5个绵羊群体微卫星标记的遗传多态性分析

为评估5个绵羊群体(160只个体)的遗传多态性,对5个微卫星座位(BM2113,ILSTS0004,GC101,Bulge5,471U)进行研究.结果表明,共发现了82个等位基因,其中在471 U座位的等位基因数最多(23个),ILSTS0004座位的等位基因数最少(13个).多态信息含量、期望杂合度和观察杂合度分析表明,5个绵羊群体均存在遗传多态性,各座位等位基因均较丰富.德国美利奴羊和滩羊的遗传多样性比较丰富,蒙古羊的遗传多样性较低.以Nei氏遗传距离(DA)构建NJ系统发育树,蒙古羊和滩羊、德国美利奴羊和德美寒杂交一代分别先聚为一类,最后才与小尾寒羊聚为一类.     

7个绵羊微卫星DNA标记在绵(山)羊群体中的多态性检测

选择分别位于绵羊2,4,6,9,17和19号染色体上的7个微卫星标记OarFCB11,OarFCB128,MAF70,OarAE101,MAF33,OarFCB48和OarFCB304,对湖羊、同羊及长江三角洲白山羊的典型群随机样本相应位点进行了PCR检测。结果表明,每个微卫星座位发现7个以上等位基因,且均存在多态;湖羊、同羊、长江三角洲白山羊群体杂合度(H)分别为0.9092,0.9177和0.8867,相应的群体基因平均多态信息含量(PIC)分别为0.9024,0.9116和0.8774,有效等位基因数(Ne)分别为11.7723,12.4538和11.6104,均以同羊为最高;以产生有效条带为标志,随机样本的PCR扩增效率存在明显的群体间和座位间差异,以长江三角洲白山羊和OarFCB128座位为最高;以7个座位微卫星等位基因频率推算群体间标准遗传距离,山羊与绵羊群体间遗传距离远大于绵羊群体之间,湖羊、同羊间亲缘距离系数R为0.1998。研究结果提示:选择的7个微卫星DNA座位均为多态座位;以7个微卫星标记为测度,湖羊、同羊及长江三角洲白山羊具有丰富的遗传多态性;7个微卫星标记可进一步用于绵山羊近缘种间遗传分析研究。     

五个绵羊品种遗传多样性的微卫星分析

利用8个微卫星标记对5个绵羊品种(藏羊、兰州大尾羊、蒙古羊、小尾寒羊、无角道赛特)进行了遗传多样性分析.结果表明:5个绵羊群体的8个微卫星位点共发现105个等位基因,平均每个位点有13个,其中,在OarFCB128位点检测到的等位基因数最多,在BM8125位点检测到的等位基因数最少.多态信息含量、有效等位基因数及群体杂合度数据表明,小尾寒羊遗传变异最大,其次是蒙古羊、兰州大尾羊、藏羊,无角道赛特变异最小.根据Nei氏遗传距离用NJ法和UPGMA法构建的聚类图表明,兰州大尾羊与蒙古羊聚为一类,遗传距离最近,与无角道赛特距离最远.     

利用微卫星标记分析滩羊群体的遗传多样性及遗传分化

利用10对微卫星标记对小尾寒羊、宁夏牧区园区白滩羊、保种场滩羊和黑滩羊的遗传多样性和遗传分化进行分析,根据DC和DA遗传距离构建系统树。结果表明:10个微卫星座位中共检测到167个等位基因,每个座位在每个群体均检测到5个以上的等位基因。座位和群体期望杂合度均在0.8左右;座位平均多态信息含量为0.722 3～0.838 5;园区白滩羊平均多态信息含量为0.794 6,保种场白滩羊平均多态信息含量为0.778 4,黑滩羊平均多态信息含量为0.772 3,群体平均多态信息含量为0.772 3～0.794 6。群体间表型分化系数为0.089 3,总群体近交系数为0.674 0,群体内近交系数为0.642 0。运用DC和DA遗传距离,分别采用UPGMA和NJ法构建的系统聚类图基本一致。综合遗传分化系数、遗传距离及聚类图,结果提示,滩羊与小尾寒羊群体间遗传分化程度最大,黑滩羊与白滩羊群体间存在一定的遗传分化,2个白滩羊群体间遗传分化程度相对最小。     

六个绵羊品种遗传距离的微卫星分析

利用30个微卫星DNA标记对乌珠穆沁羊、苏尼特羊、内蒙古细毛羊、宁夏滩羊、小尾寒羊和无角多赛特羊共237个体进行遗传距离分析,结果表明：（1）苏尼特羊和乌珠穆沁羊遗传距离为0．0833最小,无角多赛特羊与其它5个绵羊品种的遗传距离在1．0146-1．2876之间,滩羊、内蒙古细毛羊和小尾寒羊彼此之问及与乌珠穆沁羊和苏尼特羊的遗传距离为0．3623-0．6020;（2）聚类图将6个绵羊品种划分为4个分支,即苏尼特羊和乌珠穆沁羊,内蒙古细毛羊和小尾寒羊,宁夏滩羊,无角多赛特羊。（3）6个绵羊品种间遗传距离和聚类图与品种的历史演化和资料记载基本符合,为绵羊种质资源的利用和保护提供了科学依据。     

5个绵羊群体微卫星多态性分析

采用微卫星DNA标记对甘肃高山细毛羊、滩羊等5个绵羊群体(125只个体)的5个微卫星座位等位基因进行检测,分析5个绵羊群体的遗传多样性和亲缘关系。结果表明:5个绵羊群体中共发现78个等位基因,其中在甘肃高山细毛羊肃南县群体中发现的等位基因数最多(57个),滩羊最少(43个)。多态信息含量、期望杂合度和观察杂合度的数据分析表明:在研究的5个绵羊群体中甘肃高山细毛羊天祝县群体和肃南县群体的遗传多样性较丰富,而滩羊的遗传多样性相对较低。DA遗传距离和DS遗传距离构建的邻接(NJ)系统发育树均表明:甘肃高山细毛羊天祝县群体和肃南县群体与藏羊的亲缘关系较近,为一类;而滩羊与小尾寒羊的遗传距离较近,为另一类。     

国内主要绵羊品种基于染色体核型进化距离的聚类分析

收集国内10个主要绵羊品种包括藏系绵羊、乌珠穆沁羊、青海细毛羊、河南小尾寒羊、蒙古羊、东北细毛羊、同羊、兰州大尾羊、湖羊、滩羊(贺兰山及盐池)等的染色体核型数据,进行核型进化距离的计算,并采用最短距离聚类分析法进行聚类分析.结果表明:10个绵羊品种可分为2大类群,即蒙古羊系和藏羊系,两大类群在最短距离为1.131 2处相聚.归人蒙古羊系的有8个品种.蒙古羊与兰州大尾羊在最短距离为0.095 2处首先聚为一类;贺兰山滩羊、盐池滩羊、河南小尾寒羊在最短距离为0.120 0处相聚;湖羊在最短距离为0.242 4处与前述品种相聚而成为一小类,东北细毛羊、同羊、乌珠穆沁羊分别在最短距离为0.339 8、0.339 8、0.568 7处与前述品种聚为一大类.归入藏羊系的有青海细毛羊和藏系绵羊,青海细毛羊在最短距离为0.967 5处与藏系绵羊聚为一大类.所得结果与品种起源进化、产地分布大致相同,为国内主要绵羊品种的分类和演化提供了细胞学依据.     

利用结构基因座分析小尾寒羊、滩羊群体遗传分化水平

 采用中心产区典型群随机抽样方法和多种电泳技术检测60只小尾寒羊、73只滩羊编码血液蛋白17个结构基因座上的变异,引用国内外14个绵羊群体相同资料进行比较分析,探讨其遗传分化水平。研究表明：(1) 小尾寒羊、滩羊结构基因座平均杂合度分别为0.2360和0.2587;平均多态信息含量分别为0.1974、0.2102;平均有效等位基因数分别为1.5723和1.5751。(2) 4个组合（分别为4个、6个、13个及16个绵羊群体）的基因分化系数分别为0.049323、0.059987、0.1728和0.201256,说明湖羊、同羊、小尾寒羊和滩羊4个绵羊群体结构基因座的基因分化程度低;这4种绵羊与蒙古国绵羊的基因分化程度次之;蒙古羊系绵羊和南亚羊及欧洲羊之间基因传分化程度较高。(3) 前人关于小尾寒羊、滩羊由蒙古羊分化而来的考证得到遗传学实验的进一步证明,湖羊、同羊、小尾寒羊和滩羊受蒙古羊血统的影响递减。群体间亲缘关系远近与其所处地理位置远近并未表现出紧密相关。     

湖羊与同羊血液蛋白多型及其系统地位

应用中心产区典型群简单随机抽样的方法,检测湖羊、同羊控制血液酶和其他蛋白质变异的14个结构基因座上40个等位基因频率,引用国内外15个群体10个座位的研究资料,构建合成了描述群体间遗传相似性的模糊等价关系矩阵,对17个群体进行亲缘关系聚类。结果表明:湖羊、同羊与蒙古羊群体亲缘关系密切,是蒙古羊向南迁徙与驯化的产物,相对于中亚东南和欧洲绵羊而言,蒙古羊群体构成相对独立的系统。     

中国蒙系6个绵羊品种的遗传分化和基因流

利用7对微卫星引物对中国蒙系6个绵羊品种(同羊、小尾寒羊、湖羊、滩羊、乌珠穆沁羊和巴音布鲁克羊)的遗传分化、基因流、遗传分化程度与地理距离间的关系进行分析,并利用遗传距离构建系统树。结果表明:6个绵羊群体中,总群体近交系数Fit最高的位点为OarAE101(0.1670),最低的为MAF33(0.0440);群体间分化系数Fst最高的位点为OarAE101(0.1000),最低的为OarFCB48(0.0220);群体内近交系数Fis最高的位点为OarFCB304(0.0890),最低的为MAF33(0.0050)。Fst平均为3.9%,即由各群体内个体间的差异引起的遗传变异是96.1%,说明遗传变异绝大部分存在于品种内,品种间的遗传分化水平很低。6个绵羊群体每世代两群体间有效迁移个体数为2.7369(滩羊和巴音布鲁克羊)~44.3928(同羊和湖羊),平均为11.2521,均反映出品种间的基因流通畅。品种间的遗传差异与地理距离呈显著相关。初步推断我国蒙系绵羊品种间的遗传分化主要是自然选择(生境异质性)作用的结果。     

Study on polymorphisms of microsatellite DNA of six Chinese indigenous sheep and goat breeds

The genomes of six populations were screened using microsatellites as molecular markers, including Ujmuqin sheep, small-tailed Han sheep, Tan sheep, Hu sheep, Tong sheep and Yangtse River Delta (YRD) white goat. A total of seven microsatellite markers were used and genetic diversity and genetic distance were also determined. The results showed that there were 224 alleles in six populations, all seven loci showed polymorphism in all populations. The average heterozygosity of all populations was 0.949 9, and the mean polymorphism information content (PIC) of all six populations was 0.842 5–0.929 4. The six sheep (goat) popualtions were lowly differentiated with all loci, and the coefficient of phaenotype differentiation (Fst) was 2.6%, which was consistent with the coefficient of gene differentiation (Gst). The global heterozygote deficit across of all populations (Fit) amounted to 0.5%. The overall significant deficit of heterozygotes because of inbreeding within breeds (Fis) amounted to −2.2%. Two Unweighted Pair-group Method using Arithmetic Averages (UPGMA) dendrograms were constructed on the basis of Nei’s standard genetic distance (DS) and Nei’s genetic distance (DA) respectively. Hu sheep and Tong sheep were grouped at first, Ujmuqin sheep and small-tailed Han sheep clustered and then clustered with Tan sheep. Finally, Yangtse River Delta white goat joined in with all above. From this study, Ujmuqin sheep belongs to “Mongolia sheep” group, which corresponds with the historical records exactly. Ujmuqin sheep and small-tailed Han sheep, Tan sheep, Hu sheep and Tong sheep all vest in the “Mongolia sheep” group.     

利用结构基因座分析中国蒙古羊系统内部分不同生态型绵羊品种遗传分化关系及距离隔离机制

 【目的】探讨蒙古羊系统内部分品种间的遗传分化关系,揭示蒙古羊系统内主要不同生态型地方绵羊品种形成的地理距离隔离机制。【方法】以中国蒙古羊系统内5个地方绵羊品种,湖羊、同羊、小尾寒羊、滩羊和洼地绵羊为研究对象,以多座位电泳法检测5个地方绵羊品种的20个结构基因座（包括Al、Gc、Tf、Cp、Alp、Ary-Es、Lap、Hb-α、Hb-β、Xp、CA、Dia-I、Dia-Ⅱ、MDH、GPI、EsD、α2-M、Cat、Ly和Ke）的变异。【结果】5个绵羊群体间的系统发生关系不满足距离隔离模式,绵羊群体间的遗传分化关系的远近与其地理分布并未表现出紧密的线性相关。【结论】5个绵羊品种分别起源于不同时期的蒙古羊始祖群体,同时在品种间存在一定程度的基因交流,并在各自特有的生态环境中经历不同程度的自然选择和人为选择培育而成。     

The Levels of Genetic Differentiation of Small-Tailed Han Sheep and Tan Sheep Populations Using Structural Loci

Using the method of ＂random sampling in typical colonies of the central area of the habitat＂ and several electrophoresis techniques, the variations of 17 structural loci encoding blood proteins in 60 Small-Tailed Han sheep and 73 Tan sheep were examined and compared with those of 14 other sheep populations in China and other countries to investigate their levels of genetic differentiation. The average heterozygosities of Small-Tailed Han sheep and Tan sheep were 0.2360 and 0.2587, respectively. The average polymorphic information content values were 0.1974 and 0.2102, respectively. The average effective numbers of alleles were 1.5723 and 1.5751, respectively. The coefficients of gene differentiation in the four groups （including 4, 6, 13, and 16 sheep populations, respectively） were 0.049323, 0.059987, 0.1728, and 0.201256, respectively, indicating that the degree of gene differentiation at the structural loci was the least in Hu sheep, Tong sheep, Small-Tailed Hart sheep, and Tan sheep; followed by the above-mentioned four sheep populations and two Mongolian sheep populations; and was the highest in sheep populations belonging to the Mongolian sheep group, South Asian sheep, and European sheep. The earlier researchers＇ conclusions that both Small-Tailed Han sheep and Tan sheep evolved from Mongolian sheep were further verified by the results of this study. Hu sheep, Tong sheep, Small-Tailed Han sheep, and Tan sheep were decreasingly affected by the bloodline of Mongolian sheep to different degrees. The relationships among sheep populations were not closely related to the geographical distances among sheep populations.     

中亚以东南绵羊群体钾座位遗传分化的研究

利用中心产区典型群简单随机抽样法,以湖羊、同羊、滩羊、小尾寒羊、洼地羊为研究对象,引用我国周边国家、地区绵羊品种资料为分析背景,采用Na/K/Cl离子分析仪对红细胞中钾的含量进行测定。结果表明:1)各群体钾座位中立性检测的观察值除Bhyanglung绵羊外,其余均在L95和U95之间,表明钾座位作为中立性座位,基本未受到选择等因素的影响;2)血钾基因频率在地理分布上存在一定规律,基本符合将中亚以东南固有绵羊系统划分为“蒙古羊”、“藏羊”和“南亚-东南亚羊”三大集团;3)中亚以东南16个绵羊群体的钾座位基因分化系数为0.185 9。这说明上述16个绵羊群体钾座位81.41%的变异是由群体内的遗传多态现象引起的,只有18.59%的变异来自于群体间的差异。     

中国及中亚以东南部分绵羊群体的遗传多样性

以中心产区典型群简单随机抽样的方法,抽样检测65只同羊、63只湖羊的5项体尺、5项形态特征、6项生态特征及10个血液蛋白质结构基因座基因频率,引用国内外相关研究资料。(1)对国内的6个绵羊群体的17项指标进行主成分分析,并根据各群体主成分值进行聚类,结果表明,国内6个绵羊群体被明显分为牧区、农牧交错区的蒙古羊、滩羊和农区的大尾寒羊、小尾寒羊、湖羊、同羊两大类。(2)在10个结构基因座基因频率基础上,计算中亚以东南12个绵羊群体平均座位纯合度和杂合度,比较其遗传多样性,依杂合度的高低分为3类。结果表明:绵羊的4项生态指标、2项形态特征及4项体尺对绵羊品种间差异起决定作用;湖羊、同羊及蒙古羊系统的另两个群体Kh和Ub具丰富的遗传多样性。     

Genetic differentiation and gene flow among six sheep breeds of Mongolian group in China

The level of genetic differentiation, gene flow and the relationship between geographical distance and genetic differentiation among six sheep populations of Mongolian group in China (Tong sheep, small-tailed Han sheep, Hu sheep, Tan sheep, Ujumuqin sheep and Bayinbuluk sheep) were analyzed using seven microsatellites. The trees were constructed from diversity coefficient (DC) distances among the six sheep populations. The overall heterozygote deficit across all the populations (F _it) was between 0.167 (OarAE101) and 0.044 (MAF33). The over-all significant deficit of heterozygote, because of inbreeding within breeds, (F _is) was between 0.089 (OarFCB304) and 0.005 (MAF33). The coefficient of genetic differentiation (F _st) was between 0.100 (OarAE101) and 0.022 (OarFCB48). It indicated that 3.9% of the total genetic variation could be explained by breed differences and the remaining 96.1% by differences among individuals for each population. This illustrated that most variations existed within breeds and genetic differentiation level were very low among sheep breeds of the Mongolian Group in China. The average number of effective migrants exchanged per generation (Nem) ranged from 2.7369 (Tan sheep and Bayinbuluk sheep) to 44.3928 (Tong sheep and Hu sheep), and the mean value was 11.25213. Significantly positive relationships between the level of genetic differentiation and geographical distance and genetic distances were detected. It is concluded that genetic differentiation of sheep breeds of Mongolian group in China is mainly the result of natural selection (different living conditions). __________ Translated from Journal of Yangzhou University (Agricultural and Life Science Edition), 2007, 28(3): 22–26 [译自: 扬州大学学报(农业与生命科学版)]     

绵羊和山羊基于两种标记的遗传分化初步研究

对湖羊、同羊及长江三角洲白山羊的随机样本分别进行14个结构基因座和7个微卫星标记的遗传检测,比较由2种遗传标记获得的群体基因平均杂合度、多态信息含量及群体有效等位基因数,分别根椐2种类型的基因频率资料,计算3个群体间的标准遗传距离并加以比较.结果表明由微卫星等位基因频率获得的群体基因平均杂合度、多态信息含量及有效等位基因数显著大于由结构基因座获得的,由结构基因座资料计算的3个群体间标准遗传距离为0.026 8～0.248 7,微卫星标记资料计算的3个群体间标准遗传距离为0.232 1～1.231 3,绵山羊群体间显著大于绵羊间.结构基因座和微卫星标记一致揭示了3群体内的遗传变异程度由大到小依次为同羊、湖羊、长江三角洲白山羊;微卫星标记较结构基因座标记更能表达近缘种间进化趋异水平;可将FCB11、MAF33、AE101、FCB128及FCB304位点作为研究绵山羊近缘种间遗传分化的标志性位点.