Genetic Variation of Sesleria albicans Kit. ex Schultes (Poaceae): Lack of Evidence for Glacial Relict Endemism in Central Europe

Abstract: The populations of Sesleria albicans Kit. ex Schultes in central Europe are supposed to be glacial relicts. The genetic structure of these populations can provide evidence for the reliability of this hypothesis, since a distinct geographical clustering of populations and high levels of interpopulational genetic variation can be expected in the case of relict endemism. In the study presented here, we used Random Amplified Polymorphic DNA (RAPD) analysis to describe the genetic structure of 25 Sesleria albicans populations from west and central Germany, southwest Germany and the Alps. In the RAPD analysis 344 fragments could be amplified, of which 95.9 % were polymorphic. The percentage of polymorphic bands per population ranged from 29.7 % to 56.7 % and correlated significantly with population size, indicating a higher level of inbreeding in small and isolated populations. An analysis of molecular variance (AMOVA) revealed that 61.22 % of the variation resided within populations, whereas 34.25 % was found among populations within regions and only 4.53 % among regions. Genetic distance among populations (φ_ST) varied between 0.63 and 0.13, and in a Neighbour-Joining dendrogram based on the genetic distances between the investigated individuals, all populations could be discriminated from each other. The populations from the northern and southern Alps were clearly separated from the central European populations. These, however, did not form geographic clusters. Considering these results, Sesleria albicans showed a high level of intra- but only a low level of interpopulational genetic variation and very weak regional differentiation. The genetic pattern detected in this study, therefore, gives no evidence for glacial relict endemism of Sesleria albicans in central Europe.     

Diversity,variability and habitat characteristics of the communities dominated by Sesleria species (Poaceae) in the Western Carpathians

This study provides a view of vegetation types dominated by Sesleria species in the Western Carpathians (Sesleria caerulea, S. heufleriana, S. tatrae, and S. uliginosa). We also took into account characteristics/traits such as plant life forms, autochthonous status, endemism, and ploidy level occurring within each relevé in our data set. Altitude and Ellenberg indicator values derived for each relevé were considered as well. Eight vegetation types/formations/habitats were recognized in a data set of 942 phytosociological relevés: (1) synanthropic vegetation, (2) rock fissures and screes, (3) alpine grasslands and heaths, (4) mesic grasslands and pastures, (5) springs and fens, (6) forests, (7) xerophilous shrubland, and (8) high-mountain scrubs (krummholz). Results corroborated and clearly emphasized that Sesleria caerulea has the widest ecological amplitude of all studied species because the species occurred among all studied vegetation types. Sesleria tatrae was present only in several vegetation types occurring from montane to alpine vegetation belts in the highest mountains. Sesleria heufleriana and S. uliginosa were recorded only in low-altitude areas. The difference between them lies mainly in the dampness of each locality. Sesleria heufleriana was frequently found in xerophilous communities, whereas S. uliginosa preferred humid habitats of springs and fens.     

Analysis of genetic diversity and structure in Ethiopian populations of Phytolacca dodecandra using RAPD

The genetic diversity and structure in 17 wild populations (249 individuals) of Phytolacca dodecandra (endod) sampled along altitudinal gradients of 1600-3000 meters above sea level (m.a.s.l.) in Ethiopia was studied using random amplified polymorphic DNA (RAPD). A total of 70 polymorphic loci (P) scored from 12 RAPD primers were used to calculate different diversity indices within and between populations, habitats, geographical regions, climatic zones and altitude groups. The number of polymorphic loci and overall Shannon information measure (H) in the populations varied from 30 to 55 and from 0.228 to 0.418, respectively. In general, differences in population variability were found significantly correlated to effective population size. Both P and H were significantly higher in an undisturbed than in a disturbed habitat, and in the lowland and central-highland than in the highland altitude group. However, for both parameters the differences were not statistically significant between regions and climatic zones. Genetic distance between populations varied from 0.301 to 0.628. Cluster analysis performed using the genetic distance matrix revealed a clear separation of the highland populations (2501-3000 m.a.s.l.) from those of the lowland/central-highlands (1600-2500 m.a.s.l.) irrespective of their geographical regions and climatic zones. Analysis of molecular variance (AMOVA) indicated that differences in habitat, geographical regions and climatic zones explained 4.6%, 2.5% and 4.6%, respectively. But none of these differences were significant. Altitude explained 17.2% of the total variance and was highly significant. The data, therefore, clearly indicated the association of genetic structure in endod with altitude. The proportion of RAPD variation found among populations (21.2-35.0%) was somewhat intermediate between values reported for selfing and outcrossing species. The fixation index (FST) values (0.350 to 0.384) indicated very high genetic differentiation among populations.     

濒危灌木长叶红砂遗传多样性的RAPD分析

应用RAPD分子标记对濒危灌木长叶红砂(Reaumuria trigyna)种群遗传多样性进行了分析.应用18条随机引物对长叶红砂5个种群的95个个体进行扩增,检测到118个位点,其中多态位点105个.结果表明:长叶红砂种群的多态位点比率(P)为88.98%,显示出长叶红砂种群存在较高的遗传多样性.Shannon多样性指数(0.4966)、Nei基因多样性指数(0.3303)和基因分化系数(Gst=0.1425)的分析结果显示,长叶红砂种群遗传变异大多存在于种群内,种群间的遗传分化占14.25%.聚类分析表明,长叶红砂种群遗传距离与地理距离之间无直接相关关系.遗传多样性水平与物种特性和所处不同群落有关,濒危植物并不一定表现为遗传变异水平的降低.     

羊草种群遗传分化的RAPD分析I：扩增片段频率的变化

松嫩草原上分布着灰绿型和黄绿型两种叶色类型的羊草,为进一步明确这两种叶色类型种群分化的分子基础 ,对其9个种群105个个体进行了RAPD分析。15个随机引物（10nt)共扩增出149个多态性片段,总的多态位点为96．1％,黄绿型与灰绿型种群间在10．1％的多态性片段上明显不同,相似生境或同一地域的种群在一些位点上表现出相似或相同的变化;9个种群都检测到了特异性扩增。     

Divergent selection along climatic gradients in a rare central European endemic species,Saxifraga sponhemica

Background and Aims The effects of habitat fragmentation on quantitative genetic variation in plant populations are still poorly known. Saxifraga sponhemica is a rare endemic of Central Europe with a disjunct distribution, and a stable and specialized habitat of treeless screes and cliffs. This study therefore used S. sponhemica as a model species to compare quantitative and molecular variation in order to explore (1) the relative importance of drift and selection in shaping the distribution of quantitative genetic variation along climatic gradients; (2) the relationship between plant fitness, quantitative genetic variation, molecular genetic variation and population size; and (3) the relationship between the differentiation of a trait among populations and its evolvability.Methods Genetic variation within and among 22 populations from the whole distribution area of S. sponhemica was studied using RAPD (random amplified polymorphic DNA) markers, and climatic variables were obtained for each site. Seeds were collected from each population and germinated, and seedlings were transplanted into a common garden for determination of variation in plant traits.Key Results In contrast to previous results from rare plant species, strong evidence was found for divergent selection. Most population trait means of S. sponhemica were significantly related to climate gradients, indicating adaptation. Quantitative genetic differentiation increased with geographical distance, even when neutral molecular divergence was controlled for, and Q_ST exceeded F_ST for some traits. The evolvability of traits was negatively correlated with the degree of differentiation among populations (Q_ST), i.e. traits under strong selection showed little genetic variation within populations. The evolutionary potential of a population was not related to its size, the performance of the population or its neutral genetic diversity. However, performance in the common garden was lower for plants from populations with reduced molecular genetic variation, suggesting inbreeding depression due to genetic erosion.Conclusions The findings suggest that studies of molecular and quantitative genetic variation may provide complementary insights important for the conservation of rare species. The strong differentiation of quantitative traits among populations shows that selection can be an important force for structuring variation in evolutionarily important traits even for rare endemic species restricted to very specific habitats.     

Molecular Genetic Variation in a Clonal Plant Population of Leymus chinensis (Trin.) Tzvel.

Randomly amplified polymorphic DNA (RAPD) analysis was used to investigate the genetic variation among populations, between populations, and within populations, relationships between genetic distance and geographic distance, and the molecular variation and population size. The effects of geographic and genetic distances, as well as of genetic differentiation and population size, on genetic variations of Leymus chinensis (Trin.) Tzvel. are discussed. The present study showed that there was significant RAPD variation between the Baicheng region population and the Daqing region population, with a molecular variance of 6.35% (P ＜ 0.04), and for differentiation among area populations of the Daqing region, with a molecular variance of 8.78% (P ＜ 0.002). A 21.06% RAPD variation among all 16 populations among two regions was found (P ＜ 0.001), as well as 72.59% variation within populations (P ＜ 0.001). Molecular variation within populations was significantly different among 16 populations.     

内蒙古典型草原羊草种群遗传分化的RAPD分析

运用 RAPD技术对内蒙古典型草原不同生境 8个羊草种群进行分析。采用 2 4个随机引物 (10 nt)在 8个种群中共检测到2 2 4个扩增片断 ,其中多态性片断 173个 ,总的多态位点百分率达 77.2 % ,特异性片断 2 2个 ,占 9.82 % ,平均每个引物扩增的DNA带数为 9.3 3条。利用 Nei指数和 Shannon指数估算了 8个种群的遗传多样性 ,并计算种群相似系数和遗传距离 ,运用UPGMA法进行聚类分析。结果表明 :羊草大部分的遗传变异存在于种群内 ,只有少部分的遗传变异存在于种群间 ,Nei指数和Shannon指数计算结果分别为 85.4%和 66.8% ;羊草不同种群的遗传多样性存在差异 ;8个羊草种群平均遗传距离为 0 .2 3 16,变异范围为 0 .1587～ 0 .2 70 0 ,说明 8个羊草种群间的遗传变异不大 ,即 :在较小地理范围内羊草的遗传分化程度较小 ;8个种群可聚为 3个类群 ,聚类结果显示生境相似的种群能够聚在一起 ,而地理距离最近的种群不一定归为一类 ,说明小范围内羊草种群间的遗传分化与地理距离不存在相关性 ,而与其生境间的相似度相关。影响遗传相似性的不是单一因子而是各种因子的综合作用 ,较小地理范围内羊草种群间的遗传分化主要是由环境的异质性所引起的     

RAPD Profiling in Detecting Genetic Variation in Endemic Coelonema (Brassicaceae) of Qinghai-Tibet Plateau of China

Random amplified polymorphic DNA (RAPD) markers were used to measure genetic diversity of Coelonema draboides (Brassicaceae), a genus endemic to the Qilian Mountains of the Qinghai-Tibet Plateau. We sampled 90 individuals in 30 populations of Coelonema draboides from Datong and Huzhu counties of Qinghai Province in P.R. China. A total of 186 amplified bands were scored from the 14 RAPD primers, with a mean of 13.3 amplified bands per primer, and 87% (161 bands) polymorphic bands (PPB) was found. Analysis of molecular variance (AMOVA) shows that a large proportion of genetic variation (84.2%) resides among individuals within populations, while only 15.8% resides among populations. The species shows higher genetic diversity between individuals than other endemic and endangered plants. The RAPDs provide a useful tool for assessing genetic diversity of rare, endemic species and for resolving relationships among populations. The results show that the genetic diversity of this species is high, possibly allowing it to adapt more easily to environmental variations. The main factor responsible for the high level of differentiation within populations and the low level of diversity among populations is probably the outcrossing and long-lived nature of this species. Some long-distance dispersal, even among far separated populations, is also a crucial determinant for the pattern of genetic variation in the species. This distributive pattern of genetic variation of C. draboides populations provides important baseline data for conservation and collection strategies for the species. It is suggested that only populations in different habitats should be studied and protected, not all populations, so as to retain as much genetic diversity as possible.     

拟环纹豹蛛种群遗传多样性与其生境的关系

对分布于我国中南、西南和海南岛8个不同生境的拟环纹豹蛛种群进行RAPD分析.筛选出10对引物扩增出清晰稳定的200～2 500 bp片段84条,其中多态性片段62条(占73.8%).表明种群存在明显多态现象.Shannon指数、相似系数和遗传距离测定以及聚类分析的结果表明：拟环纹豹蛛种群总的遗传多样性指数为0.5177,而且种群内遗传变异（64.24%)大于种群间（35.76%）;8个狼蛛种群平均遗传距离为0.2426,变异范围为0.0753～0.3725,表明8个种群由于所处生境条件不同而产生了一定的适应性变异.多元回归统计结果表明,制约拟环纹豹蛛成为稻田优势种的主要因子是年平均气温和农药的长期施用.     

Genetic variation in wild sorghum (Sorghum bicolor ssp. verticilliflorum (L.) Moench) germplasm from Ethiopia assessed by random amplified polymorphic DNA (RAPD)

The extent and distribution of genetic variation in wild sorghum (Sorghum bicolor ssp. verticilliflorum (L.) Moench) collected from five different geographical regions in Ethiopia were analyzed using random amplified polymorphic DNA (RAPD) markers for 93 individuals representing 11 populations. Nine decamer primers generated a total of 83 polymorphic bands with 8-12 bands per primer and a mean of 9 bands across the 93 individuals. The amount of genetic variation among the populations (H = 0.37) and among the geographical region (H = 0.44) was low to moderate, despite the high degree of polymorphic bands per primer. Similarly, the mean genetic distance (0.08) among populations as well as among regions of origin (0.04) of the population was found to be low. The low genetic variation may be due to the reduced population size of the wild sorghum in Ethiopia because of habitat change. Partitioning of the genetic variation into between and within the population as well as between and within the regions of origin revealed that 75% and 88% of the variation was found within the populations and within the regions, respectively. Cluster analysis of genetic distance estimates further confirmed low level of differentiation of wild sorghum populations both on population and regional bases. The implications of the results for genetic conservation purposes are discussed.     

Comparison of multivariate methods for the analysis of genetic resources and adaptation in Phytolacca dodecandra using RAPD

The extent of genetic differentiation among 17 Ethiopian populations (249 individuals) of Phytolacca dodecandra (Endod) sampled along altitudinal gradients that varied from 1600 to 3000 m was investigated using random amplified polymorphic DNA (RAPD). The populations were classified into three altitude groups: lowland (1600–2100 m), central-highland (2101–2500 m) and highland (2500–3000 m). Seventy polymorphic loci scored from 12 RAPD primers, singly or in combination with ecogeographical variables (altitude, longitude, latitude, temperature and rainfall), were used for principal component, discriminant, correlation, and stepwise multiple regression analyses. Principal component analysis (PCA) clearly differentiated lowland and the central-highland populations from those of the highlands independent of their geographical regions. Canonical discriminant analysis separated the lowland plants from those of the highlands with the central-highland plants being intermediate. Classificatory discriminant analysis corrected classification of 92.8% of the 249 plants into their respective three altitude groups. Multiple regression analysis identified a strong association between some RAPDs and altitude, temperature and rainfall, while the variation in most RAPDs was explained by combinations of the different ecogeographical variables. It is hypothesised that the different altitude groups may be (1) chemical and/or physiological ecotypes produced as a result of complex interactions of altitude with climatic and/or edaphic factors, or (2) different in ploidy levels. The significant correlations obtained between population means from some RAPDs and altitude and temperature as well as the strong association of some RAPDs with the ecogeographical variables in the multiple regression analysis suggest that part of the RAPD polymorphism could be adaptive, and responsive to environmental selection. Received: 15 December 1999 / Accepted: 12 February 2000     

濒危植物三棱栎遗传多样性的RAPD分析

用随机扩增多态DNA (RAPD)标记对 5个三棱栎 (Trigonobalanusdoichangensis)居群共 99个个体进行遗传多样性和居群遗传结构分析。 16个引物共检测到 15 7个位点 ,其中多态位点 83个 ,占 5 2 87%。物种水平Shannon多样性指数I =0 2 4 31,Nei基因多样度h =0 15 95 ,种内总遗传变异量Ht=0 16 0 0 ,居群内遗传变异量Hs =0 0 74 9,居群间变异量大于居群内变异量 ,表明三棱栎的遗传变异主要存在于居群之间。与同科植物相比 ,三棱栎遗传多样性较低 ,遗传分化系数Gst =0 5 32 0 ,说明居群间的遗传变异占 5 3 2 0 % ,居群间已出现强烈的遗传分化。当地人的强烈活动造成的生境破碎化和居群隔离 ,以及三棱栎演化过程中的地史变化对其种群发展的影响等 ,可能是造成其居群间强烈的遗传分化和较低遗传多样性的原因。基于本研究结果 ,提出了三棱栎遗传多样性的保护策略。     

凉水国家自然保护区天然红松林遗传变异的RAPD分析

采用随机扩增多态DNA(random amplified polymorphic DNA RAPD)技术,研究了凉水国家自然保护区天然红松林的遗传变异水平及其分布规律。16个10bp长度的随机引物在8个实验样地内72个个体中共检测了96个位点,其中55个是多态位点。在种水平上,红松的多态位点比率为P=58.51%;Nei's遗传变异指数为H=0.2868;Shannon's指数为I=0.3654,所有的变异中,有92.47%的变异存在于样地内, 7.53%的变异存在于样地间。根据各遗传多样性在不同样地的分布,初步探讨了红松遗传变异水平与其种群发生及生境的相互关系。     

内蒙古中东部不同草原地带羊草种群遗传分化

运用 RAPD技术对内蒙古不同草原地带分布的 5个羊草种群 (共 10 0个个体 )的遗传多样性进行分析。 31个随机引物(10 nt)在 5个羊草种群中共检测到 4 96个扩增片断 ,其中多态性片断 4 89个 ,总的多态位点百分率达 98.6 %。利用 Nei指数和Shannon指数估算了 5个种群的遗传多样性 ,并计算种群相似系数和遗传距离运用 UPGMA法进行聚类分析。结果表明 :无论是种群内还是种群间 ,羊草均存在较高的遗传变异 ,大部分的遗传变异存在于种群内 (Nei指数和 Shannon指数估算结果分别为 85 .4 %和 72 .5 % ) ,只有少部分的遗传变异存在于种群间 ;不同种群的遗传多样性存在差异 ,各种群的遗传多样性与其所处的地理位置具有显著的相关性 ;5个种群的平均遗传距离为 0 .5 0 95 ,变异范围为 0 .4 6 84～ 0 .5 4 76 ;聚类分析结果显示地理距离较近的种群遗传距离较小 ,首先聚在一起 ,而地理距离较远的种群遗传距离较大 ,说明羊草种群间的遗传分化与地理距离存在一定相关性 ;但地理距离最近的两个种群并未最先聚集 ,说明羊草种群间的分化还与其生境的异质性有关     

地被植物鹅绒委陵菜的遗传多样性研究

采用RAPD方法对不同海拔和植被盖度下的鹅绒委陵菜（Potentilla anserinaL.）6个种群的遗传多样性水平进行了研究。结果表明,13条随机引物共扩增出132条带,多态性位点为117;鹅绒委陵菜物种水平的遗传多样性较高,且种群间分化明,多态位点比率（88.64%）、Nei’s基因多样性（0.3180）、Shannon信息多样性指数（0.4732）均远高于多数克隆植物。6个种群内的遗传多样性水平变化较大,多态位点在各种群中分布不均衡,种群间分化系数（GST）为47.7%,基因流较低（Nm=0.5482）,多态位点比率在31.06%～74.24%之间,Shannon信息指数在0.1711～0.3625之间,Nei基因多样性指数在0.1164～0.2425之间。多样性水平的变化与海拔没有明显的相关性,而与生境盖度呈显著正相关。从而推断在盖度低、资源丰富的环境中,该物种可能更倾向于克隆繁殖。     

AFLP and RAPD analyses of genetic diversity of wild and/or weedy Guizotia (Asteraceae) from Ethiopia

Amplified fragment length polymorphism (AFLP) and random amplified polymorphic DNA (RAPD) markers were used to provide estimates of the comparative genetic variation within and among populations of various Guizotia taxa with the goal of conserving and utilizing their genetic diversity. The percentage of polymorphic loci (P(S)) ranged from 28.5%-90% (AFLP) and 85.6%-99.6% (RAPD). The overall gene diversity estimate () has shown slight variation among taxa ranging from 0.32-0.37 (AFLP) and from 0.22 to 0.28 (RAPD). The within population diversity of "Chelelu" and "Ketcha" was found to be unexpectedly high. Both parameters used to estimate population differentiation (G(ST) and F(ST)) revealed the highest population differentiation G. zavattarii in followed by G. arborescens. Genetic variation among populations within a taxon was highly significant for all the five taxa as revealed by AMOVA (P<0.0001). The need for immediate conservation activities for G. arborescens and G. zavattarii, and factors that contribute to the existing genetic variability and population genetic structures are discussed.     

Genetic variation in an apomictic grass, heteropogon contortus, in the hawaiian islands

Random amplified polymorphic DNA (RAPD) markers were used to assess genetic variation within and among Hawaiian populations of an apomictic grass, Heterogopon contortus (pili grass). From among 56 individuals sampled from six populations on O'Ahu and Hawai'i, 55 unique genotypes were detected using 33 polymorphic markers. This lack of uniformity among individuals may indicate frequent sexual reproduction in these populations. Analysis of molecular variance (AMOVA) revealed significant variation among populations (30.2%), but higher levels of variation within populations (68.1%). Cluster analysis revealed a high degree of clustering for most populations, but populations from different islands did not cluster together. The presence of among-population differentiation but lack of between-island differentiation may suggest that H. contortus was an early Polynesian introduction to the Hawaiian Islands.     

Population variation and phylogeny in the endangered <Emphasis Type="BoldItalic">Chamaesyce skottsbergii</Emphasis> (Euphorbiaceae) based on RAPD and ITS analyses

Chamaesyce skottsbergii var. skottsbergii is federally listed as an endangered taxon, and is found in small and isolated populations restricted to calcareous soils in dry shrubland habitats on the Hawaiian islands of Oahu and Molokai. Concern over the genetic relationship among these disjunct populations arose as a result of threats to the habitat of the Oahu population. The populations were examined using random amplified polymorphic DNA (RAPD) markers and sequence analysis of the internal transcribed spacer (ITS) region of the rDNA cistron. Chamaesyce skottsbergii var. vaccinioides, a closely related variety found in several small populations on Molokai, was used for baseline comparison of the genetic divergence among populations. RAPD analysis demonstrated that variation within and among populations is the highest for any Hawaiian species examined. Polymorphism was greater than 95% within populations and was 99.4% at the species level. Similarly, measures of genetic similarity indicate that differentiation among these populations is higher than is known for some species. Both RAPD and ITS sequence analysis indicate that populations of C. skottsbergii var. skottsbergii on Oahu and Molokai are genetically distinct, and the extent of this genetic differentiation supports the recognition of these populations as distinct varieties. The Molokai population is in fact much more closely related to var. vaccinioides than to var. skottsbergii on Oahu, and thus should be recognized by the previously used variety name, C. skottsbergii var. audens. Further conservation measures for each of the varieties are addressed.     

Diversity and genetic structure in populations of Pseudotsuga menziesii (Pinaceae) at chloroplast microsatellite loci.

Genetic variation was compared between uniparentally-inherited (chloroplast simple sequence repeats, cpSSRs) vs. biparentally-inherited (isozyme and random amplified polymorphic DNA, RAPD) genetic markers in Douglas-fir (Pseudotsuga mensiezii) from British Columbia. Three-hundred twenty-three individuals from 11 populations were assayed. In Douglas-fir, the cpSSR primer sites were well-conserved relative to Pinus thunbergii (11 of 17 loci clearly amplified), but only 3 loci were appreciably polymorphic. At these cpSSR loci, we found an unexpectedly low level of polymorphism within populations, and no genetic differentiation among populations. By contrast, the nuclear markers showed variation typical of conifers, with significant among-population differentiation. This difference is likely the outcome of both historical factors and high pollen dispersal.