湖羊、同羊12个同工酶座位的检测

Gene frequencies of Hu sheep and Tong sheep were obtained with “Random sampling in typical colonies of a central area“.of the 12 loci tested in Hu shpee.11 loci were polymorphic.Reliability of the estimated frequencies of 27 alleles reached 0.95 except for Po^F,Tf^A,Tfd,Hb-β^A and CAF which had reliabilities of 0.5222,0.7478,0.5222,0.6212 and 0.899,respectively,Of the 12 loci tested in Tong sheep,11 loci were polymorphic.Reliability of the estimated frequency of 25 alleles reached 0.95 except for Tf^A,Tf^E and CAF which had reliabilities of 0.931,0.6922 and 0.7924,respectively.The average heterozygosity(H) and average homozygosity(J) was computed and the J of the two sheep colonies was 0.6619 and 0.6448,respectively.Consistent with our conclusions based on genetic data,previous research divided the native sheep populations of East and South Central Asia into three group:the “mongolian group“ “south-Asian group“and “European group“ .Consequently,the degree of genetic similarity between populations and known groups would seem to provide a reliable means of determining the genetic relationships between populations and may reflect the true genetic origin of Hu sheep and Tong sheep in China.     

柴达木山羊群体遗传结构及系统地位的研究

采用简单随机抽样法对柴达木山羊群体进行了遗传检测,从毛色、外型特征、体尺、血液蛋白与DNA多态性等5个方面进行了群体遗传结构分析,并探索了其系统地位.结果表明:(1)毛色和形态特征的平均表型异质度分别为0.3419、0.5207;(2)血液蛋白在6个基因座上具有多态性,平均基因座杂合度为0.2584;(3)DNA-RAPD标记表现丰富的多样性,基因多样度为0.4085～0.5318.通过对柴达木山羊与国内其他山羊的系统聚类,初步探索了该山羊群体的形成及所属,这些都表明柴达木山羊是一个比较古老的未经较高强度选育的地方山羊品种,其基因资源是我国山羊遗传资源中宝贵的一部分.     

柴达木山羊群体遗传结构及系统地位的研究

采用简单随机抽样法对柴达木山羊群体进行了遗传检测,从毛色、外型特征、体尺、血液蛋白与DNA多态性等5个方面进行了群体遗传结构分析,并探索了其系统地位。结果表明：(1)毛色和形态特征的平均表型异质度分别为0.3419、0.5207;(2)血液蛋白在6个基因座上具有多态性,平均基因座杂合度为0.2584;(3)DNA-RAPD标记表现丰富的多样性,基因多样度为0.4085～0.5318。通过对柴达木山羊与其他国内山羊的系统聚类,初步探索了该山羊群体的形成及所属,这些都表明柴达木山羊是一个比较古老的未经较高强度选育的地方山羊品种,其基因资源是我国山羊遗传资源中宝贵的一部分。Abstract: Genetic structure and character of Chaidamu Goats were studied through simple random sampling. Genetic structure was analysed from five aspects, and phylogeny status was also investigated. The results indicated that: (1) the average phenotypic heterogeneity degree of coat color and morphological character were 0.3419 and 0.5207,respectively; (2) polymorphous blood albumen existed in 6 loci and the average loci heterozygosity was 0.2584; and (3) polymorphism existed in marked genes by DND-RAPD with diversity of 0.4085～0.5318. Phylogeny status was investigated through clustering by Ward’s method on Chaidamu Goats and other domestic goats. All these indicated that Chaidamu Goats was a domestic goat with less intensively selected breed.     

基于SSR标记的8个山荆子居群遗传多样性和遗传关系分析

采用10对SSR引物对8个山荆子[Malus baccata (L.) Borkh.]居群140个单株的基因组总DNA进行PCR扩增,并据此对8个居群的遗传多样性和遗传关系进行了分析.结果表明:用10对SSR引物共扩增出91条带,多态性条带百分率达100.00％.8个居群的遗传多样性参数差异较大,有效等位基因数为1.437 9～1.535 0,Nei's基因多样性指数为0.256 0～0.309 2,Shannon信息指数为0.376 7～0.459 2,多态性条带百分率为64.84％～85.71％.居群间的有效等位基因数为1.616 9,Nei's基因多样性指数为0.355 1,Shannon信息指数为0.528 5,均明显高于居群内;8个居群间的基因流为1.739 5,基因分化系数为0.223 3,显示居群间的基因交换较多.UPGMA聚类分析结果表明:在Nei's遗传距离0.148 6处,8个居群被分为3组,河北塞罕坝居群单独为一组,山西五台山居群和北京东灵山居群为一组,其余5个居群为一组.据此推测:山荆子起源于中国华北和东北地区,山西灵空山、黑龙江小兴安岭、吉林长白山和山西中条山居群可能是其遗传多样性的核心居群.     

利用RAPD和ISSR标记分析烤烟品种间遗传关系

利用RAPD和ISSR标记对22份烤烟(Nicotiana tabacumL.)品种进行了遗传关系研究。在RAPD分析中筛选到13个引物,共扩增出167条带,其中多态性带50条,多态性比率为29.9%;在ISSR分析中筛选出7个引物,共扩增出96条带,其中多态性带44条,多态性比率为45.8%。两种标记相结合估算出的品种间遗传相似系数在0.881~0.979之间,平均为0.933。单独基于RAPD标记和ISSR标记的聚类结果有一定差异;两种标记结合起来的聚类分析结果与系谱信息吻合程度更高。定向选择可能对烤烟品种间遗传关系有较大影响;国外引进品种与国内育成品种并未完全分开,表明分子水平的遗传关系和地理来源间缺乏必然联系。     

利用微卫星和线粒体标记分析北京鸭的起源与驯化

北京鸭是我国最著名的家鸭品种. 为分析北京鸭的起源与驯化, 本实验采集了包括北京鸭在内的6个中国地方鸭种共190只个体, 及来自辽宁省的14只野生绿头鸭(Anas platyrhynchos)血样. 利用15个微卫星标记对这6个家鸭品种的186只个体进行遗传关系分析. 并对其中88只家鸭个体及14只野鸭个体线粒体的DNA控制区(mtDNA D-loop)的部分序列进行了扩增和测序. 利用微卫星标记计算出的遗传距离(D_S)及FST分析结果表明, 北京鸭与其他中国地方鸭种的遗传分化程度较高. 聚类结果显示巢湖鸭、高邮鸭、绍兴鸭和金定鸭聚为一类, 建昌鸭、北京鸭聚为另一类. 线粒体DNA D-loop区序列分析结果表明, 虽然北京鸭群体含有所有鸭种所共有的单倍型H01, 但北京鸭的单倍型与其他家鸭仍有较大差异, 北京鸭群体具有12个独特的家鸭单倍型, 其中H02, H04, H08及H22等4个单倍型比较相近并形成单倍群A, 且H02和H22单倍型与中国绿头野鸭共有, 说明北京鸭起源于绿头鸭, 其驯化过程也基本独立于其他家鸭品种.     

六个中国固有绵羊品种遗传关系的微卫星标记分析

用5对微卫星引物对哈萨克羊、蒙古羊、兰州大尾羊、同羊、大尾寒羊、小尾寒羊共6个中国固有绵羊品种基因组进行了扩增, 对扩增结果用7种聚类方法(欧氏距离、欧氏平方距离、夹角余弦、皮尔逊相关、车贝雪夫距离、街区距离、明考夫斯基距离)进行了分析.结果表明, 分布在我国西北部的绵羊品种哈萨克羊、蒙古羊、兰州大尾羊在大多数情况下总是先聚在一起, 相对分布于中原地带的同羊、小尾寒羊和西北部的3个品种间距离较远,同羊和小尾寒羊之间距离也很远,大尾寒羊和小尾寒羊之间没有明显的遗传关系, 此结果与《中国绵羊品种志》中记载的情况基本一致.同时, 本研究结果也说明7种聚类方法均适用于微卫星标记资料的遗传分析.     

中国及其他分布区域野生虎的系统地理学和遗传起源研究进展

世界野生虎(Pantheratigris)传统上被划分为8个亚种,其中3个亚种已于20世纪灭绝,而剩余种群的生存仍然受到偷猎、栖息地丧失和片断化的威胁。作为唯一栖息着4个现存虎亚种的国家,中国在世界虎的保护事业中负有重要责任,然而其野生和圈养虎的分类地位却仍然不确定。最近一项研究(Luoetal,2004)从所有现存野生虎分布地区(包括中国)采集了134份“基准样品”(即原产野外或有确定地域起源的个体生物样品),对虎的系统地理学、种群结构以及遗传起源进行了全面分析。所用的分子标记包括四千碱基对的线粒体DNA、30个核基因组微卫星位点,以及MHC-DRB基因。研究结果表明,虽然虎的整体遗传多态性较低,但是种群分化程度很高,它们被划分为6个,而不是5个现存亚种(1)西伯利亚虎(P.t.altaica);(2)苏门答腊虎(P.t.sumatrae);(3)孟加拉虎(P.t.tigris);(4)华南虎(P.t.amoyensis);(5)印支虎(P.t.corbetti);(6)新定义的亚种马来虎,暂命名为P.t.Jacksoni。由于所研究样本量有限,目前暂定的华南虎亚种还需进一步确定。现有华南虎圈养种群包括遗传关系相距较远的两支一支与印支虎(P.t.corbetti)无异;而另一支则与其他种群均相距甚远,可能代表了真正的华南虎(P.t.amoyensis)。利用分子生物学方法对中国动物园中圈养虎的遗传起源调查亟待进行,以确认该圈养种群整体的遗传独特性或者非独特性。换言之,这将是确认华南虎是否仍然存在的关键。     

利用微卫星DNA标记法进行5个肉用绵羊品种的多态性分析和杂种优势率预测

研究选择了与产肉性能相关的5个微卫星标记,分析了这5个位点在杜泊、德国美利奴、特克塞尔、道塞特和右玉本地绵羊5个品种中的遗传多态性.通过遗传分析预测了4个国外引进肉用绵羊品种与右玉本地绵羊的杂种优势,并与实际测定结果进行了比较分析.结果表明,利用微卫星DNA多态性进行品种间杂种优势预测是可行的,杜泊羊与右玉本地绵羊杂交的杂种优势最大,与实际测定结果一致.     

牦牛的分类学地位及起源研究:mtDNA D-loop序列的分析

牦牛的起源与属级分类学地位至今仍然存在一定的争议。我们测定了家养牦牛和野生牦牛线粒体控制区(D-loop)序列,并以此构建牦牛和牛属、野牛属、水牛属以及非洲水牛属相关种的系统发育树。研究结果表明线粒体D-loop区与Cytb基因序列在构建牛族的系统发育具有同样重要的价值。系统发育关系显示野牛属的灭绝种草原野牛与现存种美洲野牛先聚合为一单系群,然后再和牦牛形成一单系分支,表明牦牛与野牛属的草原野牛、美洲野牛亲缘关系最近,具有最近的共同祖先,而与牛属的其它亚洲物种亲缘关系较远。因此,本研究不支持将牦牛独立为牦牛属———Poephagus,牛属与野牛属在分类上也应合并为一个属。基于上述研究结果和化石证据,我们进一步对牦牛起源的历史背景进行了讨论,认为牦牛与野牛属的分化是由于第四纪气候变化在欧亚大陆发生的,野牛通过白令陆桥进入北美;冰期结束后,由于欧亚大陆其它地区温度升高,牦牛只能局限分布在较为寒冷的青藏高原;而野牛属在北美先后分化为草原野牛和美洲野牛,前者可能是后者的直接祖先。     

Phylogenetic Relationships among Indigenous Sheep Populations in East Asia based on Five Informative Blood Protein and Nonprotein Polymorphisms

The phylogenetic relationships among 23 local sheep breeds and varieties in East Asia were determined based on allele frequency data for five informative and polymorphic blood protein and nonprotein loci (transferrin, arylesterase, hemoglobin-β, X-protein, and potassium transport) using electrophoresis and ion-densitometric techniques. Comparatively striking differences in the frequency distributions, especially of hemoglobin-β and X-protein alleles, were seen between the northern population of the Khalkhas, Bhyanglung, Baruwal, Jakar, Sakten, and five Chinese breeds and the southern population of the Bengal, Kagi, Lampuchhre, Myanmar, and Sipsu breeds. Clustering analyses using UPGMA and NJ methods and principal component analysis, using the data of all populations and loci tested, clearly demonstrated the difference. The two population groups are divided by the boundary of the Himalayas, and each diverged into three subgroups: Mongolian, Tibetan, and Himalayan groups in the north and Indian groups I, II, and III in the south. It was noted that the genetic differentiation of populations was more distinct in the northern group. These findings strongly suggest the existence of at least two large and phylogenetically different gene pools of sheep in East Asia.     

新疆8个绵羊品种遗传多样性和系统发生关系的微卫星分析

为分析新疆北疆地区主要绵羊品种的遗传多样性和系统发生关系,利用10个微卫星标记,采用PCR扩增,12％非变性聚丙烯酰胺凝胶电泳、Sanguinetti银染法显色,对新疆北疆地区8个品种、1个杂交一代绵羊群体遗传多样性进行了检测,统计了各群体的等位基因组成、平均有效等位基因数(E)和平均基因纯合率,利用等位基因频率计算出各群体的平均遗传杂合度(h)、多态信息含量(PIC)和群体间的遗传距离。利用分子进化遗传分析软件,采用邻结法构建系统发生树;同时根据等位基因频率,利用PHYLIP(3．6)分析软件,采用最大似然法构建系统发生树,应用白举检验估计系统树中结点的白引导值,并进行了系统发生分析。结果表明：10个微卫星位点在9个绵羊群体中的多态信息含量除BMI824、MAF65为低、中度多态外,其余8个微卫星均为高度多态,可作为有效的遗传标记用于各绵羊品种的遗传多样性和系统发生关系的分析;所有绵羊群体的平均PIC(0．5631)、h(0．5721)和E(2．9)均低于国外其他品种的绵羊,其基因多态性和遗传多样性相对贫乏;新疆本地土种阿勒泰羊、哈萨克羊和巴什拜羊与国外引进绵羊品种及混有外血的本地培育品种遗传距离较远,他们聚为不同的两类,各绵羊品种的分子系统发生关系与其来源、育成史、分化及地理分布基本一致。     

应用ACGM标记分析禾本科几个物种间的系统发生关系

为了验证水稻基因组数据的通用性,利用ACGM标记分析了禾本科几个不同种属植物的亲缘关系。选用10份材料,它们分别代表禾本科的5个属（Oryza,Zea, Setaria ,Triticum,和Phyllostachys）。根据遗传距离建立了一个聚类树。这5个属的亲缘关系可以简单地表示为：（（Oryza＋（Zea＋Setaria））＋Triticum）＋Phyllostachys。研究结果表明,水稻与玉米或水稻与粟之间的遗传距离比水稻和小麦或水稻与竹子之间的遗传距离近。     

用mtDNA D-环序列探讨蒙古和中国绵羊的起源及遗传多样性

为了在分子水平上探讨绵羊的起源,对中国和蒙古共20个绵羊群体、314只绵羊mtDNA D-环的部分序列进行了测定,结果表明：中国绵羊和蒙古绵羊mtDNA D-环区的部分序列中A、T、G、C含量没有明显的差别;蒙古绵羊的多态位点数（28．85％）略高于中国绵羊（24．22％）;中国绵羊群体的单倍型多样度在青海藏羊、甘肃藏羊、甘肃高山细毛羊、青海细：色羊、甘南藏羊、小尾寒羊和滩羊群体中较高,但在湖羊和岷县黑裘皮羊中较低;蒙古绵羊的单倍型多样度在Bayad和Baidrag群体中最高,但在Gobi—Altai群体中最低。从总体上看,蒙古绵羊的遗传多样性要略高于中国绵羊,例如单倍犁比例的平均值为86．06％（142／185）：78．83％（108／137）,单倍型多样度（Hd）的甲均值为0．976：0．936,核苷酸多样度（Pi（π））的平均值为0．036：0．034,平均核苷酸差异数（k）的平均值为23．50：22．48～217个中国和蒙古绵羊的单倍型序列的系统发生分析表明,中国和蒙古绵羊均有3个母系起源,被定义为A、B和C3类主要的单倍型。其中A类单倍型在所有中国绵羊群体及绝大多数蒙古绵羊群体（9／11）中占优势,平均比例为58．73％;B类单倍型居中,为24．88％;C类单倍型最少,仅为16．59％。进一步从GenBank获得的91个绵羊D-环区的序列与中国和蒙古绵羊D-环区的单倍型的进行网络关系分析,发现欧洲摩弗仑羊（European mouflon,O．musimon）与中国和蒙古绵羊具有较近的亲缘关系,但没有发现塬羊（Argali．O．ammon）、盘羊（0．rignei bochariensis）和东方盘羊（0．ammon nigrimontana）对中国和蒙古绵羊起源有贡献的证据。     

绵羊Cytb基因序列多态性及系统进化研究

以8个中国地方绵羊品种和1个外来品种的20个个体为研究对象,通过对Cytb基因的全序列测定,结果表明：绵羊的单倍型多样性为97．1％。所有序列的平均碱基组成为27．1％T,28．5％C,31．4％A及13．0％G,G＋C含量为41．5％,核苷酸多样性为0．602％。在所有序列中共检测到43个变异位点,其中包括40处转换和3处颠换。Fu’S中性检验表明差异不显著（0．10〉P〉0．05）,说明绵羊群体未发生群体扩张事件。NJ、ME及UPGMA聚类结果均表明,我国绵羊可分为3个单倍型组,这提示我国绵羊有3个母系起源。     

我国6个地方绵羊品种微卫星DNA多态性研究

利用聚丙烯酰胺凝胶电泳技术研究了我国蒙古羊、乌珠穆沁羊、哈萨克羊、阿勒泰羊、滩羊和藏绵羊 6个地方绵羊品种 17个微卫星标记的多态性 ,以探讨其遗传多样性、起源分化及群体间的遗传亲缘关系。结果表明微卫星标记不同位点间遗传多样性差异极显著 (P <0 0 1) ,群体间多态信息含量 (PIC)、近交程度 (Fis)和观察杂合度 (Obs .Het)差异不显著 ,但基因多样性 (genediversity)和期望杂合度 (Exp .Het)差异显著 (P <0 0 5 )。所研究的我国 6个地方绵羊品种与欧洲品种具有相似的遗传多样性 ,但具有较高的近交系数。个体和群体的聚类分析结果提示我国地方绵羊品种可能起源于两类祖先。群体间的聚类分析结果还表明 ,蒙古羊与乌珠穆沁羊分化不明显且具有较近的遗传亲缘关系 ,蒙古羊与藏绵羊间分化明显且具有较远的遗传亲缘关系。滩羊、阿勒泰羊以及藏绵羊间也具有较近的遗传亲缘关系。所研究的我国 6个地方绵羊品种的遗传分化 (Fst)与西班牙绵羊品种接近 ,但明显小于欧洲其他绵羊品种     

Analysis of the Phylogenetic Relationships Among Several Species of Gramineae Using ACGM Markers

To study the transferability of rice (Oryza sativa L.) genome data, we used amplified consensus genetic markers to analyze the phylogenetic relationships among several species and genera in Gramineae. Ten accessions representing five grass genera (Oryza, Zea, Setaria, Triticum, and Phyllostachys) were used. According to the genetic distances, a cluster tree was constructed. The relationships among the five genera could be simply described as ((Oryza + (Zea + Setaria)) + Triticum) + Phyllostachys. The results suggest that the genetic distance between rice and maize (Z. mays L.) or rice and millet (Setaria italica L.) is closer than that between rice and wheat (Triticum aestivum L) or rice and bamboo.     

Genetic Differentiation of Native Sheep Populations in East and South Asia

Variations of four sheep populations in China were examined by multiloci electrophoresis, and similar data are quoted to analyze the degree of genetic differentiation of native sheep populations in East and South Asia. Among 15 populations, the average heterozygosity is 0.2746, and the effective number of alleles is 1.559. Mongolian sheep possess the highest genetic diversity, and diversity decreases sequentially in the Chinese, Vietnamese, Bangladeshi, and Nepalese populations. Coefficients of genetic differentiation are 0.0126–0.3083, with an average of 0.148, demonstrating that the major genetic variation (85%) exists within populations. Genetic identity and genetic distance all show relatively low genetic differentiation. No relationship was found between geographic distance and genetic distance. Gene flow is common among the mass of populations, which leads to the inconsistency between geographic distance and genetic distance. The 15 native sheep populations in East and South Asia can be divided into two groups, one group including part of the Chinese and Mongolian populations and another including theYunnan population of China and part of the Nepalese and Bangladeshi populations. Other populations did not separate into groups, merging instead into the two main groups.     

Origin and phylogenetic analysis of Tibetan Mastiff based on the mitochondrial DNA sequence

At present, the Tibetan Mastiff is the oldest and most ferocious dog in the world. However, the origin of the Tibetan Mastiff and its Phylogenetic relationship with other large breed dogs such as Saint Bernard are unclear. In this study, the primers were designed according to the mitochondrial genome sequence of the domestic dog, and the 2,525 bp mitochondrial sequence, containing the whole sequence of Cytochrome b, tRNA-Thr, tRNA-Pro, and control region of the Tibetan Mastiff, was obtained. Using grey wolves and coyotes as outgroups, the Tibetan Mastiff and 12 breeds of domestic dogs were analyzed in phylogenesis. Tibetan Mastiff, domestic dog breeds, and grey wolves were clustered into a group and coyotes were clustered in a group separately. This indicated that the Tibetan Mastiff and the other domestic dogs originated from the grey wolf, and the Tibetan Mastiff belonged to Carnivora, Canidae, Canis, Canis lupus, Canis lupus familiaris on the animal taxonomy. In domestic dogs, the middle and small breed dogs were clustered at first; German Sheepdog, Swedish Elkhound, and Black Russian Terrier were clustered into one group, and the Tibetan Mastiff, Old English Sheepdog, Leonberger, and Saint Bernard were clustered in another group. This confirmed the viewpoint that many of the famous large breed dogs worldwide Such as Saint Bernard possibly had the blood lineage of the Tibetan Mastiff, based on the molecular data. According to the substitution rate, we concluded that the approximate divergence time between Tibetan Mastiff and grey wolf was 58,000 years before the present (YBP), and the approximate divergence time between other domestic dogs and grey wolf was 42,000 YBP, demonstrating that the time of origin of the Tibetan Mastiff was earlier than that of the other domestic dogs.