欧洲刺槐种源群体遗传结构和多样性

对来自欧洲和美国的 18个刺槐种源子代进行了等位酶分析。可进行遗传分析的 7个酶系统 (Amy,Fe,L ap,Idh,Mdh,6 Pgd,Skd)中有 14个基因位点 ,其中 12个位点具有多态性。每个多态位点平均等位基因数 (A/L )变化在 1.5 6～ 3.6 7之间 ,平均基因型数 (G/L)变化在 1.6 1～ 7.11之间 ,平均等位基因有效数目 (Ae)变化在 1.0 2～ 2 .5 0之间 ,预期杂合度 (H e)变化在0 .0 2～ 0 .5 6之间。不同种源群体之间也存在较大的遗传差异 ,在 8个德国种源中 ,各群体的 A、Ae、和 H e等相对较小 ,但不同群体间差异较大。各位点等位基因频率在不同种源群体间变化也较大 ,表明德国各种源群体内遗传变异相对较小 ,但群体间差异较大。来自匈牙利和斯洛伐克的 8个种源群则相反 ,各群体的 A、Ae、和 H e等相对较大 ,而不同种源群体间差异则较小 ,各位点等位基因频率在种源群体间变化相对一致 ,表明这两个国家的种源群体内变异较大 ,但不同种源群体间差异较小。欧洲的刺槐种源并未形成明显的地理变异模式 ,而且欧洲的种源和来自原产地的美国种源相比 ,没有发现明显的差异。经过 Hardy-Weinberg平衡检测证明 ,88.4 1%位点符合 H- W遗传平衡 ,表明各群体基因频率和基因型频率保持较高的稳定性 ,且种源内的变异大于种源间变异 ,94 .     

孑遗植物银杏群体遗传多样性的ISSR分析

采用ISSR分子标记技术,对江苏泰兴、美国纽约的栽培银杏(Ginkgo biloba)群体和中国3个可能为野生的银杏自然群体(浙江西天目山、贵州务川、湖北大洪山区)的遗传多样性水平和群体遗传结构进行了研究。用13个引物对5个群体共66个样品进行扩增,共得到88个清晰的扩增位点,其中多态性位点62个,多态位点百分率(PPB)为70．45％。POPGENE分析结果表明：与其他裸子植物相比,银杏具有丰富的遗传变异(He=0．2408;Ho=0．3599)。贵州务川群体的遗传多样性水平最高(PPB=56．82％,He=0．2089,Ho=0．3087),江苏泰兴栽培群体(PPB=34．09％,,Ho=0．1269,Ho=0．1858)和美国纽约的栽培群体(PPB=23．86％,,He=0．0884,Ho=0．1312)的遗传多样性水平较低。Nei′s遗传多样性分析和AMOVA分析表明,3个可能的自然群体间出现了一定程度的遗传分化(Gst=0．1476,Φst=14．26％)。群体间一定程度的遗传分化可能是人为选择压力和基因流障碍引起的。根据Nei′s遗传距离矩阵分别构建了群体间和个体间的遗传关系树状图。由UPGMA聚类分析可知,贵州务川群体与浙江西天目山群体优先聚类;美国纽约群体与湖北大洪山群体具有较近的亲缘关系,它们可能为同一野生群体的后裔。通过对银杏群体遗传结构的分析并结合群落学调查研究,结果表明：贵州务川银杏群体很可能为野生自然群体。基于银杏群体遗传学和生态学的研究结果,建议在自然银杏群体最适生境和遗传多样性最高的贵州务川建立银杏保护区。由于银杏群体间出现了一定程度的分化,建议3个自然群体间可进行植株和幼苗相互移栽,以提高群体间的基因交流,以最大限度地保护银杏的遗传多样性。     

利用ＲＡＰＤ标记技术对白桦种源遗传变异的分析及种源区划

用随机扩增多态ＤＮＡ（ＲＡＰＤ）分子标记方法对白桦１７个种源１５２个个体进行了遗传变异的比较分析,通过１４个随机引物 共检测到２３３个位点,各种源多态位点百分率差异明显,范围在２０．１７％－３２．１９％之间,多态位点百分率最高的是帽儿山种源和清源种源,最低的是绰尔种源。遗璺变异在种源间占４３．５３％,在种源内个体间占５６．４７％。根据种源间的遗传距离,构建了白桦１７个种源的遗传关系聚类图,结果将东北地区的白桦为一类,华北、西北地区的白桦聚为另一类,同时根据地理气候因子和遗传距离对白桦群体进行了种源区的划分。     

两个封闭群SPF级昆明小鼠遗传背景调查

目的：比较两个封闭群SPF级昆明（ KM）小鼠的遗传差异,调查引进的SPF级KM小鼠封闭繁殖6年后,其遗传构成是否发生变化。方法应用微卫星DNA标记方法对18个位点在两个群体中的遗传差异进行分析,主要包括观察等位基因数（ Na）、有效等位基因数（ Ne）、观察杂合度（ Ho）、期望杂合度（ He）、多态信息含量（PIC ）、Shan-non信息指数、遗传分化系数（ F st）、遗传距离等遗传参数。结果两个封闭群SPF 级KM小鼠在18个微卫星位点共发现67个等位基因,Na为2～8个,平均3．7222个;Ne为1．9459～6．5442,平均2．7966个;Ho为0．4225～1．0000,平均0．8823;He为0．4892～0．8527,平均0．6162;Shannon信息指数0．6792～1．9526,平均1．0598;PIC为0．3680～0．8301,平均0．5317;Fst平均值为0．0159,表明群体间的遗传差异仅1．59％,二者间的遗传距离（DA）为0．0499。结论两个封闭群SPF级KM小鼠遗传结构相似度极高,它们与原引进群体的分化差异极小。     

缙云山特有植物缙云黄芩的遗传多样性研究

采用聚丙烯酰胺凝胶垂直平板电泳检测了重庆缙云山特有植物缙云黄芩7个居群70个个体的过氧化物酶、细胞色素氧化酶、超氧化物歧化酶、淀粉酶和酯酶5种等位酶,获得20个基因位点的资料,由此分析了其遗传分化水平。结果表明：缙云黄芩遗传多样性水平较高,多态位点比率P为45．7％,平均每个多态位点的等位基因数目A为1．46,平均预期杂合度He为0．205,平均观察杂合度Ho为0．352。缙云黄芩居群间遗传分化水平较高,Gst=0．401,居群间遗传一致度和遗传距离的平均值分别为0．741和0．300。缙云黄芩遗传多样性的40．1％来源于居群之间的基因差异,59．9％属于居群内的遗传分化,因而对缙云黄芩遗传多样性的保护,应保护其较多的居群。     

东北大口鲇2个群体的微卫星DNA多态分析

利用磁珠富集法克隆制备的24个大口鲇（Silurus meriaionalis Chen）微卫星标记,对黑龙江野生群体与松花江养殖群体2个东北大口鲇（S．soldatovi）的地理种群的等位基因频率（P）、观测杂合度（Ho）、期望杂合度（He）、多态信息含量（PIC）和有效等位基因数（Ne）等进行了遗传检测,以遗传偏离指数（d）检验Hardy—Weinberg平衡,并以Nei氏遗传分化系数（GST）和AMOVA分析（ФST）群体遗传变异的来源。同时,使用PHYLIP3．63软件绘制基于Nei氏遗传距离的个体间UPGMA系统树。结果表明：24个微卫星标记在东北大口鲇的2个群体中共扩增出1357条多态性片段,片段长度为1024385bp,总体平均等位基因8．875个,可以用于东北大口鲇遗传多样性的评估。并发现8个可区分这2个种群的遗传标记;黑龙江群体的P、Ho、He、PIC和Ne依次为0．165、0．435、0．758、0．742和5．019,松花江群体为0．147、0．299、0．847、0．764和5．944,在这些多样性参数上,方差分析也显示2地理种群差异不显著,在大多数位点并无显著差异,仅HLJcf37位点具有显著差异：在多个位点偏离Hardy—Weinberg平衡,2群体呈现不同程度的杂合体过度,纯合体完全缺失现象,其原因有待证实;群体遗传变异分析证实2群体间遗传分化较弱,其98％以上的变异是由群体内个体间的遗传变异引起的,群体间的变异对总变异影响不显著。UPGMA系统树也显示出个体间遗传距离小,亲缘关系很近。结果表明,人工繁殖没有对东北大口鲇的遗传多样性产生影响,该种群遗传分化小,种质资源状况良好。     

中国沿海中部珊瑚菜居群等位酶变异及其遗传多样性

利用聚丙烯酰胺凝胶电泳方法,分析了我国沿海中部（江苏、山东和浙江）海滨沙滩珊瑚菜（Glehnia littoralis Fr.Schmidt ex Miq.)7个居群8种酶系统19个位点的等位基因遗传变异特征,结果表明居群内多态位点比率平均为82．4％,每一位点平均等位基因数为2．77,有效等位基因数为2．24,固定指数F的平均值为－0．091。珊瑚菜居群基因多样性80．9％产生于居群内,19．1％产生于居群间。居群间的遗传距离平均为0．317,遗传一致性为0．728,居群内维持着较高水平的遗传多样性。据此,对珊瑚菜的渐危机理进行了讨论。     

濒危植物长叶榧群体遗传多样性的RAPD分析

借助随机扩增多态DNA方法,分析了中国特有的濒危植物长叶榧的遗传多样性和遗传分化．结果表明：12个随机引物在9个长叶榧自然群体180个样品中可检测到180个可重复位点,其中多态位点119个．长叶榧物种水平的遗传多样性较高,多态位点百分率（P）为66．11％,Shannon信息指数（,）为0．3087,Nei指数（h）为0．2015;而群体水平的遗传多样性较低,P、I和h分别平均为23．76％、0．1221和0．0813．AMOVA分子变异显示,42．57％变异来源于群体内,57．43％变异来源于群体间,群体间的遗传分化系数（Gst）为0．5965,群体间的遗传分化程度高．长叶榧群体间的基因流很低,为0．3382．瓶颈效应、群体隔离和群体间基因流低等因素都加剧了长叶榧群体间的遗传分化．9个长叶榧群体间的平均遗传距离为0．1630．通过UPGMA进行聚类,可将9个长叶榧群体分为浙江和福建两大类群．建议在迁地保护时应尽量避免在群体之间实施种质迁移．     

光叶珙桐的等位酶分析及其生物地理学意义

采用水平切片淀粉凝胶电泳实验方法,对光叶珙桐(Davidia involucrata var．vilmoriniana)4个居群、78个个体、11种酶系统,21个位点进行了等位酶分析。各居群的多态位点百分比P＝28.6％-47．6％,实际杂合度Ho＝0．177—0.308,平均每个位点的等位基因数A=1．3-1．6。在变种水平上,多态位点百分比P＝52．4％,平均每个位点的等位基因数A＝1．8,实际杂合度Ho＝0．272,期望杂合度Ho＝0．216。基因分化系数FST＝0．1928,说明光叶珙桐居群问分化小。各项指标表明光叶珙桐的遗传多样性水平较高,比原变种珙桐高,外部环境可能是影响珙桐分布格局的重要因素。昭通与宝兴的遗传多样性明显高于其他两个居群,遗传多样性保存较为完整,而处于分布区东、西两侧边缘(云南西北部和湖北五峰)遗传多样性则相对较低,推测四川盆地边缘山地是该种的遗传多样化中心,可能是在地质灾难中(如第四纪冰期),光叶珙桐真正的避难所。分布区东西两侧的居群可能是从四川盆地边缘山地扩散而来。     

小毛莨居群的遗传分化及其与空间隔离的相关性

采用聚丙烯酰胺凝胶电泳技术,对分布于华中地区的11个小毛莨（Ranunculus ternatus）居群的遗传分化进行了检测。对8个酶系统17个酶位点上的分析结果表明,该种各居群的各项遗传多样性指标处于一个相对较低的水平：多态位点比率（P）为0－53.0％,平均每位点等位基因数（A）为1－1.647,平均预期杂合度（He）和观察杂合度（Ho）分别为0－0.108和0－0.102。居群间遗传一致度甚高（I＝0.9754-0.9991）。根据Nei 's传距离所作出的聚灰分析表明,豫南信阳地区3个居群与湖北省武汉地区8个居群之间关系较远。而在武汉地区,长江以北的居群及长江以南的部分居群分别相聚在一起,用GPS定位方法得到居群间空间距离并据此聚类,该种的遗传分化与地理因素的相关性,并推测出长江的隔离作用加强了两岸居群间的遗传分化,同时发现一个生于独特生境的居群在表型和遗传结构上都已与其他邻近居群有了很大分异,由于该居群在所检测的酶位点上均无特有等位基因出现,作者认为不宜将其作为新种或新变种处理。     

Postglacial distribution area expansion of Polyommatus coridon (Lepidoptera: Lycaenidae) from its Ponto-Mediterranean glacial refugium

The genetic population structure of Polyommatus coridon (Poda, 1761) (Chalk-hill blue) was studied by means of allozyme electrophoresis in north-eastern Germany, the Czech Republic, Slovakia and Hungary. All analysed parameters showed high genetic diversity within populations (number of alleles: 2.61; observed and expected heterozygosity: 18.6% and 19.7%, respectively; percentage of polymorphic loci: total: 73.6%, on 95% level: 56.1%), whereas genetic differentiation between populations was comparatively low (F(ST) = 0.028 +/- 0.005 s.d.). Hierarchical variance analysis revealed significant structuring among five regional population clusters. A significant isolation-by-distance structure exists (r = 0.39; P < 0.05). The mean number of alleles per locus declined significantly from south to north and showed a strong correlation with the geographical latitude (r = -0.88, P < 0.0001). We suggest that this reflects the loss of alleles during the postglacial colonisation of eastern Central Europe from an ice-age refugium in the Balkans. A possible scenario for the postglacial expansion process in eastern Central Europe is discussed using these data: coming from the north-western part of the Balkans, P. coridon may have reached the western tip of Hungary, and consecutively colonised eastern Central Europe using two alternative expansion routes.     

Isolation and characterization of microsatellite markers from Robinia pseudoacacia L

Microsatellite markers were isolated from Robinia pseudoacacia L. using an enrichment method. Eleven of the 23 primer pairs designed successfully amplified unambiguous and polymorphic single loci among 39 individual R. pseudoacacia L. from northeastern Japan. The observed and expected heterozygosities of the 11 microsatellite markers ranged from 0.333 to 0.821 and from 0.489 to 0.867, respectively. The polymorphisms observed at the 11 microsatellite loci are useful genetic data for forest ecological studies involving R. pseudoacacia L.     

Strong genetic impoverishment from the centre of distribution in southern Europe to peripheral Baltic and isolated Scandinavian populations of the pearly heath butterfly

Aim Climatic changes and fluctuations in the past have strongly influenced the distribution of animal and plant species. Such fluctuations are also reflected in the patterns of genetic diversity on both local and global scales. The genetic pattern of the pearly heath butterfly, Coenonympha arcania, was used to evaluate the genetic differentiation of isolated (in north‐western Europe), peripheral (in north‐eastern Europe) and central (in southern Europe) populations in the context of post‐glacial distributional changes of the species. Location Europe (Sweden, Germany, the Baltic states, Italy, Slovenia, Hungary, Romania, Bulgaria). Thus, samples were collected from large parts of the species’ distribution representing the three categories mentioned above. Methods We analysed 18 loci of 569 individuals from 28 populations by allozyme electrophoresis. We used both individual‐based and population‐based analyses, including F‐statistics, various clustering methods and Markov chain Monte Carlo simulations. Results All loci, except Fum, were polymorphic. The mean F_ST for all samples was 0.18. The mean genetic distance among populations was 0.046. Two major genetic lineages were distinguished. Populations from the centre of the distributional range in southern Europe and the northern periphery of the distributional range differed significantly in their level of genetic variability. The central populations of south‐eastern Europe showed high levels of genetic diversity and no differentiation among populations. Main conclusions Most probably the two major genetic lineages evolved during glacial isolation in two disjunct Mediterranean refugia. The lack of genetic differentiation across south‐eastern Europe implies a continuous Würm ice age distribution in this area, thus supporting the functional existence of steppe forests throughout this region. The peripheral‐isolated populations in Sweden seem to have suffered from one or more severe bottlenecks, resulting in substantial genetic impoverishment. The peripheral‐connected eastern Baltic populations, on the other hand, are affected by post‐glacial and possibly recurrent gene flow from more central parts of the distribution.     

Genetic variation of chloroplast and nuclear markers in natural populations of hazelnut (Corylus avellana L.) in Germany

Corylus avellana L. (hazel) is a long-lived, monoecious and wind-pollinated shrub species, widespread all over Europe. In Germany, hazel is intensively traded and planted, and thus is of central interest from a nature conservancy point of view. To assess the within- and between-population differentiation of hazel, 20 natural populations (18 from Germany, one from Italy and one from Hungary) were investigated genetically. Seven isozyme systems comprising 11 gene loci were analysed in up to 100 samples (average 92.6) per population, amplified fragment length polymorphisms (AFLP) were analysed in up to 50 samples (average 47.4) and nine cpDNA-SSR markers were assessed in 20 samples per population. Results for overall isozyme variability with Na 2.46 alleles per locus, allelic diversity (Ne) 1.39, expected heterozygosity He 21 % and 79 % polymorphic loci were in accordance with the findings of previous studies. The respective values for AFLPs were lower, but both marker systems revealed the same level of about 3.5 % differentiation between populations. For cpSSR only the Italian sample showed within-population variation and the two haplotypes were completely differentiated from all other populations expressing a unique genetic structure with one single haplotype. Among the three marker systems AFLPs showed the best ability to differentiate between populations. While only one isozyme locus revealed significant differentiation, 41 AFLP loci showed highly significant differentiation between all populations, but 26 loci when only German populations were considered. Consequently geographic differentiation analyses focused mainly on molecular markers. Mantel tests showed significant correlations between genetic and geographic distance, but in the unweighted pair-group method with arithmetic mean analyses, adjacent populations did not always form clusters. While chloroplast markers were able to clearly distinguish only the Hungarian population, the nuclear markers revealed clear spatial genetic structures. The correlations between geographic and genetic distance was high for AFLPs. The correlograms illustrate this effect for all populations as well as for the German populations.     

Isozyme variation among teak (Tectona grandis L.f.) provenances

Fourteen enzyme systems were analysed in leaf parenchyma of nine native and introduced populations of teak. These enzyme systems were encoded by 20 putative loci of which 18 were polymorphic. Populations showed a general lack of heterozygosity (average F_IS = 0.11). On average over the 18 polymorphic loci, the genetic differentiation among provenances varied according to the estimator: 0.09 for G_ST, 0.12 for F_ST and 0.19 for . The cluster analysis showed two main gene pools, the first consisted of the Indian provenances and the second of African, Indonesian and Thai provenances. Genetic distances among populations of the same group were similar, and lower than the genetic distances between populations from different groups. The factorial analysis on genotypes of seedlings also showed the same geographic differentiation into two major groups. The possible natural distribution of teak in Java is discussed.     

Low genetic variability and strong differentiation among isolated populations of the rare steppe grass Stipa capillata L. in Central Europe

Stipa capillata L. (Poaceae) is a rare grassland species in Central Europe that is thought to have once been widespread in post‐glacial times. Such relict species are expected to show low genetic diversity within populations and high genetic differentiation between populations due to bottlenecks, long‐term isolation and ongoing habitat fragmentation. These patterns should be particularly pronounced in selfing species. We analysed patterns of random amplified polymorphic DNA (RAPD) variation in the facultatively cleistogamous S. capillata to examine whether genetic diversity is associated with population size, and to draw initial conclusions on the migration history of this species in Central Europe. We analysed 31 S. capillata populations distributed in northeastern, central and western Germany, Switzerland and Slovakia. Estimates of genetic diversity at the population level were low and not related to population size. Among all populations, extraordinarily high levels of genetic differentiation (amova : φ_ST = 0.86; Bayesian analysis: θ^B = 0.758) and isolation‐by‐distance were detected. Hierarchical amova indicated that most of the variability was partitioned among geographic regions (59%), or among populations between regions when the genetically distinct Slovakian populations were excluded. These findings are supported by results of a multivariate ordination analysis. We also found two different groups in an UPGMA cluster analysis: one that contained the populations from Slovakia, and the other that combined the populations from Germany and Switzerland. Our findings imply that S. capillata is indeed a relict species that experienced strong bottlenecks in Central Europe, enhanced by isolation and selfing. Most likely, populations in Slovakia were not the main genetic source for the post‐glacial colonization of Central Europe.     

利用RAPD 标记技术对白桦种源遗传变异的分析及种源区划

用随机扩增多态DNA(RAPD)分子标记方法对白桦17 个种源152 个个体进行了遗传变异的比较分析, 通过14 个随机引物扩增共检测到233 个位点, 各种源多态位点百分率差异明显, 范围在20.17%~32.19%之间, 多态位点百分率最高的是帽儿山种源和清源种源, 最低的是绰尔种源。遗传变异在种源间占43.53%, 在种源内个体间占56.47%。根据种源间的遗传距离, 构建了白桦17 个种源的遗传关系聚类图, 结果将东北地区的白桦聚为一类, 华北、西北地区的白桦聚为另一类。同时根据地理气候因子和遗传距离对白桦群体进行了种源区的划分。     

Characterization of highly polymorphic nuclear microsatellite loci in Juniperus communis L.

We developed five nuclear microsatellite markers in Juniperus communis L. using an enriched library method. Samples from 28 juniper individuals were collected in Spain, Germany and Slovakia and were analysed at the five loci. A high level of allelic diversity with values ranging from nine to 23 alleles was found. These highly polymorphic markers will be used in ongoing population genetic studies to evaluate the genetic resources and to contribute to the maintenance of genetic diversity of juniper in Middle Europe.     

Intraspecific allozymatic differentiation reveals the glacial refugia and the postglacial expansions of European Erebia medusa (Lepidoptera: Nymphalidae)

Allozyme analysis of Erebia medusa over large regions of Europe revealed a significant population differentiation (F_ST: 0.149 ± 0.016). A UPGMA-analysis showed a division into four major lineages with mean inter-group genetic distances ranging from 0.051 (±0.010) to 0.117 (±0.024). An AMOVA revealed that rather more than two-thirds of the variance between samples was being between these lineages and less than one-third within lineages. An eastern group included the samples from the Czech Republic, Slovakia and north-eastern Hungary. This genetic lineage expressed significantly higher genetic diversity than the other three. A second lineage was formed by the samples from France and Germany. The two samples from western Hungary represent a third delimited lineage and the sample from northern Italy a fourth. We suppose that this genetic differentiation took place during the last ice-age in four disjunct refugia. The genetically more diverse eastern genetic lineage might have evolved in a relatively large refugium in south-eastern Europe. We assume that the other three lineages developed in relatively small relict areas around the Alps. It is likely for the western lineage that its ice-age distribution showed at least one disjunction in late Würm with the consequence of further genetic differentiation. Most probably, the eastern lineage colonized postglacial Central Europe using two alternative routes: one north and one south of the Carpathians. Up to now, neither similar glacial refugia, nor comparable secondary disjunctions in late Würm, are reported for any other animal or plant species.