Slatkin’s Paradox: when direct observation and realized gene flow disagree. A case study in Ficus

In flying insects, there is frequently a lack of congruence between empirical estimates of local demographic parameters and the prediction that differentiation between populations should decrease with increasing dispersal, a puzzling phenomenon known as Slatkin’s Paradox. Here, we generalize Slatkin’s Paradox to other taxa, drawing from available information on dispersal to predict the relative importance of pollen vs. seed migration in structuring broad‐scale patterns of genetic variation in Ficus hirta, a dioecious fig whose pollen is dispersed by minute, species‐specific fig wasps and whose seeds are disseminated by a variety of vagile vertebrates (especially bats and birds). Local‐scale observational and genetic studies of dioecious understory figs suggest comparable rates of pollen and seed migration. In contrast, we found unusually low nuclear differentiation (F_ST = 0.037, R_ST = 0.074) and high chloroplast differentiation (G_ST = 0.729, N_ST = 0.798) among populations separated by up to 2850 km, leading us to reject the hypothesis of equal pollen and seed migration rates and to obtain an equilibrium estimate of the ratio of pollen to seed migration of r = 16.2–36.3. We reconcile this example of Slatkin’s Paradox with previously published data for dioecious figs and relate it to the picture of exceptionally long‐distance wasp‐mediated pollen dispersal that is emerging for large monoecious fig trees. More generally, we argue that Slatkin’s Paradox is a general phenomenon and suggest it may be common in plants and animals.     

Spatial genetic structure and restricted gene flow in a functionally dioecious fig, <Emphasis Type="Italic">Ficus pumila</Emphasis> L. var. <Emphasis Type="Italic">pumila</Emphasis> (Moraceae)

The mutualism between fig plants and fig wasps has been recognized as one of the most specialized systems of symbiosis. Figs are pollinated by their highly specific pollinating fig wasps, and the pollinating fig wasps are raised within the syconia of figs. Recent studies indicated a difference between monoecious and dioecious figs in the dispersal range of pollinating wasps, which has potential consequences for gene flow. In this study, we detected the gene-flow pattern of the dioecious climbing fig, Ficus pumila L. var. pumila, at both local and regional scales. At the local scale, spatial autocorrelation analysis indicated strong genetic structure at short distances, a pattern of limited gene flow. This result was also supported by a high inbreeding coefficient (F _IS = 0.287) and significant substructuring (F _ST = 0.060; P < 0.001). Further analysis indicated that the effective gene dispersal range was 1,211 m, and the relative contribution of seed dispersal was smaller than that of pollen dispersal. The inferred effective range of pollen dispersal ranged from 989 to 1,712 m, while the effective seed dispersal range was less than 989 m. Lack of long-distance dispersal agents may explain the limited seed dispersal. The high density of receptive fig trees was the most likely explanation for limited pollen dispersal, and the position of syconia and relatively low wind speed beneath the canopy may contribute to this phenomenon. At the regional scale, significant negative correlations (kinship coefficient F _ij ranging from −0.038 to −0.071) existed in all comparisons between the studied population and other populations, and the assignment test grouped almost all individuals of the studied population into a distinct cluster. Asynchronous flowering on the regional scale, which provides a barrier for the pollinating wasps to fly from the studied population to the other populations, is probably responsible for the limited gene flow on the regional scale.     

Genetic diversity and differentiation of the extremely dwarf Ficus tikoua in Southwestern China

Fig wasps are short-lived, weak fliers, and their long-distance dispersal depends on the ability to enter fast-flowing air above the canopy. Therefore, growth form of fig species may affect fig wasps’ dispersal. We employed six microsatellite markers to examine gene flow in Chinese populations of the dioecious Ficus tikoua, a prostrate shrub with figs partially buried in the soil. Moderate genetic diversity was found within populations of F. tikoua. Differentiation among six F. tikoua populations (F_ST = 0.196, p < 0.001) was higher than those of other dioecious figs, and significant differentiation was found between each pair of populations, indicating potential restricted gene flow. This was further demonstrated by significant isolation-by-distance pattern (p = 0.039), because low gene flow among population was needed to balance the minor effect of genetic drift, given F. tikoua was locally common. Restricted gene flow suggests that growth form may determine differences in gene flow between fig species.     

Paternity analysis of the breeding structure of strangler fig populations: evidence for substantial long-distance wasp dispersal

Abstract. The mutualistic interaction of figs with their species-specific wasp pollinators and the role of figs as 'keystone' plant resources in tropical communities has received substantial attention from both plant and animal ecologists. Despite this focus on the reproductive biology of figs, the minute size of the wasps has effectively precluded our ability to monitor patterns of wasp dispersal and fig mating relationships in natural forest habitats. In this paper we use genetic markers and genealogy reconstruction techniques to examine the breeding structure of populations of four strangler fig species occurring in central Panama. The natural history of figs facilitates the genetic analysis of full-sib progeny arrays from which the genotypes of successful pollen donors can be reconstructed precisely. Paternity reconstruction in the four study species reveals that individual flowering trees may routinely receive pollen from numerous donors despite characteristically low densities of co-flowering individuals. These data indicate not only that breeding populations of these figs are larger than the minimum critical sizes predicted to be necessary to support populations of their species-specific pollinators, but are more extensive in size and area than has been described for any plant species yet examined. Further, the fig wasps are shown to be efficient agents of long-distance dispersal, routinely moving up to 10 km between flowering trees. In accord with the potential for substantial long-distance gene flow and large effective population sizes, ten of eleven species of Panamanian figs assayed were found to maintain exceptionally high levels of genetic variation within their populations. Combined with other reports of occasional long-distance dispersal, the results of this study suggest that fig wasps may be more effective at colonizing isolated fig populations than previously thought.     

Fine-scale spatial genetic structure and gene dispersal in Silene latifolia

Plants are sessile organisms, often characterized by limited dispersal. Seeds and pollen are the critical stages for gene flow. Here we investigate spatial genetic structure, gene dispersal and the relative contribution of pollen vs seed in the movement of genes in a stable metapopulation of the white campion Silene latifolia within its native range. This short-lived perennial plant is dioecious, has gravity-dispersed seeds and moth-mediated pollination. Direct measures of pollen dispersal suggested that large populations receive more pollen than small isolated populations and that most gene flow occurs within tens of meters. However, these studies were performed in the newly colonized range (North America) where the specialist pollinator is absent. In the native range (Europe), gene dispersal could fall on a different spatial scale. We genotyped 258 individuals from large and small (15) subpopulations along a 60 km, elongated metapopulation in Europe using six highly variable microsatellite markers, two X-linked and four autosomal. We found substantial genetic differentiation among subpopulations (global F_ST=0.11) and a general pattern of isolation by distance over the whole sampled area. Spatial autocorrelation revealed high relatedness among neighboring individuals over hundreds of meters. Estimates of gene dispersal revealed gene flow at the scale of tens of meters (5–30 m), similar to the newly colonized range. Contrary to expectations, estimates of dispersal based on X and autosomal markers showed very similar ranges, suggesting similar levels of pollen and seed dispersal. This may be explained by stochastic events of extensive seed dispersal in this area and limited pollen dispersal.     

Limited seed dispersal shapes fine-scale spatial genetic structure in a Neotropical dioecious large-seeded palm

Seed and pollen dispersal contribute to gene flow and shape the genetic patterns of plants over fine spatial scales. We inferred fine-scale spatial genetic structure (FSGS) and estimated realized dispersal distances in Phytelephas aequatorialis, a Neotropical dioecious large-seeded palm. We aimed to explore how seed and pollen dispersal shape this genetic pattern in a focal population. For this purpose, we genotyped 138 seedlings and 99 adults with 20 newly developed microsatellite markers. We tested if rodent-mediated seed dispersal has a stronger influence than insect-mediated pollen dispersal in shaping FSGS. We also tested if pollen dispersal was influenced by the density of male palms around mother palms in order to further explore this ecological process in large-seeded plants. Rodent-mediated dispersal of these large seeds occurred mostly over short distances (mean 34.76 ± 34.06 m) while pollen dispersal distances were two times higher (mean 67.91 ± 38.29 m). The spatial extent of FSGS up to 35 m and the fact that seed dispersal did not increase the distance at which male alleles disperse suggest that spatially limited seed dispersal is the main factor shaping FSGS and contributes only marginally to gene flow within the population. Pollen dispersal distances depended on the density of male palms, decreasing when individuals show a clumped distribution and increasing when they are scattered. Our results show that limited seed dispersal mediated by rodents shapes FSGS in P. aequatorialis, while more extensive pollen dispersal accounts for a larger contribution to gene flow and may maintain high genetic diversity. Abstract in Spanish is available with online material.     

The role of gene flow in shaping genetic structures of the subtropical conifer species Araucaria angustifolia

The morphological features of pollen and seed of Araucaria angustifolia have led to the proposal of limited gene dispersal for this species. We used nuclear microsatellite and AFLP markers to assess patterns of genetic variation in six natural populations at the intra- and inter-population level, and related our findings to gene dispersal in this species. Estimates of both fine-scale spatial genetic structure (SGS) and migration rate suggest relatively short-distance gene dispersal. However, gene dispersal differed among populations, and effects of more efficient dispersal within population were observed in at least one stand. In addition, even though some seed dispersal may be aggregated in this principally barochorous species, reasonable secondary seed dispersal, presumably facilitated by animals, and overlap of seed shadows within populations is suggested. Overall, no correlation was observed between levels of SGS and inbreeding, density or age structure, except that a higher level of SGS was revealed for the population with a higher number of juvenile individuals. A low estimate for the number of migrants per generation between two neighbouring populations implies limited gene flow. We expect that stepping-stone pollen flow may have contributed to low genetic differentiation among populations observed in a previous survey. Thus, strategies for maintenance of gene flow among remnant populations should be considered in order to avoid degrading effects of population fragmentation on the evolution of A. angustifolia.     

A model for genetic relationship in areally continous plant populations.

Representations are based on plant populations, continuously distributed over their habitats according to specified density patterns. Migration of genetic material takes place via pollen and seed dispersal. Monoecious plants with arbitrary rates of self-fertilization and dioecious plants are considered. The model was constructed with the intention of determining coefficients of inbreeding and kinship for all locations within the seed population after its dispersal over the habitat, assuming the respective genetic relationships of the parental generation to be known. To display the consequences of single components hidden in the general result, the following specifications have been treated: finite population size combined with random dispersal of seed, equilibrium states for hypothetically infinite population size with “limited” dispersal of pollen and seed, random dispersal of pollen, and random dispersal of seed.     

Effect of gene flow on spatial genetic structure in the riparian canopy tree Cercidiphyllum japonicum revealed by microsatellite analysis

Few studies have analyzed pollen and seed movements at local scale, and genetic differentiation among populations covering the geographic distribution range of a species. We carried out such a study on Cercidiphyllum japonicum; a dioecious broad-leaved tree of cool-temperate riparian forest in Japan. We made direct measurement of pollen and seed movements in a site, genetic structure at the local scale, and genetic differentiation between populations covering the Japanese Archipelago. Parentage analysis of seedlings within a 20-ha study site indicated that at least 28.8% of seedlings were fertilized by pollen from trees outside the study site. The average pollination distance within the study site was 129 m, with a maximum of 666 m. The genotypes of 30% of seedlings were incompatible with those of the nearest female tree, and the maximum seed dispersal distance within the study site was over 300 m. Thus, long-distance gene dispersal is common in this species. The correlation between genetic relatedness and spatial distance among adult trees within the population was not significant, indicating an absence of fine-scale genetic structure perhaps caused by high levels of pollen flow and overlapping seed shadows. Six populations sampled throughout the distribution of C. japonicum in Japan showed significant isolation-by-distance but low levels of genetic differentiation (F(ST) = 0.043), also indicating long-distance gene flow in C. japonicum. Long-distance gene flow had a strong influence on the genetic structure at different spatial scales, and contributes to the maintenance of genetic diversity in C. japonicum.     

The functional implications of active and passive pollination in dioecious figs

Fig-pollinating wasps lay their eggs in fig flowers. Some species of fig-pollinating wasps are active pollinators, while others passively transfer pollen. In dioecious fig species, the ovules of male figs produce wasps but no seeds. By observations and experiments on four dioecious Ficus species we show that (i) passive pollinators distribute pollen haphazardly within figs, but fertilization of female flowers in male figs is inhibited. Consequently, wasp larvae will develop in nonfertilized ovules: they cannot benefit from pollination; (ii) active pollinators efficiently fertilize flowers in which they oviposit. Lack of pollination increases larval mortality. Hence, fig pollinators are not obligate seed eaters but ovule gallers. Active pollination has probably evolved as a way to improve progeny nourishment.
Comparison of pollination and oviposition process in male and female figs, suggests that stigma shape and function have coevolved with pollination behaviour, in relation to constraints linked with dioecy.     

Contemporary pollen and seed dispersal in a Prunus mahaleb population: patterns in distance and direction

Pollination and seed dispersal determine the spatial pattern of gene flow in plant populations and, for those species relying on pollinators and frugivores as dispersal vectors, animal activity plays a key role in determining this spatial pattern. For these plant species, reported dispersal patterns are dominated by short-distance movements with a significant amount of immigration. However, the contribution of seed and pollen to the overall contemporary gene immigration is still poorly documented for most plant populations. In this study we investigated pollination and seed dispersal at two spatial scales in a local population of Prunus mahaleb (L.), a species pollinated by insects and dispersed by frugivorous vertebrates. First, we dissected the relative contribution of pollen and seed dispersal to gene immigration from other parts of the metapopulation. We found high levels of gene immigration (18.50%), due to frequent long distance seed dispersal events. Second, we assessed the distance and directionality for pollen and seed dispersal events within the local population. Pollen and seed movement patterns were non-random, with skewed distance distributions: pollen tended moved up to 548 m along an axis approaching the N-S direction, and seeds were dispersed up to 990 m, frequently along the SW and SE axes. Animal-mediated dispersal contributed significantly towards gene immigration into the local population and had a markedly nonrandom pattern within the local population. Our data suggest that animals can impose distinct spatial signatures in contemporary gene flow, with the potential to induce significant genetic structure at a local level.     

Fragmentation reduces regional‐scale spatial genetic structure in a wind‐pollinated tree because genetic barriers are removed

Gene flow strongly influences the regional genetic structuring of plant populations. Seed and pollen dispersal patterns can respond differently to the increased isolation resulting from habitat fragmentation, with unpredictable consequences for gene flow and population structuring. In a recently fragmented landscape we compared the pre‐ and post‐fragmentation genetic structure of populations of a tree species where pollen and seed dispersal respond differentially to forest fragmentation generated by flooding. Castanopsis sclerophylla is wind‐pollinated, with seeds that are dispersed by gravity and rodents. Using microsatellites, we found no significant difference in genetic diversity between pre‐ and post‐fragmentation cohorts. Significant genetic structure was observed in pre‐fragmentation cohorts, due to an unknown genetic barrier that had isolated one small population. Among post‐fragmentation cohorts this genetic barrier had disappeared and genetic structure was significantly weakened. The strengths of genetic structuring were at a similar level in both cohorts, suggesting that overall gene flow of C. sclerophylla has been unchanged by fragmentation at the regional scale. Fragmentation has blocked seed dispersal among habitats, but this appears to have been compensated for by enhanced pollen dispersal, as indicated by the disappearance of a genetic barrier, probably as a result of increased wind speeds and easier pollen movement over water. Extensive pollen flow can counteract some negative effects of fragmentation and assist the long‐term persistence of small remnant populations.     

Fine-Scale Spatial Genetic Structure in Emmer Wheat and the Role of Population Range Position

The extent of spatial genetic structure (SGS) within plant populations depends on seed and pollen dispersal distance, breeding type, level of self-fertilization and effective plant density. Self-fertilizing species with gravity-dispersed seeds are expected to have both small effective population sizes and low pollen movement leading to high genetic structure. Higher SGS can be expected in more patchy and peripheral populations because of lower plant density and population sizes, and lower intensity of gene flow. We tested these predictions analyzing SGS in two core and two peripheral populations of predominantly self-fertilizing emmer wheat. Analysis of SGS with 11 nuclear microsatellites revealed (1) a negative linear relationship between kinship coefficients, calculated for pairs of individuals, and the logarithm of geographical distance between members of the pairs, in all studied populations; and (2) a significant autocorrelation for a distance up to 5 m (core populations) or 20 m (peripheral populations). Pollen flow, estimated from comparison of nuclear and chloroplast variation, was spatially limited, as was seed dispersal. Our results support a hypothesized relationship between SGS intensity and breeding system, the mode of seed dispersal and the population range position (core vs. periphery).     

Movements of genes between populations: are pollinators more effective at transferring their own or plant genetic markers?

The transfer of genes between populations is increasingly important in a world where pollinators are declining, plant and animal populations are increasingly fragmented and climate change is forcing shifts in distribution. The distances that pollen can be transported by small insects are impressive, as is the extensive gene flow between their own populations. We compared the relative ease by which small insects introduce genetic markers into their own and host-plant populations. Gene flow via seeds and pollen between populations of an Asian fig species were evaluated using cpDNA and nuclear DNA markers, and between-population gene flow of its pollinator fig wasp was determined using microsatellites. This insect is the tree''s only pollinator locally, and only reproduces in its figs. The plant''s pollen-to-seed dispersal ratio was 9.183–9.437, smaller than that recorded for other Ficus. The relative effectiveness of the pollinator at introducing markers into its own populations was higher than the rate it introduced markers into the plant''s populations (ratio = 14 : 1), but given the demographic differences between plant and pollinator, pollen transfer effectiveness is remarkably high. Resource availability affects the dispersal of fig wasps, and host-plant flowering phenology here and in other plant–pollinator systems may strongly influence relative gene flow rates.     

Habitat loss and fragmentation reduce effective gene flow by disrupting seed dispersal in a neotropical palm

Habitat loss and fragmentation often reduce gene flow and genetic diversity in plants by disrupting the movement of pollen and seed. However, direct comparisons of the contributions of pollen vs. seed dispersal to genetic variation in fragmented landscapes are lacking. To address this knowledge gap, we partitioned the genetic diversity contributed by male gametes from pollen sources and female gametes from seed sources within established seedlings of the palm Oenocarpus bataua in forest fragments and continuous forest in northwest Ecuador. This approach allowed us to quantify the separate contributions of each of these two dispersal processes to genetic variation. Compared to continuous forest, fragments had stronger spatial genetic structure, especially among female gametes, and reduced effective population sizes. We found that within and among fragments, allelic diversity was lower and genetic structure higher for female gametes than for male gametes. Moreover, female gametic allelic diversity in fragments decreased with decreasing surrounding forest cover, while male gametic allelic diversity did not. These results indicate that limited seed dispersal within and among fragments restricts genetic diversity and strengthens genetic structure in this system. Although pollen movement may also be impacted by habitat loss and fragmentation, it nonetheless serves to promote gene flow and diversity within and among fragments. Pollen and seed dispersal play distinctive roles in determining patterns of genetic variation in fragmented landscapes, and maintaining the integrity of both dispersal processes will be critical to managing and conserving genetic variation in the face of continuing habitat loss and fragmentation in tropical landscapes.     

A model for genetic relationship in areally continuous plant populations

Representations are based on plant populations, continuously distributed over their habitats according to specified density patterns. Migration of genetic material takes place via pollen and seed dispersal. Monoecious plants with arbitrary rates of self-fertilization and dioecious plants are considered. The model was constructed with the intention of determining coefficients of inbreeding and kinship for all locations within the seed population after its dispersal over the habitat, assuming the respective genetic relationships of the parental generation to be known. To display the consequences of single components hidden in the general result, the following specifications have been treated: finite population size combined with random dispersal of seed, equilibrium states for hypothetically infinite population size with “limited” dispersal of pollen and seed, random dispersal of pollen, and random dispersal of seed.     

Fine-scale genetic structure and gene flow within Costa Rican populations of mahogany (Swietenia macrophylla)

Fine-scale structure of genetic diversity and gene flow were analysed in three Costa Rican populations of mahogany, Swietenia macrophylla. Population differentiation estimated using AFLPs and SSRs was low (38.3 and 24%) and only slightly higher than previous estimates for Central American populations based on RAPD variation (20%). Significant fine-scale spatial structure was found in all of the surveyed mahogany populations and is probably strongly influenced by the limited seed dispersal range of the species. Furthermore, a survey of progeny arrays from selected mother trees in two of the plots indicated that most pollinations involved proximate trees. These data indicate that very little gene flow, via either pollen or seed, is occurring between blocks of mahogany within a continuous or disturbed forest landscape. Thus, once diversity is removed from a forest population of mahogany, these data suggest that recovery would be difficult via seed or pollen dispersal, and provides an explanation for mahogany's apparent susceptibility to the pressures of logging. Evidence is reviewed from other studies of gene flow and seedling regeneration to discuss alternative extraction strategies that may maintain diversity or allow recovery of genetic resources.     

The impact of pollination syndrome and habitat on gene flow: a comparative study of two Streptocarpus (Gesneriaceae) species

Gene flow through pollen and seed dispersal is important in terms of population differentiation and eventually speciation. Seed and pollen flow are affected in turn by habitats and pollen vectors. We examined the effect of different pollinators and habitats on gene flow by comparing two species of Streptocarpus, using microsatellite and chloroplast RFLP markers. Populations of the forest-dwelling S. primulifolius were highly differentiated according to nuclear microsatellite data and had mutually exclusive chloroplast haplotypes. This result is congruent with infrequent seed dispersal and limited between-population foraging by the long-tongued fly pollinator Stenobasipteron wiedemanni. In contrast, populations of S. dunnii growing in exposed crags had lower levels of population differentiation according to both nuclear and chloroplast data, congruent with a hypothesis of more effective between population seed dispersal and greater pollen-mediated gene flow due to the sunbird pollinator Nectarinia famosa. The population genetic behavior of these species is reflected in their taxonomy and phylogenetic position; S. primulifolius belongs to a taxonomically complex clade in which recent speciation is evident, while the clade containing S. dunnii is characterized by taxonomically well-defined species on longer phylogenetic branches. Our study shows that pollinator movements and seed dispersal patterns are a major determinant of the evolutionary trajectories of these species.     

Direct evidence for the cycling of fig wasps within one male fig tree

Fig wasps can only survive when flowering fig trees are present all the year around. Ficus trees can only reproduce if they are pollinated by highly specific wasps. In highly seasonal habitats, when only few trees occur at a specific site, gaps in fruiting may lead to the extinction of the local pollinator population. This paper demonstrates that in a dioecious fig tree, Ficus hirta , the fig wasp population can be maintained successfully within an individual plant, through the strong intra-tree asynchrony in flowering. By experimentally bagging trees, we showed that the pollinating wasps ( Blastophaga javana hilli ) could live for two generations, and the non-pollinating wasps ( Sycoscapter sp.) for up to three generations in a closed intra-tree system. However, there was a sharp decline in wasp abundance, deviating sex ratios and decreasing flower occupancy before their ultimate extinction, indicating that the wasp populations were not sustainable. This phenological strategy may enable dioecious figs, which are not constrained by the cost of selfing, to occupy a wider breadth of niches in both tropical and seasonal habitats.     

Spatial Segregation of the Sexes and Nutrients affect Reproductive Success in a Dioecious Wind-pollinated Grass

In many dioecious plant species in which spatial distributions of males and females have been examined, the sexes are spatially segregated – usually along an environmental gradient. Unless pollen is uniformly distributed in a population, spatial segregation of the sexes should reduce the average mating success of individuals. In three Californian populations of Distichlis spicata – a wind-pollinated grass species that exhibits spatial segregation of the sexes – I examined patterns of pollen movement and the effects of pollen load and nutrient availability on seed set to determine whether spatial segregation of the sexes actually reduces mating success for both males and females. In two of the populations, pollen dispersal was restricted, and pollen augmentation consistently, significantly increased seed set. However, in the third population – which had the lowest seed set – I found that although there were some indications of pollen limitation, pollen dispersal was not restricted, and seed production was limited primarily by nutrient availability. These results imply that in some populations of D. spicata nutrient limitation on the production of seeds by females may be sufficiently strong that spatial segregation of the sexes causes a fairly low cost to reproductive success compared with a more random distribution of the sexes. However, in other populations, pollen does limit mating success, and the spatial segregation of males and females in these populations is reducing the fecundity of both males and females.