云南山羊线粒体DNA遗传多样性与聚类分析

为了对云南省5个地方山羊品种67个个体的遗传多样性及其起源进化进行探讨,提取总DNA,扩增mtDNA D-loop区,并测序。结果发现:云南地方山羊67个个体共定义47种单倍型,单倍型多样性(Hd)为0.985;平均核苷酸多样性(Pi)为0.019 15;平均核苷酸差异数(K)为23.195;群体平均遗传距离0.20。系统发生树分析共聚为两个支系,A支系和B支系。其中A支系个体较多,与野山羊(胃石山羊)聚为一类;B支系个体较少,分为B1和B2分支,均没有野山羊与其聚为一类。以上表明:云南山羊遗传多样性丰富,品种间基因交流频繁,至少有两个母系起源,其中之一为胃石山羊。     

中国山羊mtDNA D-loop遗传多样性及其起源研究

采用DNA测序技术分析了中国9个山羊品种（板角山羊、成都麻羊、贵州黑山羊、贵州白山羊、黔北麻羊、马头山羊、陕南白山羊、黄淮山羊和雷州山羊）共计128个个体的mtDNA D-loop全序列。结果表明：山羊mtDNA D-loop全序列长度为1212-1213bp,检测到102个变异位点,约占分析位点总数的8．42％,可变位点中转换占99个,颠换2个,1个转换/颠换共存;界定了92种单倍型,有78种为各品种独享单倍型,另外14种为群体内或群体间共享单倍型。9个山羊品种单倍型多样度为0．9333-1．0000,核苷酸多样度为0．7062％-1．8265％,表明中国山羊品种遗传多样性丰富。根据92种mtDNA单倍型构建了中国山羊的NJ分子系统树,聚类表明,中国山羊mtDNA D-loop序列单倍型分为支系A和支系B两大类。支系A包括75种单倍型,代表95个样本,占总数的74．22％;支系B包括17种单倍型,代表33个样本,占总数的25．78％,说明中国山羊存在支系A和支系B两大母系起源。对中国山羊mtDNA D-loop的支系A和支系B进行核苷酸不配对分布曲线分析和Fu的Fs中性检验,分析表明,支系A的分布曲线呈单峰形,Fs值为-24．6491,P值为0．0000,显著偏离中性,表明山羊支系A曾经历群体扩张;支系B呈近似双峰分布,Fs值为-3．3947,P值为0．0980,中性检验差异不显著,表明山羊支系B没有经历群体扩张,群体大小保持相对稳定。山羊支系B可能起源于中国。     

中国荷斯坦牛和鲁西黄牛mtDNAD-loop序列多态性分析

为了从母系遗传角度深入阐明中国荷斯坦牛和鲁西黄牛的群体遗传多样性以及起源进化,本研究采用PCR测序法测定了9头中国荷斯坦牛和11头鲁西黄牛的线粒体DNA D-loop区的部分序列,并经剪切后进行生物信息学软件分析.结果表明,20个个体D-loop区共711 bp,共检测到19种单倍型和50个多态位点.核苷酸多样性(Pi)为0.021 33±0.004 54,单倍型多样性(Hd)为0.994±0.019,平均核苷酸差异数(k)为15.146 20.构建的NJ网络进化树共分为两大支系,其中部分鲁西黄牛与瘤牛聚为一支,而中国荷斯坦牛和部分鲁西黄牛与普通黄牛聚为一支.说明中国荷斯坦牛和鲁西黄牛群体遗传多样性均较高;鲁西黄牛同时含有瘤牛和普通黄牛的血统,而中国荷斯坦牛只含有普通黄牛的血统.     

鹅喉羚线粒体D-loop和Cytb序列遗传多样性分析

[目的]为了探讨新疆天山野生动物园17只鹅喉羚的遗传多样性、亲缘关系和群体来源。[方法]对这些鹅喉羚线粒体控制区(D-loop)和细胞色素b(Cytb)基因序列进行了PCR扩增和测序,分别计算了群体单倍型多样度(H)和核苷酸多样性(Pi)以及个体间的遗传距离,并构建了单倍型NJ(Neighbor-Joining)聚类图。[结果]D-loop区和Cytb分别获得了978bp和543bp的基因片段,D-loop区识别出11个单倍型,Cytb区识别出3个单倍型;D-loop和Cytb的单倍型多样度(H)和核苷酸多样性(Pi)分别为0.863、0.01028和0.330、0.00077;个体之间最大遗传距离为0.024,最小遗传距离为0.001;17只鹅喉羚分别聚为2个分支。[结论]该鹅喉羚群体D-loop区多态性较丰富,在野外可能来自两个不同的群体。     

中国部分地方黄牛品种线粒体D-loop区的遗传变异与进化分析

测定了13个黄牛品种125个个体的线粒体D-loop区段的全序列,包括12个中国地方黄牛品种的123个个体和德国黄牛2个个体,并进行了分析。结果显示,共检测到93个变异位点,57个单倍型,平均核苷酸差异(average number ofnucleotide differences,k)为22.708,核苷酸多样度(nucleotide diversity,π)为0.0251±0.00479,单倍型多样度(haplotypediversity,Hd)为0.888±0.026,表明我国黄牛品种遗传多样性非常丰富。构建的Neighbor-Joining进化树显示这13个品种主要分成两大类型:普通牛和瘤牛;新发现的特殊类型Ⅲ只有一个西藏阿沛甲咂牛的个体,它与牦牛D-loop序列最相近,证明西藏地区的黄牛与牦牛之间存在基因渗入现象。普通牛和瘤牛在日喀则驼峰牛中占的比例分别是64.3%和35.7%,在阿沛甲咂牛中占的比例分别是50.0%和50.0%,证明了西藏的黄牛也有瘤牛类型。云南牛品种的单倍型非常丰富证明了云南在中国黄牛起源上的重要地位;在27个中国黄牛品种中(本研究11个品种以及GenBank上的16个品种)找到了中国瘤牛的核心单倍型i1,并且对它进行了讨论。同时证明了西藏瘤牛独立于中国瘤牛核心类群的特殊性。     

福建省家鸭的遗传多样性及其与野鸭的亲缘关系

为了探讨福建省家鸭的遗传多样性和品种进化, 本研究测定了福建省4个地方家鸭品种的32个个体和野生斑嘴鸭(Anas zonorhyncha) 8个个体的线粒体DNA D-loop区667 bp序列, 分析了这4个家鸭品种的遗传多态性及品种进化。结果显示4个家鸭D-loop区序列的A、G、C、T碱基平均含量分别为25.5%, 15.2%, 33.5%, 25.8%。确定了8种单倍型, 其中Hap2(与GenBank中的A7一致)为家鸭的主体单倍型, 平均单倍型多样度为0.75202, 平均核苷酸多样度为0.21%。4个品种中金定鸭单倍型多样度、核苷酸多样度最高。4个品种之间的Kimura双参数遗传距离为0.00094–0.00395, 其中连城白鸭和金定鸭之间遗传距离最大。结合GenBank上已发表的绿头鸭(A. platyrhynchos)和斑嘴鸭序列, 共有30种单倍型, 共同构建系统发生树和网络关系图, 结果表明4个家鸭品种单一地起源于绿头鸭。     

普氏野马线粒体DNA D-loop区序列多态性研究

目的:了解中国新疆吉木萨尔野马繁殖中心的普氏野马(Equus przewalskii)遗传多样性及其遗传背景.方法:采用PCR产物直接测序法,对15匹普氏野马线粒体DNA D-loop高变区进行测序分析.结果:测定15个个体的线粒体DNA D-loop高变区15464～15866片段序列402bp.检测到12种单倍型,包括37个多态位点,占全部序列的9.2%,其中转换位点24个、颠换位点20个、转换位点和颠换并存位点8个、缺失位点3个.A%+T%含量(56.1%)高于G%+C%含量(43.9%),平均A含量为28.4%,T含量为27.7%,C含量为29%,G含为14.9%.单倍型间平均遗传距离为0.030,单倍型多态性(h)为1±0.00116,核苷酸多态性(π)为2.90%.15匹普氏野马线粒体DNA D-loop高变区之间平均核苷酸变异率为2.48%.结论:研究表明我国新疆吉木萨尔野马繁殖中心的普氏野马线粒体DNA D-loop区序列存在丰富的多态性.     

基于线粒体D-loop区和Cyt b基因分析广西禾花鲤三个群体遗传结构

研究基于线粒体D-loop区和Cyt b基因部分序列, 分析了广西全州、融水、环江三地禾花鲤(Cyprinus carpio)的遗传结构。在3个群体中共鉴定到19种D-loop与Cyt b序列的组合单倍型, 总的单倍型多样性(H_d)和核苷酸多样性(π)分别为0.916±0.010和0.008±0.004, 禾花鲤线粒体DNA具有单倍型多样性高和核苷酸多样性低的特点。环江群体单倍型多样性最低, 但是核苷酸多样性却最高, 反映了环江群体存在最明显的谱系混杂。分子方差分析(AMOVA)显示遗传变异主要来源于群体内部(71.54%), 禾花鲤三个群体间有显著的遗传分化(F_st=0.285; P<0.01)。系统发育分析表明, 多数样本的线粒体单倍型属于华南鲤(C. carpio rubrefusus)类型, 同时三地禾花鲤中均检测到一定频率的西鲤(C. carpio carpio)或远东鲤(C. carpio haematopterus)类型的线粒体单倍型, 提示禾花鲤可能受到鲤养殖品种的杂交渐渗。研究初步揭示了广西禾花鲤的种质资源现状, 为禾花鲤这一地方特色“稻鱼共作”品种的选育和苗种管理提供了必要的依据。     

东帕米尔高原喜马拉雅雪鸡遗传多样性及系统发育地位

东帕米尔高原作为生物多样性丰富的区域之一,喜马拉雅雪鸡和藏雪鸡在此混群分布。以东帕米尔高原喜马拉雅雪鸡为研究对象,采用PCR和测序技术,研究了mt DNA D-loop区序列特征,下载Gen Bank已提交的雪鸡序列,利用最大似然法构建系统发育树和中介网络关系,以阐明东帕米尔高原喜马拉雅雪鸡遗传多样性水平和系统进化地位。结果表明:东帕米尔高原喜马拉雅雪鸡mt DNA D-loop序列富含A、T碱基,含量为59.8%,存在64个多态位点,占核苷酸总数的5.5%,其中单一多态位点29个,简约信息位点33个,两处插入或缺失,转换发生的频率远远高于颠换;25个个体存在23种单倍型,平均单倍型多样度(Hd)为0.92±0.0001,平均核苷酸多样度(π)为0.00958,平均核苷酸差异度(K)为11.067,说明东帕米尔高原喜马拉雅雪鸡核苷酸多样性较低,单倍型多样性高,具有较为丰富的遗传多样性;系统发育分析显示喜马拉雅雪鸡与藏雪鸡分为明显的两大簇群,本研究涉及的东帕米尔高原喜马拉雅雪鸡出现遗传分化,呈现明显的2个进化支;中介网络关系分析显示雪鸡具有明显的地理分布特征,本研究雪鸡84%的单倍型聚在以东帕米尔高原为中心的进化支上。因此,建议扩大塔什库尔干野生动物自然保护区(位于东帕米尔高原境内)范围,建立国家级自然保护区,恢复生态环境,以提高雪鸡栖息地的生存适宜性。     

贵州黄牛mtDNA D-loop 遗传多样性研究

对贵州4个地方黄牛品种共计82个个体的线粒体DNA D-loop区全序列910 bp进行分析,检测到31种单倍型,其核苷酸多态位点65个,约占所测核苷酸总长的7.14%,其中有62个转换,2个颠换,1个转换/颠换共存。贵州4个黄牛品种mtDNA D-loop区核苷酸多样度（π值）为2.16%～2.61%,单倍型多样度（H）为0.695～0.909,表明贵州黄牛mtDNA遗传多样性比较丰富。根据单倍型构建了贵州4个黄牛品种的NJ分子系统树。聚类表明,贵州黄牛有普通牛和瘤牛2大母系起源,其影响较为均一。并探讨了用核苷酸多样度π值的大小来衡量黄牛群体遗传分化程度的可行性。      

Mitochondrial DNA diversity of Pakistani goats

Pakistan contains numerous domestic goat breeds, but until now there has been no comprehensive study on genetic diversity or a phylogenetic analysis of Pakistani goats. In this study, we analysed the complete mitochondrial DNA D-loop and the cytochrome b gene of 13 Pakistani domestic goat breeds (Capra hircus) and one wild goat, the Sindh Ibex (Capra aegagrus blythi). The phylogenetic analyses and sequence divergence (SD) established four distinct mt-lineages termed as A, B and C (previously reported) and a new lineage D. The Sindh Ibex appeared as an outgroup of domestic goats. The estimated divergence times between the most recently evolved mt-lineages A and D were from 260,483 to 371,052 YA. This suggested that at least four different strains of wild Capra might have been the source of the modern domestic goats. The new mt-lineage D revealed high SD from mt-lineage A and may be the oldest branch under domestication, while mt-lineages B and C showed lower SD and might have been domesticated during an advanced stage of the domestication process.     

The Genetic Diversity of mtDNA D-loop and the Origin of Chinese Goats

The complete sequences of mitochondrial DNA D-loop of 128 individuals in nine Chinese goat (Capra hircu) breeds were analyzed by DNA sequencing technology. The results show that the length of mtDNA D-loop in Chinese goats is 1 212-1 213 bp. There are 102 polymorphic sites, accounting for 8.42% of 1 212 bp sequence. Ninety-two mtDNA haplotypes were determined. The haplotype diversity and nucleotide diversity are 0.9333-1 .0000 and 0.7062%-1.8265%, respectively. The results indicate that the genetic diversity of Chinese goats is very abundant. The NJ tree indicates that Chinese goats have two types of maternal origins from lineage A and lineage B. The possibility of lineage B originating from China is also discussed.     

Genetic diversity and molecular phylogeography of Chinese domestic goats by large-scale mitochondrial DNA analysis

Mitochondrial DNA (mtDNA) D-loop sequences of 666 individuals (including 109 new individuals, 557 individuals retrieved from GenBank) from 33 Chinese domestic goat breeds throughout China were used to investigate their mtDNA variability and molecular phylogeography. The results showed that all goat breeds in this study proved to be extremely diverse, and the average haplotype diversity and nucleotide diversity were 0.990 ± 0.001 and 0.032 ± 0.001, respectively. The 666 sequences gave 326 different haplotypes. Phylogenetic analyses revealed that there were 4 mtDNA haplogroups identified in Chinese domestic goats, in which haplogroup A was predominant and widely distributed. Our finding was consistent with archaeological data and other genetic diversity studies. Amova analysis showed there was significant geographical structuring. Almost 84.31 % of genetic variation was included in the within-breed variance component and only 4.69 % was observed among the geographic distributions. This genetic diversity results further supported the previous view of multiple maternal origins of Chinese domestic goats, and the results on the phylogenetic relationship contributed to a better understanding of the history of goat domestication and modern production of domestic goats.     

Mitochondrial DNA diversity and origins of domestic goats in Southwest China (excluding Tibet)

Southwest China contains about one third Chinese indigenous goat breeds representing special economic and ecological characteristics. Mitochondrial DNA (mtDNA) D-loop sequences of 312 individuals (including 109 new individuals, 203 individuals retrieved from GenBank) from 18 Chinese domestic goat breeds were used to investigate breed genetic diversity, origin and phylogeography. All goat breeds in this study proved to be extremely diverse, average haplotype diversity and nucleotide diversity being 0.9829 ± 0.0027 and 0.03615 ± 0.03257, respectively. The 312 sequences gave 148 different haplotypes. Phylogenetic analyses revealed that there were two mtDNA haplogroups identified in domestic goats in Southwest China, in which haplogroup A was predominant. Mismatch analysis showed haplogroup A had experienced population expansion events, whereas haplogroup B did not. Amova analysis showed there was no significant geographical structuring. Almost 86.23% of genetic variation was included in the within-breed variance component and only 3.5% was observed among the four geographic provinces. The results of this study contribute to the knowledge of the genetic structure and origin of domestic goats in Southwest China.     

Genetic Variations of 13 Indigenous Chinese Goat Breeds Based on Cytochrome b Gene Sequences

Phylogenetic relationships among and genetic variability within 13 Chinese indigenous goat breeds and Boer goat were analyzed using cytochrome b gene sequences. There were 44 variable sites found in a 642 bp sequence, and 46 Cyt b haplotypes were subsequently defined. The phylogeny analysis of haplotypes in combination with goat Cyt b sequences from GenBank shows that Chinese goats are obviously separated from wild goats and might come from Capra aegagrus. Further analysis indicated that indigenous Chinese goats might descend from at least two lineages; most of the individuals analyzed could be classified into lineage A as defined by Luikart, but five other goats were of uncertain lineage. The Tibet plateau is a possible place of origin for Chinese goats. The neighbor-joining tree based on pairwise differences among populations shows that most Tibetan goats, except the Middle Tibet type, cluster closely with North China goats, and then with South China goats. This result confirms that differences in genetic structure exist among goats in different geographic locations. Nucleotide diversity varied among populations. Tibet and North China goats had higher genetic diversity than South China goats. The fixation index (F _st=87.72%) suggested that most of the total genetic variation was due to variation within populations. In addition, the results indicate that Cyt b gene sequence information alone might not be enough for phylogeny analysis among breeds within species, as shown by fewer polymorphic sites and lower bootstrap values on the neighbor-joining tree.     

Bezoar (Capra aegagrus) Is a Matriarchal Candidate for Ancestor of Domestic Goat (Capra hircus): Evidence from the Mitochondrial DNA Diversity

The leading hypothesis on the ancestor of domestic goats (Capra hircus) is that it is the wild goat called the bezoar or pasang (Capra aegagrus). To verify this hypothesis, we sequenced and compared the cytochrome b gene of mitochondrial DNA from six domestic goats and a bezoar. A further sequence for the markhor was taken from the database. In total we detected 51 nucleotide substitutions among the domestic goats, bezoar and markhor. However, only one specific nucleotide substitution was found between the domestic goats and the bezoar. On the other hand, 43 nucleotide substitutions were specific for the markhor. This result suggested a close relationship between the domestic goats and the bezoar. A neighbor-joining and parsimony phylogenetic tree constructed using the sequences showed that the domestic goats and the bezoar belong to the same cluster, while the markhor showed a distinct cluster separate from that of the domestic/bezoar cluster. This result was confirmed by trees based on the sequence of the mitochondrial displacement loop regions. These results suggest that the strongest candidate for a matriarchal ancestor of domestic goats is the bezoar.     

Mitochondrial diversity and phylogeographic structure of Chinese domestic goats

China has numerous native domestic goat breeds, but so far there has been no extensive study on genetic diversity, population demographic history, and origin of Chinese goats. Here, we examined the genetic diversity and phylogeographic structure of Chinese domestic goats by determining a 481-bp fragment of the first hypervariable region of mitochondrial DNA (mtDNA) control region from 368 individuals representing 18 indigenous breeds. Phylogenetic analyses revealed that there were four mtDNA lineages (A-D) identified in Chinese goats, in which lineage A was predominant, lineage B was moderate, and lineages C and D were at low frequency. These results further support the multiple maternal origins of domestic goats. The pattern of genetic variation in goat mtDNA sequences indicated that the two larger lineages A and B had undergone population expansion events. In a combined analysis of previously reported sequences and our sequences belonging to lineage B, we detected two subclades, in which one was unique to eastern Asia and another was shared between eastern and southern Asia. A larger genetic variation in eastern Asia than southern Asia and the pattern of phylogeographic variation in lineage B suggest that at least one subclade of lineage B originated from eastern Asia. There was no significant geographical structuring in Chinese goat populations, which suggested that there existed strong gene flow among goat populations caused by extensive transportation of goats in history.     

A dig into the past mitochondrial diversity of Corsican goats reveals the influence of secular herding practices

The goat (Capra hircus) is one of the earliest domesticated species ca. 10,500 years ago in the Middle-East where its wild ancestor, the bezoar (Capra aegagrus), still occurs. During the Neolithic dispersal, the domestic goat was then introduced in Europe, including the main Mediterranean islands. Islands are interesting models as they maintain traces of ancient colonization, historical exchanges or of peculiar systems of husbandry. Here, we compare the mitochondrial genetic diversity of both medieval and extant goats in the Island of Corsica that presents an original and ancient model of breeding with free-ranging animals. We amplified a fragment of the Control Region for 21 medieval and 28 current goats. Most of them belonged to the A haplogroup, the most worldwide spread and frequent today, but the C haplogroup is also detected at low frequency in the current population. Present Corsican goats appeared more similar to medieval goats than to other European goat populations. Moreover, 16 out of the 26 haplotypes observed were endemic to Corsica and the inferred demographic history suggests that the population has remained constant since the Middle Ages. Implications of these results on management and conservation of endangered Corsican goats currently decimated by a disease are addressed.     

Phylogenetic analysis of Sicilian goats reveals a new mtDNA lineage

The mitochondrial hypervariable region 1 (HVR1) sequence of 67 goats belonging to the Girgentana, Maltese and Derivata di Siria breeds was partially sequenced in order to present the first phylogenetic characterization of Sicilian goat breeds. These sequences were compared with published sequences of Indian and Pakistani domestic goats and wild goats. Mitochondrial lineage A was observed in most of the Sicilian goats. However, three Girgentana haplotypes were highly divergent from the Capra hircus clade, indicating that a new mtDNA lineage in domestic goats was found.