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Limited pollen dispersal and biparental inbreeding in Symphonia globulifera in French Guiana.

In this paper, we report a study of the mating system and gene flow of Symphonia globulifera, a hermaphroditic, mainly bird-pollinated tree species with a large geographic distribution in the tropical Americas and Africa. Using three microsatellites, we analysed 534 seeds of 28 open pollinated families and 164 adults at the experimental site 'Paracou' in French Guiana. We observed, compared to other tropical tree species, relatively high values for the effective number of alleles. Significant spatial genetic structure was detected, with trees at distances up to 150 m more genetically similar than expected at random. We estimated parameters of the mating system and gene flow by using the mixed mating model and the TwoGener approach. The estimated multilocus outcrossing rate, tm, was 0.920. A significant level of biparental inbreeding and a high proportion of full-sibs were estimated for the 28 seed arrays. We estimated mean pollen dispersal distances between 27 and 53 m according to the dispersal models used. Although the adult population density of S. globulifera in Paracou was relatively high, the joint estimation of pollen dispersal and density of reproductive trees gave effective density estimates of 1.6 and 1.3 trees/ha. The parameters of the mating system and gene flow are discussed in the context of spatial genetic and demographic structures, flowering phenology and pollinator composition and behaviour.

French Guiana↗

To self, or not to self... a review of outcrossing and pollen-mediated gene flow in neotropical trees.

Despite the typically low population densities and animal-mediated pollination of tropical forest trees, outcrossing and long-distance pollen dispersal are the norm. We reviewed the genetic literature on mating systems and pollen dispersal for neotropical trees to identify the ecological and phylogenetic correlates. The 36 studies surveyed found >90% outcrossed mating for 45 hermaphroditic or monoecious species. Self-fertilization rates varied inversely with population density and showed phylogenetic and geographic trends. The few direct measures of pollen flow (N=11 studies) suggest that pollen dispersal is widespread among low-density tropical trees, ranging from a mean of 200 m to over 19 km for species pollinated by small insects or bats. Future research needs to examine (1) the effect of inbreeding depression on observed outcrossing rates, (2) pollen dispersal in a wide range of pollination syndromes and ecological classes, (3) and the range of variation of mating system expression at different hierarchical levels, including individual, seasonal, population, ecological, landscape and range wide.

Genetic Markers↗

Seasonal atopic dermatitis in dogs sensitive to a major allergen of Japanese cedar (Cryptomeria japonica) pollen.

Three dogs were examined because of episodes of recurrent pruritic dermatitis in the spring, the season of Japanese cedar (Cryptomeria japonica, CJ) pollination in Japan. The dogs were shown to be sensitive to CJ pollen allergen using intradermal testing and antigen-specific IgE measurement. Fluorometric enzyme-linked immunosorbant assay (ELISA) showed increased concentrations of IgE specific to Cry j 1 and a negative result for Cry j 2 in the three dogs. The concentrations of IgE specific to Cry j 1 during the season of CJ pollination were higher than the concentrations found during the off-season in all the dogs, and the variation in the concentrations correlated with the variation in clinical signs. Peripheral blood mononuclear cells showed apparent proliferative responses to crude CJ pollen antigen and Cry j 1 during CJ pollination season. These findings indicated that Cry j 1 was the major allergen recognized by IgE and lymphocytes and resulted in the development of type I hypersensitivity to CJ pollen allergen in these atopic dogs.

Allergens↗

Quantification of group 5 grass pollen allergens in house dust.

BACKGROUND: It is widely known and accepted that grass pollen is a major outdoor cause of hay fever. However, it is of virtual importance for grass pollen allergic patients with symptoms all the year round to know the concentration of grass pollen allergens in their homes. OBJECTIVE: The main objective of this study was to quantify the amount of grass pollen allergen in mass units (microg Phl p 5) in dust settled indoors and to detect the distribution of allergenic activity in different sampling locations of homes. Furthermore, we studied the seasonal fluctuation of allergen content in dust samples. METHODS: We adapted the two site binding assay for detection of group 5 grass pollen allergens in samples from randomly selected homes in Hamburg (n = 371), Erfurt (n = 396), Hettstedt (n = 353), Zerbst (n = 289) and Bitterfeld (n = 226), Germany. Dust samples were collected from floor of living room (LR), bedroom (BR) or children's room (CR) and mattress (MA) during period of June 1995 to August 1998. The amount of the major grass group 5 allergens was detected in microg/g dust. RESULTS: Phl p 5 was detected in 67% of the samples analysed (n = 4760). The range was between undetectable (< 0.03 microg/g dust) and 81 microg/g dust. Phl p 5 levels were significantly higher in the dust from LR (geometric mean 0.117 microg/g dust) or BR/CR floors (geometric mean 0.098 microg/g dust) than in mattresses (geometric mean 0.043 microg/g dust). We observed seasonal fluctuation of indoor Phl p 5 levels with peak in June but also annual differences. Thus Phl p 5 content indoors reflects also the different quantities of pollen counts of annual courses. During pollination period we found two times higher Phl p 5 levels (0.172 microg/g dust, P < 0.001) than outside of grass pollination season (0.095 microg/g dust). The indoor Phl p 5 levels outside of season seem to be independent of pollination before. We suppose that settled pollen grains or allergenic material from outdoor particles carried indoors via footwear and clothes accumulates in house dust. CONCLUSION: Although we not known how the allergens in settled dust are equilibrated with those in the air, the considerable high level of Phl p 5 in indoor dust even during periods when no grass pollen is present in the atmosphere may be an important cause of pollen-allergy symptoms outside of season.

Air Pollution, Indoor↗

The effect of ecological factors on the mating system of a South American shrub species (Helicteres brevispira).

Mating systems are influenced by several ecological factors, including plant density, number of flowers per plant, and pollinator movements. In this study, we investigated the simultaneous effects of these three factors on the mating system of a self-compatible Brazilian shrub species: Helicteres brevispira St. Hil. Outcrossing rate is directly correlated with plant density. Changes in the number of flowers per plant affect outcrossing rate through their effect on the density of flowers. Variation in foraging behaviour of hummingbird pollinators is a consequence of the interaction between plant density and number of flowers per plant. Territorialist pollinators are common in high density areas but visit few flowers on each plant, thereby promoting outcrossing. In areas of low plant density, trapliners and rare territorialists visit several flowers per plant, thus increasing selfing. Our results indicate that outcrossing rate is a dynamic parameter, with the extent of variation depending on a number of ecological factors. In successional species such as those in the genus Helicteres, demographic changes may be accompanied by alterations in mating system parameters, with concomitant effects on the genetic structure of populations.

Crosses, Genetic↗

High resolution microsatellite based analysis of the mating system allows the detection of significant biparental inbreeding in Caryocar brasiliense, an endangered tropical tree species.

In this work we investigate the mating system of four populations of the endangered tropical tree species Caryocar brasiliense, using genetic data from 10 microsatellite loci. Eight to 10 open-pollinated progeny arrays of 16 individuals, together with their mother tree, were sampled per population. Mating system parameters were estimated under the mixed mating model, implemented by the software MLTR. The single-locus outcrossing rate (ts) varied among loci and populations, but multilocus outcrossing rates (tm) were equal to one for all four populations. Nevertheless, biparental inbreeding (tm - ts) was different from zero for all populations, indicating that outcrossing events may occur between relatives. Our results also indicate that the high polymorphism of microsatellite markers provide an extraordinary resolution to discriminate precisely selfing events from outcrossing events between close relatives. Our results indicate that, although highly outcrossed, C. brasiliense shows high levels of biparental inbreeding, most likely due to the limited flight range of pollinators and restriction in seed dispersal. Furthermore, these results suggest that Cerrado fragmentation could limit gene flow by isolating seed dispersers and territorial small sized bat pollinators inside fragments, increasing the rate of mating between close relatives. The conservation of nonisolated populations in large preserved areas may be necessary to foster outcrossing events between unrelated individuals and thus maintain species viability.

Crosses, Genetic↗

Field and allozyme studies investigating optimal mating success in two sympatric spring-ephemeral plants, Trillium erectum and T. grandiflorum.

A combination of field experiments and allozyme studies was used to test whether two spring-ephemeral plants growing in eastern North America, Trillium erectum and T. grandiflorum (Liliaceae), exhibited an optimal outcrossing distance for fruit and seed production. Furthermore, the spatial genetic structure of the Trillium populations was examined in light of the outcrossing results. In field experiments, recipient plants were pollinated with either self pollen or with pollen from donors growing 1 m, 10 m, 100 m, and 1500 m away. These distances represented crosses between individuals growing within the same population (1 m, 10 m, and 100 m crosses) and between individuals growing in different populations (1500 m crosses). Self-pollinated T. erectum and T. grandiflorum produced 71% and 89% fewer seeds, respectively, than all other outcrossing treatments. However, there were no significant differences among outcrossing treatments for fruit or seed production. As neither T. erectum nor T. grandiflorum exhibited an optimal outcrossing distance for fruit or seed production, it was predicted that populations of the two would not demonstrate strong spatial genetic structure or isolation by distance. The allozyme results only partially supported the patterns revealed in the outcrossing treatments. Populations of T. erectum and T. grandiflorum showed moderate spatial genetic differentiation based on F-statistics, and only T. grandiflorum exhibited significant isolation by distance based on spatial autocorrelation analyses. The lack of optimal outcrossing distances and the patterns of allozyme variation in T. erectum and T. grandiflorum populations may be attributed to a number of factors, including active seed dispersal by ants, rare long-distance gene-flow events, post-pollination and post-fertilization selection, and/or the severity of inbreeding depression.

Isoenzymes↗

'Floral' scent production by Puccinia rust fungi that mimic flowers.

Crucifers (Brassicaceae) in 11 genera are often infected by rust fungi in the Puccinia monoica complex. Infection causes a 'pseudoflower' to form that is important for attracting insect visitors that sexually outcross the fungus. 'Pollinator' attraction is accomplished through visual floral mimicry, the presence of a nectar reward and floral fragrances. Here we used gas chromatography and mass spectrometry to identify and quantify fragrance production by these rust fungi on several Arabis hosts, and by co-occurring true flowers that share insect visitors. Fungal pseudoflowers produced distinctive floral fragrances composed primarily of aromatic alcohols, aldehydes and esters. Pseudoflower fragrances were chemically similar to noctuid-moth-pollinated flowers, such as Cestrum nocturnum and Abelia grandiflora, but were very different from host flowers, host vegetation and the flowers of coblooming, nonhost angiosperms. There was variation in the quantity and composition of fragrance profiles from different fungal species as well as within and among hosts. The evolution of scent chemistry is relatively conservative in these fungi and can be most parsimoniously explained in three steps by combining chemical data with a previously determined rDNA ITS sequence-based phylogeny. Pseudoflower scent does not appear to represent a simple modification of host floral or vegetative emissions, nor does it mimic the scent of coblooming flowers. Instead, we suspect that the unique fragrances, beyond their function as pollinator attractants, may be important in reducing gamete loss by reinforcing constancy among foraging insects.

Basidiomycota↗

Reproductive biology and genetic diversity of a cryptoviviparous mangrove aegiceras corniculatum (Myrsinaceae) using allozyme and intersimple sequence repeat (ISSR) analysis

Mangroves consist of a group of taxonomically diverse species representing about 20 families of angiosperms. However, little is known about their reproductive biology, genetic structure, and the ecological and genetic factors affecting this structure. Comparative studies of various mangrove species are needed to fill such gaps in our knowledge. The pollination biology, outcrossing rate, and genetic diversity of Aegiceras corniculatum were investigated in this study. Pollination experiments suggested that the species is predominantly pollinator-dependent in fruit setting. A quantitative analysis of the mating system was performed using progeny arrays assayed for intersimple sequence repeat (ISSR) markers. The multilocus outcrossing rate (tm) was estimated to be 0.653 in a wild population. Both allozyme and ISSR were used to investigate genetic variation within and among populations. The combined effects of founder events and enhanced local gene flow through seedling dispersal by ocean currents apparently played an important role in shaping the population genetic structure in this mangrove species. Both allozyme variation (P = 4.76%, A = 1.05, HE = 0.024) and ISSR diversity (P = 16.18%, A = 1.061, HE = 0.039) were very low at the species level, in comparison with other woody plants with mixed-mating or outcrossing systems. Gene differentiation among populations was also low: allozyme GST = 0.106 and ISSR GST = 0.178. The unusually high genetic identities (0.997 for allozyme and 0.992 for ISSR loci), however, suggest that these populations are probably all descended from a common ancestral population with low polymorphism.

Journal Article↗

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

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

Alleles↗

Foundress re-emergence and fig permeability in fig tree-wasp mutualisms.

Some female pollinating fig wasps (foundresses) re-emerge from figs after oviposition/pollination. We investigated why this occurs in the mutualism between the gynodioecious Ficus montana and Liporrhopalum tentacularis. Re-emergence increased with foundress density in figs and some foundresses oviposited in two male figs, indicating that they re-emerge because of oviposition site limitation. Re-emergence was independent of fig diameter, indicating that permeability is not because of fig age at entry. Rather, as some foundresses also pollinate two female figs we suggest permeability is selected for because it increases pollinator production and/or efficiency (although, potentially opposing these hypotheses, we also found between-tree differences in permeability in male figs). In addition, we show that re-emergence is much more common than previously suspected, and more common from gynodioecious than monoecious fig species. We argue that our findings in F. montana could explain this pattern of incidence.

Animals↗

Reproductive biology of the epiphytic bromeliad Werauhia gladioliflora in a premontane tropical forest.

The floral phenology, fruit and seed production, and self-compatibility of Werauhia gladioliflora, an epiphytic bromeliad with a wide distribution, were studied in a premontane forest in the Monteverde area in Costa Rica. The species presents the pollination syndrome of chiropterophily, and it is visited by the small bats Hylonycteris underwoodi and Glossophaga commissarisi (Glossophaginae). The population flowering period extended from October to early December (end of rainy season) and seed dispersal occurred from February to April (dry season). Most plants opened a single flower per night, either every day or at one-day intervals during the flowering period. In natural conditions, the average fruit set amounted to almost half of the potential output, but individual fecundity (number of seeds) remained high. Seed number per fruit and germination capacity after artificial selfing and out-crossing treatments did not differ from natural pollination conditions. Werauhia gladioliflora exhibited high levels of autonomous self-pollination and self-compatibility at the individual and population level, characters associated with the epiphytic habitat. These reproductive traits are also associated with early colonizer species, yet life history traits, such as seed dispersal, seedling establishment success, and growth, are likely to have a major role in determining the presence of this species in the successional vegetation patches scattered over the studied premontane area.

Bromeliaceae↗

Nectar resorption and translocation in Cucurbita pepo L. and Platanthera chlorantha Custer (Rchb.).

Nectar resorption and sugar translocation were studied in Cucurbita pepo (Cucurbitaceae) and Platanthera chlorantha (Orchidaceae) by micro-autoradiography. In both species, nectar was resorbed in pollinated and unpollinated flowers and ovules developing into seeds were found to be the main sugar sink. In C. pepo, the mobility of resorbed sugars in pollinated female flowers was higher than in unpollinated ones; male flowers showed lower mobility of resorbed sugar. In P. chlorantha, radioactivity was detected in pollinated flowers below and above labelled unpollinated ones: the nearer the flower, the stronger the accumulation of label in developing fruits.

Autoradiography↗

Critical review of host specificity and its coevolutionary implications in the fig/fig-wasp mutualism.

Figs (Ficus spp., Moraceae) and their pollinating wasps (Agaonidae, Chalcidoidea) constitute perhaps the most tightly integrated pollination mutualism that is known. Figs are characterized by extraordinarily high global and local species diversity. It has been proposed that the diversification of this mutualism has occurred through strict-sense coadaptation and cospeciation between pairs of fig and wasp species that are associated in highly specific one-to-one relationships. However, existing studies cast doubt on the generality of this proposition. Here, we review our current knowledge of the evolutionary history of the fig/fig-wasp mutualism. We critically examine the idea that codivergence between figs and their pollinators has been dominated by strict-sense cospeciation. We present phylogenetic and population genetic data from neotropical fig and fig wasp species that suggest that a more accurate model for diversification in this mutualism is that of groups of genetically well defined wasp species coevolving with genetically less well defined (frequently hybridizing) groups of figs. Last, we use our results to assess previously proposed hypotheses on models of speciation in this mutualism.

Animals↗

Expression and parent-of-origin effects for FIS2, MEA, and FIE in the endosperm and embryo of developing Arabidopsis seeds.

The promoters of MEA (FIS1), FIS2, and FIE (FIS3), genes that repress seed development in the absence of pollination, were fused to beta-glucuronidase (GUS) to study their activity pattern. The FIS2GUS product is found in the embryo sac, in each of the polar cell nuclei, and in the central cell nucleus. After pollination, the maternally derived FIS2GUS protein occurs in the nuclei of the cenocytic endosperm. Before cellularization of the endosperm, activity is terminated in the micropylar and central nuclei of the endosperm and subsequently in the nuclei of the chalazal cyst. MEAGUS has a pattern of activity similar to that of FIS2GUS, but FIEGUS protein is found in many tissues, including the prepollination embryo sac, and in embryo and endosperm postpollination. The similarity in mutant phenotypes; the activity of FIE, MEA, and FIS2 in the same cells in the embryo sac; and the fact that MEA and FIE proteins interact in a yeast two-hybrid system suggest that these proteins operate in the same system of control of seed development. Maternal and not paternal FIS2GUS, MEAGUS, and FIEGUS show activity in early endosperm, so these genes may be imprinted. When fis2, mea, and fie mutants are pollinated, seed development is arrested at the heart embryo stage. The seed arrest of mea and fis2 is avoided when they are fertilized by a low methylation parent. The wild-type alleles of MEA or FIS2 are not required. The parent-of-origin-determined differential activity of MEA, FIS2, and FIE is not dependent on DNA methylation, but methylation does control some gene(s) that have key roles in seed development.

Arabidopsis↗

Bumblebee preference for symmetrical flowers.

Fluctuating asymmetry, which represents small random deviations from otherwise bilateral symmetry, is a measure of the phenotypic quality of individuals indicating the ability of controlled development under given environmental and genetic conditions. I tested whether floral symmetry reliably reflects phenotypic quality measured in terms of pollinator rewards and whether pollinators respond to floral symmetry in a series of observations and experiments on Epilobium angustifolium (Onagraceae). Lower petal asymmetry was negatively related to mean lower petal length, whereas asymmetry in leaf width was positively related to mean leaf width. Flowers visited by bumblebees were larger and more symmetrical than the nearest neighboring flower. This relationship between pollinator preference for large and symmetrical flowers was demonstrated to be causal in experiments in which the lower petals were manipulated symmetrically or asymmetrically. Nectar production was larger in symmetrical flowers, and this may explain the bumblebee preference for flower symmetry. Floral symmetry therefore reliably reflects nectar production and hence enhances pollen transport. Extensive embryo abortion has been documented in E. angustifolium and other outcrossing plant species. Floral fluctuating asymmetry, which reflects general developmental homeostasis, may explain such developmental selection in these plants.

Journal Article↗

Developmental selection within the angiosperm style: using gamete DNA to visualize interspecific pollen competition.

Among flowering plants, females often have little control over the genetic relatedness of pollen deposited on stigmas. Thus, postpollination processes are of primary importance for mate discrimination. The ability to screen and select among male gametes during pollen tube growth within the female tissues of the stigma, style, and ovary is critical to the process of mate choice and reproductive isolation. However, direct evidence of the mechanistic/developmental processes associated with mate choice in flowering plants is sparse. We studied the contribution of postpollination, prefertilization processes to reproductive isolation in two wind-pollinated species of birch (Betula) that commonly hybridize in nature. We exploited the 3-fold difference in ploidy level between these two species to determine the paternity of individual pollen tubes growing within female reproductive tissues. We then tracked their developmental fate in conspecific, heterospecific, and mixed-species crosses. This design allowed a direct comparison of developmental processes responsible for reproductive isolation in both single-species and mixed-species pollinations. Our results have important implications for the population genetic outcome of hybridization dynamics in natural populations. Paternity analysis of progeny from mixed-species pollinations revealed that conspecific pollen tubes sired more than 98% of seedlings. Biased siring success was not the result of differential embryo abortion. We detected strong, early postpollination barriers such as pollen tube incompatibility, slower pollen tube growth, and delayed generative cell mitosis. Conspecific fertilization precedence was mediated by favorable or unfavorable male-female interactions, but there was no evidence for antagonistic male-male interactions.

Crosses, Genetic↗

Patterns of endangerment in the hawaiian flora.

The Hawaiian flora, because of its great isolation, high levels of endemism, known lineages, and high rates of endangerment, offers unique opportunities to explore patterns of endangerment related to phylogeny, ecological and life history traits, and geographic patterns. Nine percent of the native flora of 1159 taxa are already extinct, and 52.5% are at risk (extinct, endangered, vulnerable, or rare). Risk is strongly associated with limited geographic distribution at several scales: endemic taxa (native only to the Hawaiian Islands) are at far greater risk than indigenous taxa (with both Hawaiian and extra-Hawaiian ranges); single-island endemics are more at risk than multi-island endemics; small islands have the highest proportion of endemic taxa at risk; and endemics with more limited habitat distributions (elevation, community type) are more at risk. Historic population density is a strong predictor of risk, and taxa with low historic population densities are at greatest risk with rapid anthropogenic changes. Among the major islands, Maui Nui has the highest percent of taxa that are extinct. Kaua'i has the lowest percent of extinct taxa and the highest proportion of single-island endemic taxa that are rare. Endemic taxa at risk are associated with distributions in shrublands, forests, bogs, and cliff habitats. Endemic taxa with distributions in low elevation dry habitats have the highest proportion of taxa at risk, but the greatest absolute numbers of taxa at risk have distributions in mesic lowland and montane forests, and in wet montane forests. The life history patterns associated with risk are complicated, and inclusion of the effects of evolutionary relationships (lineages) changes some of these patterns. Species level analyses without respect to lineage shows risk associated with monomorphic (hermaphroditic) breeding systems and bird pollination because of the large number of hermaphroditic, bird-pollinated species in the Campanulaceae. Analyses incorporating the effect of lineage greatly reduce the impact of large lineages and result in an association of risk with insect pollination, and no effect of breeding system. There is no association of lineage size and the percent of taxa at risk within the lineage; endemic taxa from lineages with large radiations are at no greater risk than endemic single-taxon lineages. The percentages of taxa at risk at the family level in the Hawaiian Islands and worldwide (excluding Hawaiian taxa) are positively correlated, although flowering plant families in the Hawaiian Islands have a much greater proportion of taxa at risk. Some of the approaches described here may be useful to predict geographical and biological patterns of endangerment in island and island-like ecosystems under increasing pressures of endangerment and extinction.

Cell Lineage↗