Long-and short-billed hummingbirds as pollinators of Palicourea demissa, a distylous treelet of Neotropical cloud forests.

Distyly has been interpreted as a mechanism that promotes cross-pollination between floral morphs. According to this hypothesis, pollen from anthers positioned at different heights could adhere to different body parts of the pollinator that would correspond to those points where stigmas of compatible morph contact the animal. In this regard, hummingbird species with different bill sizes may play different roles as pollinators of each morph. If pollinators mobilize more legitimate pollen towards one of the two morphs, gender specialization may occur. This work aimed to assess experimentally the role of long- and short-billed hummingbirds as pollinators of short-style (SS) and long-style (LS) flowers of Palicourea demissa, a distylous, hummingbird-pollinated treelet in Venezuelan cloud forests. Flowers were emasculated and exposed to a single visit of the hummingbird Coeligena torquata (long-billed), Heliangelus spencei (short-billed) or Adelomyia melanogenys (short-billed). Later, stigmas were removed, and pollen load counted under a microscope to calculate the probability of legitimate- and illegitimate-pollen transfer by hummingbirds. The probability analyses of pollen transference showed that short-billed hummingbirds have higher pollination probabilities from SS-anthers to LS- and SS-stigmas, and from LS-anther to LS-stigmas than from LS-anther to SS-stigmas. In contrast, long-billed hummingbirds have higher probabilities of pollen transference from LS-anthers to SS-stigmas than in other directions. A deeper view of the sexual expression of each morph in P. demissa will depend on future studies that determine possible morpho-differences in the biological function of male and female floral structures, and the role played by less frequent floral visitors as mediators of legitimate pollination between floral morphs.

Direct tracking of pollen with quantum dots reveals surprising uniformity in dispersal distance across 11 populations of an annual plant.

PREMISE: Pollen movement is a crucial component of dispersal in seed plants. Although pollen dispersal is well studied, methodological constraints have made it challenging to directly track pollen flow within multiple populations across landscapes. We labeled pollen with quantum dots, a new technique that overcomes past limitations, to evaluate the spatial scale of pollen dispersal and its relationship with conspecific density within 11 populations of Clarkia xantiana subsp. xantiana, a bee-pollinated annual plant. METHODS: We used experimental arrays in two years to track pollen movement across distances of 5-35 m within nine populations and across distances of 10-70 m within two additional populations. We tested for distance decay of pollen dispersal, whether conspecific density modulated dispersal distance, and whether dispersal kernels varied among populations across an environmentally complex landscape. RESULTS: Labeled pollen receipt did not decline with distance over 35 m within eight of nine populations or over 70 m within either of two populations. Pollen receipt increased with conspecific density. Overall, dispersal kernels were consistent across populations. CONCLUSIONS: The surprising uniformity in dispersal distance within different populations was likely influenced by low precipitation and plant density in our study years. This suggests that spatiotemporal variation in the abiotic environment substantially influences the extent of gene flow within and among populations.

Complementary roles of hummingbirds and bees: Pollen pickup, pollen deposition, and fruit production in the distylous Palicourea rigida.

PREMISE: Distyly is a condition in which individual plants in a population express two floral morphs, L- and S-morph, characterized by reciprocal placements of anthers and stigmas between morphs. The function of distyly requires that pollinators collect pollen from L- and S-morphs on different parts along their bodies to then deposit it on the stigmas of the opposite morph, known as legitimate pollination. However, different pollinator groups might differ in the ability to transfer pollen legitimately. METHODS: We investigated patterns of pollen pickup along the body of different functional groups (hummingbirds and bees) using preserved specimens to analyze their role in the reproductive success of Palicourea rigida. We measured pollen deposition on the body of pollinators, on stigmas, and fruit production after a single visit. RESULTS: Pollen from L- and S-flowers appeared segregated on different body parts of the hummingbird and bee used in the study. S-pollen was deposited primarily on the proximal regions (near the head), and L-pollen was placed in the distal regions (tip of the proboscis and bill). Hummingbirds were more efficient at legitimate pollination than bees, particularly to S-stigmas. However, fruit formation after single visits by both pollinators was similar. CONCLUSIONS: The morphology of distylous flowers allows the segregated placement of L-and S-pollen on different body parts of the animal specimens used, facilitating the promotion of legitimate pollen transfer, an observation consistent between the two functional pollinator groups. Also, the results show that full fruit set requires more than one visit.

Population Structure and Genetic Diversity in the Natural Distribution of Neolamarckia cadamba in China.

Neolamarckia cadamba (Roxb.) Bosser is a fast-growing deciduous tree species and belongs to the Neolamarckia genus of the Rubiaceae family. This species has great economic and medical values in addition to being an important timber species for multiple industrial purposes. However, few studies have examined the genetic diversity and population structure in the natural distribution of this species in China. Here, we applied both the haploid nrDNA ITS (619 bp for aligned sequences) and mtDNA (2 polymorphic loci) markers to investigate 10 natural populations (239 individuals in total) that covered most of the distribution of the species in China. The results showed that the nucleotide diversity was π = 0.1185 ± 0.0242 for the nrDNA ITS markers and π = 0.00038 ± 0.00052 for the mtDNA markers. The haplotype diversity for the mtDNA markers was h = 0.1952 ± 0.2532. The population genetic differentiation was small (Fstn = 0.0294) for the nrDNA ITS markers but large (Fstm = 0.6765) for the mtDNA markers. There were no significant effects of isolation by distance (IBD), by elevation, and by two climatic factors (annual average precipitation and tem perature). A geographic structure among populations (Nst

Touch-sensitive stamens enhance pollen dispersal by scaring away visitors.

Animal-pollinated plants have to get pollen to a conspecific stigma while protecting it from getting eaten. Touch-sensitive stamens, which are found in hundreds of flowering plants, are thought to function in enhancing pollen export and reducing its loss, but experimental tests are scarce. Stamens of Berberis and Mahonia are inserted between paired nectar glands and when touched by an insect's tongue rapidly snap forward so that their valvate anthers press pollen on the insect's tongue or face. We immobilized the stamens in otherwise unmodified flowers and studied pollen transfer in the field and under enclosed conditions. On flowers with immobilized stamens, the most common bee visitor stayed up to 3.6× longer, yet removed 1.3× fewer pollen grains and deposited 2.1× fewer grains on stigmas per visit. Self-pollen from a single stamen hitting the stigma amounted to 6% of the grains received from single bee visits. Bees discarded pollen passively placed on their bodies, likely because of its berberine content; nectar has no berberine. Syrphid flies fed on both nectar and pollen, taking more when stamens were immobilized. Pollen-tracking experiments in two Berberis species showed that mobile-stamen-flowers donate pollen to many more recipients. These results demonstrate another mechanism by which plants simultaneously meter out their pollen and reduce pollen theft.

Pollen transfer and patterns of reproductive success in pure and mixed populations of nectariferous Platanthera bifolia and P. chlorantha (Orchidaceae).

Plant species evolution is driven by many factors that have different roles in space and time. Using different field and laboratory methods, we studied reproductive patterns and their determinants in pure and mixed P. bifolia and P. chlorantha populations in different habitats. We also considered the probability of hybridisation between these two species and the role of intra-population processes in maintaining species integrity. Generally, we found a high level of reproductive success in both Platantherans. In both species, male (MRS) and female (FRS) reproductive success depended on floral display, and male reproductive success additionally on population structure. The flower traits were only weakly related to reproductive success. Moths' assemblages varied spatially and temporally, and their diversity and numbers were correlated with MRS in the year, when their abundance was markedly lower. Analysis of patterns of pollen transfer showed that pollen was transported up to 25 m (average 8.2 ± 4.83 m) and showed gene exchange between these two Platanthera species. The germination level of both species was significantly lower than seed viability, although P. bifolia seed germinated with higher frequency than P. chlorantha seeds. We noted differences in viability and germination of seeds developed as an effect of experimental interspecies crossings and those developed from natural pollination. The presence of intermediate ecotypes together with observations of spontaneous interspecies crosses in the field and viability of seeds produced in interspecies crossing suggest that both pre- and postzygotic reproductive barriers are not complete and do not prevent hybrid production.

Limited-Distance Pollen Dispersal and Low Paternal Diversity in a Bird-Pollinated Self-Incompatible Tree.

Bird pollination in Asia is regarded as an uncommon phenomenon and, therefore, only a few investigations on mating pattern and paternity in fruits of Asian bird-pollinated plants have been conducted. Here, we examined spatial genetic structure, pollen dispersal, and multiple paternity in a natural population of Bombax ceiba (B. ceiba) (Malvaceae) in Hainan Island, South China, using simple sequence repeat (SSR) markers. A low genetic diversity (H e = 0.351 ± 0.0341 and 0.389 ± 0.043, respectively, for adults and offspring) and bottleneck effects were observed. Genetic kinship was significant within 400 m or in 1,800-3,800 m. Both the mating pattern and paternity analysis confirmed obligate xenogamy and a low multiple paternity in B. ceiba. There was a strongly negative relationship between the frequency of matings and the distance between mating pairs. The average pollen dispersal distance was 202.89 ± 41.01 m (mean ± SE) and the farthest distance of > 1 km was recorded. Realized mating events showed an extremely leptokurtic distribution within 1,200 m, suggesting that the pollen dispersal distance was consistent with the optimal foraging theory of generalist birds such as Zosterops spp. and Pycnonotus spp. Paternity per tree ranged from two to six and the average effective number of pollen donors per maternal plant was 3.773, suggesting a low level of paternity diversity as compared to other bird-pollinated plants. We concluded that optimal foraging behavior by generalist birds could explain the leptokurtic pollen dispersal distribution and predominantly near-neighbor matings in B. ceiba. The limited pollen dispersal distance and low multiple paternity were consistent with low fruit setting rate (3.27 ± 0.93%) in this self-incompatible tree, which was caused mainly by the restricted flight distance of birds and human disturbances. Low genetic diversity and significant spatial genetic structure might have largely resulted from logging and human collection of fruits.

Strongly unbalanced gender attractiveness in a dioecious mass flowering tropical tree pollinated by stingless bees.

In dioecious, bee-pollinated tree species, male and female flowers offer different resources. It is unclear how this unbalanced quantity and quality of floral resources affects flower visits and pollen transfer to female flowers. We asked, what characteristics of flowering and dynamics of resource production by trees favour flights of bees between male and female trees. We quantified the floral resources produced by individual flowers and entire trees of Myracrodruon urundeuva (Anacardiaceae), measured pollen flow to female flowers, fruit set of naturally pollinated flowers and determined the effective pollinators. Crown volume of male trees was four-fold, flower number 15-fold and nectar volume 60 times higher than in female trees. While ~70% of male flowers opened in the morning, ~70% of female flowers opened in the afternoon. Fruit set was 27%. Stingless bee species were the main pollinators, while honeybees were common only on male flowers. Strongly unbalanced production of floral resources, high potential lifespan of female flowers and anticipated opening of male flowers favour pollinator movement and pollen deposition on female flowers, albeit low (0.003% of pollen that reached stigmas), but sufficient to produce thousands of fruits per tree. Besides being an astonishing pollen and nectar source for numerous social bee species, only a few of them were effective pollinators. Our quantitative approach to floral resource production of each gender provides new insights, such as the proportion of resources allocated to each gender and the corresponding behaviour of flower visitors, for understanding the reproductive strategy of dioecious tropical mass-flowering trees.

Phylogeography of a gypsum endemic plant across its entire distribution range in the western Mediterranean.

PREMISE: Gypsum soils in the Mediterranean Basin house large numbers of edaphic specialists that are adapted to stressful environments. The evolutionary history and standing genetic variation of these taxa have been influenced by the geological and paleoclimatic complexity of this area and the long-standing effect of human activities. However, little is known about the origin of Mediterranean gypsophiles and the factors affecting their genetic diversity and population structure. METHODS: Using phylogenetic and phylogeographic approaches based on microsatellites and sequence data from nuclear and chloroplast regions, we evaluated the divergence time, genetic diversity, and population structure of 27 different populations of the widespread Iberian gypsophile Lepidium subulatum throughout its entire geographic range. RESULTS: Lepidium subulatum diverged from its nearest relatives ~3 million years ago, and ITS and psbA/matK trees supported the monophyly of the species. These results suggest that both geological and climatic changes in the region around the Plio-Pleistocene promoted its origin, compared to other evolutionary processes. We found high genetic diversity in both nuclear and chloroplast markers, but a greater population structure in the chloroplast data. These results suggest that while seed dispersal is limited, pollen flow may be favored by the presence of numerous habitat patches that enhance the movement of pollinators. CONCLUSIONS: Despite being an edaphic endemic, L. subulatum possesses high genetic diversity probably related to its relatively old age and high population sizes across its range. Our study highlights the value of using different markers to fully understand the phylogeographic history of plant species.

Effective gene flow patterns across a fragmented landscape in southern Costa Rica for Symphonia globulifera (Clusiaceae); a species with mobile seed and pollen dispersers

In tropical trees, forest fragmentation has been shown to affect mating and gene flow patterns. Mobile dispersal vectors should be less sensitive to fragmented landscapes and may ameliorate the genetic effects of forest fragmentation on plant populations. To test this hypothesis, we analyzed gene flow patterns in Symphonia globulifera, a tropical tree species with highly mobile pollinators and seed dispersers in the Osa Peninsula in southern Costa Rica. We used microsatellites to study genetic diversity and realized gene flow patterns between a continuous forest and a forest fragment. We found high levels of genetic diversity in adults and seedlings at both sites. Parentage analyses suggest near-neighbor matings and frequent long-distance gene flow events. Half the progeny beneath an adult was not sired by that tree and the majority of established seedlings were the result of long-distance gene dispersal. Gene flow from the forest into the fragment was more common than from the fragment into the continuous forests. Despite long distance gene flow events, seedling spatial genetic structure was stronger and extended further in the forest fragment likely due to limited seed dispersal. We conclude that fragmentation affects gene flow in this tropical tree and may compromise its genetic diversity in forest fragments even for a species with mobile pollen and seed vectors.

En árboles tropicales se ha demostrado que la fragmentación del bosque afecta los patrones de apareamiento y reduce el flujo génico. Las especies forestales cuyos dispersores y polinizadores son capaces de moverse largas distancias sobre ambientes alterados, serán menos sensibles a los efectos de la fragmentación y podrán intercambiar genes regularmente entre fragmentos. Para probar esta hipótesis, en la Península de Osa en el sur de Costa Rica, estudiamos los patrones de flujo génico de Symphonia globulifera, una especie de árbol tropical con polinizadores y dispersores de semillas altamente móviles. Utilizamos microsatélites para estudiar la diversidad genética y los patrones de flujo génico entre un bosque continuo y un fragmento de bosque. Encontramos altos niveles de diversidad genética tanto en adultos como en plántulas. Los análisis de parentesco indican apareamiento entre vecinos cercanos, sin embargo el flujo génico a larga distancia es frecuente. La mitad de la progenie debajo de un adulto no es engendrada por ese árbol y la mayoría de las plántulas establecidas son el resultado del flujo génico a larga distancia. El flujo de genes desde el bosque continuo hacia el fragmento era más frecuente que el movimiento inverso. A pesar del flujo génico a larga distancia, hay diferencias en frecuencias alélicas entre plántulas y la estructura espacial era más fuerte y se extendía a mayor distancia en el fragmento, probablemente por una dispersión limitada de semillas. Concluimos que la fragmentación afecta los patrones de flujo génico en esta especie de árbol tropical y puede afectar su diversidad genética en paisajes fragmentados, inclusive para una especies con polinizadores y dispersores con alta movilidad.

Estimating seed and pollen dispersal kernels from genetic data demonstrates a high pollen dispersal capacity for an endangered palm species.

PREMISE OF THE STUDY: Seed and pollen dispersal are key processes shaping plant population dynamics and maintaining genetic diversity. The essence of these processes is the movement of propagules from a parental tree to the site of propagule establishment. The estimation of plant dispersal kernels has remained challenging due to the difficulty of making direct observations. We estimated the dispersal capacity of the endangered palm Euterpe edulis, whose seeds are dispersed by vertebrates and pollen by insects. METHODS: We used a hierarchical Bayesian model with genetic data from reproductive plants, juveniles, and embryos to estimate dispersal kernels. Our analyses account for genotyping error and uncertainty in parental assignment. KEY RESULTS: We found that seeds were dispersed at most a few hundred meters, but pollen was dispersed up to several kilometers. We hypothesize that this long-distance pollen dispersal is generated mainly by euglossine bees, whereas the main dispersal vectors for short-distance seed dispersal are thrushes. The long-distance dispersal of pollen suggests a high level of gene flow that should maintain genetic diversity of E. edulis. Despite the relation between long-distance dispersal and genetic diversity, we observed low genetic diversity and inbreeding within the local population, which are probably due to restricted gene flow due to the low density of this population and its aggregated spatial distribution. CONCLUSIONS: We conclude that if conservation actions are able to restore the population density of E. edulis, the recovery of its genetic diversity will be facilitated because of its high dispersal capacity, especially with regard to pollen.

Pollen and seed dispersal of Brazil nut trees in the southwestern Brazilian Amazon / Dispersão de pólen e sementes da castanheira no sudoeste da Amazônia brasileira

Pollen and seed dispersal patterns greatly influence the spatial distribution of plant genetic diversity. Microsatellite-based parentage analysis provides accurate estimates of contemporary gene dispersal. Although most tropical trees have been shown to exhibit widespread pollen dispersal, few studies have estimated contemporary gene dispersal after seedling establishment. Bertholletia excelsa (Lecythidaceae) is pollinated by large-bodied bees, while previous seed-tracking experiments suggest their seeds are mainly dispersed across very short distances by scatter-hoarding rodents, who primarily act as seed predators. Here we used parentage analysis to provide contemporary estimates of pollen and seed dispersal in B. excelsa recruits. We examined six 25-ha plots located in two natural stands in the Acre River valley, in the southwestern Brazilian Amazon. We used 11 microsatellite markers to estimate genetic diversity and fixation index parameters in adults, seedlings and saplings. Genetic diversity was moderate and did not differ across size classes or sampling locations. We assigned pollen and seed parents for < 20% of the recruits, indicating that most events of realized gene flow occurred beyond our 25-ha plots. Only 10 parentage assignments were confirmed with 80% confidence. Pollen distance ranged from 33 to 372 m and seed dispersal from 58 to 655 m. Actual seed-dispersal distances were far greater than the estimates obtained in previous seed-tracking experiments. Thus, studies encompassing larger sampling areas are necessary to determine a more representative spatial scale of B. excelsas pollen and seed dispersal capacity in natural stands.

Os padrões de dispersão de pólen e sementes influenciam a distribuição espacial da diversidade genética. Muitas espécies arbóreas tropicais apresentam ampla dispersão de pólen, mas poucos estudos avaliaram fluxo gênico a partir de plântulas. Bertholletia excelsa (Lecythidaceae) é polinizada por abelhas e as sementes são dispersas por roedores do tipo scatter-hoarders (que estocam recursos em diferentes pontos de sua área de vida), que atuam primariamente como predadores de sementes. Experimentos de remoção de sementes tem mostrado que a dispersão de sementes por esses roedores é espacialmente limitada. Nosso objetivo foi obter estimativas de dispersão de pólen e sementes em B. excelsa a partir da análise de parentesco de regenerantes. Nós estudamos seis parcelas de 25 ha, em duas áreas de floresta nativa no vale do Rio Acre, no sudoeste da Amazônia brasileira. Parâmetros de diversidade genética e índice de fixação foram estimados em adultos, varetas e plântulas com 11 marcadores microssatélites. A diversidade genética foi moderada e não diferiu entre classes de tamanho ou entre localidades. A paternidade foi determinada em menos de 20% dos regenerantes, indicando que a maioria dos eventos de fluxo gênico ocorreu em distâncias maiores que as encontradas nas parcelas de 25 ha. As distâncias de pólen variaram de 33 a 372 m e as de dispersão de sementes variaram de 58 a 655 m. As distâncias de dispersão obtidas neste estudo excedem em muito as estimativas obtidas em experimentos de remoção de sementes. Estudos envolvendo áreas maiores são necessários para que possamos aprofundar nosso conhecimento sobre capacidade de dispersão de pólen e sementes em populações naturais de B. excelsa.

The relative contributions of seed and pollen dispersal to gene flow and genetic diversity in seedlings of a tropical palm.

Seed and pollen dispersal shape patterns of gene flow and genetic diversity in plants. Pollen is generally thought to travel longer distances than seeds, but seeds determine the ultimate location of gametes. Resolving how interactions between these two dispersal processes shape microevolutionary processes is a long-standing research priority. We unambiguously isolated the separate and combined contributions of these two dispersal processes in seedlings of the animal-dispersed palm Oenocarpus bataua to address two questions. First, what is the spatial extent of pollen versus seed movement in a system characterized by long-distance seed dispersal? Second, how does seed dispersal mediate seedling genetic diversity? Despite evidence of frequent long-distance seed dispersal, we found that pollen moves much further than seeds. Nonetheless, seed dispersal ultimately mediates genetic diversity and fine-scale spatial genetic structure. Compared to undispersed seedlings, seedlings dispersed by vertebrates were characterized by higher female gametic and diploid seedling diversity and weaker fine-scale spatial genetic structure for female gametes, male gametes and diploid seedlings. Interestingly, the diversity of maternal seed sources at seed deposition sites (Nem ) was associated with higher effective number of pollen sources (Nep ), higher effective number of parents (Ne ) and weaker spatial genetic structure, whereas seed dispersal distance had little impact on these or other parameters we measured. These findings highlight the importance maternal seed source diversity (Nem ) at frugivore seed deposition sites in driving emergent patterns of fine-scale genetic diversity and structure.

Florally rich habitats reduce insect pollination and the reproductive success of isolated plants.

Landscape heterogeneity in floral communities has the potential to modify pollinator behavior. Pollinator foraging varies with the diversity, abundance, and spatial configuration of floral resources. However, the implications of this variation for pollen transfer and ultimately the reproductive success of insect pollinated plants remains unclear, especially for species which are rare or isolated in the landscape. We used a landscape-scale experiment, coupled with microsatellite genotyping, to explore how the floral richness of habitats affected pollinator behavior and pollination effectiveness. Small arrays of the partially self-compatible plant Californian poppy (Eschscholzia californica) were introduced across a landscape gradient to simulate rare, spatially isolated populations. The effects on pollinator activity, outcrossing, and plant reproduction were measured. In florally rich habitats, we found reduced pollen movement between plants, leading to fewer long-distance pollination events, lower plant outcrossing, and a higher incidence of pollen limitation. This pattern indicates a potential reduction in per capita pollinator visitation, as suggested by the lower activity densities and richness of pollinators observed within florally rich habitats. In addition, seed production reduced by a factor of 1.8 in plants within florally rich habitats and progeny germination reduced by a factor of 1.2. We show this to be a consequence of self-fertilization within the partially self-compatible plant, E. californica. These findings indicate that locally rare plants are at a competitive disadvantage within florally rich habitats because neighboring plant species disrupt conspecific mating by co-opting pollinators. Ultimately, this Allee effect may play an important role in determining the long-term persistence of rarer plants in the landscape, both in terms of seed production and viability. Community context therefore requires consideration when designing and implementing conservation management for plants which are comparatively rare in the landscape.

Corridors restore animal-mediated pollination in fragmented tropical forest landscapes.

Tropical biodiversity and associated ecosystem functions have become heavily eroded through habitat loss. Animal-mediated pollination is required in more than 94% of higher tropical plant species and 75% of the world's leading food crops, but it remains unclear if corridors avert deforestation-driven pollination breakdown in fragmented tropical landscapes. Here, we used manipulative resource experiments and field observations to show that corridors functionally connect neotropical forest fragments for forest-associated hummingbirds and increase pollen transfer. Further, corridors boosted forest-associated pollinator availability in fragments by 14.3 times compared with unconnected equivalents, increasing overall pollination success. Plants in patches without corridors showed pollination rates equal to bagged control flowers, indicating pollination failure in isolated fragments. This indicates, for the first time, that corridors benefit tropical forest ecosystems beyond boosting local species richness, by functionally connecting mutualistic network partners. We conclude that small-scale adjustments to landscape configuration safeguard native pollinators and associated pollination services in tropical forest landscapes.

A direct assessment of realized seed and pollen flow within and between two isolated populations of the food-deceptive orchid Orchis mascula.

Gene flow can counteract the loss of genetic diversity caused by genetic drift in small populations. For this reason, clearly understanding gene flow patterns is of the highest importance across fragmented landscapes. However, gene flow patterns are not only dependent upon the degree of spatial isolation of fragmented populations, but are also dependent upon the life-history traits of the species. Indeed, habitat fragmentation effects appear especially unpredictable for food-deceptive orchid species, because of their highly specialised seed and pollen dispersal mechanisms. In this study we used amplified fragment length polymorphism markers and subsequent parentage and spatial autocorrelation analysis to quantify the extent and the patterns of realized gene flow within and between two adjacent fragmented populations of the food-deceptive Orchis mascula. We observed considerable gene flow between both populations, occurring mainly through pollen dispersal. Seed dispersal, on the other hand, was mainly limited to the first few meters from the mother plant in both populations, although at least one among-population seed dispersal event was observed. This, in turn, resulted in a significant spatial genetic structure for both populations. Although genetic diversity was high in both populations and mainly outcrossing occurred, reproductive output was strongly skewed toward a limited number of successful adult plants. These observed patterns are likely due to the different pollinator behaviour associated with food-deceptive plants. We conclude that these populations can be considered viable under their current fragmented state.

Pollination by sexual deception promotes outcrossing and mate diversity in self-compatible clonal orchids.

The majority of flowering plants rely on animals as pollen vectors. Thus, plant mating systems and pollen dispersal are strongly influenced by pollinator behaviour. In Australian sexually deceptive orchids pollinated by male thynnine wasps, outcrossing and extensive pollen flow is predicted due to floral deception, which minimizes multiple flower visitations within patches, and the movement of pollinators under mate-search rather than foraging behaviours. This hypothesis was tested using microsatellite markers to reconstruct and infer paternity in two clonal, self-compatible orchids. Offspring from naturally pollinated Chiloglottis valida and C. aff. jeanesii were acquired through symbiotic culture of seeds collected over three seasons. In both species, outcrossing was extensive (tm  = 0.924-1.00) despite clone sizes up to 11 m wide. The median pollen flow distance based on paternity for both taxa combined was 14.5 m (n = 18, range 0-69 m), being larger than typically found by paternity analyses in other herbaceous plants. Unexpectedly for orchids, some capsules were sired by more than one father, with an average of 1.35 pollen donors per fruit. This is the first genetic confirmation of polyandry in orchid capsules. Further, we report a possible link between multiple paternity and increased seed fitness. Together, these results demonstrate that deceptive pollination by mate-searching wasps enhances offspring fitness by promoting both outcrossing and within-fruit paternal diversity.

Mating system as a barrier to gene flow.

Understanding mating system as one of reproductive isolating barriers remains important although this barrier is classified in a different sense from behavioral, ecological, and mechanical isolating barriers. Selfing enhances incipient speciation while outcrossing facilitates species integrity. Here, I study how mating system affects gene exchanges between genetically diverging species in a hybrid zone. Results show that a predominant selfing species has a greater barrier to selective gene flow than does a predominant outcrossing species. Barrier to neutral gene flow convexly changes with the selfing rate due to linkage disequilibrium, with a maximum at around intermediate selfing rate. Asymmetric transient or steady-state barriers to neutral gene flow occur between two sides of a hybrid zone when the neutral gene is affected by its linked selective gene whose alternative alleles are adaptive to heterogeneous habitats. Selfing interacts with both a physical barrier and a density-dependent ecological regulation (a logarithmic model) to strengthen the barriers to neutral and selective gene flow. This theory helps to interpret incipient speciation driven by selfing or to explain the asymmetric gene flow or unequal genomic mixtures between closely related species caused by their asymmetric mating systems in natural hybrid zones.

Pollen flow of wheat under natural conditions in the Huanghuai River Wheat Region, China.

The transgenic pollen spread is the main pathway of transgenic plant gene flow. The maximum distance of pollen dispersal (horizontal), the spatial dynamics of pollen movement (vertical), and the patterns of pollen dispersal are important considerations in biosafety assessments of genetically modified crops. To evaluate wheat (Triticum aestivum) pollen dispersal, we measured the pollen suspension velocity and analyzed pollen dispersal patterns under natural conditions in the Huanghuai River wheat-growing region in 2009. The pollen suspension velocity was 0.3-0.4 m/s. The highest pollen densities were detected in the north, northwest, and south of the pollen source. Pollen was dispersed over distances greater than 245 m in the northwest and northeast directions. At the pollen source center, pollen density decreased with increasing vertical height. In the north of the pollen source, the pollen density from 65 m to 225 m showed a wave-mode decrease with increasing height. The horizontal transport of pollen over longer distances fitted polynomial equations. In the north, the pollen density was highest at 45 m from the pollen source, and decreased with increasing distance. In the northwest, the pollen density showed a double-peak trend. In the northeast, pollen density was highest from 45 m to 125 m from the source. Wind speeds greater than the pollen suspension velocity and the duration of continuous gusts were the main factors affecting pollen dispersal. This information will be useful for determining the spatial isolation distances for hybrid seed production and for the commercial production of transgenic wheat.

orchard: Paternity program for autotetraploid species.

Advances in molecular marker technology have provided new opportunities to study the population genetics of polyploid taxa. Paternity analysis using microsatellite markers can be used in detection of gene flow between individuals and populations, in mating system analysis, to identify factors that influence fecundity and fertility, to identify behaviour of parent-offspring relationships and in the analysis of the reproductive success of different ecological groups. As there is no specific program for carrying out paternity analysis in tetraploid species, specialized software was designed for the assignment of paternity for autotetraploid species. orchard is a novel implementation of exclusion and likelihood statistics for carrying out paternity analysis of autotetraploids. First, the program performs an exclusion method, and then, a likelihood statistic is used with nonexcluded candidate fathers. Optional features include estimation of allele dosage of known mother trees and the estimation of pollen flow distances. orchard was tested using a data set of microsatellite data of Dipteryx odorata, a tetraploid Amazonian tree species.