Long-distance pollinator flights and pollen dispersal between populations of Delphinium nuttallianum

Spatial processes in pollination biology are poorly understood, especially at levels above that of the local population. For example, little is known about how pollinators and pollen move among populations, although there is evidence that such movement can exceed what is predicted from intrapopulational movement. We explored pollination success in experimental isolates of the bumblebee- and hummingbird-pollinated wildflower Delphinium nuttallianum. We established a total of 15 arrays of potted plants isolated by 50-400 m from ten natural "source" populations, as well as control arrays embedded within each source. Flowers on potted plants were emasculated, so any pollen received could be assumed to come from source populations. A total of 69 h of observation suggested that pollinators were somewhat less abundant in isolates than in controls, but visited more plants and flowers once within an isolate. Consistent with this, 82.1% of all flowers in isolated arrays received pollen, versus 87.7% in controls. Mean receipt was more than 100 pollen grains per flower in most arrays, and seed set in isolates and controls respectively averaged 69.8% and 74.3% of ovules. Furthermore, pollen receipt in isolates declined relatively slowly with distance from the source. We conclude that pollinators of D. nuttallianum often will fly up to 400 m among populations, and that substantial pollination ensues. Thus isolated populations of this species often belong to metapopulations in terms of pollen dispersal, with important consequences for genetic differentiation, and potential implications for the management of endangered plant species.     

Acid-Fastness of pine pollen

This is a study of the acid-fast staining characteristics of pine pollen and an investigation of factors causing loss of acid-fastness after pine pollen has been in contact with tisues or mucous membranes. Intact loblolly pine pollen was readily stained with cold carbol-fuchsin, and retained its acid-fastness after decolorization with 3% HC1 in 95% ethyl alcohol for 2 min, followed by methylene blue counterstain. Pine pollen resembles spermatozoa in ease of staining and resistance to decolorization. Acid-fastness was destroyed by crushing or by germination of the pollen grain, and by contact for several hours with serum or saline solutions, but was unchanged by exposure to 0.1% solution of streptomycin in water. Nonviable pine pollen did not lose acid-fastness after suspension for several days in serum or water. When counterstain was omitted, crushed or germinated pollen appeared red to pink after staining with carbol-fuchsin and decolorization with acid alcohol, thus indicating that lipids of an acid-fast nature were still present. The large size and biologic properties of pine pollen provide a unique means of studying the chemical and physical aspects of the acid-fast phenomenon.     

Long-distance dispersal of tree seeds by wind

Some mechanisms that promote long-distance dispersal of tree seeds by wind are explored. Winged seeds must be lifted above the canopy by updrafts to have a chance of further dispersal in high velocity horizontal winds aloft or in landscape-scale convection cells. Shear-induced turbulent eddies of a scale up to one-third of canopy height provide a lifting mechanism. Preliminary data suggest that all seeds of a given species may be viable candidates for uplift and long-distance dispersal, despite the evidence that slow-falling seeds are dispersed farther under any given wind conditions. Turbulence is argued more often and more extensively to advance long-distance dispersal than to retard it. Seeds may take advantage of Bernoulli sailing to move with faster than average winds. Elasticity of branches and trees may play a role in regulating the release of seeds into unusually favorable winds. Dispersal is at least biphasic, and the study of long-distance dispersal calls for mixed models and mixed methods of gathering data.     

Long-distance dispersal: a framework for hypothesis testing

Tests of hypotheses about the biogeographical consequences of long-distance dispersal have long eluded biologists, largely because of the rarity and presumed unpredictability of such events. Here, we examine data for terrestrial (including littoral) organisms in the Pacific to show that knowledge of dispersal by wind, birds and oceanic drift or rafting, coupled with information about the natural environment and biology of the organisms, can be used to generate broad biogeographic predictions. We then examine the predictions in the context of the origin, frequency of arrival and location of establishment of dispersed organisms, as well as subsequent patterns of endemism and diversification on remote islands. The predicted patterns are being increasingly supported by phylogenetic data for both terrestrial and littoral organisms.     

Heat-shock response in germinating pine pollen

Summary The in vitro culture of pine pollen at various temperatures reveals only a moderate degree of thermotolerance, with considerably reduced levels of growth at and above 35° C. Unlike the pollen of many previously studied species, pine pollen shows some ability to recover from short periods of growth at temperatures as high as 40° C, especially when such exposures occur during the early stages of pollen germination. The pollen of Pinus taeda, unlike that of most other species, shows both quantitative and qualitative changes in the proteins synthesized during germination in vitro following a switch to elevated temperatures (37° C). This response, which can be elicited both during the very early stages of germination as well as during the later stages of pollen tube growth, is reversible following a shift back to the lower temperatures. As previously shown with vegetative tissue of other plant species, the heat-shock response not only involves the induction of high-molecular-weight proteins (most notably 82 kDa and 70 kDa proteins), but also a number of low-molecular-weight (10–20 kDa) species. Two-dimensional gel electrophoretic analysis reveals a small number of qualitative differences in the types of low-molecular-weight heat-shock proteins synthesized in pollen versus vegetative tissue.     

Long-distance transport ofBetula pollen grains and allergic symptoms

Summary An example of the potential importance of air masses as carriers of aero-allergens from distant source areas is provided. Considerable amount ofBetula pollen is relatively often transported to Fennoscandia before the local birch flowering period, mainly by southeastern air masses from eastern part of central Europe. Although the distance and the transport time in some cases can be extensive, the pollen grains seem to cause allergic reactions among sensitive persons.A comparison between the clinical results andBetula pollen counts from the time before the local flowering season in Stockholm in 1989 is presented.     

Air pollen dispersal in a tropical area

Volumetric data on airborne pollen have been gathered for two consecutive years at a neotropical location (Caracas). Among the 65 taxa which were identified, pollen from aCupressus species (introduced) and from aCecropia species (indigenous) were dominant. Less numerous but also abundant (daily averages ≥5 grains/m³ air) were pollen from Gramineae, Urticaceae,Alcalypha, Pinus, Piperaceae andMimosa. Pollen grains were recorded daily throughout the year. They increased in numbers during April–May and again during November–December. The first peak was contributed mainly by indigenous species, the second peak mainly by introduced species.     

Long-distance dispersal of seeds in the fire-tolerant shrub Banksia attenuata

Long-distance dispersal (LDD) of seeds enables alleles, individuals and species of plants to (re)colonize suitable but remote habitats. Banksia attenuata is a long-lived resprouting shrub restricted to dune crests in fire-prone sclerophyll shrublands of the Eneabba sandplain, southwestern Australia. Highly polymorphic microsatellite DNA genetic markers and population assignment tests were employed to identify LDD immigrants among 788 individuals from 27 stands of B. attenuata comprising a metapopulation. Of the 487 (61.8% of the total) individuals unambiguously assigned to a unique source population, 27 (5.5%) were identified as immigrants by assignment to a known population other than that from which they were sampled, while the remaining 460 were assigned to the population from which they were sampled. The distance between source and sink populations for these immigrants ranged from 0.2 to 2.6  km, averaging 1.4  km, and broadly trending in the direction of seasonal winds. These results suggest that B. attenuata has similar long-distance seed dispersal properties as its co-occurring shorter-lived and fire-sensitive congener, B. hookeriana , despite fewer, larger and less mobile seeds. The frequency and distance of LDD for seeds observed in both species (5.5–6.8%) helps explain the persistence of populations on these geographically isolated dunes, where they are subject to local extinction from recurrent fire and severe summer drought, and will remain important under predicted climate change conditions. Analysis also revealed that species richness of the functional group to which B. attenuata belongs was positively correlated with the number of immigrants identified per dune, and such correlation was likely driven by environmental properties of the dunes, particularly water availability.     

Marking live conifer pollen for long-distance dispersal experiments

Long-distance dispersal (LDD) theory requires a method for marking live LDD pollen. Such a method must complement more intensive sampling methods inclusive of molecular cytogenetics, proteomics and genomics. We developed a new method for marking live Pinus taeda pollen using two dyes, rhodamine 123 and aniline blue, dissolved in a sucrose solution. Marked and unmarked pollen were compared with respect to in vitro germination, storage, terminal velocity, and in vivo pollen tube penetration of ovules. We found that: (1) both types of marked pollen retained their capacity for germination, (2) both types of marked pollen had similar aerodynamic properties when compared to unmarked pollen controls, (3) marked pollen retained its germination capacity for 48 h, and (4) of the marked pollen, only the aniline-marked pollen penetrated ovules during pollination. Germination declined rapidly for both types of marked pollen after 48 h and before 37 days at -20°C storage, while unmarked pollen lots retained 93% germination at all stages. This method for marking live P. taeda pollen is feasible for tracing LDD pollen only if released and deposited within 48 h of dye treatment.     

Long-distance seed and pollen dispersal inferred from spatial genetic structure in the very low-density rainforest tree, Baillonella toxisperma Pierre, in Central Africa

We analysed the spatial distribution of genetic diversity to infer gene flow for Baillonella toxisperma Pierre (Moabi), a threatened entomophilous pollinated and animal-dispersed Central African tree, with typically low density (5-7 adults trees/km(2)). Fifteen nuclear and three universal chloroplast microsatellites markers were used to type 247 individuals localized in three contiguous areas with differing past logging intensity. These three areas were within a natural forest block of approximately 2886 km(2) in Gabon. Expected heterozygosity and chloroplast diversity were He(nuc) = 0.570 and H(cp) = 0.761, respectively. F(IS) was only significant in one area (F(IS) = 0.076, P < 0.01) and could be attributed to selfing. For nuclear loci, Bayesian clustering did not detect discrete gene pools within and between the three areas and global differentiation (F(STnuc) = 0.007, P > 0.05) was not significant, suggesting that they are one population. At the level of the whole forest, both nuclear and chloroplast markers revealed a weak correlation between genetic relatedness and spatial distance between individuals: Sp(nuc) = 0.003 and Sp(cp) = 0.015, respectively. The extent of gene flow (σ) was partitioned into global gene flow (σ(g)) from 6.6 to 9.9 km, seed dispersal (σ(s)) from 4.0 to 6.3 km and pollen dispersal (σ(p)) from 9.8 to 10.8 km. These uncommonly high dispersal distances indicate that low-density canopy trees in African rainforests could be connected by extensive gene flow, although, given the current threats facing many seed disperser species in Central Africa, this may no longer be the case.     

LAT52 protein is essential for tomato pollen development: pollen expressing antisense LAT52 RNA hydrates and germinates abnormally and cannot achieve fertilization

The LAT52 gene of tomato is expressed in a pollen-specific manner. It is shown that LAT52 encodes a heat-stable, glycosylated protein that traverses the secretory pathway when expressed in a baculovirus expression system. The LAT52 protein shows some similarity with Kunitz trypsin inhibitors and with pollen proteins from maize, rice and olive, but the biological function of these pollen proteins is unknown. To test whether the LAT52 protein plays an important role during pollen development, tomato plants were transformed with an antisense LAT52 gene driven by the LAT52 promoter. Because the LAT52 gene is expressed gametophytically, only 50% of the pollen of the primary transformants would be expected to express the antisense construct. Self-progeny of 19 of the primary transformants showed the predicted 3:1 segregation for a single locus insertion of the linked kanamycin-resistance gene. However, the self-progeny of the other 32 primary transformants showed a 1:1 segregation pattern and could not transmit the linked kanamycin-resistance gene through the male. A subset of these 1:1 segre. gation class plants was examined in detail. The pollen showed lower levels of LAT52 mRNA and LAT52 protein when compared with wild-type. In vitro , approximately 50% of the pollen grains appear to hydrate abnormally; this anomaly is not present when the same pollen grains are Incubated in a medium with higher water potential. In vivo pollination experiments showed that the growth of around 50% of the pollen tubes is arrested in the style. The 3:1 segregation class plants showed no significant differences from untransformed control plants. Taken together, the results show a direct correlation between the reduced expression of LAT52 protein and abnormal pollen function, and suggest that the LAT52 protein plays a role in pollen hydration and/or pollen germination.     

Fluorescent dye particles as pollen analogues for measuring pollen dispersal in an insect-pollinated forest herb

In flowering plants, pollen dispersal is often the major contributing component to gene flow, hence a key parameter in conservation genetics and population biology. A cost-effective method to assess pollen dispersal consists of monitoring the dispersal of fluorescent dyes used as pollen analogues. However, few comparisons between dye dispersal and realized pollen dispersal have been performed to validate the method. We investigated pollen dispersal in two small populations of the insect-pollinated herb Primula elatior from urban forest fragments using direct (paternity analyses based on microsatellite DNA markers) and indirect (fluorescent dyes) methods. We compared these methods using two approaches, testing for the difference between the distance distributions of observed dispersal events and estimating parameters of a dispersal model, and related these results to dye dispersal patterns in three large populations. Dye and realized (based on paternity inference) pollen dispersal showed exponential decay distributions, with 74.2?C94.8% of the depositions occurring at <50?m and a few longer distance dispersal events (up to 151?m). No significant difference in curve shape was found between dye and realized pollen dispersal distributions. The best-fitting parameters characterizing the dye dispersal model were consistent with those obtained for realized pollen dispersal. Hence, the fluorescent dye method may be considered as reliable to infer realized pollen dispersal for forest herbs such as P. elatior. However, our simulations reveal that large sample sizes are needed to detect moderate differences between dye and realized pollen dispersal patterns because the estimation of dispersal parameters suffers low precision.     

Delayed seed dispersal in Pinus torreyana (Torrey pine)

Summary Torrey pine exhibits a pattern of seed release intermediate between an open and closed-cone species. It was found that the cones open at maturity, but that seed fall from some cones continues for up to 13 years. Approximately 77% of the total seed crop was present in age classes one or more years past the time of seed maturity.Serotiny is thought to evolve in response to fire, and partial serotiny at the population level has been attributed to circumstances in which the strong selective pressure of fire is relaxed. We hypothesize that delayed seed dispersal in Torrey pine is a stable trait which is the result of a pattern of large relatively rare crown fires separated by intervals longer than one generation. The reduced serotiny observed in other conifers on headlands and islands in California seems consistent with our hypothesis.     

Realized pollen and seed dispersal within a continuous population of the dioecious coniferous Brazilian pine [Araucaria angustifolia (Bertol.) Kuntze]

Extensive realized pollen and seed flow across populations reduces inbreeding and spatial genetic structure (SGS) and increases the genetic diversity and effective size within populations. Inbreeding, SGS and realized patterns of pollen and seed dispersal of the dioecious, wind pollinated Araucaria angustifolia were investigated based on microsatellite loci. The study was conducted in a 7.2 ha plot established within a continuous Araucaria Forest in Southern Brazil. In the plot, all 290 adult trees were mapped, measured (diameter at breast height, dbh), sexed, sampled and genotyped. We also sampled, measured (total height), mapped and genotyped 223 juveniles. A total of 86 alleles were detected in the sample (n = 513). Adults and juveniles showed a positive and significant fixation index (minimum of 0.096), suggesting inbreeding or Wahlund effect. Juveniles presented a significant aggregated distribution which was associated with a genetic aggregation (significant SGS up to 20 m), indicating that near neighbor trees may be related due the limited seed dispersal. However, the intensity of SGS was not significantly higher among juveniles (Sp = 0.0041) than adults (Sp = 0.0026). Realized pollen and seed immigration into the plot was low (pollen = 6 %; seeds = 5 %) and the patterns of pollen and seed dispersal were similar. Pollen was dispersed over long distances (343 m), but 50 % was dispersed up to 124 m. Seeds also reached long distances (318 m), with 50 % being dispersed up to 133 m. Our results are discussed in terms of auto-ecology and the genetic conservation of A. angustifolia populations.     

Long-distance dispersal helps germinating mahogany seedlings escape defoliation by a specialist caterpillar

Herbivores and pathogens with acute host specificity may promote high tree diversity in tropical forests by causing distance- and density-dependent mortality of seedlings, but evidence is scarce. Although Lepidoptera larvae are the most abundant and host-specific guild of herbivores in these forests, their impact upon seedling distributions remains largely unknown. A firm test of the mechanism underpinning the Janzen–Connell hypothesis is difficult, even for a single tree species, because it requires more than just manipulating seeds and seedlings and recording their fates. Experimental tests require: (1) an insect herbivore that is identified and highly specialised, (2) linkage to an in situ measure (or prevention) of herbivory, and (3) evaluation and confirmation among many conspecific adult trees across years. Here we present experimental evidence for a spatially explicit interaction between newly germinating seedlings of a Neotropical emergent tree, big-leaf mahogany (Swietenia macrophylla, Meliaceae), and caterpillars of a noctuid moth (Steniscadia poliophaea). In the understory of a southeastern Amazon forest, the proportion of attacks, leaf area lost, and seedling mortality due to this specialised herbivore peaked near Swietenia trees, but declined significantly with increasing distance from mature fruiting trees, as predicted by the Janzen–Connell hypothesis. We conclude that long-distance dispersal events (>50 m) provided an early survival advantage for Swietenia seedlings, and propose that the role of larval Lepidoptera as Janzen–Connell vectors may be underappreciated in tropical forests.     

Interactions during in vitro germination of Scots pine pollen

Pre-zygotic pollen competition is believed to play an important role in nonrandom mating, i.e., the unequal success of different pollen donors. We studied pollen–pollen interactions of Scots pine in vitro using multiwell plates with freely permeable inserts. Six genotypes were included in our experiments: three from northern Finland and three from southern Finland. We conducted control experiments by placing pollen of each genotype in both the well and its insert. In competition experiments each southern genotype was incubated in insert with each northern genotype in well, and vice-versa. Samples for the germinability observations were taken from inserts. The mean germination percentage of northern genotypes was lower when incubating with southern genotypes in competition experiments than in control experiments. In one case, a northern genotype showed lower mean germination percentage in competition experiment than in control experiment and in another case a southern genotype showed a higher value. Our results suggest a chemically mediated interaction between pollen from different genotypes and one which can vary among genotypes.     

Enhanced airborne radioactivity during a pine pollen release episode

 A single episode of pine pollen release in the highly contaminated area of Novozybkov, Russian Federation, which led to enhanced atmospheric concentrations of ¹³⁷Cs is discussed. The pollen grains were sampled by a rotating arm impactor and analysed by gamma-spectrometry for ¹³⁷Cs activity and by image analysis for their size. In the vicinity of a forest, a maximum concentration of 4.5±0.4 mBq m^–3 was measured, and a mean activity per pollen grain of 260±80 nBq was determined. The emission rate of the Novozybkov mixed pine forest was estimated to be approximately 400 Bq m^–2 per year. Because of the large size of pine pollen grains (about 50 μm) and the short emission period of 5–8 days per year, the estimated potential annual inhalation doses are very low. Biological emissions including pollen release may be a source of increased airborne radionuclide concentrations at larger distances from the source areas as well. Received: 2 October 1998 / Accepted in revised form: 30 January 1999     

Spatially explicit modelling of transgenic maize pollen dispersal and cross-pollination

Modelling of pollen dispersal and cross-pollination is of great importance for the ongoing discussion on thresholds for the adventitious presence of genetically modified material in food and feed. Two different modelling approaches for pollen dispersal are used to simulate the cross-pollination rate of pollen emerged from an adjacent transgenic crop field. The models are applied to cross-pollination data from field experiments with transgenic maize (Zea mays). The data were generated by an experimental setup specifically designed to suit the demands of mathematical modelling. First a Gaussian plume model is used for the simulation of pollen transport in and from plant canopies. This is a semiempirical approach combining the atmospheric diffusion equation and Lagrangian methodology. The second model is derived from the localised near field (LNF) theory and based on the physical processes in the canopy. Both modelling approaches prove to be appropriate for the simulation of the cross-pollination rates at distances of about 7.5m and more from the transgene source. The simulation of the cross-pollination rate is less precise at the edge of the source plot especially with the LNF theory. However, the simulation results lie within the range of variability of the observations. Concluding can be pointed out that both models might be adapted to other pollen dispersal experiments of different crops and plot sizes.     

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.