Clonal diversity of <Emphasis Type="Italic">Clintonia udensis</Emphasis> Trautv. <Emphasis Type="Italic">et</Emphasis> Mey. populations and its correlation with ecological factors

The clonal diversity of Clintonia udensis Trautv. et Mey. was detected by ISSR markers among 16 populations, and its correlation with ecological factors was analyzed as well in this work. Results showed that individuals (clonal ramets) per genotype were 1.12 and 1.149 at population and species levels, respectively, and that the 16 populations were all multiclonal. The detected genotypes were localized, without exception, within populations but demonstrated relatively high clonal differentiation among populations. The clonal diversity of the studied populations was high, with the average Simpson’s index of 0.975, while the genets showed a clonal architecture of “guerilla”. The population genetic diversities revealed by genet were consistent with those by ramet, further confirming their genetic differentiation among populations. And its genotype diversity within populations probably resulted largely from the frequent seedling regeneration and self-compatibility. In addition, the correlation analysis further revealed that, among the ecological factors, Simpson’s index of C. udensis had a significant positive correlation (P<0.05) with pH values in the soil but not others.     

贺兰山丁香自然居群克隆生长格局及遗传多样性的ISSR分析

运用ISSR分子标记技术,通过制定挖掘采样、"+"形采样及"垂直"采样3种采样方案,对贺兰山丁香(Syringa pinnatifolia var.alanshanica)不同居群的克隆多样性、克隆生长格局及其遗传多样性进行了分析.克隆多样性分析表明:挖掘采样方式采到的3个克隆系内,各自所包含的单株间具有完全相同的基因型;"垂直"采样及"+"形采样的7个居群、 239个样品表现出136个不同的基因型或克隆,显示贺兰山丁香具有较高的克隆多样性(D=0.994)及基因型分布均匀性(E=0.985).克隆生长空间格局分析表明,贺兰山丁香为密集型克隆植物.每个居群都由多克隆组成,克隆生长只发生在同一丛内,多数基株只含有1个分株,最多可达8个.物种水平上的平均克隆大小(N_C)和平均基因型比例(PD)分别为1.757和0.569.对贺兰山丁香遗传多样性分析的结果显示,在居群水平和物种水平上都保持着较高的遗传多样性,其遗传变异主要存在于居群内;但居群间分化程度较低(G_ST=0.320),表明自然居群间基因交流有限.     

七筋菇自然居群的遗传结构分析

采用ISSR分子标记,对七筋菇(Clintonia udensis)17个居群的遗传多样性与遗传结构进行了研究。结果表明:七筋菇不同居群的多态位点百分率PPB为11.90%~59.52%,总的多态位点百分率PPB为98.8%,具有高的遗传多样性。Shannon多样性指数(0.6903)和基因分化系数(GST=0.6944)均揭示出七筋菇居群间存在明显的遗传差异,AMOVA分析结果也显示遗传变异主要发生在居群之间(81.47%),而居群内部的遗传变异仅为18.53%。七筋菇居群间的遗传距离从0.1871~0.6632,平均为0.3838,大于同一物种居群间的平均遗传距离值(0.05),同样表明七筋菇居群间的遗传多样性存在较大差异。七筋菇居群间的基因流Nm=0.2200,远远低于一般广布种植物的基因流(Nm=1.881)。Mantel检测显示居群间的遗传距离与地理距离之间没有显著相关性(r=0.029,P=0.3196)。七筋菇分布范围广以及其进化历史是其具有高遗传多样性的原因;居群间存在较高遗传变异可能是由于七筋菇本身的生物学特性、有限的基因流以及遗传漂变等原因造成的。     

Clonal diversity and structure within a population of the pondweed Potamogeton pectinatus foraged by Bewick's swans

Clonal diversity within plant populations is affected by factors that influence genet (clone) survival and seed recruitment, such as resource availability, disturbance, seed dispersal mechanism, propagule predation and the age of the population. Here we studied a population of Potamogeton pectinatus, a pseudo-annual aquatic macrophyte. Within populations reproduction appears to be mainly asexually through subterranean propagules (tubers), while recruitment via seeds is believed to be relatively unimportant. RAPD markers were used to analyse clonal diversity and genetic variation within the population. Ninety-seven genets were identified among 128 samples taken from eight plots. The proportion of distinguishable genets (0.76) and Simpson's diversity index (0.99) exhibited high levels of clonal diversity compared to other clonal plants. According to an analysis of molecular variance (amova) most genetic variation occurred between individuals within plots (93-97%) rather than between plots (8-3%). These results imply that sexual reproduction plays an unexpectedly important role within the population. Nevertheless, autocorrelation statistics revealed a spatial genetic structure resulting from clonal growth. In contrast to genetic variation, clonal diversity was affected by several ecological factors. Water depth and silt content had direct negative effects on clonal diversity. Tuber predation by Bewick's swans had an unexpected indirect negative effect on clonal diversity through reducing the tuber-bank biomass in spring, which on its turn was positively correlated to clonal diversity. The disturbance by swans, therefore, did not enhance seed recruitment and thus clonal diversity; on the contrary, heavily foraged areas are probably more prone to stochastic loss of genets leading to reduced clonal diversity.     

Clonal diversity and genetic differentiation in Ilex leucoclada M. patches in an old-growth beech forest

We investigated clonal diversity within patches of Ilex leucoclada and genetic variation within and among patches using random amplified polymorphic DNA (RAPD) markers in a 1-ha plot within an old-growth beech forest. We found 38 patches that exhibited a clumped distribution in the middle of the plot. We identified a total of 166 RAPD phenotypes among the 215 stems sampled from 27 patches that were completely within the plot. The population showed high clonal diversity within patches (mean number of genets relative to number of stems = 0.79; mean Simpson's D = 0.89). Variation in RAPD phenotypes among patches was highly significant (PhiST in the molecular variance analysis = 0.316, P < 0.001), indicating genetic differentiation among patches. Pairwise genetic distances, PhiST, among patches did not correlate with geographical distances among patches. The cluster analysis based on the genetic distances showed few clear clusters of patches, indicating no spatial genetic structure among patches. High levels of clonal diversity both within patches and within the population may be explained by multiple founders, seedling recruitment during patch-formation, and somatic mutation. The significant genetic differentiation among patches may be caused by separate founding events and/or kin structuring within patches.     

RAPD analysis of genetic diversity and population genetic structure of Stipa krylovii Reshov. in Inner Mongolia steppe

Random amplified polymorphic DNA (RAPD) analysis was used to characterize the genetic diversity and population genetic structure of Stipa krylovii populations in Inner Mongolia steppe of North China. Thirteen 10 bp oligonucleotide primers, which generated 237 RAPD bands, were used to analyze 90 plants of five populations from three regions, meadow steppe, typical steppe and desert steppe, from the east to the west. The genetic diversity of Stipa krylovii that was revealed by observed number of alleles (na), expected number of alleles (ne), Nei's diversity index (h), Shannon's diversity index (H), amplificated loci, polymorphic loci and the percentage of polymorphic loci (PPB) increased from the east to the west. The Pearson's correlation analysis between genetic diversity parameters and ecological parameters indicated that the genetic diversity of Stipa krylovii was associated with precipitation and cumulative temperature variations along the longitude (humidity were calculated by precipitation and cumulative temperature). Dendrogram based on Jaccard's genetic distance showed that the individuals from the same population formed a single sub-group. Although most variation (56.85%) was within populations, there was high genetic differentiation among populations of Stipa krylovii, high differentiation within and between regions by AMOVA analysis. Either Nei's unbiased genetic distance (G(ST)) or gene flow (Nm) among pairwise populations was not correlated with geographical distance by Mantel's test (P > 0.05), suggesting that there was no consistency with the isolation by distance model in these populations. Natural selection may have played a role in affecting the genetic diversity and population structure, but habitat destruction and degradation in northern grassland in China may be the main factor responsible for high genetic differentiation among populations, within and among regions.     

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.     

内蒙古小花棘豆遗传多样性的ISSR分析

应用ISSR分子标记对内蒙古西部地区小花棘豆（Oxytropis glabra DC.）6个地理种群进行了遗传多样性研究.结果表明：小花棘豆具有较高的遗传多样性;14个引物共扩增出327个位点,物种水平上Nei＇s基因多样性指数为0.1972,Shannon多样性指数为0.3287;种内总基因多样性为0.1976,种群内基因多样性为0.1653,83.67%的遗传变异存在于种群内,16.33%的遗传变异存在于种群间,种群间的遗传分化系数为0.1633,基因流为2.5613,6个种群间有一定遗传分化,但遗传漂变不会引起遗传分化.UPGMA遗传距离聚类结果表明,生态地理条件相似的种群优先聚集.     

Demographic and random amplified polymorphic DNA analyses reveal high levels of genetic diversity in a clonal violet

We performed demographic and molecular investigations on woodland populations of the clonal herb Viola riviniana in central Germany. We investigated the pattern of seedling recruitment, the amount of genotypic (clonal) variation and the partitioning of genetic variation among and within populations. Our demographic study was carried out in six violet populations of different ages and habitat conditions. It revealed that repeated seedling recruitment takes place in all of these populations, and that clonal propagation is accompanied by high ramet mortality. Our molecular investigations were performed on a subset of three of these six violet populations. Random amplified polymorphic DNA analyses using six primers yielded 45 scorable bands that were used to identify multilocus genotypes, i.e. putative clones. Consistent with our demographic results and independent of population age, we found a large genotypic diversity with a mean proportion of distinguishable genotypes of 0.93 and a mean Simpson's diversity index of 0.99. Using AMOVA we found a strong genetic differentiation among these violet populations with a PhiST value of 0.41. We suggest that a high selfing rate, limited gene flow due to short seed dispersal distances and drift due to founder effects are responsible for this pattern. Although Viola riviniana is a clonal plant, traits associated with sexual reproduction rather than clonality per se are moulding the pattern of genetic variation in this species.     

大别山山核桃种群遗传多样性研究

为了更有效地保护和合理开发大别山山核桃(Carya dabieshanensis)资源,该文利用RAPD分子标记技术,对3个天然大别山山核桃种群的90个单株的遗传多样性、种群内和种群间的遗传变异进行了研究,结果表明:20对10 bp随机引物共检测到238条谱带,其中多态带为162条,占68.1%。遗传多样性分析结果显示: Shannon多样性指数为0.476 1,58.18%的变异分布于群体内,而种群间变异占了41.82%;Nei指数群体总基因多样度为0.314 5,群体内平均基因多样度(H_S)为0.186 5,群体间的基因多样度(H_ST)为0.128 0,群体Nei基因分化系数(G_ST)为0.406 7,说明40.67%的变异存在于种群间,群体内的变异占了总变异的59.33%,与Shannon多样性指数相比基本一致,均表明种群内有较丰富的遗传变异,这为优良品种选育提供广阔前景;种群间的基因流(N_m)为0.730 6,证明种群间遗传交换较小,这与环境适应性和高山阻隔有一定的关系。     

海南岛海菖蒲种群克隆多样性和遗传结构

海草是生长在潮间带和潮下带的单子叶植物,由海草植物组成的海草床是生态系统服务价值最高的生态系统之一.然而,近几十年人类活动干扰、全球气候变化等因素导致海草床衰退严重.海菖蒲是分布于热带、体型最大的雌雄异株海草,我国位于该物种的分布北缘,本文对其克隆多样性和遗传结构进行研究,以期为该海草的保护提供参考.采用4对多态微卫星标记对采自海南岛4个地点的现存海菖蒲种群的样品进行基因型分型.结果表明:海菖蒲种群克隆多样性和遗传多样性较低,这与所研究种群处于分布区北缘有关;种群间遗传分化值范围较大(0.073~0.309),这可能是由于分布于不同港湾的种群间距离范围较大以及局域绝灭/再拓殖的遗传漂变效应所致;各种群未发现近期经历种群瓶颈的信号,很可能是由于种群内遗传多样性已经很低,种群减小未能导致遗传多样性明显降低.根据种群遗传特征,提出了重点保护种群的建议,鉴于目前我国海菖蒲等海草快速衰退的局面,应强化海草保护并实施海草床生态恢复.     

Clonal genetic diversity and populational genetic differentiation in Phragmites australis distributed in the Songnen Prairie in northeast China as revealed by amplified fragment length polymorphism and sequence-specific amplification polymorphism molecular markers

Genetic variation within and between four naturally occurring Phragmites australis land populations, DBS, QG, SS1 and SS2 (named after locality), which colonise distinct habitats (different edaphic conditions) in the Songnen Prairie in northeast China, were investigated by amplified fragment length polymorphism (AFLP) and sequence‐specific amplification polymorphism (S‐SAP) markers. It was found that the selected primer combinations of both markers were highly efficient in revealing the inter‐clonal genetic diversity and inter‐populational genetic differentiation in P. australis from a molecular ecological perspective. Cluster analysis categorised the plants into distinct groups (DBS, QG and SS groups), which were in line with their localities, albeit the two SS group populations (SS1 and SS2) showed a lower degree of inter‐populational differentiation. These results were strongly supported by multiple statistical analysis including Mantel’s test, principal coordinate analysis, allocation test and analysis of molecular variance, which further suggested that gene flow, genetic drift and differences in as yet unidentified edaphic factors may all underpin the inter‐clonal genetic diversity and inter‐populational differentiation at the nucleotide sequence level. Analysis of intra‐population clonal diversity also revealed that the QG population harboured a strikingly lower amount of within‐population variation compared with those of the other three populations, presumably being caused by genetic drift and followed by physical and/or biological isolation. Homology analysis of a subset of population‐specific or population‐private AFLP and S‐SAP bands suggested that regulatory genes and retroelements might play important roles in the ecological adaptation and differentiation of the P. australis populations. Possible causes for and implications of the extensive genetic variability in P. australis were discussed for its future genetic conservation and use in ecological revegetation.     

Genetic diversity and genetic structure of populations of Ranunculus japonicus Thunb. (Ranunculaceae)

Abstract In order to clarify the genetic diversity and population structure of Ranunculus japonicus , allozymic analysis was conducted on 60 populations in southwestern Japan. Considerable genetic variati ons were detected among the populations of R. japonicus . The genetic diversities within species ( H _es = 0.215) and within populations ( H _ep = 0.172) were slightly higher than those of other perennial herbs with widespread distribution and outcrossing plants. Significantly higher values of fixation index were detected in some populations, which might have arisen from restricted mating partners. The majority of genetic variation (approx. 80%) resided within a population and a moderate level of genetic differentiation ( G _ST = 0.203) was observed among populations. The F _ST value (0.203) suggests the existence of a substantial population structure in this species. The highly significant correlation between geographic distance and F _ST values indicates that isolation by distance has played an important role in the construction of the genetic structure of this species.     

竹叶眼子菜居群遗传多样性和克隆结构

采用ISSR技术对长江中游南岸豹澥湖和大冶湖不同生境中的竹叶眼子菜(Potamogeton malaianus)居群的遗传多样性及克隆结构进行了研究.结果表明,在两居群的106株个体中,利用6条ISSR引物共得到40条符合3/N标准并无连锁不平衡的清晰位点,竹叶眼子菜具有较高的遗传多样性,其多态位点百分率为75.0％,Shannon多样性指数为0.3736, 两居群的遗传分化很小.竹叶眼子菜的克隆多样性很高(D=0.9917),两居群间的克隆分化很大,不具有共有的基因型.竹叶眼子菜的基株分布为游击型构型,位于湖心的大冶居群克隆距离(3.0～31.5 m)明显地大于湖岸豹澥居群(2.4～6.7 m).     

Genetic Diversity and Geographic Differentiation in the Threatened Species Dysosma pleiantha in China as Revealed by ISSR Analysis

Dysosma pleiantha, an important threatened medicinal plant species, is restricted in distribution to southeastern China. The species is capable of reproducing both sexually and asexually. In this study, inter-simple sequence repeat marker data were obtained and analyzed with respect to genetic variation and genetic structure. The extent of clonality, together with the clonal and sexual reproductive strategies, varied among sites, and the populations under harsh ecological conditions tended to have large clones with relatively low clonal diversity caused by vegetative reproduction. The ramets sharing the same genotype show a clumped distribution. Across all populations surveyed, average within-population diversity was remarkably low (e.g., 0.111 for Nei’s gene diversity), with populations from the nature reserves maintaining relatively high amounts of genetic diversity. Among all populations, high genetic differentiation (AMOVA: Φ_ST = 0.500; Nei’s genetic diversity: G _ST = 0.465, Bayesian analysis: Φ_B = 0.436) was detected, together with an isolation-by-distance pattern. Low seedling recruitment due to inbreeding, restricted gene flow, and genetic drift are proposed as determinant factors responsible for the low genetic diversity and high genetic differentiation observed.     

Populations do not become less genetically diverse or more differentiated towards the northern limit of the geographical range in clonal Vaccinium stamineum (Ericaceae)

Geographically peripheral populations are expected to exhibit lower genetic diversity and higher differentiation than central populations because of their smaller size and greater spatial isolation. In plants, a shift from sexual to clonal asexual reproduction may further reduce diversity and increase differentiation. Here, these predictions were tested by assaying 36 inter-simple sequence repeat (ISSR) polymorphisms in 21 populations of the woody, clonal plant Vaccinium stamineum in eastern North America, from the range center to its northern limit where it has 'threatened' status. Populations decline in frequency, but not size or sexual reproductive output, across the range. Within-population diversity did not decline towards range margins. Modest genetic differentiation among populations increased slightly towards range margins and in small populations with high clonal propagation and low seed production, although none of these trends was significant. Low seed production and high clonal propagation were not associated with large-scale clonal spread. By combining demographic and genetic data, this study determined that increased population isolation, rather than reduced population size, can account for the weak increase in genetic differentiation at range margins.     

RAPD analysis of genetic diversity and population genetic structure of Stipa krylovii reshov. in Inner Mongolia steppe

Random amplified polymorphic DNA (RAPD) analysis was used to characterize the genetic diversity and population genetic structure of Stipa krylovii populations in Inner Mongolia steppe of North China. Thirteen 10-bp oligonucleotide primers, which generated 237 RAPD bands, were used to analyze 90 plants of five populations from three regions, meadow steppe, typical steppe and desert steppe, from the east to the west. The genetic diversity of Stipa krylovii that was revealed by observed number of alleles (na), expected number of alleles (ne), Nei’s diversity index (h), Shannon’s diversity index (H), amplificated loci, polymorphic loci and the percentage of polymorphic loci (PPB) increased from the east to the west. The Pearson’s correlation analysis between genetic diversity parameters and ecological parameters indicated that the genetic diversity of Stipa krylovii was associated with precipitation and cumulative temperature variations along the longitude (humidity were calculated by precipitation and cumulative temperature). Dendrogram based on Jaccard’s genetic distance showed that the individuals from the same population formed a single subgroup. Although most variation (56.85%) was within populations, there was high genetic differentiation among populations of Stipa krylovii, high differentiation within and between regions by AMOVA analysis. Either Nei’s unbiased genetic distance (G _ST) or gene flow (Nm) among pairwise populations was not correlated with geographical distance by Mantel’s test (P > 0.05), suggesting that there was no consistency with the isolation by distance model in these populations. Natural selection may have played a role in affecting the genetic diversity and population structure, but habitat destruction and degradation in northem grassland in China may be the main factor responsible for high genetic differentiation among populations, within and among regions. The text was submitted by the authors in English.     

Genetic differentiation among Hemarthria compressa populations in south China and its genetic relationship with H. japonica

Within and among populations genetic variance of twelve Hemarthria compressa populations and one Hemarthria japonica population from China were analyzed using inter simple sequence repeat (ISSR). Twelve primers amplified a total of 165 genomic DNA fragments across a total of 148 individuals of which 156 were polymorphic (94.55%). 75.76% of the bands were unique to each species, while the average genetic distance (GD) between one population of H. japonica and twelve populations of H. compressa was 0.44, which suggest that there was distinct differentiation between these two species. In H. compressa, twelve primers produced 145 bands across 145 individuals. High genetic diversity was observed at species level. The percentage of polymorphic loci (P) was 86.21% and Shannon's information index of diversity (I) was 0.357. In contrast, there were relatively low levels of genetic diversity within population (P=32.93%, I=0.174). Analysis of molecular variance (AMOVA) showed that a considerable proportion of genetic variation (48.02%) resided among populations. The coefficient of gene differentiation (G(ST)=48.6%) also suggested that there was strong genetic differentiation among H. compressa populations in southern China. An indirect estimate of the number of migrants per generation (N(m)=0.264) indicated that gene flow was low among populations of this species. Relative high clonal diversity was found, and all local genotypes were found.     

Genetic diversity and multilocus genetic structure in the relictual endemic herb<Emphasis Type="Italic"> Japonolirion osense</Emphasis> (Petrosaviaceae)

Plant clonality may greatly reduce effective population size and influence management strategies of rare and endangered species. We examined genetic diversity and the extent of clonality in four populations of the monotypic herbaceous perennial Japonolirion osense, which is one of the most rare flowering plants in Japan. Allozyme analysis revealed moderate levels of genetic variation, and the proportion of polymorphic loci (P=66.7%) was higher than the value for species with similar life-history traits. With four polymorphic loci, 19 multilocus genotypes were observed among 433 aerial shoot samples and 10 (52%) were found only in single populations. The proportion of distinguishable genotypes (PD=0.10) and Simpson's index of diversity (D=0.52) also exhibited moderate levels of genotypic diversity compared to other clonal plants, with genotype frequencies at Hardy-Weinberg equilibrium. The distributions of genotypes were often localized and they were mostly found within a radius of 5 m. Spatial autocorrelation analysis showed that shoot samples located 4 m apart were expected to be genetically independent. The results suggest that the spatial extent of genets was relatively narrow and thus the clonality was not extensive.     

山东省长岛县南部四岛藜（Chenopodium album L.）天然种群遗传多样性的ISSR分析

采用ISSR分子标记对分布于山东省长岛县南部的北长山、南长山、大黑山和小黑山4个岛屿上的藜天然种群共81个个体进行了遗传多样性及遗传结构的研究。13个引物共检测到157个可重复的位点。遗传多样性研究结果表明：种内的多态位点比率为66.24％,具有较高的遗传多样性（Shannon（I）指数在物种水平上为0.332 0）;种群间有一定的遗传分化,根据G_st值,种群间的遗传多样性占总群体的9.27％。遗传距离分析表明,XHS种群和NCS种群的遗传一致度最高,与地理距离无相关性。