云南4个地区高山姬鼠线粒体细胞色素b遗传分化的研究

基于云南昆明、中甸、丽江和剑川4个种群(n=43)的线粒体细胞色素b(Cyt b)基因全序列的遗传变异分析,探讨了该地区高山姬鼠种群的遗传分化。在1 140 bp Cyt b基因序列中,有50个变异位点(占全变异的4.38%),定义了22个单倍型。在4个种群中,昆明种群单倍型多样性和核苷酸多样性最高。分子变异分析表明,种群间的遗传变异占30.2%,种群内的遗传变异占69.8%。FST分析表明,除中甸种群和丽江种群之间差异不显著(P>0.05),其它种群间的差异均极显著(P<0.01)。22个单倍型在系统发生树中明显聚为两支(A和B),进化网络关系显示昆明种群处于进化分支最末端,推测种群进化方向可能从横断山地区到昆明地区,支持了姬鼠属从北向南扩散的理论。     

长江上游干流及赤水河蛇遗传多样性与种群历史分析

通过分析76尾来自长江上游干流宜宾(14尾)、合江(30尾)及支流赤水河(32尾)蛇(Saurogobio dabryi)种群的线粒体细胞色素b(Cyt b)基因序列,研究蛇3个地理种群的遗传多样性及种群历史。用于分析的Cyt b序列长1 097 bp,含变异位点28个,其中简约信息位点18个。76尾个体共检测到26个单倍型,整体呈现较高的单倍型多样性(Hd=0.872)和较低的核苷酸多样性(Pi=0.004 0)。三个种群共享较多的单倍型。基于单倍型构建的系统发育树及NETWORK网络关系图显示:所有来自长江上游干流和赤水河的单倍型不能按照地理分布各自聚类,而是相互混杂聚在一起;网络关系图呈星状分布,未检测到原始单倍型和进化中心。此外,基于单倍型频率分析得到的三个种群间的遗传分化指数FST值较低(分别为﹣0.0293、0.0280和0.0258)。分子方差分析(AMOVA)显示,整体上长江上游干流及赤水河蛇种群变异主要来源于种群内,种群内的变异占总变异的80.00%,表明长江上游干流及赤水河蛇种群属于同一种群,蛇各地理种群间基因交流频繁。中性检验、错配分析及BSP(Bayesian skyline plot)分析显示,整体上长江干流及赤水河蛇种群在距今0~0.025 Ma(百万年)期间发生过种群扩张现象。     

基于COⅠ基因探讨北京及其周边地区跳钩虾种群遗传结构

跳钩虾Platorchestia japonica栖息于湖泊、河流岸边,是重要的环境指示生物。本研究以线粒体COⅠ基因片段为分子标记,对北京及其周边地区22个采样点的128个样本进行种群遗传多样性和遗传结构研究。结果显示,623 bp的COⅠ基因序列中有567个保守位点、56个变异位点和38个简约信息位点。128个样本检测到43个单倍型,单倍型多样性为0.938,核苷酸多样性为0.011 72。最大似然法和贝叶斯法构建的系统发育树以及单倍型网络图表明,研究区域跳钩虾没有明显的地理种群结构,但所有单倍型形成2个遗传进化支。分子变异分析结果证实,跳钩虾2个进化支间的遗传变异显著高于进化支内的,进化支间固定系数为0.852 19,表明2个进化支遗传分化明显。本研究为进一步研究中国区域跳钩虾的遗传结构提供了有意义的基础数据。     

海陵岛近岸2种虾虎鱼线粒体COⅠ基因和D-loop区序列变异及遗传分析

以海陵岛近岸优势物种舟山缰虾虎鱼(Amoya chusanensis)、纵带鹦虾虎鱼(Exyrias puntang)为研究对象,获取2种虾虎鱼的线粒体COⅠ基因序列45条,控制区(D-loop)序列48条,分析其序列变异情况和遗传特征。结果表明:获得的638 bp COⅠ序列中,舟山缰虾虎鱼存在变异位点20个,纵带鹦虾虎鱼变异位点占18个。467 bp控制区中,舟山缰虾虎鱼变异位点10个,纵带鹦虾虎鱼占19个。碱基组成上,2种虾虎鱼COⅠ基因和控制区序列均存在A+T含量大于C+G含量现象,且COⅠ基因序列中碱基T偏向密码子第一位。最大似然法估算的COⅠ序列转换颠换R比值为2.89,控制区序列R值为2.66。聚类分析揭示舟山缰虾虎鱼在进化上较原始,更接近祖先物种,而纵带鹦虾虎鱼种群分化较晚。单倍型分析指明2种虾虎鱼单倍型多样性均处在较高水平。中性检验的Fu-Li D分析发现,基于COⅠ基因和控制区序列的舟山缰虾虎鱼的检验结果均显著,不符合中性进化假设,纵带鹦虾虎鱼Fu-Li D test的结果均呈现不显著,符合中性进化假设。舟山缰虾虎鱼种群间可能存在群体扩张或持续增长现象。     

基于线粒体Cyt b基因序列变异的克孜河塔里木裂腹鱼和斑重唇鱼遗传多样性

采用线粒体细胞色素b基因(Cyt b)序列,分析了采自新疆克孜河3个群体(斯木哈纳SM、牙师YS、卡拉贝利KL)的塔里木裂腹鱼(Schizothorax biddulphi)41尾个体及1个斑重唇鱼(Diptychus maculates)群体(斯木哈纳)23尾个体的种群遗传多样性和遗传结构.结果显示,塔里木裂腹鱼检测到6个碱基变异位点,定义了4种单倍型,平均单倍型多样性指数及核苷酸多样性指数分别为0.525 4和0.001 16.分子变异分析(AMOVA)结果提示,塔里木裂腹鱼的遗传变异全部发生于群体内部;群体间Kimura-2-parameter遗传距离、分化指数(F<,st><0.085 25)和基因流(N<,m>>3.18)都显示3个群体没有种群分化,属于单一种群.斑重唇鱼检测出7个变异位点,定义了8个单倍型,平均单倍型多样性指数与核苷酸多样性指数分别为0.830 1和0.001 13.研究表明,克孜河的塔里木裂腹鱼和斑重唇鱼均处于很低的遗传多样性水平,物种维持力较弱.     

安徽长江流域黄鳝6个地理种群的遗传变异研究

采用线粒体细胞色素b(Cyt b)为分子标记,研究了安徽长江流域黄鳝6个地理种群(当涂、无为、繁昌、贵池、怀宁和望江)共180尾个体的遗传变异关系。结果显示,在1087 bp序列中共检测到变异位点101个,单倍型68个,碱基组成中A+T的含量大于G+C的含量。6个地理种群的单倍型多样性(0.623~0.940)较高,核苷酸多样性(0.001 78~0.019 04)较低。群体分化指数(Fst:0.026 12~0.947 07)、基因流(Nm:0.027 94~18.6400)和分子变异分析(AMOVA)结果表明,安徽长江流域黄鳝一些地理种群间存在着明显的遗传分化。单倍型系统进化树和进化网络图表明:6个地理种群分为2个大的进化支,当涂与繁昌种群为一支,其余4个种群为另一支。地理隔离和黄鳝有限的迁移能力可能是造成种群遗传分化的原因。     

喜马拉雅旱獭线粒体DNA控制区遗传多样性及系统发育

喜马拉雅旱獭是青藏高原的优势种,数量多、分布广,全面了解其遗传背景对该地区旱獭资源的保护与合理利用具有重要的意义。本研究以青藏高原云南、西藏和青海三省区共13个地理种群计258只旱獭为研究对象,PCR扩增获得线粒体DNA控制区基因部分序列(887 bp),并运用种群遗传学方法进行遗传多样性分析。结果显示:258份样品共发现了84个变异位点(9.40%),定义了68种单倍型,其单倍型多样性(h)平均值为0.968±0.003、核苷酸多样性(π)平均值为0.017 25±0.016 37,种群总体遗传多样性较高。AMOVA方差分析显示13个地理种群间存在着明显的遗传分化(Fst=0.620 67,P<0.001),种群间基因交流多数较低(Nm<1)。基于单倍型构建的系统发育树中13个地理种群的喜马拉雅旱獭聚为两支,其中来自青藏高原西南地区(西藏安多、青海格尔木、青海囊谦、云南迪庆)的18个单倍型聚成一个大的分支(A支),其余50个单倍型聚为一个大的分支(B支),在NETWORK网络图中也可见到相似网络拓扑结构。研究结果显示青藏高原喜马拉雅旱獭种群以唐古拉山脉为界分为两个大的种群,说明地理隔离是影响喜马拉雅旱獭种群动态变化的主要因素。     

江西井冈山地区灰胸竹鸡的遗传多样性研究

灰胸竹鸡Bambusicola thoracica是我国特有鸟类.本文采用聚合链式反应和直接测序的方法测定井冈山地区灰胸竹鸡3个种群线粒体DNA(mtDNA)控制区1142 bp的序列,分析其序列变异和种群遗传多样性.30个样本共发现16个变异位点和10种单倍型,其中hapl广泛分布,占所分析样本的23.33%,是其祖先单倍型.3个种群的平均单倍型多样性和核苷酸多样性分别为0.815和0.00243.青原区种群与其它两个种群遗传分化显著,基因交流受限制.受隔离影响,青原区种群遗传多样性最低.在系统发生树上,10种单倍型形成两支,井冈山种群和永新县种群聚在一起,与其地理位置相一致.     

长江上游干流及赤水河蛇鮈遗传多样性与种群历史分析

通过分析76尾来自长江上游干流宜宾(14尾)、合江(30尾)及支流赤水河(32尾)蛇(鮈)(Saurogobio dabryi)种群的线粒体细胞色素b(Cyt b)基因序列,研究蛇(鮈)3个地理种群的遗传多样性及种群历史.用于分析的Cyt b序列长1 097 bp,含变异位点28个,其中简约信息位点18个.76尾个体共检测到26个单倍型,整体呈现较高的单倍型多样性(Had=0.872)和较低的核苷酸多样性(Pi=0.004 0).三个种群共享较多的单倍型.基于单倍型构建的系统发育树及NETWORK网络关系图显示:所有来自长江上游干流和赤水河的单倍型不能按照地理分布各自聚类,而是相互混杂聚在一起;网络关系图呈星状分布,未检测到原始单倍型和进化中心.此外,基于单倍型频率分析得到的三个种群间的遗传分化指数FST值较低(分别为-0.0293、0.0280和0.0258).分子方差分析(AMOVA)显示,整体上长江上游干流及赤水河蛇(鮈)种群变异主要来源于种群内,种群内的变异占总变异的80.00％,表明长江上游干流及赤水河蛇(鮈)种群属于同一种群,蛇(鮈)各地理种群间基因交流频繁.中性检验、错配分析及BSP(Bayesian skyline plot)分析显示,整体上长江干流及赤水河蛇(鮈)种群在距今0～ 0.025 Ma(百万年)期间发生过种群扩张现象.     

三化螟COⅠ序列分析及DNA条形码研究

三化螟Scirpophaga incertulas是水稻的重要害虫之一。本研究通过对三化螟3个地理种群线粒体COⅠ基因的序列分析,初步探讨了中国三化螟种群间的遗传分化。36个三化螟样本共得到了9个单倍型,其中有17个变异位点,占总位点数的3.13%。系统进化分析表明,来源于各地理种群的单倍型散布在不同的进化支上,缺乏明显的地理分布格局。联合网上公布的相关物种序列进行的遗传分析表明,线粒体COⅠ基因能够对三化螟进行准确的物种鉴定。     

中国内蒙古鄂伦春族线粒体DNA高变区Ⅰ和高变区Ⅱ遗传多态性

收集50份鄂伦春族无关人群外周血样本, 用ABI PRISM377测序仪对其mtDNA HVRⅠ和HVRⅡ进行测序, 计算多态性位点数、单倍型数目、单倍型频率、平均核苷酸差异数目等多态性指标; 结合已发表的其他民族mtDNA遗传资料, 根据Nei法计算鄂伦春族与各群体之间的遗传距离, 进行聚类分析, 绘制系统发生树。鄂伦春族群体mtDNA两个高变区与CRS序列比对, 分别发现52和24个多态性位点, 分别界定了38和27种单倍型, 单倍型多态性分别为0.964±0.018和0.929±0.019; 平均核苷酸差异分别为7.379和2.408; 用HVRⅠ序列多态性数据计算Fst和dA两种遗传距离, 相关系数r为0.993(P<0.01); 基于HVRⅠ序列的系统树显示鄂伦春族与中国台湾、南方汉族和中国香港人群遗传距离较近, 与北方汉族、蒙古族及其国外人群遗传距离相对较远。我国鄂伦春族人群mtDNA具有相对独特的遗传特征, 其遗传多态性和个体识别力较高, 可用于民族起源、迁徙、法医学个体识别等领域研究。     

Mitochondrial DNA coding region sequences support the phylogenetic distinction of two Indian wolf species

Two small endangered populations of Indian wolves were recently shown to be distant from other wolf and dog mtDNA lineages characterized so far. None of the inner branches in the tree of canid species based on partial hypervariable D-loop sequences were, however, statistically supported by the data raising the question whether the two Indian wolf lineages represent two new species, occupying an intermediate position between Canis latrans and C. lupus or have diverged from the sub-species of C. lupus due to isolation and drift. Here we report complete D-loop, cytochrome b, and 16S rRNA sequences data for 23 additional wolves from India analysed in the context of other canid species. Extended analyses of D-loop data and partial sequences of 16S rRNA showed highly reticulated pattern and were unable to resolve unambiguously the phylogenetic relationship of Indian wolves among other canid species. The phylogenetic reconstructions of cytochrome b sequences, however gave significant statistical support for the inner branches supporting genetic distinction of the two Indian wolf lineages within themselves as well as from all other wolves of the world, including individuals belonging to subspecies C. lupus chanco and C. lupus pallipes to which the two Indian wolf populations have been traditionally assigned. Their genetic differentiation relative to worldwide variation of wolves supports the suggestion to treat them as separate wolf species, C. himalayensis and C. indica .     

A new mitochondrial haplotype confirms the distinctiveness of the Italian wolf (Canis lupus) population

In the past century the Italian wolf has been repeatedly indicated as a distinct subspecies, Canis lupus italicus, due to its unique morphology and its distinctive mtDNA control region (CR) monomorphism. However, recent studies on wolf x dog hybridization in Italy documented the presence of a second mtDNA CR haplotype (W16), previously found only in wolves from Eastern Europe, casting doubts on the genetic uniqueness of the Italian wolves. To test whether this second haplotype belongs to the Italian wolf population, we genotyped 92 wolf DNA samples from Italy, Slovenia, Greece and Bulgaria at four mtDNA regions (control-region, ATP6, COIII and ND4 genes) and at 39 autosomal microsatellites. Results confirm the presence of two mtDNA multi-fragment haplotypes (WH14 and WH19) in the Italian wolves, distinct from all the other European wolves. Network analyses of the multi-fragment mtDNA haplotypes identified two strongly differentiated clades, with the Italian wolf WH14 and WH19 multi-fragment haplotypes rooted together. Finally, Bayesian clustering clearly assigned all the wolves sampled in Italy to the Italian population, regardless of the two different multi-fragment haplotypes. These results demonstrate that the W16 CR haplotype is part of the genetic pool of the Italian wolf population, reconfirming its distinctiveness from other European wolves. Overall, considering the presence of unique mtDNA and Y-linked haplotypes, the sharply different frequencies of genome-wide autosomal alleles and the distinct morphological features of Italian wolves, we believe that this population should be considered a distinct subspecies.     

Ancient wolf lineages in India

All previously obtained wolf (Canis lupus) and dog (Canis familiaris) mitochondrial (mt) DNA sequences fall within an intertwined and shallow clade (the 'wolf-dog' clade). We sequenced mtDNA of recent and historical samples from 45 wolves from throughout lowland peninsular India and 23 wolves from the Himalayas and Tibetan Plateau and compared these sequences with all available wolf and dog sequences. All 45 lowland Indian wolves have one of four closely related haplotypes that form a well-supported, divergent sister lineage to the wolf-dog clade. This unique lineage may have been independent for more than 400,000 years. Although seven Himalayan wolves from western and central Kashmir fall within the widespread wolf-dog clade, one from Ladakh in eastern Kashmir, nine from Himachal Pradesh, four from Nepal and two from Tibet form a very different basal clade. This lineage contains five related haplotypes that probably diverged from other canids more than 800,000 years ago, but we find no evidence of current barriers to admixture. Thus, the Indian subcontinent has three divergent, ancient and apparently parapatric mtDNA lineages within the morphologically delineated wolf. No haplotypes of either novel lineage are found within a sample of 37 Indian (or other) dogs. Thus, we find no evidence that these two taxa played a part in the domestication of canids.     

The evolutionary history of grey wolf Y chromosomes

Analyses of Y chromosome haplotypes uniquely provide a paternal picture of evolutionary histories and offer a very useful contrast to studies based on maternally inherited mitochondrial DNA (mtDNA). Here we used a bioinformatic approach based on comparison of male and female sequence coverage to identify 4.7 Mb from the grey wolf (Canis lupis) Y chromosome, probably representing most of the male‐specific, nonampliconic sequence from the euchromatic part of the chromosome. We characterized this sequence and then identified ≈1,500 Y‐linked single nucleotide polymorphisms in a sample of 145 resequenced male wolves, including 75 Finnish wolf genomes newly sequenced in this study, and in 24 dogs and eight other canids. We found 53 Y chromosome haplotypes, of which 26 were seen in grey wolves, that clustered in four major haplogroups. All four haplogroups were represented in samples of Finnish wolves, showing that haplogroup lineages were not partitioned on a continental scale. However, regional population structure was indicated because individual haplotypes were never shared between geographically distant areas, and genetically similar haplotypes were only found within the same geographical region. The deepest split between grey wolf haplogroups was estimated to have occurred 125,000 years ago, which is considerably older than recent estimates of the time of divergence of wolf populations. The distribution of dogs in a phylogenetic tree of Y chromosome haplotypes supports multiple domestication events, or wolf paternal introgression, starting 29,000 years ago. We also addressed the disputed origin of a recently founded population of Scandinavian wolves and observed that founding as well as most recent immigrant haplotypes were present in the neighbouring Finnish population, but not in sequenced wolves from elsewhere in the world, or in dogs.     

Low genetic diversity in the vulnerable Goitred Gazelle,Gazella subgutturosa (Cetartiodactyla: Bovidae), in Iran: potential genetic consequence of recent population declines

The Goitered Gazelle, Gazella subgutturosa, is the most widespread gazelle species in the Middle East and central Asia inhabiting desert and semi-desert habitats. Today it is threatened and its geographic range and population size have experienced significant decline in the last decades. In Iran, the remnant populations are confined to fragmented habitats. We aimed to characterise genetic diversity and phylogenetic status of the populations of Goitered Gazelle in Central Iran and to evaluate the potential effect of a historic population bottleneck on the genetic variation of today’s population. We used noninvasive sampling to uncover structure and level of genetic variation in a fragment of the cytochrome-b gene from 170 samples. Genealogical analyses were performed using HKY+I model and phylogenetic trees reconstructed using Bayesian inference and maximum likelihood. We found extremely low levels of genetic variation, with altogether only five haplotypes in samples from different populations. Overall haplotype diversity was 0.081 and nucleotide diversity 0.0003. The mean observed mismatch between any two sequences was 0.093 with the largest peak for small numbers. The mismatch distribution fit the model of population expansion and suggested that gazelles had experienced a sudden expansion. An unrooted median-joining network analysis of mtDNA haplotypes showed a star-like structure which few mutations steps separating the haplotypes from other regions. Our findings strengthen the urgency of preserving the species’ genetic diversity to prevent local extinction.     

东北地区棕背鼠平种群遗传结构及亚种分类

Using partial sequences of mtDNA control region,we sought genetic diversity,population genetic structure and subspecies classification in 5 populations of Myodes rufocanus from Northeast China and Hokkaido,Japan. After sequencing a 616 bp segment of control region,we detected 45 haplotypes based on 57 variable sites among 55 individuals. Our results revealed that high-level haplotype diversity and slightly less nucleotide diversity occurred in these populations. AMOVA analysis showed that high-level genetic differentiation happened between populations due to 64.70% genetic diversity and Fst ranging from 0. 240 to 0. 814. In a phylogenetic analysis,the haplotypes from Northeast China split into two groups:Daxing’anling-Xiaoxing’anling-Wandashan and Hailin-Dahailin. This suggested that we can separate the populations from Northeast China into two subspecies,M . r. irkutensis and M. r. Changbaishanensis respectively corresponding with the former and latter areas.     

Characterization of cytochrome b diversity in Chinese domestic horses

Previous mitochondrial DNA (mtDNA) D‐loop and microsatellite studies have shown that Chinese horses have multiple maternal origins and high genetic diversity. To better characterize maternal genetic origins and diversity of Chinese domestic horses, we conducted a comprehensive analysis of 407 complete 1140 bp sequences of the horse mitochondrially encoded cytochrome b (CYTB) gene, including 323 horses from 13 Chinese indigenous breeds and 84 reference sequences from GenBank. A total of 114 haplotypes were identified, of which 73 appeared among the 13 Chinese horse breeds. The high mitochondrially encoded cytochrome b haplotypic diversity suggests multiple maternal origins in Chinese horses.     

Phylogenetic relationships among Chinese indigenous goat breeds inferred from mitochondrial control region sequence

Although it is generally accepted that a ‘multiple origins’ hypothesis could explain the origin of Chinese goats, little supportive evidence from mtDNA control region sequencing analysis has been collected. We assessed the phylogenetic relationships among 84 individuals representing 13 Chinese indigenous goat breeds and Boer, using a hypervariable segment of mtDNA control region. A total of 49 haplotypes defined by 85 polymorphic sites were found in the study. Combining with the published mtDNA control region sequences, the phylogenetic analysis classified these goats into four distinctive groups corresponding to lineage A–D, implying that Chinese goats have multiple maternal origins. Qinghai-Tibet Plateau is an important cradle of Chinese indigenous goats. As far as the major lineage A was concerned, it was possibly derived from Tibetan founders and was further subject to domestication in North China; some of them were dispersed to South China while the others remained. The hierarchical analysis indicated that a large percentage (73.9%) of total mtDNA variation existed within populations and a minority was due to differences among geographical types, suggesting that Chinese goats have relatively weak phylogeographic structure.     

Phylogeographic history of grey wolves in Europe

Background  

While it is generally accepted that patterns of intra-specific genetic differentiation are substantially affected by glacial history, population genetic processes occurring during Pleistocene glaciations are still poorly understood. In this study, we address the question of the genetic consequences of Pleistocene glaciations for European grey wolves. Combining our data with data from published studies, we analysed phylogenetic relationships and geographic distribution of mitochondrial DNA haplotypes for 947 contemporary European wolves. We also compared the contemporary wolf sequences with published sequences of 24 ancient European wolves.