生境片段化对千岛湖岛屿上黄足厚结猛蚁遗传多样性的影响

利用相关序列扩增多态性(SRAP)分子标记法,对千岛湖片段化生境中14个岛屿上的黄足厚结猛蚁(Pachycondyla luteipes)种群遗传结构和多样性进行研究。利用5对SRAP引物对42份材料的基因组进行扩增,共得到大小在50-800 bp之间的71个可重复位点,其中63个为多态性位点,多态性比率达88.73%。AMOVA分析结果显示,65.03%的遗传变异存在于种群间,34.97%的遗传变异来自种群内(P < 0.001)。利用PopGene Version 1.32软件对SRAP多态性数据进行分析,不同岛屿种群的多态位点比例和Nei's基因多样性指数变化范围分别介于35.21%-91.55%和0.2662-0.4905之间,平均值分别为58.25%和0.3729,其中多态位点比率最高的岛屿为面积最大的JSE岛。多态位点比例和Nei's基因多样性指数与岛屿面积、海拔均无显著相关性,但与隔离度呈显著正相关关系。种群间遗传分化指数介于0.0777-0.9328之间,平均值为0.4419,基因流值介于0.0360-5.9350之间,平均值为1.0451,种群间遗传分化程度较高,基因流较低。利用UPGMA聚类分析法对14岛屿上的42个个体进行遗传聚类分析,表明地理距离较近的个体和岛屿具有优先聚在一起的趋势。Mantel 检验表明黄足厚结猛蚁各种群间地理距离与遗传距离间存在显著相关性(r=0.7757,P < 0.01)。以上结果表明地理隔离是影响千岛湖黄足厚结猛蚁种群遗传结构和多样性的主要因素。     

基于SSR标记的大明松天然群体遗传多样性分析

大明松是广西和贵州特有的高山松类,具有很高的经济价值和生态价值,其自然种群长期没有得到充分的保护和利用,不利于该树种长期稳定发展。为了合理保护和开发利用大明松天然遗传资源,该文利用12个SSR分子标记对大明松3个天然群体进行遗传多样性研究,分析其群体间的遗传分化和基因流,为该物种的保护策略提供参考。研究结果表明:(1)12对引物共检测到37 个等位基因,多态位点百分率为100%。每个位点平均观察等位基因数(Na)为3.08,平均有效等位基因数(Ne)为1.68, 不同位点间有效等位基因数差异较大; 每个位点平均观察杂合度(Ho)为0.35,平均期望杂合度(He)为0.40,平均多态信息含量(PIC)为0.31。(2)3 个群体的Shannon''s多样性指数变化范围为0.48～0.65,Nei''s基因多样度的变化范围为0.27～0.39,与其他近缘种松类相比遗传多样性偏低。群体平均观察杂合度为0.40,平均期望杂合度为0.33,平均有效等位基因数为1.58。群体间基因分化系数(Gst)为0.10,群体间的遗传分化水平较低,大部分变异均存在群体内。群体间的基因流(Nm)为2.74,说明大明松群体间的基因交流比较充分。该研究可为大明松生物多样性保护提供重要参考依据,为科学利用大明松资源打下基础。     

千岛湖岛屿社鼠的种群年龄结构和性比

2009年7月至11月以及2010年3月至11月在千岛湖地区2个岛屿上对社鼠(Niviventer confucianus)种群进行标志重捕,通过对社鼠种群年龄结构和性比的研究,探讨陆桥岛屿环境下生境片段化对社鼠种群结构的影响。结果显示:社鼠种群年龄结构整体上均表现为由上半年幼年组和亚成年组占优势逐渐过渡到下半年成年组和老年组占绝对优势,岛屿环境对社鼠年龄结构变化的影响与陆地环境有所不同;两岛社鼠种群数量与性比(♀:♂)之间均存在显著的负相关性(B岛2010年7-11月除外),当种群数量处于高峰期时,雌性少于雄性,而种群数量处于低潮期时,雌性多于雄性,这可能是社鼠种群数量变动内部调节机制的一种表现;由于9-10月和2-3月的繁殖高峰以及高的胎仔数,翌年上半年种群数量就能恢复并达到全年最高峰。     

我国西南部喀斯特森林特有濒危树种掌叶木新的微卫星分子标记的开发

采用基因组DNA富集文库法FIASCO(Fast isolation by AFLP of sequences containing repeats)从掌叶木(Handeliodendron bodinieri)基因组中分离和筛选了10个新的微卫星位点,进而对掌叶木茂兰自然分布居群的遗传多样性进行了分析。结果表明, 每个位点在30株掌叶木个体上的平均等位基因数(A)为3.5(3~5个),平均观察杂合度(H_O)为0.650(0.267~0.900),平均预期杂合度(H_E)为0.494(0.224~0.652)。每个位点的第一排除概率值P_r(Ex₁)为0.029~0.240,位点综合值为0.7496。单个位点的第二排除概率P_r(Ex₂)为0.123~0.419,位点综合值为0.9517。这些信息预示着这些微卫星标记可以为研究喀斯特特有濒危树种掌叶木的基因流及居群遗传结构提供有效的遗传工具。     

舟山群岛黄毛鼠种群的遗传多样性及多重父权研究

岛屿常被生态学家和进化生物学家视为研究生物进化的天然实验室。本研究利用微卫星分子标记技术,分析了舟山群岛优势鼠种——黄毛鼠(Rattus losea)刺山岛种群、六横岛种群、桃花岛种群3个种群的遗传多样性,并探讨该物种是否存在多重父权现象。研究结果表明,(1)黄毛鼠种群的平均观测等位基因数(NA)和平均有效等位基因数(Ne)分别为8.792和5.494,平均期望杂合度(He)和平均观测杂合度(Ho)分别为0.821和0.956,多态信息含量(PIC)均大于0.5,说明3个种群均具有较高的遗传多样性;(2)3个种群的平均遗传分化系数(Fst)为0.040,其中桃花岛种群与刺山岛种群的分化最大,分化系数为0.052,呈中度分化水平,不同种群间所贡献的变异百分数为3.98%,说明遗传变异主要发生在种群内,而种群间仅存在很小的遗传变异;(3)14只孕鼠中,有6只孕鼠的胚胎存在多重父权现象,占43%,最小父本值为2,其余8个孕鼠的胚胎均只有唯一父本。本研究提出黄毛鼠存在多重父权现象,为该物种繁殖策略及鼠害控制的研究提供依据。     

用微卫星标记分析皱纹盘鲍群体的遗传变异

利用微卫星标记技术, 对皱纹盘鲍山东长岛和辽宁獐子岛的两个野生群体以及山东崆峒岛一个养殖群体的遗传变异进行了分析。对6个微卫星基因座的多态性进行了评估, 各基因座的多态信息含量(PIC)值均大于0.5, 适合对鲍群体遗传结构的分析。结果表明, 这6个基因座在3个皱纹盘鲍群体中共获得57个等位基因, 等位基因数(A)平均为9.50, 有效等位基因数(N_e)平均为5.8572, 平均杂合度观测值(H_o)和期望值(^H_e)分别为0.6925和0.7966; 两个野生群体的杂合度观测值(H_o)和期望值(H_e)均高于养殖群体。上述结果为保护和利用皱纹盘鲍的遗传多样性提供了依据。     

北京地区野生大豆种群SSR标记的遗传多样性评价

 使用40对SSR引物分析了北京地区野生大豆(Glycine soja)天然种群的遗传结构与遗传多样性。10个种群共检测到526个等位变异, 平均每对引物等位基因数为13.15个, 种群平均Shannon指数(I)为0.658, 群体平均位点预期杂合度(H_e)为0.369, 群体平均位点杂合度(H_o)为1.29 %。平均种群内遗传多样度(H_s)为0.362, 平均种群间遗传多样度(D_ST)为0.446, 基因分化程度(G_ST)为0.544。该研究显示, 中-西部生态区种群比北部和东部山区种群有较高的遗传多样性。在地理上, 环绕北京地区的太行山和燕山两大余脉区域野生大豆种群遗传分化表现出地理差异。可能是经过干旱选择而形成的有抗旱潜力的种群在遗传上表现单一化。期待该种群提供耐旱基因。     

基于COⅢ和HNF-1α序列研究龟鳖类的系统进化特征

用PCR扩增和测序的方法,获得小鳄龟(Chelydra serpentina)的COⅢ和HNF-1α序列,并分别结合NCBI中其他龟鳖的同源性序列进行比对分析。比对后得到757 bp的COⅢ一致序列和769 bp的HNF-1α一致序列。其中,COⅢ一致序列含有可变位点324个,序列总变异率为42.8%,简约信息位点230个;T、C、A、G的平均含量分别为27.5%、26.6%、30.8%、15.1%,A+T含量(58.3%)高于G+C含量(41.7%),转换/颠换比率(R)为2.62。HNF-1α一致序列有变异位点112个,变异率为14.6%,简约信息位点67个;T、C、A、G的平均含量为26.1%、23.1%、28.3%、22.6%,A+T含量为54.4%,G+C含量为45.7%,转换/颠换比率(R)为1.42。基于Kimura双参数模型计算龟鳖类种间、属间、科间遗传距离,并采用邻接法、最大简约法和最大似然法构建分子系统进化树。结果显示:基于COⅢ序列的淡水龟科4个属间的遗传距离为0.090～0.153,平均遗传距离为0.129;曲颈龟亚目5个科之间的遗传距离为0.150～0.207,平均遗传距离为0.177;基于HNF-1α序列的龟科9属间的遗传距离为0.003～0.051,平均为0.016;鳄龟科、龟科、淡水龟科3科间的遗传距离为0.044～0.067,平均为0.053。由遗传距离和构建的系统进化树可知,淡水龟科与陆龟科具有较近的亲缘关系,而与龟科的亲缘关系较远;支持龟科重新划分为两个分支;鳄龟科和海龟科亲缘关系较近,大鳄龟(Macroclemys temminckii)和小鳄龟可能同为一属。     

秦巴山区中华蜜蜂种群微卫星DNA遗传分析

【目的】中华蜜蜂Apis cerana cerana是一种兼有生态价值和经济价值的授粉昆虫。本研究拟揭示秦巴山区中华蜜蜂种群遗传多态性现状,探讨中华蜜蜂的种群分化机制及其影响因素。【方法】使用8个微卫星DNA标记评估秦巴山区17个样点共979个蜂群的中华蜜蜂遗传多态性,以长白山中华蜜蜂和阿坝中华蜜蜂作为外群进行种群遗传分化分析。【结果】秦巴山区中华蜜蜂95%的差异来源于样点内,样点间遗传分化系数(Fst)为0.002~0.037,基因流参数Nm为6.51~124.75,与外群的遗传分化分析中均显示秦巴山区中华蜜蜂不存在遗传分化。平均期望杂合度(He)为0.6877±0.1098,平均观察杂合度(Ho)为0.6364±0.1367,平均有效等位基因数(Ne)为3.7488±1.6201个,平均等位基因数(Na)为7.71±2.52个,多态信息含量(PIC)为0.6418±0.1152,香农指数(I)为1.5026±0.3754,这些参数均表明微卫星遗传多态性丰富。中蜂囊状幼虫病流行过的地区,其杂合度、多态信息含量以及等位基因数均显著低于未发病地区。【结论】秦巴山区中华蜜蜂种群数量大,分布均匀,基因流水平高,在650 km距离范围内没有发生种群分化;秦巴山区中华蜜蜂微卫星遗传多态性丰富,仅部分地区中华蜜蜂遗传多态性受中蜂囊状幼虫病的选择压而降低。     

黑琴鸡北方亚种mtDNA D-loop遗传多样性初步研究

黑琴鸡(Tetrao tetrix)为国家Ⅱ级保护动物,但近年来种群数量不断减少。因此,了解不同种群的遗传变异情况,可为制定保护管理策略提供依据,以便恢复野外种群的数量。本文测定加格达奇(42个个体)和呼伦贝尔市扎兰屯(76个个体)两个黑琴鸡种群的共118个个体的mtDNA D-loop 序列,共发现25个变异位点,定义了33个单倍型,整体的平均核苷酸差异数(K)为2.608,核苷酸多样性(π)为0.228%,种群的遗传多样性偏低,两个种群有一定的基因交流,N_m为14.63,群体间无显著的遗传分化。Tajima's D和Fu&Li's D的估算结果表明,这两个黑琴鸡种群相对于中性的歧异度并没有明显的偏离(P>0.1),两个种群可能未经过大规模的种群扩张过程。     

Genetic diversity and population structure of the endangered species Psammosilene tunicoides revealed by DALP analysis

Psammosilene tunicoides W. C. Wu et C.Y. Wu (Caryophyllaceae) is a rare plant endemic to China. We used the direct amplification of length polymorphism (DALP) method to analyse the genetic diversity of 17 natural populations of this species from Yunnan, Sichuan and Guizhou provinces. The percentage of polymorphic bands (PPB) was 99.1% at the species level and 19.9% at the population level. The total genetic diversity (Ht) was 0.2827; the genetic diversity within populations (H_s) was 0.0594 and the coefficient of gene differentiation (G_st) was 0.7899. The results indicated low levels of genetic diversity within populations and extremely high genetic differentiation among populations. This pattern might be caused by genetic drift resulting from intensive harvesting and habitat fragmentation. It might also be a result of local adaptation. We further performed experiments on its breeding system and found a high rate of selfing in P. tunicoides, which may be partially responsible for its population structure.     

Effects of geographic distance,sea barriers and habitat on the genetic structure and diversity of all-hybrid water frog populations

The history of population size and migration patterns leaves its mark in the genetics of populations. We investigate the genetic structure of the edible frog, Pelophylax esculentus in the Danish archipelago and adjacent countries. This frog is of particular interest because it is a hybrid that, in this area, forms all-hybrid populations of diploid (LR) and triploid (LLR and LRR) genomotypes with no (or very few) adults of the parental species (LL and RR). This study is the first to cover the entire geographic range of Danish, Swedish and German all-hybrid populations, documenting their extent and providing a broad picture of their diversity of neutral genetic markers and genomotype proportions. With 18 microsatellite markers, we found that genetic diversity declines northwards in agreement with the glacial refuge and central-marginal hypotheses; however, populations on small and medium-sized islands are no less diverse than those on large islands and continental peninsulas. Isolation by distance exists across the archipelago with limited influence of fragmentation by brackish seawater. The extremely low genetic diversity in all-hybrid populations, compared with adjacent populations, may be responsible for the maintenance of their special breeding system. We also show large variation among ponds in proportions of LLR, LR and LRR genomotypes, but little geographic pattern in their distribution. Instead, we found relationships between the genomotype proportions and some of 15 habitat parameters monitored. Body size differences among LLR, LR and LRR further suggest ecological differences.     

Genetic diversity of Stipa grandis P.Smirn populations across the species’ range in the Inner Mongolia Plateau of China

In order to investigate the genetic diversity of dominant species under the background of climate change and grassland utilization in the Inner Mongolia Plateau of China, we sampled seven Stipa grandis populations along an increasing aridity gradient in the present study. The Nei’s gene diversity of populations (He) was estimated to be 0.15 and the percentage of polymorphic loci (PPL) was 49.28%. The genetic differentiation among populations (Φ_ST) was 0.2431. There was a significant relationship between genetic distance and geographic distance among the S. grandis populations by Mantel's test. The genetic diversity was significantly correlated with longitude and annual mean precipitation, which suggested that a combination of climatic factors affected the genetic diversity. The populations in the marginal habitat should be paid more attention because of their low genetic diversity and its significance for conservation of the whole species.     

Exploring the legacy of goat grazing: signatures of habitat fragmentation on genetic patterns of endemic weevil populations in Northern Isabela Island,Galápagos (Ecuador)

Together with reduction in habitat area and quality, reduction in habitat connectivity is one of the major factors influencing species’ persistence in fragmented landscapes. We explore the consequences of recent habitat fragmentation on volcanoes across Isabela Island, Galápagos by analyzing genetic patterns of populations of endemic weevils whose host plants have been depleted by indiscriminate goat grazing. We predicted that if grazing on the weevils’ host plants has caused habitat fragmentation on the weevils populations themselves, then the effects on the genetic architecture of populations should be more severe on Galapaganus conwayensis populations from volcanoes on Northern Isabela than on those from Santa Cruz or Pinta islands where vegetation destruction was not as extensive. We used mitochondrial sequences to reveal historical colonization patterns and microsatellite variation to understand more contemporary genetic changes. We found significantly lower microsatellite genetic diversity and population size within localities and increased genetic differentiation at a small geographic scale with a stronger isolation by distance pattern and larger numbers of genetic clusters on Isabela. In the absence of long-standing mitochondrial structure within each volcano, we interpret the microsatellite results as suggesting that recent host plant habitat fragmentation may indeed influence the genetic patterns of plant feeding insects and highlight the importance of controlling the spread of introduced herbivores in the Galápagos Islands.     

Genetic diversity within and among populations of the endangered and endemic species Primula merrilliana in China

Primula merrilliana Schltr. is an endangered and narrowly-distributed endemic species of southern Anhui Province in China. In this study, the level of genetic variation and the pattern of genetic structure in six natural populations of P. merrilliana were assessed by using ISSR (inter-simple sequence repeats) markers. Based on ten primers, 137 clear and reproducible DNA fragments were generated, of which 109 were polymorphic. The statistical results indicated that there was a relatively low genetic diversity within populations, and a high genetic differentiation among populations (G_ST = 0.53, Φ_ST = 0.49). The level of population genetic diversity was correlated to habitat type and the gene flow (N_m) was low with only 0.45. The unexpected genetic structure of P. merrilliana may be explained by limited gene flow that was caused by habitat fragmentation and limited seeds and pollen dispersal ability, self-compatible breeding system and biennial life form.     

基于COI基因全长序列的假眼小绿叶蝉地理种群遗传分化研究

假眼小绿叶蝉Empoasca vitis(Gthe)是我国大陆茶区分布最广、为害最重的茶树害虫,其种群世代重叠严重,数量大。从我国13个主要产茶省份各选出一个重点产茶县(市),采集假眼小绿叶蝉标本,首次扩增得到其线粒体CO I基因全长序列,并以此探讨了假眼小绿叶蝉13个地理种群间的遗传多样性、分子变异、遗传分化程度及基因流水平。在13个地理种群中共得到了176条CO I基因序列,发现了113个变异位点,形成了105个CO I单倍型。总群体单倍型多样性Hd为0.9720,种群内单倍型多样性在0.804—1.000范围内,总群体和各种群的Tajima's D检验结果均不显著,说明茶园假眼小绿叶蝉的进化符合中性模型,种群数量较为稳定。AMOVA分子变异分析结果表明,茶园假眼小绿叶蝉的遗传分化主要来自于种群内部。Mantel检验显示各地理种群的遗传距离与地理距离之间不具有显著的相关性。总群体的遗传分化系数Gst为0.03652,固定系数Fst为0.10876,基因流Nm为4.097,表明地理种群间存在较频繁的基因交流,遗传差异较小。推测,20世纪90年代以来的省际之间频繁的茶树鲜叶贩运、异地茶苗的大批量调拨等因素促进了假眼小绿叶蝉的长距离迁移,加强了不同种群间的基因交流。     

Genetic variation and conservation of the endangered endemic Anagyris latifolia Brouss. ex Willd. (Leguminosae) from the Canary Islands

Anagyris latifolia is an endemic and endangered species from the Canary Islands, whose distribution is limited to four islands, with less than 400 individuals in fragmented and isolated localities. RAPD markers have been used to assess the genetic diversity and genetic differentiation of its populations, in order to formulate appropriate management and conservation genetics strategies. Nine polymorphic primers generated 74 polymorphic DNA fragments. Genetic variation levels detected in Anagyris latifolia were significant high (H = 0.200; P% = 97.3), principal coordinates analysis and genetic differentiation coefficient showed a high degree of genetic differentiation between islands, without a define east-to-west stepping stone colonization route. AMOVA analysis showed that of the total genetic variation detected, 32.43% was maintained among islands, 20.73% contained among population within islands, and 46.84% resided within populations. According to these results, management strategies should be focused on each island separately.     

Population structure of an endangered frog (<Emphasis Type="Italic">Babina subaspera</Emphasis>) endemic to the Amami Islands: possible impacts of invasive predators on gene flow

The otton frog (Babina subaspera) is an endangered species endemic to the Amami Islands, Japan. High predation pressure from an introduced carnivore, the mongoose, has caused declines in the frog populations and created a large habitat gap around an urban area. To promote effective conservation, we investigated the genetic status of the species and examined the effect of the habitat gap on gene flow among populations. Using five polymorphic microsatellite loci and mitochondrial DNA sequences, we investigated genetic diversity, genetic structure and gene flow in B. subaspera populations on the islands of Amami-Oshima and Kakeroma-jima. The expected heterozygosity (H _E) within each locality was generally high (range: 0.67–0.85), indicating that B. subaspera maintains high genetic diversity. However, genetic differentiation was observed, and the two populations, TAG and KAR, showed little gene flow with other populations. The clustering and F _ST analyses also predicted that these two populations were clearly distinct. According to the mitochondrial DNA analysis, the observed genetic differentiation occurred relatively recently. Possible barriers such as mountain ridges, rivers or roads did not result in genetic separation of the populations. These data support the hypothesis that the habitat gap created by an introduced predator prevented the gene flow among B. subaspera populations. When developing conservation strategies for B. subaspera, focus should be directed to these two isolated populations; careful monitoring of population size and genetic diversity should be conducted along with the mongoose elimination project ensues.     

Host-associated genetic differentiation in rice grasshopper, Oxya japonica, on wild vs. cultivated rice

Rice grasshopper, Oxya japonica, is one of the most important pests in south China, mainly inhabiting fields of wild rice (Oryza rufipogon) and cultivated rice (Oryza sativa). In this study, we used AFLP marker to investigate the genetic diversity and population structure of rice grasshoppers collected from south China, with emphasis on testing the hypothesis that there was significant genetic differentiation among grasshopper populations associated with different hosts (i.e. wild vs. cultivated rice). Seven populations consisting of 104 individuals were sampled from Hainan Island and the mainland of south China. Eight primer combinations produced 564 reliable bands, of which 563 were polymorphic. O. japonica showed considerable genetic variation at population level, with gene diversity (H_E) ranging from 0.1103 to 0.2035. Genetic diversity were studied on seven populations, and generally three populations from wild rice had higher levels of genetic diversity (H_E = 0.1635) than the other four populations feeding on cultivated rice (H_E = 0.1327). We observed high population differentiation, with F_st ranging from 0.4172 to 0.7652 among the seven populations. However, Mantel test detected no significant correlation between genetic distance and geographical distance (r = 0.3541; p = 0.0689). By contrast, we found significant genetic differentiation between groups collected from different hosts. These data suggested that the anthropogenic activity in cultivated rice fields (i.e. pesticides, fertilization and cultivation) could have played an important role in shaping the genetic structure of O. japonica.