内蒙古地区西门塔尔牛亲子鉴定微卫星标记筛选

研究旨在筛选适合于我国肉牛亲子鉴定的微卫星标记,以内蒙古乌拉盖地区368头西门塔尔牛为实验材料,利用荧光引物PCR和ABI3730测序仪对12个微卫星标记进行基因型检测,应用Cervus3.0软件统计12个微卫星标记的遗传多态性。结果表明:其中4个微卫星标记扩增效率较低,被剔除;其他8个微卫星的多态性丰富,平均等位基因数为11.38,平均期望杂合度为0.6686,平均多态信息含量为0.6354,适合于我国肉牛亲子鉴定。     

草原红牛及其杂种牛群体遗传变异与肉用性能的微卫星标记研究

选用草原红牛、草原红牛与利木赞杂交后代作为试验牛群体，经过基因组DNA的提取、微卫星引物的PCR扩增、扩增产物的电泳分型、各座位等位基因分析以及基因频率、多态信息含量（PIC）和杂合度（H）的计算等步骤，从分子水平上分析了草原红牛及其杂交牛群体的遗传变异。在此基础上，以体重、体尺作为衡量牛生长发育的指标，以肉牛线性体型评分方法中的肌肉度线性评分性状和屠宰肉用性状作为衡量牛肉用性能的指标，运用SPSS软件分析了21个性状与8个微卫星标记的关系，发现了6个微卫星标记对试验牛群体某些生长发育性状和肉用性状存在正面或负面影响。     

家兔群体遗传变异的微卫星标记

选用欧洲野兔的 5个微卫星位点 ,分析了 5个品种 (系 )家兔群体的遗传变异。结果表明 :Vc- 系獭兔的群体内平均多态信息含量和平均杂合度最大 ,分别为 0 .5 5 2 6和 0 .6 2 0 2 ;新西兰兔群体内平均多态信息含量和平均杂合度最小 ,分别为 0 .4 5 15和 0 .5 2 6 1。由此说明 ,5个品种 (系 )家兔中 ,新西兰兔群体内变异较小 ,相对较纯 ;Vc- 系獭兔群体内变异较大 ,纯度相对较小。但总的来看 ,5个品种 (系 )家兔的平均多态信息含量和平均杂合度相差都不大 ,说明Vc獭兔在遗传性能上已接近其他优良品种兔。聚类结果表明 :Vc- 系獭兔与 Vc- 系獭兔亲缘关系最近 ,与日本大耳白兔、青紫蓝兔亲缘关系较近 ,与新西兰兔亲缘关系最远 ,这与 Vc獭兔育种历史事实相符。另外 ,在 Sat4和 Sat8位点上分别检测出了 1条 Vc獭兔所特有的条带 ,从而为进一步研究 Vc獭兔的遗传特性打下良好的基础     

微卫星标记及其在牛遗传育种中的应用

微卫星标记技术是目前在动物遗传标记研究中运用较广泛的一种分子标记技术，本文介绍了微卫星标记技术的基础理论及分析方法，并对其在牛遗传育种研究中的应用进行阐述。     

家兔群体遗传变异的微卫星标记

选用欧洲野生穴兔的5个微卫星座位,分析了5个品种(系)家兔群体的遗传变异.结果表明:Vc-Ⅱ系獭兔的群体内平均多态信息含量和平均杂合度均最大,分别为0.552 6和0.620 2;新西兰兔群体内平均多态信息含量和平均杂合度都最小,分别为0.451 5和0.526 1,说明在5个品种(系)家兔中,新西兰兔群体内变异较小,相对较纯;Vc-Ⅱ系獭兔群体内变异较大,纯度相对较小.但总的来看,5个品种(系)家兔的平均多态信息含量和平均杂合度相差都不大,说明Vc獭兔在遗传性能上已接近其他优良品种兔.聚类结果表明:Vc-Ⅰ系獭兔与Vc-Ⅱ系獭兔亲缘关系最近,与日本大耳兔、青紫蓝兔亲缘关系较近,与新西兰兔亲缘关系最远,这与Vc獭兔育种事实相符.在Sat4和Sat8座位分别检测出了一条Vc獭兔所特有的条带,从而为进一步研究Vc獭兔的遗传特性打下良好的基础.     

微卫星标记与牛的遗传育种

微卫星标记技术由于其众多的优点在目前的动物遗传标记研究中已经成为一种应用较广泛的分子标记技术。本文在简介微卫星标记技术的原理和微卫星标记结构的基础上，对其在牛遗传育种中的应用作了概述。     

微卫星DNA标记及其在牛遗传育种研究中的应用

就微卫星DNA标记特征、优点及其在构建牛基因图谱、牛群体遗传结构分析、寻找与生产性状位点相连锁的分子遗传标记、制作DNA指纹图,进行亲子鉴定和血缘控制、标记辅助选择等方面应用的综述。     

塞北兔群体遗传变异的微卫星标记

检测了125只塞北兔在6个微卫星座位的基因型,分析了等位基因数、基因频率、杂合度和多态信息含量等参数值,并对塞北兔在6个微卫星座位Hardy-Weinberg平衡检验。结果显示:塞北兔在6个微卫星座位共检测到的等位基因30个,每个基因座位为4~6个,等位基因大小在129~244bp之间,塞北兔除了在SOL03基因座位呈中度多态性变异以外,在SOL33、SOL44、7L1B10、L7C11和SAT12等5个微卫星座位呈高度多态性变异;除了在基因座位SOL33和SAT12以外,塞北兔在其他4个基因座位SOL03、SOL44、7L1B10、L7C11均处于Hardy-Weinberg平衡状态。     

新疆草原褐牛体尺性状微卫星DNA标记研究

以新疆褐牛为素材,采用体尺测量、微卫星DNA标记,研究不同位点基因与体尺性状间的相关关系。结果表明,在牛的3条染色体上有4种微卫星DNA标记位点,19个等位基因分别与不同体尺性状间构成紧密相关,从而为该品种资源的开发与利用,提供科学参考依据。     

中国西门塔尔牛产奶性状与微卫星标记相关分析

利用微卫星技术对中国西门塔尔牛育种核心群中152头母牛第6号染色体上9个微卫星位点进行了研究,检测其基因型,利用SAS程序下的GLM,对8个微卫星位点进行产奶性状关系的最小二乘方差分析,找出各位点对产奶性状的影响差异。同时,在一些位点上找到对产奶性状有利的基因型。     

种群遗传变异及基因多样度分析

阐述了种群遗传学和生态遗传学研究的基本单位－种群的定义和意念含义,其一是种系变异含义,其二是生态适应含义,并概述了遗传变异的研究历史。从３个方面及质量性状变异,数量性状变异与基金变异,介绍了种群遗传变异的研究方法,重点介绍了基因变异的测量和亚种群基因多样度的测量,并且依据研究所得的遗传参数对中国苜蓿不同种群遗传结构和多样度进行了例证分析。     

A multiplex PCR assay for differentiating economically important gastrointestinal nematodes of cattle

A multiplex polymerase chain reaction (PCR) test was developed for identifying gastrointestinal (GI) nematodes that commonly infect cattle. This assay was developed using adult-derived genomic DNA and shown capable of discriminating parasite eggs from the feces of experimentally-infected animals at both the species and genus levels. Sequence data from internal (ITS) and external (ETS) transcribed spacers of the ribosomal DNA (rDNA) repeats as well as the 3'-end of the small subunit rDNA and 5'-end of the large subunit rDNA were used to generate five primer sets which, when used simultaneously in a multiplex PCR, produce a unique electrophoretic DNA banding pattern characterized by a single DNA fragment for Ostertagia ostertagi (257bp), Haemonchus placei (176bp), Oesophagostomum radiatum (329bp), Trichostrongylus colubriformis (243bp) and Cooperia oncophora (151bp). In a similar manner, the constructed primer sets amplified DNA from Ostertagia lyrata, Haemonchus contortus, Trichostrongylus axei, Cooperia surnabada and Cooperia punctata. With respect to H. contortus, a closely migrating doublet was generated suggesting size heterogeneity in the ETS which is consistent with multiple rDNA repeat units within this species. PCR analyses using mixtures of monospecifically-purified nematode eggs indicated a sensitivity of less than 0.5 egg-DNA equivalent per species. Although, not designed as a quantitative technique, relative PCR signal intensities corresponded to relative egg burdens within the DNA samples from mixed species of eggs.     

Pedigree analysis of genetic subdivision in a population of Japanese Black cattle

Twenty‐five subpopulations (i.e. populations of prefectures) of more than 2000 Japanese Black cows younger than or equal to 10 years of age were analyzed to evaluate the genetic relationships in the current population. The total number of cows analyzed was 392 346 and their pedigrees were traced back to 1944 or before. Using the pedigree records, the genetic relationships among the subpopulations were estimated by the two different measurements: (i) the average additive relationship coefficients and (ii) Nei's standard genetic distances. Principal component analysis (PCA) was performed to the matrix of the average additive relationship coefficients, and the factor loadings of subpopulations were plotted on the plane to visualize the genetic configuration of subpopulations. To understand the grouping process of the subpopulations, cluster analysis was applied to the matrix of the Nei's genetic distances, and a dendrogram was constructed. There was a high consistency between the results from PCA and cluster analysis. Eight subpopulations with relatively low migration rates showed their unique genetic compositions, and the other 17 subpopulations with high migration rates formed a single cluster. The major cause of the genetic similarity among the 17 subpopulations was inferred to be the strong genetic influence from one subpopulation (Hyogo prefecture) with prominent characteristics for meat quality.     

Evaluation of genetic diversity in Japanese Brown cattle population by pedigree analysis

The Japanese Brown is the second most common domestic beef breed in Japan. However, nowadays this breed is facing reduction in numbers because of pressure from a profitable domestic breed. This breed is uniformly characterized by its brown coat colour, but is comprised of two isolated sub‐breeds, Kumamoto and Kouchi, each possessing a different gene pool. Pedigree analyses were carried out for the two sub‐breeds using the pedigree records of animals born from 1970 to 2000. The effective population size has been found to be consistently reducing during the last three decades in both sub‐breeds. The current effective sizes were estimated to be 25.5 and 6.0 for the Kumamoto and Kouchi sub‐breeds, respectively. The estimate of the effective number of founders (N_ef) in the Kumamoto sub‐breed decreased from 152.1 to 74.4; that of non‐founders (N_enf), from 41.7 to 5.3; and that of founder genome equivalents (N_ge), from 32.7 to 4.9. The corresponding changes in the Kouchi sub‐breed were from 108.2 to 79.4, 16.2 to 4.1, and 14.1 to 3.9. Increasing differences between the two genetic diversity indices in the sub‐breeds indicate that the greater part of the reduction of genetic diversity can be attributed to genetic drift that accumulated in the non‐founder generations. A comparison with published estimates for several cattle breeds suggests the extremely limited genetic diversity of Japanese Brown. In addition to the avoidance of further reduction of genetic diversity, it will be important to counteract the process of breed decline by establishing a production system to efficiently utilize the unique characteristics of this breed and by developing links between the breed and products with market value.     

Molecular genetic approaches for identifying the basis of variation in resistance to tick infestation in cattle

In recent years there has been renewed interest in the adaptation of cattle to challenging environments, largely driven by advances in genomic methods. The current interest in tick resistance is understandable given the major production and welfare implications of tick infestation in tropical and subtropical zones where around 70% of beef cattle are located. Heritability for tick burden in cattle has been shown to range about 0.30, which is sufficient to result in the success of some programs of selection for tick resistance in cattle. Gene-expression studies strongly indicate that both immune and non-immune mechanisms are associated with tick resistance in cattle. Recent quantitative-trait mapping studies have identified chromosome segments and single nucleotide polymorphisms associated with tick burden, but no causal variant has been identified so far. Most of the genetic markers identified for tick burden explain a relatively small proportion of the variance, which is typical of markers for quantitative traits. This leads to the conclusion that panels of multiple markers for tick resistance rather than a single marker will most likely be developed, possibly involving specific panels for zebu or taurine breeds, which could be used for future selection and breeding programs in cattle.     

多重PCR检测圈养牛、猪和羊源魏氏梭菌

采用多重PCR对圈养源魏氏梭菌的α，β，ε，ι毒素基因进行了检测，结果证实该方法具有很高的特异性。通过对山东德州、枣庄、泰安、蒙阴曾经流行过魏氏梭菌病的猪场、牛场和羊场的162个粪便样品进行检测，检出率为19．1％，均为A型。应用该方法鉴别魏氏梭菌血清型快速、简便，结果对于预防治疗均具有重要的指导作用。     

Estimates of genetic diversity in the brown cattle population of Switzerland obtained from pedigree information

The study investigates the genetic diversity present as well as its development in the Brown Cattle population of Switzerland from pedigree information. The population consisted of three subpopulations, the Braunvieh (BV), the original Braunvieh (OB) and the US‐Brown Swiss (BS). The BV is a cross of OB with BS where crossing still continues. The OB is without any genetic influence of BS. The diversity measures effective population size, effective number of ancestors (explaining 99% of reference genome) and founder genome equivalents were calculated for 11 reference populations of animals born in a single year from 1992 onwards. The BS‐subpopulation consisted of animals and their known ancestors which were used in the crossing scheme and was, therefore, quite small. The youngest animals were born in 2002, the oldest ones in the 1920s. Average inbreeding was by far the highest in BS, in spite of the lowest quality of pedigrees, and lowest in OB. Effective population size obtained from the difference between average inbreeding of offspring and their parents was, mostly due to the heavy use of few highly inbred BS‐sires, strongly overestimated in some BV‐reference populations. If this parameter was calculated from the yearly rate of inbreeding and a generation interval of 5 years, no bias was observed and ranking of populations from high to low was OB – BV – BS, i.e. equal to the other diversity parameters. The high genetic diversity found in OB was a consequence of the use of many natural service sires. Rate of decrease of effective number of ancestors was steeper in BV than OB was, however, equal for founder genome equivalents. Founder genome equivalents were more stable than effective population sizes calculated from the difference between average inbreeding of offspring and parents. The five most important ancestors contributed one‐third of the 2002‐reference genomes of BV and OB, in BV all were BS‐sires. The relative amount of BS‐genes in the BV‐genome increased from 59.2% to 78.5% during the 11 years considered.     

德南牛全基因组变异解析和功能基因挖掘

[目的]研究德南牛的群体遗传结构和解析影响其重要性状的关键基因。[方法]通过主成分分析、群体进化树分析、群体遗传结构分析等方法,研究德南牛的遗传结构和与其他品种的亲缘关系;同时,利用全基因组SNP数据,计算德南牛与德国黄牛、南阳牛群体固定指数(Fst)和核苷酸多样性比值(π ratio)、复合似然比(CLR)、XP-EHH等多种选择信号指标,以筛选出德南牛基因组上受到选择的信号,鉴定出重要的候选基因。[结果] 德南牛具有欧洲普通牛(0.789)、东亚牛(0.176)、中国瘤牛(0.035)的混合血统。在θπ和CLR方法中鉴定出93个共有基因,使用FST和XP-EHH方法鉴定出17个共有基因。进行富集以及基因功能注释后,发现这些基因可能与生殖性能(TUBB3、NSD2、GRM1、ERBB4)、生长性状(RFX3、FGFR3、GABRB1、PDE4D)、环境适应性(PGR、GRIA4)和免疫系统反应(P2RY12、P2RY13、GPR87、P2RY14、GPR171)有关。[结论]本项研究发现德南牛血统受德国黄牛影响较多,且挖掘了德南牛基因组内可能具有经济意义的性状相关的选择特征,为德南牛种群的选育提高提供理论支撑。     

Monitoring of genetic diversity in the Japanese Black cattle population by the use of pedigree information

The gene pool of the Japanese Black cattle has been completely closed to foreign breeds during the last 100 years. Genetic diversity of the Japanese Black cattle from 1960 to 2000 was monitored with three estimates of effective number of ancestors. Founder genome equivalent (N_ge) accounts for all the causes of reduction of diversity. Effective number of founders (N_ef) and non‐founders (N_enf) explain reduced diversity because of unequal genetic contributions of founders and random genetic drift in non‐founders, respectively. Further examination using gene dropping simulation was conducted to obtain information on survival of founder alleles. Unique founder alleles were dropped down along the actual pedigree with Monte Carlo procedure following Mendelian segregation rules, and generated genotypes of all the current live animals (612 959 heads). Pedigree records consisted of 2 075 188 animals was used for these analysis. The estimates of three effective numbers (N_ef, N_ge, and N_enf) decreased from 418.6 to 50.3, 86.6 to 7.3, and 109.2 to 8.5, respectively, during the period 1960–2000. The increasing differences between two kinds of genetic diversity indices derived from N_ge and N_ef showed that large part of the reduced diversity from 1980 was attributed to genetic drift caused by the intensive use of particular limited number of sires. In gene dropping analysis, probabilities of extinction of founder alleles were derived from their distributions of frequency in the current animals. Several founders showed low probabilities of allele extinction, irrespective of their relatively low genetic contributions. This suggests that these founders have lineages through which their alleles are surely transmitted to the current breed. The use of these founders as a strategy for recovering the genetic diversity was discussed.     

Indicators of genetic erosion in an endangered population: the Alentejana cattle breed in Portugal

A study was conducted to characterize genetic diversity in the Alentejana breed of cattle based on its demographic trends and to investigate the major factors affecting genetic erosion in this breed. Herdbook information collected between 1940 and 2004, including pedigree records on 100,562 animals in 155 herds, was used to estimate demographic parameters. The mean generation intervals were 6.0 +/- 2.4 yr and 6.8 +/- 3.2 yr for sires and dams of calves, respectively. Average inbreeding increased steadily over the period analyzed, with an annual rate of inbreeding of 0.33 +/- 0.004% (P < 0.01) and an effective population size of 23.3. In the reference population (28,531 calves born between 2000 and 2003) the average inbreeding was 8.35 +/- 9.02% and nearly 80% of the calves were inbred, whereas the average relationship among all animals was 0.026 +/- 0.040. Nevertheless, the mean relationship was 0.328 +/- 0.264 and 0.022 +/- 0.026 for animals born in the same and in different herds, respectively. The computed genetic contributions to the reference population resulted in estimates for the effective number of founders, ancestors, founding herds, and herds supplying sires of 121.6, 55.0, 17.1, and 26.9, respectively, the 2 most influential herds and ancestors contributing 24.2 and 15.1%, respectively, of the current genetic pool. Of the 671 founding sires, only 24 Y-chromosomes are currently represented, but 1 sire alone contributes nearly 60% of this representation, such that the effective number of Y-chromosomes is only 2.73. The observed inbreeding per herd was, on average, 0.053 +/- 0.071 lower than expected from the relationship among the generation of parents of calves in the reference population, indicating that producers have followed breeding strategies that have kept inbreeding at lower levels than anticipated with random selection and mating. When compared with other cattle breeds, Alentejana has some of the highest levels of mean inbreeding and annual rate of inbreeding, and an effective population size that is nearly half of the minimum recommended for maintenance of genetic variability. These critical indicators demonstrate the need to adopt strategies aimed at minimizing inbreeding to avoid further losses of genetic diversity.