Sexual reproduction, clonal diversity and genetic differentiation in patchily distributed populations of the temperate forest herb Paris quadrifolia (Trilliaceae)

Clonal plant species have been shown to adopt different strategies to persist in heterogeneous environments by changing relative investments in sexual reproduction and clonal propagation. As a result, clonal diversity and genetic variation may be different along environmental gradients. We examined the regional and local population structure of the clonal rhizomatous forest herb Paris quadrifolia in a complex of forest fragments in Voeren (Belgium). Relationships between population size (the number of shoots), shoot density (the number of shoots per m²) and local growth conditions were investigated for 47 populations. Clonal diversity and genetic variation within and among 19 populations were investigated using amplified fragment length polymorphism markers. To assess the importance of sexual reproduction, seed set, seed weight and germination success were determined in 18 populations. As predicted, local growth conditions largely affected population distribution, size and density of P. quadrifolia. Populations occurring in moist and relatively productive sites contained significantly more shoots. Here, shoots were also much more sparsely distributed compared to populations occurring in dry and relatively unproductive sites, where shoots showed a strongly aggregated distribution pattern. Clonal diversity was relatively high, compared with other clonal species (G/N ratio = 0.43 and Simpson’s D=0.81). Clonal diversity significantly (P<0.01) decreased with increasing shoot density while molecular genetic variation was significantly (P<0.01) affected by population size and local environmental conditions. Lack of recruitment and out-competition of less-adapted genotypes may explain the decreased genetic variation in dry sites. Analysis of molecular variance revealed significant genetic variation among populations (Φ _ST=0.42, P<0.001), whereas pairwise genetic distances were not correlated to geographic distances, suggesting that gene flow among populations is limited. Finally, the number of generative shoots, the number of seeds per fruit and seed weight were significantly and positively related to population size and local growth conditions. We conclude that under stressful conditions populations of clonal forest plant species can slowly evolve into remnant populations characterized by low levels of genetic variation and limited sexual reproduction. Conservation of suitable habitat conditions is therefore a prerequisite for effective long-term conservation of clonal forest plant species.     

Reproductive strategies and isolation‐by‐demography in a marine clonal plant along an eutrophication gradient

Genetic diversity in clonal organisms includes two distinct components, (i) the diversity of genotypes or clones (i.e. genotypic richness) in a population and (ii) that of the alleles (i.e. allelic and gene diversity within populations, and differentiation between populations). We investigated how population differentiation and genotypic components are associated across a gradient of eutrophication in a clonal marine plant. To that end, we combined direct measurements of sexual allocation (i.e. flower and seed counts) and genotypic analyses, which are used as an estimator of effective sexual reproduction across multiple generations. Genetic differentiation across sites was also modelled according to a hypothesis here defined as isolation‐by‐demography, in which we use population‐specific factors, genotypic richness and eutrophication that are hypothesized to affect the source‐sink dynamics and thus influence the genetic differentiation between a pair of populations. Eutrophic populations exhibited lower genotypic richness, in agreement with lower direct measurements of sexual allocation and contemporaneous gene flow. Genetic differentiation, while not explained by distance, was best predicted by genotypic richness and habitat quality. A multiple regression model using these two predictors was considered the best model (R² = 0.43). In this study, the relationship between environment and effective sexual–asexual balance is not simply (linearly) predicted by direct measurements of sexual allocation. Our results indicate that population‐specific factors and the isolation‐by‐demography model should be used more often to understand genetic differentiation.     

Genetic variation and clonal structure of the rare,riparian shrub <Emphasis Type="Italic">Spiraea virginiana</Emphasis> (Rosaceae)

Genetic diversity is often considered important for species that inhabit highly disturbed environments to allow for adaptation. Many variables affect levels of genetic variation; however, the two most influential variables are population size and type of reproduction. When analyzed separately, both small population size and asexual reproduction can lead to reductions in genetic variation, although the exact nature of which can be contrasting. Genetic variables such as allelic richness, heterozygosity, inbreeding coefficient, and population differentiation have opposite predictions depending upon the trait (rarity or clonality) examined. The goal of this study was to quantify genetic variation and population differentiation in a species that resides in a highly stochastic environment and is both rare and highly clonal, Spiraea virginiana, and to determine if one trait is more influential genetically than the other. From populations sampled throughout the natural range of S. virginiana, we used microsatellite loci to estimate overall genetic variation. We also calculated clonal structure within populations, which included genotypic richness, evenness, and diversity. Gene flow was investigated by quantifying the relationship between genetic and geographic distances, and population differentiation (θ) among populations. Observed heterozygosity, genotypic richness, and inbreeding coefficients were found to be representative of high clonal reproduction (averaging 0.505, 0.1, and –0.356, respectively) and the number of alleles within populations was low (range = 2.0–3.6), being more indicative of rarity. Population differentiation (θ) among populations was high (average = 0.302) and there was no relationship between genetic and geographic distances. By examining a species that exhibits two traits that both can lead to reduced genetic variation, we may find an enhanced urgency for conservation. Accurate demographic counts of clonal species are rarely, if ever, possible and genetic exploration for every species is not feasible. Therefore, the conclusions in this study can be potentially extrapolated to other riparian, clonal shrubs that share similar biology as S. virginiana.     

Fine-scale spatial genetic structure and clonal distribution of the cold-water coral Lophelia pertusa

Determining the spatial genetic structure within and among cold-water coral populations is crucial to understanding population dynamics, assessing the resilience of cold-water coral communities and estimating genetic effects of habitat fragmentation for conservation. The spatial distribution of genetic diversity in natural populations depends on the species’ mode of reproduction, and coral species often have a mixed strategy of sexual and asexual reproduction. We describe the clonal architecture of a cold-water coral reef and the fine-scale population genetic structure (<35 km) of five reef localities in the NE Skagerrak. This study represents the first of this type of analysis from deep waters. We used thirteen microsatellite loci to estimate gene flow and genotypic diversity and to describe the fine-scale spatial distribution of clonal individuals of Lophelia pertusa. Within-population genetic diversity was high in four of the five reef localities. These four reefs constitute a genetic cluster with asymmetric gene flow that indicates metapopulation dynamics. One locality, the Säcken reef, was genetically isolated and depauperate. Asexual reproduction was found to be a highly important mode of reproduction for L. pertusa: 35 genetic individuals were found on the largest reef, with the largest clone covering an area of nearly 300 m².     

High genotypic diversity and strong spatial structure in populations of Trifolium alpestre with low seed production

Studies on clonal plants indicate that the proportion between clonal and sexual reproduction can be variable, depending on local habitat conditions and the biological characteristics of the species. In the present study, we assessed this question in Trifolium alpestre by assaying genetic diversity and spatial genotypic structure of natural populations with the use of allozyme markers. Populations revealed high genetic diversity as well as strong spatial structure of multilocus genotypes. The values of genetic diversity were moderately high. Spatially aggregated, identical genotypes spread up to 15 m along linear transects and across 4‐m² plots indicate extensive clonal propagation within populations. However, the existence of numerous unique and small‐sized clones reflects significant contribution from sexual reproduction. Spatially and temporarily stochastic soil disturbances have evidently opened new opportunities for the successful sexual recruitment from the permanent soil seed bank and thus counteracted losses of genetic and genotypic diversity. Seed production in all populations during the three study years was low, in average up to 1.5–2.4 seeds per shoot. The almost total lack of seed set for 57 bagged flower heads on genotypes grown in a common garden indicates that T. alpestre needs pollinators for seed production.     

Parentage analyses identify local dispersal events and sibling aggregations in a natural population of Millepora hydrocorals,a free‐spawning marine invertebrate

Dispersal is a critical process for the persistence and productivity of marine populations. For many reef species, there is increasing evidence that local demography and self‐recruitment have major consequences on their genetic diversity and adaptation to environmental change. Yet empirical data of dispersal patterns in reef‐building species remain scarce. Here, we document the first genetic estimates of self‐recruitment and dispersal distances in a free‐spawning marine invertebrate, the hydrocoral Millepora cf. platyphylla. Using twelve microsatellite markers, we gathered genotypic information from 3,160 georeferenced colonies collected over 27,000 m² of a single reef in three adjacent habitats in Moorea, French Polynesia; the mid slope, upper slope, and back reef. Although the adult population was predominantly clonal (85% were clones), our parentage analysis revealed a moderate self‐recruitment rate with a minimum of 8% of sexual propagules produced locally. Assigned offspring often settled at <10 m from their parents and dispersal events decrease with increasing geographic distance. There were no discrepancies between the dispersal distances of offspring assigned to parents belonging to clonal versus nonclonal genotypes. Interhabitat dispersal events via cross‐reef transport were also detected for sexual and asexual propagules. Sibship analysis showed that full siblings recruit nearby on the reef (more than 40% settled at <30 m), resulting in sibling aggregations. Our findings highlight the importance of self‐recruitment together with clonality in stabilizing population dynamics, which may ultimately enhance local sustainability and resilience to disturbance.     

CONTRIBUTIONS OF SEXUAL AND ASEXUAL REPRODUCTION TO POPULATION STRUCTURE IN THE CLONAL SOFT CORAL,ALCYONIUM RUDYI

Numerous studies of population structure in sessile clonal marine invertebrates have demonstrated low genotypic diversity and nonequilibrium genotype frequencies within local populations that are monopolized by relatively few, highly replicated genets. All of the species studied to date produce planktonic sexual propagules capable of dispersing long distances; despite local genotypic disequilibria, populations are often panmictic over large geographic areas. The population structure paradigm these species represent may not be typical of the majority of clonal invertebrate groups, however, which are believed to produce highly philopatric sexual propagules. I used allozyme variation to examine the population structure of the temperate soft coral, Alcyonium rudyi, a typical clonal species whose sexually produced larvae and asexually produced ramets both have very low dispersal capabilities. Like other clonal plants and invertebrates, the local population dynamics of A. rudyi are dominated by asexual reproduction, and recruitment of new sexually produced genets occurs infrequently. As expected from its philopatric larval stage, estimates of genetic differentiation among populations of A. rudyi were highly significant at all spatial scales examined (mean θ = 0.300 among 20 populations spanning a 1100-km range), suggesting that genetic exchange seldom occurs among populations separated by as little as a few hundred meters. Mapping of multilocus allozyme genotypes within a dense aggregation of A. rudyi ramets confirmed that dispersal of asexual propagules is also very limited: members of the same genet usually remain within < 50 cm of one another on the same rock surface. Unlike most previously studied clonal invertebrates, populations of A. rudyi do not appear to be dominated by a few widespread genets: estimates of genotypic diversity (G_o) within 20 geographically distinct populations did not differ from expectations for outcrossing, sexual populations. Despite theoretical suggestions that philopatric dispersal combined with typically small effective population sizes should promote inbreeding in clonal species, inbreeding does not appear to contribute significantly to the population structure of A. rudyi. Genet genotype frequencies conformed to Hardy-Weinberg expectations in all populations, and inbreeding coefficients (f) were close to zero. In general, the population structure of A. rudyi did not differ significantly from that observed among outcrossing sexual species with philopatric larval dispersal. Age estimates suggest, however, that genets of A. rudyi live for many decades. Genet longevity may promote high genotypic diversity within A. rudyi populations and may be the most important evolutionary consequence of clonal reproduction in this species and the many others that share its dispersal characteristics.     

Temporal fragmentation of a critically endangered forest ecosystem

Landscape change and habitat fragmentation is increasingly affecting forests worldwide. Assessments of patterns of spatial cover in forests over time can be critical as they reveal important information about landscape condition. In this study, we assessed landscape patterns across the Mountain Ash (Eucalyptus regnans) and Alpine Ash (Eucalyptus delegatensis) forests in the Central Highlands of Victoria between 1999 and 2019. These forests have experienced major disturbance over the past 20 years through a major fire (in 2009) and extensive industrial logging. We found that around 70% and 65% of the Mountain Ash and Alpine Ash forest areas, respectively, were either disturbed or within 200 m of a disturbed area. Inclusion of planned logging increased these disturbance categories to 72% and 70%, respectively. We also found that the isolation of Mountain Ash core areas (patches of undisturbed forest >1000 ha) increased significantly (P < 0.05) over our study period, with the proximity between disturbed areas conversely increasing significantly (P < 0.05). This means that continued and planned disturbance through industrial logging will have an amplified adverse effect on remaining undisturbed ash forest patches, which will become smaller and more dispersed across the landscape.     

Clonal diversity and structure related to habitat of the marine angiosperm Thalassia testudinum along the Atlantic coast of Mexico

The clonal structure of the tropical seagrass Thalassia testudinum was studied at 16 sites along the Mexican Atlantic coast, situated in back-reef, shallow coastal and lagoon habitats. Thalassia testudinum was highly clonal, with an overall average clonal richness (R) of 0.55. The largest genet found in this study extended throughout the sampling area (∼230 m), with an estimated max age almost reaching 600 years. Lagoons with higher nutrient availability reflected by nutrient content of leaves (mean leaf C:N ratio 11.4) and lower hydrodynamic regimes reflected by the percentage of fine sediments (on average 23%), sustained larger genets of T. testudinum (mean of the largest genets over populations was 167.3 m) than the shallow coastal areas (C:N 12.3, 6.2% fine sediment, mean largest genet 10.3 m) and the more oligotrophic back-reefs (C:N 16.3, 2.7% fine sediment, mean largest genet 6.5 m). Population genetic analysis showed different levels of clonality, genotypic diversity and spatial genetic relatedness for this seagrass per habitat, with the lagoons presenting much lower levels of clonal diversity than the other two habitats.     

Disturbances by human trampling alter the performance,sexual reproduction and genetic diversity in a clonal woodland herb

Disturbances play a major role in structuring the community composition in a variety of ecosystems. The potential impact of disturbances on sexual reproduction and genetic diversity of plants has so far received little attention. We examined the effects of disturbance by human trampling on the performance, sexual reproduction and clonal diversity in the woodland herb Anemone nemorosa in six sites disturbed by intensive recreational activities and in six undisturbed sites in suburban forests in Switzerland. Disturbance by human trampling decreased shoot height of A. nemorosa and altered the size-dependent flowering probability function. Furthermore, in disturbed sites an increased percentage of aborted seeds was found, resulting in a reduced sexual reproductive potential of A. nemorosa populations. Both clonal and genetic diversity of A. nemorosa were reduced in disturbed sites. Our study shows for the first time that disturbance by human trampling can decrease the genetic diversity in a clonal woodland plant. This suggests that genetic aspects of woodland plants should be considered in restoring degenerated forest sites.     

Effects of anthropogenic disturbance on plant diversity and community structure of a sacred grove in Meghalaya, northeast India

This study analyses the effects of anthropogenic disturbance on plant diversity and community attributes of a sacred grove (montane subtropical forest) at Swer in the East Khasi Hills district of Meghalaya in northeast India. The undisturbed, moderately disturbed and highly disturbed stands were identified within the sacred grove on the basis of canopy cover, light interception and tree (cbh 15 cm) density. The undisturbed forest stand had >40% canopy cover, >50% light interception and a density of 2103 trees per hectare, whereas the highly disturbed stand had <10% canopy cover, <10% light interception and 852 trees per hectare. The moderately disturbed stand occupied the intermediate position with respect to these parameters. The study revealed that the mild disturbance favoured species richness, but with increased degree of disturbance, as was the case in the highly disturbed stand, the species richness markedly decreased. The number of families of angiosperms was highest (63) in the undisturbed stand, followed by the moderately (60) and highly disturbed (46) stands. The families Rubiaceae, Asteraceae and Poaceae were the dominant families in the sacred forest. Rubiaceae was represented by 11, 14 and 10 species in the undisturbed, moderately disturbed and highly disturbed stands, respectively, whilst the family Asteraceae had 16 species in the moderately disturbed stand and 14 species in the highly disturbed stand. The number of families represented by a single species was reduced significantly from 33 in the undisturbed stand to 23 in the moderately and 21 in the highly disturbed stand. The similarity index was maximum (71%) between the undisturbed and moderately disturbed stand and minimum (33%) between the undisturbed and highly disturbed stands. The Margalef index, Shannon diversity index and evenness index exhibited a similar trend, with highest values in the moderately disturbed stand. In contrast, the Simpson dominance index was highest in the highly disturbed stand. There was a sharp decline in tree density and basal area from the undisturbed (2103 trees ha^–1 and 26.9 m² ha^–1) to the moderately disturbed (1268 trees ha^–1 and 18.6 m² ha^–1) and finally to the highly disturbed (852 trees ha^–1 and 7.1 m² ha^–1) stand. Density–girth curves depicted a successive reduction in number of trees in higher girth classes from the undisturbed to the moderately and highly disturbed stands. The log-normal dominance–distribution curve in the undisturbed and moderately disturbed stands indicated the complex and stable nature of the community. However, the short-hooked curve obtained for the highly disturbed stand denoted its simple and unstable nature.     

Genetic diversity of the expansive grass Brachypodium pinnatum in a changing landscape: Effect of habitat age

Perennial grasses constitute a major group of species showing a dramatic decline of biodiversity in successional plant communities. Using AFLP markers, we examined 12 populations of the expansive grass Brachypodium pinnatum differing in habitat age (30–50, ca. 100 and >300 years old) in order to determine whether clonal diversity of populations, genetic variation, and the relative importance of clonal propagation versus sexual reproduction change with grassland age. Five AFLP primer combinations gave a total of 517 bands, 79% of which were polymorphic. 314 different multilocus lineages were distinguished among the 453 samples analyzed. The number of genotypes (G) and clonal richness (R) decreased with habitat age, while the distribution of the frequency of genets changed from many clones of similar size to dominance by one or a few large clones. We consider these results to give evidence of significant role of sexual reproduction in the early phases of colonization and prevalence of clonal growth and competitive exclusion of less adapted genotypes in the later ones. However, habitat age had only marginal effect on genetic diversity, as percentage of polymorphic loci (PPL) within all the populations analyzed was similar, viz. 38.6–43.5%.     

A coding SNP of <Emphasis Type="Italic">LHX4</Emphasis> gene is associated with body weight and body length in bovine

Heterozygous mutations in LHX4 are associated with combined pituitary hormone deficiency. In this study, the polymorphism of LHX4-HaeIII locus was revealed in 822 individuals from four Chinese cattle breeds. The PCR–RFLP analysis showed that there were three genotypes: GG, GA, AA. The frequencies of genotype GG ranged from 0.6620 to 0.9789 in analyzed populations. The genotypic frequencies of LHX4 locus in the four populations all agreed with Hardy–Weinberg equilibrium (P > 0.05). Distributions of genotypic frequencies of different breeds (QC, NY, JX, CH) at this locus were found to be significantly different based on a χ ² test (P < 0.001). The genetic diversity analysis revealed the JX cattle possessed intermediate genetic diversity, and the other three Chinese cattle breeds belonged to poor genetic diversity. Correlation analysis with growth traits in the NY breed indicated that: the animals with genotype GA had greater body weight than those with genotype GG (P < 0.05); the animals with GA genotype owned significantly longer body length than the ones with GG genotype (P < 0.05) at 18 and 24 months.     

Habitat connectivity, more than species’ biology, influences genetic differentiation in a habitat specialist, the short-eared rock-wallaby (Petrogale brachyotis)

It is difficult to assess the relative influence of anthropogenic processes (e.g., habitat fragmentation) versus species’ biology on the level of genetic differentiation among populations when species are restricted in their distribution to fragmented habitats. This issue is particularly problematic for Australian rock-wallabies (Petrogale sp.), where most previous studies have examined threatened species in anthropogenically fragmented habitats. The short-eared rock-wallaby (Petrogale brachyotis) provides an opportunity to assess natural population structure and gene flow in relatively continuous habitat across north-western Australia. This region has reported widespread declines in small-to-medium sized mammals, making data regarding the influence of habitat connectivity on genetic diversity important for broad-scale management. Using non-invasive and standard methods, 12 microsatellite loci and mitochondrial DNA were compared to examine patterns of population structure and dispersal among populations of P. brachyotis in the Kimberley, Western Australia. Low genetic differentiation was detected between populations separated by up to 67?km. The inferred genetic connectivity of these populations suggests that in suitable habitat P. brachyotis can potentially disperse far greater distances than previously reported for rock-wallabies in more fragmented habitat. Like other Petrogale species male-biased dispersal was detected. These findings suggest that a complete understanding of population biology may not be achieved solely by the study of fragmented populations in disturbed environments and that management strategies may need to draw on studies of populations (or related species) in undisturbed areas of contiguous habitat.     

Effects of anthropogenic habitat disturbance on local pollinator diversity and species turnover across a precipitation gradient

Anthropogenic habitat disturbance can have profound effects on multiple components of forest biotas including pollinator assemblages. We assessed the effect of small-scale disturbance on local richness, abundance, diversity and evenness of insect pollinator fauna; and how habitat disturbance affected species turnover across the landscape and overall diversity along a precipitation gradient in NW Patagonia (Argentina). We evaluated the effect of disturbance on overall pollinator fauna and then separately for bees (i.e. Apoidea) and non-bee pollinators. Locally, disturbed habitats had significantly higher pollinator species richness and abundances than undisturbed habitats for the whole pollinator assemblage, but not for bees or non-bees separately. However, significant differences in species richness between habitats vanished after accounting for differences in abundance between habitat types. At a local scale Shannon–Weaver diversity and evenness did not vary with disturbance. A β diversity index indicated that, across forest types, species turnover was lower between disturbed habitats than between undisturbed habitats. In addition, rarefaction curves showed that disturbed habitats as a whole accumulated fewer species than undisturbed habitats at equivalent sample sizes. We concluded that small patches of disturbed habitat have a negligible effect on local pollinator diversity; however, habitat disturbance reduced β diversity through a homogenization of the pollinator fauna (in particular of bees) across the landscape.     

Population genetic structure of the tropical moss Acanthorrhynchium papillatum as measured with microsatellite markers

Mosses and other bryophytes are vital components of forests, because they sustain a tremendous diversity of invertebrates and influence significant ecological functions. There have been few studies on moss population diversity in Southeast Asia, despite the escalating deforestation in this region of rich biodiversity. The genetic diversity of the tropical moss Acanthorrhynchium papillatum (Harv.) Fleisch., collected from forested areas in Singapore and Peninsular Malaysia, was elucidated using eight microsatellite markers developed for this species. Significant levels of allelic and haplotypic diversity were observed among clumps of the moss. Differences in allelic richness and genotypic diversity among the populations were higher in less disturbed forests compared to the more disturbed areas, suggesting that genetic diversity is affected by habitat quality. Genetic diversity levels within the clumps studied were low, indicating that vegetative reproduction was more important within clumps than sexual reproduction. However, multilocus genotypes of samples within the clumps studied were not all alike, providing evidence of microsatellite mutation or of occasional sexuality. Despite the isolation of populations, A. papillatum can introduce genetic variability by mutation among vegetatively propagated individuals. This study provides baseline information on the genetic diversity of A. papillatum tropical rain forests.     

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.     

Habitat and sex specific differences in the dioecious weed Acetosella vulgaris (Polygonaceae)

Abstract Patterns of resource allocation, numbers of reproductive structures and sex ratios of flowering populations of the dioecious weed Acetosella vulgaris (Fourr.) were examined in the Kosciuszko alpine region of Australia. Specifically, the sex‐specific response of ramets was compared between a disturbed alpine habitat, in which weeds such as A. vulgaris are common (disturbed roadside/path‐edge), and a native alpine habitat in which weeds appear to have a limited capacity to germinate, grow and reproduce (undisturbed tall alpine herbfield). The disturbed habitat was generally more favourable for growth of A. vulgaris. Ramets of both sexes had greater total and vegetative biomass than ramets in native habitats. Although reproductive biomass was also greater in disturbed habitats, females had less reproductive biomass than males, which was not as predicted. Reproductive effort was not affected by habitat or gender. The disturbed habitat also favoured increased numbers of inflorescences per ramet and flowers per ramet, as expected. Whereas the gender of the ramet also influenced numbers of reproductive structures, again, this was not as predicted. Females had more flowers per ramet and more flowers per inflorescence than males. This may be because of factors associated with wind pollination. Females were taller in native habitats but there was no difference between the sexes in disturbed habitats. Sex ratios varied from all male populations to nearly all female populations among the 25 sites sampled irrespective of habitat. Factors such as time since last disturbance may have contributed to variation in the sex ratios of alpine populations of A. vulgaris in Australia.     

Genetic diversity of Cypripedium calceolus in Poland

In this work we assessed the genetic diversity of 32 C.?calceolus populations from Poland. Mean genetic diversity was moderate (P?=?36.4%, A?=?1.58, H _O?=?0.143, F _IS?=?0.059), and seven geographic regions did not differ significantly in their levels of polymorphism (p?>?0.05), although allele frequencies varied greatly. Only four unique alleles were found, at three sites in southern and southeastern Poland. Genetic (P, A) and genotypic diversity parameters (G, G _U) were significantly correlated with population size (p?<?0.001). In the 32 studied populations we separated 422 different multilocus genotypes; none was common to all populations. Overall population differentiation was moderate at 0.137 (p?<?0.001), but we found a significant pattern of isolation by distance for the whole dataset (r ²?=?0.65, p?<?0.001). Our chloroplast DNA (cpDNA) results suggest a single evolutionary lineage and a common origin for all Polish C.?calceolus populations. Information about the genetic health of C.?calceolus populations should be useful in developing conservation strategies.     

Genetic variation in Pueraria lobata (Fabaceae), an introduced, clonal, invasive plant of the southeastern United States

Pueraria lobata (kudzu), a clonal, leguminous vine, is invading the southeastern United States at a rate of 50 000 ha per year. Genetic variability and clonal diversity were measured in 20 southeastern U.S. populations using 14 allozyme loci. Within its U.S. range, 92.9% of the loci were polymorphic and overall genetic diversity was 0.290. Such high levels of genetic diversity are consistent with its history of multiple introductions over an extended period of time. The average proportions of polymorphic loci and genetic diversity within populations were 55.7% (range = 28.6–85.7%) and 0.213 (range = 0.114–0.317), respectively. The proportion of total genetic diversity found among populations was similar to species with equivalent life history characters (G_ST = 0.199). No regional patterns of variation were seen. The number of putative genotypes in each population ranged from 2 to 26. Mean genotypic diversity was 0.694, ranging from 0.223 to 0.955. Such high levels of genotypic diversity indicate that local sites are often colonized by several propagules (most likely seeds) and/or that sexual reproduction occurs within populations after establishment. An excess of heterozygosity was observed in populations with few unique genets, implying that selection for highly heterozygous individuals may occur in populations of P. lobata.