Mean Pollen Dispersal Distance Research Articles

Estimating genetic diversity, mating system and pollen dispersal to inform ex situ conservation of the tree Genipa americana L.

AbstractUsing microsatellite loci, we assessed the mating system and genetic diversity of the dioecious tropical tree Genipa americana in a natural population (NP) and a progeny test (PT). For NP, we also estimated the paternity correlation within and among fruits and mean pollen dispersal distance. As expected for dioecious species, all offspring originated from outcrossing (t = 1). Mating among relatives (1 − ts) and paternity correlation (rp) were variable among progenies (1 − ts = 0.03–0.19; rp = 0.04–0.40), but greater in NP than in PT. Fixation index (F) was generally significant and lower in adults than in offspring, indicating selection against inbred individuals. Paternity correlation was higher within (0.40) than among (0.26) fruits, indicating a lower effective number of pollen donors (Nep) within (2.5) than among (3.8) fruits. Due to the higher rp in NP, the effective size within progenies (Ne) was lower (2.69) than PT (3.27). The pollen dispersal pattern was strongly leptokurtic, suggesting long-distance pollen dispersal (mean of 179 m). The results show that both populations can be used for seed collection in environmental reforestation programmes; however, considering that PT is structured in maternal progenies, NP is more suitable for seed collection due to the lower probability of mating among related trees.

Optimising the conservation of genetic diversity of the last remaining population of a critically endangered shrub.

An understanding of genetic diversity and the population genetic processes that impact future population viability is vital for the management and recovery of declining populations of threatened species. Styphelia longissima (Ericaceae) is a critically endangered shrub, restricted to a single fragmented population near Eneabba, 250 km north of Perth, Western Australia. For this population, we sought to characterize population genetic variation and its spatial structure, and aspects of the mating portfolio, from which strategies that optimize the conservation of this diversity are identified. A comprehensive survey was carried out and 220 adults, and 106 seedlings from 14 maternal plants, were genotyped using 13 microsatellite markers. Levels of genetic variation and its spatial structure were assessed, and mating system parameters were estimated. Paternity was assigned to the offspring of a subsection of plants, which allowed for the calculation of realized pollen dispersal. Allelic richness and levels of expected heterozygosity were higher than predicted for a small isolated population. Spatial autocorrelation analysis identified fine-scale genetic structure at a scale of 20 m, but no genetic structure was found at larger scales. Mean outcrossing rate (tm = 0.66) reflects self-compatibility and a mixed-mating system. Multiple paternity was low, where 61 % of maternal siblings shared the same sire. Realized pollen dispersal was highly restricted, with 95 % of outcrossing events occurring at 7 m or less, and a mean pollen dispersal distance of 3.8 m. Nearest-neighbour matings were common (55 % of all outcross events), and 97 % of mating events were between the three nearest-neighbours. This study has provided critical baseline data on genetic diversity, mating system and pollen dispersal for future monitoring of S. longissima. Broadly applicable conservation strategies such as implementing a genetic monitoring plan, diluting spatial genetic structure in the natural population, genetically optimizing ex situ collections and incorporating genetic knowledge into translocations will help to manage the future erosion of the high genetic variation detected.

Stepping stones or stone dead? Fecundity, pollen dispersal and mating patterns of roadside Qualea grandiflora Mart. trees

Forest fragmentation may affect mating and pollen dispersal patterns through conversion of continuous forests into small, spatially isolated remnant patches and individual trees in an anthropogenic landscape. We investigated reproductive investment and success, pollen dispersal, mating system, and genetic diversity and spatial structure of Qualea grandiflora trees in two environmental contexts: a continuous natural Cerrado area and isolated individuals on roadsides. Roadside trees produced more flowers and more fruit than Cerrado trees. Pollen dispersal kernels were fat-tailed in both contexts, indicating long-distance dispersal, but in Cerrado the mean pollen dispersal distance (524.7 m) and the effective number of pollen donors per mother-tree (Nep = 12.7) were higher than for roadside trees (60.9 m, Nep = 4.6). The levels and structure of genetic diversity, outcrossing rates ( $$t_{m}$$ > 0.98), and mating among relatives ( $$t_{m} - t_{s}$$ < 0.1) were similar in both environmental contexts. Allelic richness and number of private alleles were similar between the two environments. The fixation index was significantly lower in adults (minimum of 0.08) than in offspring (minimum of 0.23) in both contexts, suggesting selection against inbred individuals between offspring and adult stage. Our results indicate that the spatial isolation of roadside trees, by increasing the number of flowers produced, decreased pollinator movements, thereby reducing effective pollen flow and the number of pollen donors. All these results suggest that roadside trees can be used for harvesting seeds for recovery plans, and that these trees are a biological legacy, and reservoir of Q. grandiflora genetic diversity, from the original Cerrado forest.

Patterns of Pollen Dispersal in an Invasive Population of Solanum rostratum (Solanaceae) in China

Paternity analysis, carried out on 13 microsatellite loci, was used to investigate pollen dispersal and mating patterns in an invasive population of Solanum rostratum in China. We determined a high outcrossing rate (0.81 ± 0.02) and found that all 174 offspring for which the father could be identified were outcross progeny. The mean pollen dispersal distance was 13.66 m (range 0.41–71.67 m); however, 59.2% of effective pollination events occurred at <10 m. Our results indicate substantial near-neighbor pollen dispersal and imply that successful invasion of S. rostratum is facilitated by a high level of pollen flow within the population.

Paternity analysis, pollen flow, and spatial genetic structure of a natural population of Euterpe precatoria in the Brazilian Amazon.

Euterpe precatoria, known as açaí do Amazonas, is a regionally important palm of the Amazon rainforest for the fruit production through extractive agriculture. Little information is available with regard to genetic diversity, gene flow, and spatial genetic structure (SGS) of açaí populations, which are essential for the use, management, and conservation of genetic resources of the species. This research aimed to assess the genetic diversity, inbreeding level, SGS, and gene flow in four ontogenetic stages of a natural E. precatoria population in the Brazilian Amazon, based on 18 microsatellite loci. The study was carried out in a natural population dispersed in an area of about 10 ha. Leaf tissues of 248 plants were mapped and sampled and classified into four ontogenetic stages: reproductive (59), immature (70), young (60), and seedling (59). Genetic diversity indices were high for all ontogenetic stages. The fixation index (F) for all ontogenetic stages was not significantly different from zero, indicating the absence of inbreeding. A significant SGS was found for all ontogenetic stages (68–110 m), indicating seed dispersal over short distances. Paternity analysis detected pollen immigration of 39.1%, a selfing rate of 4.2%, and a mean pollen dispersal distance within the population of 531 m. The results indicate substantial allele input in the population via pollen immigration, contributing to the maintenance of the genetic diversity of the population. However, within a population, the renewal with new progenies selected from seed plants spaced at least 110 m apart is important to avoid collecting seeds from related plants.

Breeding system, gene dispersal and small-scale spatial genetic structure of a threatened food tree species, Pentadesma butyracea (Clusiaceae) in Benin

Pentadesma butyracea Sabine, a rain forest food tree species, plays a vital role in the socio-economic livelihood of some West African rural communities due to its various products. However, its scattered populations are threatened in Benin. Defining appropriate conservation strategies requires a good knowledge of mating patterns and their consequences for population genetics. The outcrossing rate, levels of correlated paternity and fine-scale spatial genetic structure of adults and maternal sibships were estimated for one small population and three large populations in Benin using microsatellite markers. Similar outcrossing rates (88–95%) were found in all populations, showing that P. butyracea is mainly an outbreeding species. We found no evidence of inbreeding depression from a decay of inbreeding with age. The spatial genetic structure within the large populations (Sp statistic = 0.003–0.038) was consistent with isolation-by-distance expectations, showing that gene dispersal is spatially limited. Limited pollen dispersal is highlighted by the decay of the degree of correlated paternity between sibships with spatial distance. The mean pollen dispersal distance was estimated between 50 m and 450 m, but up to 21% pollen may migrate from external sources. The smallest population displayed slightly higher correlated paternity than the large populations (r p = 0.37 vs. r p = 0.17–0.30). In conclusion, our results suggest that small populations may show a reduction in sire numbers in seed, while the fragmented populations, large and small, are connected through gene flow. There is little inbreeding and no evidence of inbreeding depression.

High selfing rate, limited pollen dispersal and inbreeding depression in the emblematic African rain forest tree Baillonella toxisperma – Management implications

Mating system and gene flow are major influencing factors of species population dynamics and evolution. These factors are often not characterized in tropical tree species, yet they constitute basic information that must be considered to implement sustainable management practices. In particular, as logging implies a reduction of the density of congeneric mates, the connectivity through pollination between individuals has to be well characterized (selfing versus outcrossing rates, distances between mates). We conducted a genetic-based analysis (using 10 nuclear microsatellites) to determine the mating system and gene flow characteristics of an emblematic timber tree species from lowland rain forests of the Congo Basin, Baillonella toxisperma (Sapotaceae). The species, which is frequently exploited for its wood and for a number of non-timber forest products, naturally occurs at low densities (ca. 0.01–0.1individuals/ha). It is supposedly an entomophilous species whose seeds are probably dispersed by mammals. We have shown that the species presents a mixed-mating system (about 20–40% of selfing depending on analysis method). However, the comparison of inbreeding parameters among cohorts suggests that inbred individuals die between seedling and mature tree stages. The mean pollen dispersal distance was relatively low for such a low-density population species (estimated to be 690 or 777m depending on analysis method) and, together with a low mean number of pollen donors (NEP=2.76), it suggests a pattern of nearest-neighbour mating where allo-pollen could be a limiting factor. However, B. toxisperma presents a relatively weak genetic structure (Sp statistic=0.0095) indicative of long gene dispersal distance (σg=3–5km according to the assumed effective population density). Overall, this would indicate that gene flow occurs mainly by extensive seed dispersal in this species. These results suggest that mammals and local populations involved in the dispersal of the species play a key role by lowering biparental inbreeding effects. Sustainable population management might require assisted regeneration using unrelated planting material.

Nuclear microsatellite markers for population genetic studies in sugar maple (Acer saccharum Marsh.)

A set of seven new nuclear microsatellite markers (nSSRs) was developed for sugar maple (Acer saccharum Marsh.) using paired-end Illumina sequencing. Out of 96 primers screened in a panel of six unrelated individuals, seven markers amplified polymorphic products. The utility of these markers, in addition to six already published microsatellites, for genetic variation and gene flow studies was assessed. Out of the seven newly developed markers, three amplified multiple fragments and were interpreted as dominant (absence/presence) markers, while four markers amplified a maximum of two amplification products per sample. The six published microsatellites and three of the four newly developed markers showed regular segregation in an open-pollinated single tree progeny. Observed heterozygosity (Ho) and expected heterozygosity (He) in 48 individuals from one population ranged from 0.436 to 0.917 and from 0.726 to 0.894, respectively. Dominant markers revealed 64 variable positions and moderate genetic variation within the population (He = 0.102, Shannon’s I = 0.193). Paternity analyses in the program CERVUS at co-dominant markers showed effective dispersal of pollen in the sugar maple population both at 95% and 80% confidence levels. Dependent on the confidence level, the mean pollen dispersal distance within the population ranged from 33.25 m to 38.75 m and gene flow from utside the stand from 78% to 82%. The absence of fine-scale Spatial Genetic Structure (SGS) suggested effective dispersal of both seeds and pollen.

Evaluating the influence of different aspects of habitat fragmentation on mating patterns and pollen dispersal in the bird‐pollinated Banksia sphaerocarpa var. caesia

Habitat fragmentation can significantly affect mating and pollen dispersal patterns in plant populations, although the differential effects of the various aspects of fragmentation are poorly understood. In this study, we used eight microsatellite loci to investigate the effect of fragmentation on the mating system and pollen dispersal within one large and eight small population remnants of Banksia sphaerocarpa var. caesia, a bird-pollinated shrub in the southern agricultural region of Western Australia. The large population had a much larger neighbourhood size and lower selfing rate, maternal pollen pool differentiation and within-plot mean pollen dispersal distance than the small populations. Outcrossing was consistently high and ranged from 85.7% ± 2.6 to 98.5% ± 0.9, and mating patterns suggested nearest-neighbour pollination. Pollen immigration into small populations ranged from 2.8% ± 1.8 to 16.5% ± 3.2. Using the small populations, we tested for correlations between various fragmentation variables and mating system and pollen dispersal parameters. We found significant negative linear relationships between population isolation and outcrossing rate; population shape and neighbourhood size; and conspecific density and mean pollen dispersal distance. There were significant positive linear relationships between population shape and pollen pool differentiation and between population size and number of different fathers per seed crop. Our results suggest that birds may use a series of fragmented populations as a vegetation corridor while foraging across the landscape and that population connectivity is a critical determinant of pollinator visitation. Our results also suggest that the effect of a linear population shape on the mating system and pollen dispersal is routinely underestimated.

Ecological implications of a flower size/number trade-off in tropical forest trees.

BackgroundIn angiosperms, flower size commonly scales negatively with number. The ecological consequences of this trade-off for tropical trees remain poorly resolved, despite their potential importance for tropical forest conservation. We investigated the flower size number trade-off and its implications for fecundity in a sample of tree species from the Dipterocarpaceae on Borneo.Methodology/Principal FindingsWe combined experimental exclusion of pollinators in 11 species, with direct and indirect estimates of contemporary pollen dispersal in two study species and published estimates of pollen dispersal in a further three species to explore the relationship between flower size, pollinator size and mean pollen dispersal distance. Maximum flower production was two orders of magnitude greater in small-flowered than large-flowered species of Dipterocarpaceae. In contrast, fruit production was unrelated to flower size and did not differ significantly among species. Small-flowered species had both smaller-sized pollinators and lower mean pollination success than large-flowered species. Average pollen dispersal distances were lower and frequency of mating between related individuals was higher in a smaller-flowered species than a larger-flowered confamilial. Our synthesis of pollen dispersal estimates across five species of dipterocarp suggests that pollen dispersal scales positively with flower size.Conclusions and Their SignificanceTrade-offs embedded in the relationship between flower size and pollination success contribute to a reduction in the variance of fecundity among species. It is therefore plausible that these processes could delay competitive exclusion and contribute to maintenance of species coexistence in this ecologically and economically important family of tropical trees. These results have practical implications for tree species conservation and restoration. Seed collection from small-flowered species may be especially vulnerable to cryptic genetic erosion. Our findings also highlight the potential for differential vulnerability of tropical tree species to the deleterious consequences of forest fragmentation.

Temporal variation in pollen dispersal and breeding structure in a bee-pollinated Neotropical tree.

Variation among flowering seasons in the time of flowering, synchrony and length of flowering, and fluctuations in the abundance of pollinators may cause a variation in pollen dispersal distance. In this study, we analyzed the temporal variation in pollen dispersal and breeding structure in the Neotropical tree species Tabebuia aurea (Bignoniaceae) and evaluated pollen dispersal between a population inside the reserve and a patch of isolated individuals on the edge of the reserve, and tested the hypothesis that isolated individuals are sinking for pollen. All adult trees (260) within a population of 40 ha and 9 isolated individuals on the edge of the reserve were sampled, and from these adults, 21 open-pollinated progeny arrays were analyzed in 2 flowering seasons (309 seeds in 2004 and 328 in 2005). Genetic analyses were based on the polymorphism at 10 microsatellite loci. A high proportion of self-pollination found in both flowering seasons indicated a mixed-mating system. The mean pollen dispersal distance differed significantly between the two flowering seasons (307.78 m in 2004 and 396.26 m in 2005). Maximum pollen dispersal was 2608 m, but most pollination events (65%) occurred at distances <300 m. Our results also showed that isolated individuals are sinking for pollen, with high pollen flow between the population inside the reserve and individuals on the edge. These results are most likely due to the large pollinator species, which can potentially fly long distances, and also due to temporal variation in individual fecundity and contribution to pollen dispersal.

Short-distance pollen dispersal and high self-pollination in a bat-pollinated neotropical tree

We investigated pollen dispersal and breeding structure in the tropical tree species Caryocar brasiliense Camb. (Caryocaraceae), using genetic data from ten microsatellite loci. All adult trees (101) within a patch of 8.3 ha were sampled, and from these adults 18 open-pollinated maternal progeny arrays were analyzed (280 seeds from 265 fruits). Most fruits presented only one seed (median equal to 1.000) and mean number of ripened seeds per fruit was 1.053 (SD = 0.828). Our results showed that C. brasiliense presents a mixed-mating system, with 11.4% of self-pollination, multilocus outcrossing rate of t m = 0.891 ± 0.025, and high probability of full-sibship within progeny arrays (r p = 0.135 ± 0.032). Outcrossing rate and self-pollination varied significantly among mother trees. We could detect a maximum pollen dispersal distance of ∼500 m and a mean pollen dispersal distance of ∼132 m. However, most pollination events (80%) occurred at distances less than 200 m. Our results also indicated that pollen dispersal is restricted to a neighborhood of 5.4 ha with rare events of immigration (∼1% N e m = 0.35). C. brasiliense also presents a significant but weak spatial genetic structure (Sp = 0.0116), and extension of pollen dispersal distance was greater than seed dispersal (N b = 86.20 m). These results are most likely due to the foraging behavior of pollinators that may have limited flight range. The highly within-population synchronous flowering, high population density, and clumped distribution reinforce pollinator behavior and affect residence time leading to a short-distance pollen dispersal.

Effects of density and floral morph on pollen flow and seed reproduction of an endangered heterostylous herb,Primula sieboldii

SummaryWe assessed the effects of population density and the spatial arrangement of genetically compatible mates on the seed set and pollen flow of a heterostylous, bumblebee‐pollinated perennial,Primula sieboldiiE. Morren (Primulaceae), by using an experimental population under natural pollination conditions.We also examined the intermorph differences in the pollen dispersal distance and the frequency of self‐ and intramorph fertilization.Seed set was significantly correlated with the number of opposite‐morph flowers within 2 m in short‐styled genets, and within 3 m in long‐styled genets.Mean pollen dispersal distance within the experimental population was 5.4 m, which was slightly shorter than that in a low‐density population measured in a previous study (7.2 m).The proportions of seedlings sired by the same morph or selfed were 10.2% and 3.0%, respectively, in long‐styled mothers, compared with 1.9% and 0.0% among short‐styled mothers. However, these differences were not statistically significant and no marked differences in pollen dispersal distances were observed between the two morphs.Our results suggest that the presence of opposite‐morph genets within several metres is essential for success of seed reproduction inP. sieboldii, irrespective of population density, and that stochastic deviation of the morph ratio in small populations is therefore likely to cause considerable reduction in seed set.

Parentage versus two‐generation analyses for estimating pollen‐mediated gene flow in plant populations

Assessment of contemporary pollen-mediated gene flow in plants is important for various aspects of plant population biology, genetic conservation and breeding. Here, through simulations we compare the two alternative approaches for measuring pollen-mediated gene flow: (i) the NEIGHBORHOOD model--a representative of parentage analyses, and (ii) the recently developed TWOGENER analysis of pollen pool structure. We investigate their properties in estimating the effective number of pollen parents (N(ep)) and the mean pollen dispersal distance (delta). We demonstrate that both methods provide very congruent estimates of N(ep) and delta, when the methods' assumptions considering the shape of pollen dispersal curve and the mating system follow those used in data simulations, although the NEIGHBORHOOD model exhibits generally lower variances of the estimates. The violations of the assumptions, especially increased selfing or long-distance pollen dispersal, affect the two methods to a different degree; however, they are still capable to provide comparable estimates of N(ep). The NEIGHBORHOOD model inherently allows to estimate both self-fertilization and outcrossing due to the long-distance pollen dispersal; however, the TWOGENER method is particularly sensitive to inflated selfing levels, which in turn may confound and suppress the effects of distant pollen movement. As a solution we demonstrate that in case of TWOGENER it is possible to extract the fraction of intraclass correlation that results from outcrossing only, which seems to be very relevant for measuring pollen-mediated gene flow. The two approaches differ in estimation precision and experimental efforts but they seem to be complementary depending on the main research focus and type of a population studied.

Pollen dispersal of tropical trees (Dinizia excelsa: Fabaceae) by native insects and African honeybees in pristine and fragmented Amazonian rainforest

Tropical rainforest trees typically occur in low population densities and rely on animals for cross-pollination. It is of conservation interest therefore to understand how rainforest fragmentation may alter the pollination and breeding structure of remnant trees. Previous studies of the Amazonian tree Dinizia excelsa (Fabaceae) found African honeybees (Apis mellifera scutellata) as the predominant pollinators of trees in highly disturbed habitats, transporting pollen up to 3.2 km between pasture trees. Here, using microsatellite genotypes of seed arrays, we compare outcrossing rates and pollen dispersal distances of (i) remnant D. excelsa in three large ranches, and (ii) a population in undisturbed forest in which African honeybees were absent. Self-fertilization was more frequent in the disturbed habitats (14%, n = 277 seeds from 12 mothers) than in undisturbed forest (10%, n = 295 seeds from 13 mothers). Pollen dispersal was extensive in all three ranches compared to undisturbed forest, however. Using a twogener analysis, we estimated a mean pollen dispersal distance of 1509 m in Colosso ranch, assuming an exponential dispersal function, and 212 m in undisturbed forest. The low effective density of D. excelsa in undisturbed forest (approximately 0.1 trees/ha) indicates that large areas of rainforest must be preserved to maintain minimum viable populations. Our results also suggest, however, that in highly disturbed habitats Apis mellifera may expand genetic neighbourhood areas, thereby linking fragmented and continuous forest populations.

Paternity and gene dispersal in limber pine (Pinus flexilis James).

This study provides empirical information on intrapopulation gene dispersal via pollen, the size of genetic 'neighbourhoods', and interpopulation gene flow in a long-lived conifer, limber pine (Pinus flexilis). We used allozyme loci for a paternity analysis of 518 seeds produced in an isolated population of limber pine located in north-eastern Colorado, U.S.A., separated by 2 km from the nearest conspecific trees and nearly 100 km from populations in the Rocky Mountains. We also used indirect techniques (FST analyses) to estimate gene flow rates among subdivisions of the study population and among five widely separated populations. Within the main study population limber pine exhibited a polymorphism level of 50%, observed heterozygosity of 0.159, and 2.36 alleles per polymorphic locus. Mountain populations were slightly more variable. The main study population showed significant differentiation in allozymes among neighbouring subpopulations. The mean FST was 0.031 and the gene flow rate among subpopulations was estimated as 7.8 migrants per generation. Among widely separated populations the mean FST was 0.035 and the gene flow rate was estimated as 6.9 migrants per generation. The paternity analysis indicated a best estimate of 6.5% pollen immigration (minimum 1.1%) from populations 2 km to 100+ km away. For 4% of the seeds examined, paternity could be ascribed to a single tree in the study population. Fractional paternity and likelihood methods were used to estimate pollen dispersal distances for the remainder of the seeds. Mean pollen dispersal distance was estimated at 140 m using the fractional method, similar to results from the other techniques. This compares with a mean distance of 172 m between potential mates. These results suggest near-panmictic pollen dispersal over this population, which covers about 15 ha. The observed allozyme differences and surprisingly low estimates of among-subpopulation gene flow are ascribed to a probable restriction of gene dispersal by seeds.

Mechanisms of pollen deposition by insect pollinators

Studies of pollen dispersal in insect-pollinated plants have often documented highly leptokurtic patterns of pollen deposition that can increase the likelihood of long-distance mating. To examine potential causes of highly leptokurtic deposition, we introduce four functions that arise when (1) the duration of pollinator visits to pollen sources is limited, (2) the rate of pollen deposition varies randomly among pollinators and/or among visits, (3) the rate of pollen deposition changes monotonically over time or (4) pollen is carried in layers or compartments on the pollinator's body that differ in deposition rate. Maximum likelihood techniques were used to fit deposition functions to data obtained from honey bees (Apis mellifera L.) visiting mustard plants (Brassica campestris L.) that contained a marker gene. Each of the alternative leptokurtic functions fit the experimental data better than a simple exponential function and the best-fit function predicted a mean pollen dispersal distance more than three times greater than the exponential. We argue that studies of pollen deposition need to test a broader range of deposition models to assess outcrossing distance in plant populations accurately.

Pollen limitation and distance-dependent fecundity in females of the clonal gynodioecious herb Glechoma hederacea (Lamiaceae)

Pollen movement is often restricted in natural populations, and insufficient pollination is a potential constraint on sexual reproduction in outcrossing species. Seed-set should decrease with increased distance from the pollen source in outcrossing plants. This prediction was tested using females of the clonal, gynodioecious herb Glechoma hederacea in three natural populations. In controlled pollinations, both hermaphrodites and females had similar high percentages of fruit-set and seed-set. In a natural population where a female clone was isolated from the nearest hermaphroditic clone by c. 100 m, fruit-set was low (1%). In another population where hemaphroditic clones were rare and female clones had a patchy distribution, fruit-and seed-set in females were pollen-limited and decreased with increased distance from the nearest pollen source. The estimated mean pollen dispersal distance was 5.9 m when calculated on fruit-set and 5.3 m when calculated on seed-set. The most frequent pollinators were bumblebees. The mean and median distances moved by pollinators between ramets were 0.13 m and 0.05 m. In a third population where female clones were isolated from the nearest hermaphrodites by more than 200 m, fruit-set was 0%. After introduction of 16 hermaphroditic ramets in the center of the female clone, fruit-set varied between 0% and 100% in individual female ramets. Fruit-set decreased with increased distance from the pollen source. The mean and median pollen movement distances were 1.06 m and 0.54 m.