Field of Dreams: Restitution of Pollinator Services in Restored Bird‐Pollinated Plant Populations

Ecosystem functionality is an increasingly important objective of ecological restoration. Despite this, a few studies have rigorously assessed reproductive functionality within restored plant populations, and it is largely assumed that pollinators follow restoration of plant communities—“build it and they will come.” Here, we applied an ecological genetic approach to determine the impact of spatial separation on mating in Banksia menziesii (Proteaceae), a dominant bird‐pollinated species of Banksia woodlands of Western Australia. All plants at three post‐mining restored sites (n = 72 [13 years old], n = 21 [8 years old], and n = 20 [9 years old]), as well as a sample from an adjacent natural reference site (n = 42), were genotyped at nine microsatellite loci. Seed set, mating system parameters, realized pollen dispersal through the assignment of paternity to seed, and avian pollinator species composition, abundance and behavior, were assessed. All patches displayed equivalent heterozygosity (H_e = 0.53–0.59) and very weak genetic divergence (F_ST ≤ 0.01). Seed of plants within restored sites showed complete outcrossing and relatively high seed set, 26% of which were sired by pollen donors located beyond the local patch. Similar abundance and movement of nectar‐feeding birds was observed in restored and natural sites, despite lower bird species diversity in the restored site, where a smaller, less aggressive species was dominant. Our results demonstrate the restitution of wide outcrossing in these restored Banksia patches within an active mine‐site, and suggest that restored bird‐pollinated Banksia populations are resilient to human impacts, due largely to their generalist pollinator requirements and highly‐mobile avian pollinators.     

Does restored plant diversity play a role in the reproductive functionality of Banksia populations?

Vegetation structure and plant species diversity of restoration sites are predicted to directly affect pollinator attraction, with potential impacts on gene flow, reproduction, genetic diversity of future generations, and ultimately restoration success. We compared Banksia attenuata R.Br. (Proteaceae) in a low species diversity restoration site and an adjacent natural remnant. We assessed fecundity genetic diversity in adult plants and their offspring, mating system parameters and pollen dispersal using paternity assignment. Results were compared to an earlier study of reproductive functionality within a high species diversity restoration site that was restored in a similar manner, enabling us to investigate any association between plant species diversity and fecundity. Seed set data indicated no significant differences between restored and adjacent natural sites; however, seed set data between restoration sites was significantly different (2.08 ± 0.39 and 6.89 ± 1.12, respectively). The mean number of fruits (follicles) per inflorescence was not significantly different between restoration sites. Genetic diversity of adult plants and their offspring were comparable in all sites. Higher allelic richness and genetic differentiation in one restored site reflected sourcing beyond local provenance. Low correlated paternity indicated high levels of multiple siring of seeds and paternity assignment demonstrated strong genetic connectivity between sites. Reproductive functionality, as measured by fecundity and genetic diversity in the offspring of B. attenuata, is resilient to low species diversity within a restored plant community. We consider our results in the context of establishing seed production areas (SPAs) that maximize the quantity and genetic quality of Banksia seeds for restoration.     

Ecological divergence and evolutionary transition of resprouting types in Banksia attenuata

Resprouting is a key functional trait that allows plants to survive diverse disturbances. The fitness benefits associated with resprouting include a rapid return to adult growth, early flowering, and setting seed. The resprouting responses observed following fire are varied, as are the ecological outcomes. Understanding the ecological divergence and evolutionary pathways of different resprouting types and how the environment and genetics interact to drive such morphological evolution represents an important, but under‐studied, topic. In the present study, microsatellite markers and microevolutionary approaches were used to better understand: (1) whether genetic differentiation is related to morphological divergence among resprouting types and if so, whether there are any specific genetic variations associated with morphological divergence and (2) the evolutionary pathway of the transitions between two resprouting types in Banksia attenuata (epicormic resprouting from aerial stems or branch; resprouting from a underground lignotuber). The results revealed an association between population genetic differentiation and the morphological divergence of postfire resprouting types in B. attenuata. A microsatellite allele has been shown to be associated with epicormic populations. Approximate Bayesian Computation analysis revealed a likely evolutionary transition from epicormic to lignotuberous resprouting in B. attenuata. It is concluded that the postfire resprouting type in B. attenuata is likely determined by the fire's characteristics. The differentiated expression of postfire resprouting types in different environments is likely a consequence of local genetic adaptation. The capacity to shift the postfire resprouting type to adapt to diverse fire regimes is most likely the key factor explaining why B. attenuata is the most widespread member of the Banksia genus.     

Primary pollinator exclusion has divergent consequences for pollen dispersal and mating in different populations of a bird‐pollinated tree

Pollination by nectarivorous birds is predicted to result in different patterns of pollen dispersal and plant mating compared to pollination by insects. We tested the prediction that paternal genetic diversity, outcrossing rate and realized pollen dispersal will be reduced when the primary pollinator group is excluded from bird‐pollinated plants. Pollinator exclusion experiments in conjunction with paternity analysis of progeny were applied to Eucalyptus caesia Benth. (Myrtaceae), a predominantly honeyeater‐pollinated tree that is visited by native insects and the introduced Apis mellifera (Apidae). Microsatellite genotyping at 14 loci of all adult E. caesia at two populations (n = 580 and 315), followed by paternity analysis of 705 progeny, revealed contrasting results between populations. Honeyeater exclusion did not significantly impact pollen dispersal or plant mating at Mount Caroline. In contrast, at the Chiddarcooping site, the exclusion of honeyeaters led to lower outcrossing rates, a threefold reduction in the average number of sires per fruit, a decrease in intermediate‐distance mating and an increase in near‐neighbour mating. The results from Chiddarcooping suggest that bird pollination may increase paternal genetic diversity, potentially leading to higher fitness of progeny and favouring the evolution of this strategy. However, further experimentation involving additional trees and study sites is required to test this hypothesis. Alternatively, insects may be effective pollinators in some populations of bird‐adapted plants, but ineffective in others.     

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.     

Evaluating Ecological Restoration Success: A Review of the Literature

Assessing the success of ecological restoration projects is critical to justify the use of restoration in natural resource management and to improve best practice. Although there are extensive discussions surrounding the characteristics that define and measure successful restoration, monitoring or evaluation of projects in practice is widely thought to have lagged behind. We conducted a literature review to determine trends in evaluations of restoration projects and identify key knowledge gaps that need to be addressed. We searched the Web of Knowledge plus two additional restoration journals not found in the database for empirical papers that assessed restoration projects post‐implementation. We quantified the extent that key attributes of success, including ecological (vegetation structure, species diversity and abundance, and ecosystem functioning) and socioeconomic, were addressed by these papers along with trends in publication and restoration characteristics. Encouragingly, we found the number of empirical evaluations has grown substantially in recent years. The increased age of restoration projects and number of papers that assessed ecological functions since previous reviews of the literature is also a positive development. Research is still heavily skewed toward United States and Australia, however, and identifying an appropriate reference site needs further investigation. Of particular concern is the dearth of papers identified in the literature search that included any measure of socioeconomic attributes. Focusing future empirical research on quantifying ecosystem services and other socioeconomic outcomes is essential for understanding the full benefits and costs of ecological restoration and to support its use in natural resource management .     

Mating patterns and pollinator mobility are critical traits in forest fragmentation genetics

Most woody plants are animal-pollinated, but the global problem of habitat fragmentation is changing the pollination dynamics. Consequently, the genetic diversity and fitness of the progeny of animal-pollinated woody plants sired in fragmented landscapes tend to decline due to shifts in plant-mating patterns (for example, reduced outcrossing rate, pollen diversity). However, the magnitude of this mating-pattern shift should theoretically be a function of pollinator mobility. We first test this hypothesis by exploring the mating patterns of three ecologically divergent eucalypts sampled across a habitat fragmentation gradient in southern Australia. We demonstrate increased selfing and decreased pollen diversity with increased fragmentation for two small-insect-pollinated eucalypts, but no such relationship for the mobile-bird-pollinated eucalypt. In a meta-analysis, we then show that fragmentation generally does increase selfing rates and decrease pollen diversity, and that more mobile pollinators tended to dampen these mating-pattern shifts. Together, our findings support the premise that variation in pollinator form contributes to the diversity of mating-pattern responses to habitat fragmentation.     

Influence of pollinator grooming on pollen‐mediated gene dispersal in Mimulus ringens (Phrymaceae)

Pollinator foraging patterns and the dynamics of pollen transport influence the quality and diversity of flowering plant mating opportunities. For species pollinated by grooming pollinators, such as bees, the amount of pollen carried between a donor flower and potential recipient flowers depends on how grooming influences pollen transfer. To investigate the relationship between grooming and pollen‐mediated gene dispersal, we studied bumblebee (Bombus fervidus) foraging behavior and resulting gene dispersal in linear arrays of Mimulus ringens. Each of the 14 plants in an array had a unique multilocus genotype, facilitating unambiguous assignment of paternity to 1050 progeny. Each plant was trimmed to a single flower so that pollinator movements could be linked directly to resulting gene dispersal patterns. Pollen‐mediated gene dispersal was very limited. More than 95% of the seeds sired by a donor flower were distributed over the first three recipient flowers in the visitation sequence. However, seeds were occasionally sired on flowers visited later in the pollinator's floral visitation sequence. Intensive grooming immediately following pollen removal from a donor flower significantly increased the decay rate of the donor flower's gene dispersal curve. These results suggest that the frequency and relative intensity of grooming can have significant effects on patterns of pollen‐mediated gene dispersal from individual pollen donors.     

Mating patterns, pollen dispersal, and the ecological maternal neighbourhood in a Prunus mahaleb L. population

Gender polymorphism, plant-animal interactions, and environmental heterogeneity are the three important sources of variation in mating system and pollen dispersal patterns. We used progeny arrays and paternity analysis to assess the effects of gender type and density level on variation in mating patterns within a highly isolated population of Prunus mahaleb, a gynodioecious species. All the adult trees in the population were sampled and located. The direct estimate of long-distance insect-mediated pollination events was low (< 10%). Gender expression deeply influenced the mating system, decreasing the outcrossing rates (t(m)) and the pollen pool diversity in hermaphrodite trees. Long intermate distances (> 250 m) were significantly more frequent among female mother trees. Variation in local tree density also affected pollen pool diversity and intermate distance, with a higher effective number of fathers (k(e)) and longer intermate distances for female trees in low-density patches. A canonical correlation analysis showed significant correlations between mating variables and the maternal ecological neighbourhood. Only the first canonical variable was significant and explained 78% of variation. Outcrossing rates tended to decrease, and the relatedness among the fathers tended to increase, when mother trees grew in dense patches with high cover of other woody species and taller vegetation away from the pine forest edge. We highlight the relevance of considering maternal ecological neighbourhood effects on mating system and gene flow studies as maternal trees act simultaneously as receptors of pollen and as sources of the seeds to be dispersed.     

High paternal diversity in the self-incompatible herb Arabidopsis halleri despite clonal reproduction and spatially restricted pollen dispersal

The number of sires fertilizing a given dam is a key parameter of the mating system in species with spatially restricted offspring dispersal, since genetic relatedness among maternal sibs determines the intensity of sib competition. In flowering plants, the extent of multiple paternity is determined by factors such as floral biology, properties of the pollen vector, selfing rate, spatial organization of the population, and genetic compatibility between neighbours. To assess the extent of multiple paternity and identify ecological factors involved, we performed a detailed study of mating patterns in a small population of a self-incompatible clonal herb, Arabidopsis halleri . We mapped and genotyped 364 individuals and 256 of their offspring at 12 microsatellite loci and jointly analysed the level of multiple paternity, pollen and seed dispersal, and spatial genetic structure. We found very low levels of correlated paternity among sibs ( P _full-sib = 3.8%) indicating high multiple paternity. Our estimate of the outcrossing rate was 98.7%, suggesting functional self-incompatibility. The pollen dispersal distribution was significantly restricted (mean effective pollen dispersal distance: 4.42 m) but long-distance successful pollination occurred and immigrating pollen was at most 10% of all pollination events. Patterns of genetic structure indicated little extent of clonal reproduction, and a low but significant spatial genetic structure typical for a self-incompatible species. Overall, in spite of restricted pollen dispersal, the multiple paternity in this self-incompatible species was very high, a result that we interpret as a consequence of high plant density and high pollinator service in this population.     

Impact of selective logging on inbreeding and gene dispersal in an Amazonian tree population of Carapa guianensis Aubl

Selective logging may impact patterns of genetic diversity within populations of harvested forest tree species by increasing distances separating conspecific trees, and modifying physical and biotic features of the forest habitat. We measured levels of gene diversity, inbreeding, pollen dispersal and spatial genetic structure (SGS) of an Amazonian insect-pollinated Carapa guianensis population before and after commercial selective logging. Similar levels of gene diversity and allelic richness were found before and after logging in both the adult and the seed generations. Pre- and post-harvest outcrossing rates were high, and not significantly different from one another. We found no significant levels of biparental inbreeding either before or after logging. Low levels of pollen pool differentiation were found, and the pre- vs. post-harvest difference was not significant. Pollen dispersal distance estimates averaged between 75 m and 265 m before logging, and between 76 m and 268 m after logging, depending on the value of tree density and the dispersal model used. There were weak and similar levels of differentiation of allele frequencies in the adults and in the pollen pool, before and after logging occurred, as well as weak and similar pre- and post-harvest levels of SGS among adult trees. The large neighbourhood sizes estimated suggest high historical levels of gene flow. Overall our results indicate that there is no clear short-term genetic impact of selective logging on this population of C. guianensis.     

High Levels of Genetic Contamination in Remnant Populations of Acacia saligna from a Genetically Divergent Planted Stand

It is essential to understand the patterns of pollen dispersal in remnant vegetation occupying highly disturbed landscapes in order to provide sustainable management options and to inform restoration programs. Direct and indirect methods of paternity analysis were used to detect genetic contamination via inter‐subspecific pollen dispersal from a planted stand of nonlocal Acacia saligna ssp. saligna (ms) into remnant roadside patches of local A. saligna ssp. lindleyi (ms). Genetic contamination was detected in 25.5% (indirect paternity assignment) to 32% (direct paternity assignment) of ssp. lindleyi progeny and occurred over a distance of 1.6 km. The results support studies that suggest genetic continuity is maintained by high levels of pollen dispersal in temperate entomophilous species. The results also indicate that patchily distributed remnant populations may be exposed to substantial amounts of genetic contamination from large‐scale restoration with native taxa in the highly fragmented agricultural landscape of southern Western Australia. Management practices to reduce the risk of genetic contamination are considered.     

新疆阜康荒漠红砂种群遗传结构及其与生态因子的耦合关系

Genetic structure and differentiation of Reaumuria soongorica (Pall.) Maxim population from the desert of Fukang, Xinjiang, were assessed by means of random amplified polymorphic DNA (RAPD) markers. High genetic diversity and differentiation were revealed in the population of R. soongorica by 15 random primers. One hundred and thirty-six individuals from seven subpopulations were sampled in the study. Seventy-one loci have been detected, and among them 69 were polymorphic. The mean proportion of polymorphic loci (PPB) was 97.18%. The analyses of Shannon information index (0.307 5), Nei抯 gene diversity (0.312 7) and GST (0.312 0) indicated that there were more genetic variations within the subpopu-lations than those among the subpopulations. The results of AMOVA analysis showed that 61.58% of the genetic variations existed within subpopulations, and 38.02% among the subpopulations. The gene flow among the subpopulations of R. soongorica (Nm = 1.102 8) was much less than that of the common anemophytes (Nm = 5.24), so genetic differentiation among the subpopulations occurred to some extent. Additionally, through the use of clustering and the correlation analyses, we found that the genetic struc-ture of natural population of R. soongorica was related to some ecological factors (soil factors mainly) of the oasis-desert transition zone. The genetic diversity level of R. soongorica had negative correlation with the content of total soil P and Cl- significantly (P＜0.05). On the contrary, it had significant positive correlation with CO32- (P＜0.05), showing that the distribution of the individuals of R. soongorica in the sampled areas correlates with certain soluble salt. Furthermore, the genetic diversity of the natural population of R. soongorica increased with the decreasing of the content of soil organic matters, water, total N and total P in soil. The paper concluded that the microenvironment ecological factors played an important role in the adaptive evolution of R. soongorica population.     

新疆阜康荒漠红砂种群遗传结构及其与生态因子的耦合关系

应用RAPD标记技术对分布于新疆阜康荒漠的重要植物红砂(Reaumuria soongorica(Pall.)Maxim)种群遗传结构和遗传多样性进行了分析。15条随机引物对红砂7个亚种群的136个个体进行扩增,共检测71个位点,其中多态位点69个。研究结果表明:红砂种群的多态位点比率(PPB)为97.18％,显示出分布于过渡带的红砂种群内存在较高的遗传多样性。Shannon多样性指数(0.307 5)、Nei基因多样性指数(0.312 7)和基因分化系数(G_(ST)=0.312 0)揭示了红砂种群遗传变异多存在于亚种群内,而亚种群间的遗传分化则较小。红砂业种群间的基因流Nm=1.102 8,Nm>1但低于一般风媒传粉植物(Nm=5.24)的基因流水平,处于分化的临界状态。AMOVA分析说明红砂种群变异的61.58％存在于亚种群内,而亚种群间的变异占总变异的38.02％。另外,通过RAPD资料的聚类分析及相关性分析研究,发现红砂自然种群的遗传结构与绿洲荒漠过渡带的微生境生态因子(主要是土壤因子)相关,其中红砂亚种群遗传多样性水平与土壤中全磷和CI含量呈显著负相关,与CO_3~2含量呈显著正相关。而其它土壤生态因子则与红砂的遗传多样性指数的相关性均不显著(P>0.05)。表明红砂个体的分布可能与过渡带土壤的某些易溶性盐分相关。研究还发现,随着土壤中有机质、水分、全氮和全磷含量的减     

The incidence and selection of multiple mating in plants

Mating with more than one pollen donor, or polyandry, is common in land plants. In flowering plants, polyandry occurs when the pollen from different potential sires is distributed among the fruits of a single individual, or when pollen from more than one donor is deposited on the same stigma. Because polyandry typically leads to multiple paternity among or within fruits, it can be indirectly inferred on the basis of paternity analysis using molecular markers. A review of the literature indicates that polyandry is probably ubiquitous in plants except those that habitually self-fertilize, or that disperse their pollen in pollen packages, such as polyads or pollinia. Multiple mating may increase plants'' female component by alleviating pollen limitation or by promoting competition among pollen grains from different potential sires. Accordingly, a number of traits have evolved that should promote polyandry at the flower level from the female''s point of view, e.g. the prolongation of stigma receptivity or increases in stigma size. However, many floral traits, such as attractiveness, the physical manipulation of pollinators and pollen-dispensing mechanisms that lead to polyandrous pollination, have probably evolved in response to selection to promote male siring success in general, so that polyandry might often best be seen as a by-product of selection to enhance outcross siring success. In this sense, polyandry in plants is similar to geitonogamy (selfing caused by pollen transfer among flowers of the same plant), because both polyandry and geitonogamy probably result from selection to promote outcross siring success, although geitonogamy is almost always deleterious while polyandry in plants will seldom be so.     

Limiting inbreeding in disjunct and isolated populations of a woody shrub

Pollen movements and mating patterns are key features that influence population genetic structure. When gene flow is low, small populations are prone to increased genetic drift and inbreeding, but naturally disjunct species may have features that reduce inbreeding and contribute to their persistence despite genetic isolation. Using microsatellite loci, we investigated outcrossing levels, family mating parameters, pollen dispersal, and spatial genetic structure in three populations of Hakea oldfieldii, a fire‐sensitive shrub with naturally disjunct, isolated populations prone to reduction in size and extinction following fires. We mapped and genotyped a sample of 102 plants from a large population, and all plants from two smaller populations (28 and 20 individuals), and genotyped 158–210 progeny from each population. We found high outcrossing despite the possibility of geitonogamous pollination, small amounts of biparental inbreeding, a limited number of successful pollen parents within populations, and significant correlated paternity. The number of pollen parents for each seed parent was moderate. There was low but significant spatial genetic structure up to 10 m around plants, but the majority of successful pollen came from outside this area including substantial proportions from distant plants within populations. Seed production varied among seven populations investigated but was not correlated with census population size. We suggest there may be a mechanism to prevent self‐pollination in H. oldfieldii and that high outcrossing and pollen dispersal within populations would promote genetic diversity among the relatively small amount of seed stored in the canopy. These features of the mating system would contribute to the persistence of genetically isolated populations prone to fluctuations in size.     

Pollen dispersal and genetic structure of the tropical tree Dipteryx panamensis in a fragmented Costa Rican landscape

In the face of widespread deforestation, the conservation of rainforest trees relies increasingly on their ability to maintain reproductive processes in fragmented landscapes. Here, we analysed nine microsatellite loci for 218 adults and 325 progeny of the tree Dipteryx panamensis in Costa Rica. Pollen dispersal distances, genetic diversity, genetic structure and spatial autocorrelation were determined for populations in four habitats: continuous forest, forest fragments, pastures adjacent to fragments and isolated pastures. We predicted longer but less frequent pollen movements among increasingly isolated trees. This pattern would lead to lower outcrossing rates for pasture trees, as well as lower genetic diversity and increased structure and spatial autocorrelation among their progeny. Results generally followed these expectations, with the shortest pollen dispersal among continuous forest trees (240 m), moderate distances for fragment (343 m) and adjacent pasture (317 m) populations, and distances of up to 2.3 km in isolated pastures (mean: 557 m). Variance around pollen dispersal estimates also increased with fragmentation, suggesting altered pollination conditions. Outcrossing rates were lower for pasture trees and we found greater spatial autocorrelation and genetic structure among their progeny, as well as a trend towards lower heterozygosity. Paternal reproductive dominance, the pollen contributions from individual fathers, did not vary among habitats, but we did document asymmetric pollen flow between pasture and adjacent fragment populations. We conclude that long-distance pollen dispersal helps maintain gene flow for D. panamensis in this fragmented landscape, but pasture and isolated pasture populations are still at risk of long-term genetic erosion.     

Effects of male fecundity, interindividual distance and anisotropic pollen dispersal on mating success in a Scots pine (Pinus sylvestris) seed orchard

Quantifying the effect of pollen dispersal and flowering traits on mating success is essential for understanding evolutionary responses to changing environments and establishing strategies for forest tree breeding. This study examined, quantitatively, the effects of male fecundity, interindividual distance and anisotropic pollen dispersal on the mating success of Scots pine (Pinus sylvestris), utilizing a well-mapped Scots pine seed orchard. Paternity analysis of 1021 seeds sampled from 87 trees representing 28 clones showed that 53% of the seeds had at least one potential pollen parent within the orchard. Pronounced variation in paternal contribution was observed among clones. Variations in pollen production explained up to 78% of the variation in mating success, which was 11.2 times greater for clones producing the largest amount of pollen than for clones producing the least pollen. Mating success also varied with intertree distance and direction, which explained up to 28% of the variance. Fertilization between neighboring trees 2.3 m apart was 2.4 times more frequent than between trees 4.6 m apart, and up to 12.4 times higher for trees downwind of the presumed prevailing wind direction than for upwind trees. The effective number of pollen donors recorded in the seed orchard (12.2) was smaller than the theoretical expectation (19.7). Based on the empirical observations, a mating model that best describes the gene dispersal pattern in clonal seed orchards was constructed.     

Extensive pollen immigration and no evidence of disrupted mating patterns or reproduction in a highly fragmented holm oak stand

Aims Forest fragmentation and reduced tree population densities can potentially have negative impacts on mating patterns, offspring genetic diversity and reproductive performance. The aim of the present study is to test these hypotheses comparing an extremely fragmented, low tree density (~0.02 trees/ha) holm oak (Quercus ilex L.) stand from Central Spain with a nearby high tree density stand (~50 trees/ha).Methods We genotyped adult trees and seeds from the low-density stand (436 seeds from 15 families) and the high-density stand (404 seeds from 11 families) using nine microsatellite markers. With these data, we performed paternity analyses, determined pollen flow, mating patterns and pollen pool structure, and estimated progeny genetic diversity in both stands. We also studied seed set and production and performed a pollen supplementation experiment to determine whether reduced tree density has limited foreign pollen availability.Important findings We have found extensive pollen immigration (>75%) into the low tree density stand and Monte Carlo simulations revealed that pollen moves larger distances than expected from null models of random dispersal. Mating patterns and differentiation of pollen pools were similar in the high-density stand and the low-density stand but we found higher inter-annual differentiation of pollen pools in the former. Progeny genetic diversity and self-fertilization rates did not differ between the low-density stand and the high-density stand. Seed set rates were significantly lower in the low-density stand than in the high-density stand and experimental cross-pollen supplementation evidenced that foreign pollen availability is indeed a limiting factor in the former. However, seed crops did not differ between the low-density stand and the high-density stand, indicating that limitation of foreign pollen is not likely to be of great concern in terms of reduced seed production and potential recruitment. Poor forest regeneration due to other ecological and human factors is probably a more important threat for the persistence of fragmented and low tree density stands than reduced pollen flow and only extremely small and isolated tree populations would be expected to suffer severe loss of genetic diversity in the long term.