of gene expression. Thus the findings presented here provide strong evidence for associations of PTK2 variants with dairy production traits and may be applied in Chinese Holstein breeding program.