基于粪便DNA的东北马鹿遗传多样性

东北马鹿(Cervus canadensis xanthopygus)为国家二级重点保护野生动物,近些年其种群数量急剧下降、分布区不断退缩、种群基因交流受阻,很多地区更是难觅其踪迹。亟需对其种群的遗传变化,特别是遗传多样性和近交衰退等种群遗传信息开展进一步评价,增强保护与管理的针对性。本研究在大、小兴安岭和长白山脉的6个重点研究区域,共收集409份疑似马鹿粪便样本。首先基于mtDNA Cyt b基因测序技术进行物种鉴定,并对鉴定为马鹿的阳性样本利用微卫星技术进行个体识别。结果共识别出172只东北马鹿个体;Cyt b基因序列共检测出14个变异位点和11个单倍型,单倍型多样性为0.849 (0.105～0.732),核苷酸多样性为0.678%(0.099%～0.775%)。10个微卫星位点检测出种群平均等位基因数为5.7 (5.2～7.2),有效等位基因数为3.3 (2.5～4.1),观测杂合度为0.687 (0.644～0.725),期望杂合度为0.619 (0.564～0.689),近交系数为-0.113 (-0.160～-0.037)。结果表明,东北马鹿种群遗传多样性处于中等水平,其...     

东北梅花鹿野生种群分布与 遗传变异的分子鉴定

梅花鹿东北亚种(Cervus nippon hortulorum)曾被认为已野外灭绝,近年来在黑龙江东南部和吉林东部临近边境地区发现少量分布,其生境隔离、面积狭小,破碎化严重。亟需对其种群的遗传变化,特别是遗传多样性和近交衰退等种群遗传信息开展进一步评价,增强保护与管理的针对性。本研究在大、小兴安岭和长白山设计9个重点研究区域,共收集673份疑似梅花鹿粪样样本。首先基于线粒体DNA Cyt b基因测序技术开展物种鉴定,并对鉴定为梅花鹿的阳性样本利用微卫星技术进行个体识别。结果证实,东北梅花鹿仅在老爷岭东部山脉有分布,106份梅花鹿粪便DNA中识别出33只个体(穆棱保护区20只,老爷岭保护区13只)。33个Cyt b基因序列共检测出6个变异位点和5个单倍型,单倍型多样性指数(Hd)为0.621,核苷酸多样性指数(Pi)为0.006 7;微卫星检出种群平均等位基因数(Na)7.1个,观测杂合度(Ho)0.604,期望杂合度(He)0.712,固定系数(Fis)0.152。结果表明,东北梅花鹿种群遗传多样性丰富,但也存在一定程度的杂合度不足和近亲繁殖;种群近期经历了瓶颈效应,未发生种群扩张;群体间无遗传分化,可作为一个管理单元加以保护。建议,对东北梅花鹿稀有单倍型个体重点监测和保护,恰当时期考虑圈养种群野外放归,以提高野外个体间基因交流和快速种群恢复。     

黑龙江省完达山东部林区马鹿种群遗传多样性的微卫星分析

黑龙江省完达山东部林区是东北马鹿(Cervus elaphus xanthopygus)种群密度较高的分布区之一,本文对2年冬季采集的167份马鹿粪便进行了7个微卫星座位的个体识别,评价了马鹿种群遗传多样性,并分析近期马鹿数量的急剧下降对种群遗传结构的影响。结果表明:167份粪便DNA分属66只个体;种群平均等位基因数9.00±2.77;平均有效等位基因数3.97±0.99;平均多态信息含量0.69±0.09;平均期望杂合度0.74±0.08;平均观察杂合度0.69±0.08。整个种群显著偏离Hardy-Weinberg平衡,但种群固定系数Fis为0.060,没有显著偏离零。可见,完达山东部林区马鹿种群遗传多样性较高,数量的急剧下降近期还没有表现在种群遗传结构上。     

三江源和祁连山国家公园雪豹种群的遗传结构分析

掌握遗传信息对濒危物种的保护和管理具有重要意义。本研究在我国雪豹重要分布区祁连山和三江源国家公园分别采集粪便样品,利用mtDNA的cyt b基因、微卫星多态性位点进行了雪豹的物种鉴定、个体识别和种群遗传结构评估。在采集286份疑似雪豹粪便样品中,成功的对86份雪豹样品进行了扩增鉴定,利用微卫星位点进行个体识别获得41只雪豹个体,其中祁连山国家公园26只,三江源国家公园15只。通过等位基因数、有效等位基因数、观测杂合度、期望杂合度、多态信息含量等指标进行种群遗传多样性评估,认为雪豹种群遗传多样性相对较低,但祁连山国家公园雪豹种群遗传多样性相对较高。STRUCTURE进行群体遗传结构分析表明,4个种群可以划分为3个遗传类群,祁连山国家公园的种群（YCW和QLS）与三江源国家种群(DC和SJ)的遗传差异,可能与种群间的地理隔离存在明显的相似性。     

基于粪便DNA的马鹿种群数量和性比

为分析黑龙江省完达山林区马鹿种群生存状态,制定科学有效地保护措施,从分子水平研究了种群数量和性比。2006、2007两年冬季跟踪马鹿足迹链,于五泡林场共搜集210份粪便,以成功提取DNA的167份作为分析样本。通过多态性较高的7个微卫星位点进行了基因分型分析,显示167份粪便DNA分属66只个体。基于非损伤性标志重捕法,统计出林场内马鹿数量2a平均47(39—60)只,密度0.302(0.251—0.386)只/km2,与2002年大样方法调查结果相比有减无增。SRY基因性别鉴定显示,种群雌雄性比1.00∶2.00(22♀,44♂),存在较多雄性个体,分析认为偷猎是导致性比失衡的最主要原因。数量的持续下降和性比失衡提示完达山林区马鹿种群数量的恢复需要更好地保护工作。     

川金丝猴遗传多样性研究进展

川金丝猴目前处于生境破碎、种群隔离的濒危境地,对其进行遗传多样性研究是有效保护这一物种的必要前提.结合文献,比较了川金丝猴遗传研究中常用样品采集方法、标记方法等,总结了其遗传多样性的研究现状,并建议通过建立遗传谱系来避免或减少近交,建立生态走廊促进群间基因交流,以提高其遗传变异能力.     

马鹿阿拉善亚种遗传多样性和群体结构

马鹿阿拉善亚种（Cervus elaphus alashanicus）又称阿拉善马鹿,目前仅分布于贺兰山地区,是我国马鹿亚种分布范围最狭窄的一个隔离种群。为了解阿拉善马鹿的种群遗传多样性及遗传变异情况,以对该种群的保护提供科学参考,对在贺兰山采集的93个野生个体新鲜粪便样本的线粒体控制区部分序列（991 bp）进行扩增和分析,共检测到68个变异位点,定义16种单倍型,平均单倍型多样性为0.405,平均核苷酸多样性为0.00232,说明种群遗传多样性水平较低。中性检验和错配分布分析表明阿拉善马鹿曾出现过种群扩张,贝叶斯天际线分析（BSP）显示扩张时间约在末次冰盛期（0.028—0.010 Ma）。FST检验表明阿拉善马鹿种群内存在显著遗传分化,系统发育树和单倍型网络图分析表明群体间没有明显的系统地理格局。本研究表明阿拉善马鹿目前种群遗传多样性较低,建议加大对该亚种的关注和保护力度。       

大兴安岭南段马鹿生境适宜性分析与生态廊道构建

构建生态廊道在缓解生境破碎化对生物多样性的影响、维持濒危物种的遗传多样性、维护自然生态系统结构完整与功能稳定方面具有重要作用。以内蒙古大兴安岭南段分布的马鹿(Cervus elaphus)种群为研究对象,利用MaxEnt模型对其生境适宜性进行分析,并利用最小累积阻力模型构建潜在生态扩散廊道,探讨大兴安岭南段区域隔离马鹿种群的栖息地连通方案。结果显示,马鹿栖息地呈破碎化状态,种群有明显的隔离分布趋势,现有适宜栖息地具有海拔较低(800—1200 m)、坡度较缓(<15°)、靠近水源、植被类型多为靠近山林的灌丛或草地等特点。所构建12条生态廊道具有经过河流浅水节段、远离村落等特点,便于落实栖息地生态恢复管理措施。研究从区域尺度综合分析了大兴安岭南段马鹿栖息地现状及连通性,有助于优化适宜栖息地格局,促进马鹿扩散和栖息地连通,为该物种隔离种群及其栖息地保护规划提供现实指导和基础资料。     

卧龙圈养大熊猫遗传多样性现状及预测,

以中国最大的大熊猫圈养种群—四川卧龙中国大熊猫保护中心的圈养种群为对象,以８个大熊猫微卫星位点为分子标记, 探讨了大熊猫圈养种群的遗传多样性, 并与邛崃野生种群及其他７个濒危物种进行比较。微卫星数据表明, 圈养种群的遗传多样性水平(A＝5.5, He ＝0.620, Ho＝0.574) 低于邛崃野生种群(A＝9.8,He＝0.779,Ho＝0.581),但高于其他7 个濒危物种的种群(He＝0.13~0.46)。在此数据的基础上对未来100个世代内圈养种群遗传多样性的变化情况做出了预测。结果表明假设种群数量比现在扩大一倍, 经历100个世代后也只会使平均等位基因数少减少0.4。因此继续增加野生个体对保持遗传多样性的意义已经不大, 建议该圈养种群的保护策略应将重点放到制定更有效的繁殖计划以避免近交上。     

应用微卫星标记分析圈养大熊猫遗传多样性

以来源于成都大熊猫繁育研究基地和中国保护大熊猫研究中心的34只圈养大熊猫(分为a群体和b群体)和7只圈养野生大熊猫(圈养野生群)作为研究对象,利用AY161177～AY161218、Ame-μ5～Ame-μ70和g001～g905等30个微卫星标记对其遗传多样性现状进行分析,并探讨保持圈养大熊猫遗传多样性的方法.微卫星数据表明,30个微卫星标记多态性好(PIC=0.621～0.640),圈养大熊猫遗传多样性水平(a群体:A=5.48,Ho=0.475,He=0.696;b群体:A=5.24,Ho=0.453,He=0.719;圈养野生群:A=3.80,Ho=0.514,He=0.725)高于6个濒危物种(Ho=0.210～0.390,He=0.150～0.430)但低于3个非濒危物种(Ho=0.620～0.710),圈养大熊猫遗传多样性水平都保持在较高水平,但圈养群遗传多样性水平与圈养野生群相比有所降低.F统计量及基因流Nm分析结果证明,a、b两群体间遗传分化程度不高(Nm=2.610,Fst=0.0874,Fit=0.4116),存在个体交换和一定程度的近交,b群体近交程度高于a群体(a群体Fis=0.3221,b群体Fis=0.3983).因此,现阶段圈养大熊猫的管理重点是避免近交和遗传多样性丧失,将圈养大熊猫种群作为同一管理单元,把纠正大熊猫系谱中的错误、科学选择大熊猫个体进行群体间交流作为关键点,利用微卫星技术保持和提高大熊猫种群的遗传多样性水平.     

Genetic analysis of an isolated red deer (<Emphasis Type="Italic">Cervus elaphus</Emphasis>) population showing signs of inbreeding depression

In the present study, we analysed 18 red deer specimens from a small (N = 50) and isolated population in Schleswig-Holstein, northern Germany, with respect to variability at nine polymorphic microsatellite loci and 439 bp of the mitochondrial DNA control region. Several cases of brachygnathy (shortened lower jaw), commonly associated with inbreeding depression, have been recorded in the population. Genetic variability was very low compared with other European red deer populations including the neighbouring population from which the population under study was derived some 130 years ago. The effective population size was estimated to be seven individuals corresponding to an increase in inbreeding (or a loss of heterozygosity) of 7% each generation. This value is seven times higher than the theoretical threshold level up to which natural selection is believed to counteract the fixation of deleterious alleles in the gene pool. As a consequence, the population urgently needs genetic input from other populations to overcome the negative effects of random drift and inbreeding. To our knowledge, this study is one of the first to genetically analyse a red deer population showing strong signs of inbreeding depression.     

Past bottlenecks and current population fragmentation of endangered huemul deer (<Emphasis Type="Italic">Hippocamelus bisulcus</Emphasis>): implications for preservation of genetic diversity

Small populations in fragmented habitats can lose genetic variation through drift and inbreeding. The huemul (Hippocamelus bisulcus) is an endangered deer endemic to the southern Andes of Chile and Argentina. Huemul numbers have declined by 99% and its distribution by 50% since European settlement. The total population is estimated at less than 2,000 individuals and is highly fragmented. At one isolated population in Chilean Patagonia we sampled 56 individuals between 2005 and 2007 and genotyped them at 14 microsatellite loci. Despite low genetic variability (average 2.071 alleles/locus and average H _O of 0.341), a low inbreeding coefficient (F _IS) of 0.009 suggests nearly random mating. Population genetic bottleneck tests suggest both historical and contemporary reductions in population size. Simulations indicated that the population must be maintained at 75% of the current size of 120 individuals to maintain 90% of its current genetic diversity over the next 100 years. Potential management strategies to maintain genetic variability and limit future inbreeding include the conservation and establishment of habitat corridors to facilitate gene flow and the enlargement of protected areas to increase effective population size.     

Combining genetic structure and demographic analyses to estimate persistence in endangered Key deer (<Emphasis Type="Italic">Odocoileus virginianus clavium</Emphasis>)

Recent improvements in genetic analyses have paved the way in using molecular data to answer questions regarding evolutionary history, genetic structure, and demography. Key deer are a federally endangered subspecies assumed to be genetically unique, homogeneous, and have a female-biased population of approximately 900 deer. We used 985 bp of the mitochondrial cytochrome b gene and 12 microsatellite loci to test two hypotheses: (1) that Key deer are isolated and have reduced diversity compared to mainland deer and (2) that isolation of the Florida Keys has led to a small population size and a high risk of extinction. Our results indicate that Key deer are indeed genetically isolated from mainland white-tailed deer and that there is a lack of genetic substructure between islands. While Key deer exhibit reduced levels of genetic diversity compared to their mainland counterparts, they contain enough diversity to uniquely identify individual deer. Based on genetic identification, we estimated a census size of around 1000 individuals with a heavily skewed female-biased adult sex ratio. Furthermore, our genetic and contemporary demographic data were used to generate a species persistence model of the Key deer. Sensitivity tests within the population viability analysis brought to light the importance of fetal sex ratio and female survival as the primary factors at risk of driving the subspecies to extinction. This study serves as a prime example of how persistence models can be used to evaluate population viability in natural populations of endangered organisms.     

Genetic assessment of captive Chinese water deer populations in Zhoushan Archipelago,China

Chinese water deer is a rare and vulnerable animal in China because of the poaching for medical use and the habitat loss. In this study, the genetic diversity and population genetic structure of 40 Chinese water deer from three populations in Zhoushan Archipelago were investigated with ten highly polymorphic microsatellite loci, including 4 screened from the nuclear DNA in the study, and 6 selected from the literature. According to the results, these captive populations had a higher genetic diversity than other rare cervid species, such as forest musk deer. No signs of inbreeding were detected. Low genetic differentiation among these populations was found. The probable reasons included the isolation by distance, the exchange among islands, or the supplement of the wild Chinese water deer. We proposed the deer raisers to strengthen the exchanges from different islands or mainland, and if possible, some deer would be returned to the wild to expand the wild population.     

Landscape features affect gene flow of Scottish Highland red deer (Cervus elaphus)

Landscape features have been shown to strongly influence dispersal and, consequently, the genetic population structure of organisms. Studies quantifying the effect of landscape features on gene flow of large mammals with high dispersal capabilities are rare and have mainly been focused at large geographical scales. In this study, we assessed the influence of several natural and human-made landscape features on red deer gene flow in the Scottish Highlands by analysing 695 individuals for 21 microsatellite markers. Despite the relatively small scale of the study area (115 × 87 km), significant population structure was found using F -statistics ( F _ST = 0.019) and the program structure , with major differentiation found between populations sampled on either side of the main geographical barrier (the Great Glen). To assess the effect of landscape features on red deer population structure, the ArcMap GIS was used to create cost-distance matrices for moving between populations, using a range of cost values for each of the landscape features under consideration. Landscape features were shown to significantly affect red deer gene flow as they explained a greater proportion of the genetic variation than the geographical distance between populations. Sea lochs were found to be the most important red deer gene flow barriers in our study area, followed by mountain slopes, roads and forests. Inland lochs and rivers were identified as landscape features that might facilitate gene flow of red deer. Additionally, we explored the effect of choosing arbitrary cell cost values to construct least cost-distance matrices and described a method for improving the selection of cell cost values for a particular landscape feature.     

Recovery process of genetic diversity through seed and pollen immigration at the northernmost leading-edge population of Fagus crenata

The range expansion of a plant species begins with colonization of ecological empty patches from posterior source populations. This process involves stochastic loss of genetic diversity. However, the founder population could restore genetic diversity by gene flow from posterior populations via seeds and pollen and its recovery affects evolutionary potential for species expansion. To clarify the recovery process of genetic diversity during species range expansion, gene flow via seeds and pollen was investigated at the expansion front of Fagus crenata. Based on eight nuclear microsatellite genotypes of a total of 150 individuals and 225 seeds at the northernmost leading-edge population, genetic diversity, fine-scale spatial genetic structure (FSGS), and genetic differentiation from other five northern populations were investigated. Moreover, both seed and pollen immigration and their effects on genetic diversity at different successional stages were analyzed. The leading-edge population showed lower genetic diversity and substantial genetic differentiation, reflecting its strong genetic drift. Non-significant FSGS and a negative inbreeding coefficient for mature trees may indicate that the earliest generation consisted of founders from foreign seed sources. The significant proportion of seed and pollen immigration increased the number of different alleles for later successional stages. The effective number of pollen parents from foreign sources (20.8) was markedly higher than that from the local source (2.1). These results indicated that pollen immigration incorporated new and rare alleles and increased the genetic diversity of the population. However, the proportion of foreign gene flow decreased during succession, probably due to the increased reproductive success of local individuals as they reached maturity and grew in size.     

Genetic diversity, gene flow and drift in Bavarian red deer populations (Cervus elaphus)

The red deer (Cervus elaphus) populationof Bavaria in Southern Germany was severelyreduced during the 19th century due toover-hunting. The species has since recoveredwithin designated areas. Subsequent habitatfragmentation presumably has changed thegenetic structure of Bavarian red deer.In order to assess the genetic diversity, weanalysed samples obtained from nine differentBavarian and two adjacent (Thueringen andCzech-Republic) red deer populations,genotyping 19 microsatellite loci. Our analysesrevealed moderate and significant differencesin diversity. Referring to assignment tests,the genetic differentiation of Bavarian reddeer was sufficient to assign an individual'sorigin to the correct population at an averageof 91.6%. The correlation of genetic andgeographic distance matrices revealed noevidence for isolation by distance. Thecoalescent model analysis suggests that thegenetic structure used to be characterized by adrift–gene flow equilibrium and is nowinfluenced by drift and disruption of the geneflow. Only three of the examined populationsshowed a probability of less than 10% that twogenes within these populations share a commonancestor (F _IS-value). Twopopulations had high F _IS values,indicating the influence of drift.Additionally, the intrapopulation indicesrevealed a low variability in thesepopulations. The estimated effective populationsizes (N _e) were generally in thesame range as the actual population sizes. Theinbreeding rates, based on the estimated N _e, and the inbreeding coefficientssuggested that the Bavarian red deerpopulations are in a stable state.     

Biochemical and DNA markers yield strikingly different results regarding variability and differentiation of roe deer (Capreolus capreolus, Artiodactyla: Cervidae) populations from northern Germany

Three mainland and two island roe deer ( Capreolus capreolus ) populations with a total sample size of 105 individuals from Schleswig–Holstein, northern Germany, were analysed with regard to genetic variability within and differentiation among populations as revealed by eight allozyme loci known to be polymorphic in roe deer, eight microsatellite loci and 404 bp of the mitochondrial control region. Surprisingly, the allozymes were completely monomorphic, but microsatellite and control region variability were high. Hypotheses as to demographic reasons for the variability patterns found, including bottlenecks, founder effects and translocations, are put forward. There were no statistically significant differences between the island and the mainland populations in terms of genetic variability as measured by expected heterozygosity, inbreeding coefficient and allelic richness. The correlations of the various variability indices were not statistically significant after Bonferroni correction. Nevertheless, there was a clear tendency for differentiation indices to yield concordant results for microsatellite and mitochondrial markers.     

Genetic structure of red deer population in northeastern Poland in relation to the history of human interventions

We studied the genetic structure of a red deer (Cervus elaphus) population in 8 woodlands of northeastern Poland and 1 in western Belarus and compared it with the documented history of the population in the region. Red deer nearly went extinct in the region in the 18th and 19th centuries. In the mid-19th century, reintroductions began and continued until the mid-1960s. Animals were translocated from various sites in Poland and other European countries. We genotyped 303 individuals using 14 microsatellite loci and sequenced 253 individuals for a fragment of the control region (mitochondrial DNA [mtDNA]). The microsatellite analyses demonstrate that 3 genetically separate subpopulations exist, but 4 according to mtDNA. All haplotypes found in northeastern Poland are closely related to haplotypes from northern and northwestern Europe. The only individuals that could have originated from autochthonous red deer populations, rather than introductions, were found in Napiwoda Forest. The present regional genetic structure of the species is consistent with the known history of red deer translocations. Current patterns of genetic diversity in these populations are determined by the interaction of past human management and contemporary natural migrations. © 2012 The Wildlife Society.