Effects of flowering plant density on pollinator visitation, pollen receipt, and seed production in Delphinium barbeyi (Ranunculaceae)

Variation in flowering plant density can have conflicting effects on pollination and seed production. Dense flower patches may attract more pollinators, but flowers in those patches may also compete for pollinator visits and abiotic resources. We examined how natural and experimental conspecific flowering plant density affected pollen receipt and seed production in a protandrous, bumble bee-pollinated wildflower, Delphinium barbeyi (Ranunculaceae). We also compared floral sex ratios, pollinator visitation rates, and pollen limitation of seed set from early to late in the season to determine whether these factors mirrored seasonal changes in pollen receipt and seed production. Pollen receipt increased with natural flowering plant density, while seed production increased across lower densities and decreased across higher flower densities. Experimental manipulation of flowering plant density did not affect pollinator visitation rate, pollen receipt, or seed production. Although pollinator visitation rate increased 10-fold from early to late in the season, pollen receipt and seed set decreased over the season. Seed set was never pollen-limited. Thus, despite widespread effects of flowering plant density on plant reproduction in other species, the effects of conspecific flowering plant density on D. barbeyi pollination and seed production are minor.     

Timing of invasive pollen deposition influences pollen tube growth and seed set in a native plant

Invasive plants may threaten the reproductive success of native sympatric plants by modifying the pollination process. One potential mechanism takes place through the deposition of invasive pollen onto native stigmas when pollinators are shared among species. We explore how pollen from the invasive plant Brassica nigra influences pre- and post-fertilization stages in the native plant Phacelia parryi, through a series of hand pollination experiments. These two species share pollinators to a high degree. P. parryi flowers were hand-pollinated with either pure conspecific pollen (the control) or with B. nigra pollen applied prior to, simultaneously with, or following conspecific pollen. Application of B. nigra pollen lowered seed set, with the simultaneous application resulting in the highest reduction. Pollen tube growth was also influenced by the presence of invasive pollen, with fewer conspecific pollen tubes reaching the base of P. parryi styles in treatments where B. nigra pollen was applied prior to or simultaneously with conspecific pollen. The deleterious effects of invasive pollen on native seed set in this study are likely not due to loss of stigmatic receptivity since seed set was less affected when heterospecific pollen was applied prior to conspecific pollen, but may instead involve interactions between interspecific pollen grains on the stigma or within the style. Our study highlights the importance of timing of foreign pollen deposition on native stigmas and suggests that interspecific pollen transfer between native and exotic plants may be an important mechanism of competition for pollination in invaded plant communities.     

Reproductive interactions mediated by flowering overlap in a temperate hummingbird–plant assemblage

Pollinator‐mediated competition through shared pollinators can lead to segregated flowering phenologies, but empirical evidence for the process responsible for this flowering pattern is sparse. During two flowering seasons, we examined whether increasing overlap in flowering phenology decreased conspecific pollination, increased heterospecific pollination, and depressed seed output in the seven species composing a hummingbird–plant assemblage from the temperate forest of southern South America. Overall trends were summarized using meta‐analysis. Despite prevailing negative associations, relations between phenological overlap and conspecific pollen receipt varied extensively among species and between years. Heterospecific pollen receipt was low and presumably of limited biological significance. However, our results supported the hypothesis that concurrent flowering promotes interspecific pollen transfer, after accounting for changes in the abundance of conspecific flowers. Seed output was consistently reduced during maximum phenological overlap during the first flowering season because of limited fruit set. Responses varied more during the second year, despite an overall negative trend among species. Relations between estimated effects of phenological overlap on pollination and seed output, however, provided mixed evidence that conspecific pollen loss during pollinator visits to foreign flowers increases pollen limitation. By flowering together, different plant species might benefit each other's pollination by increasing hummingbird recruitment at the landscape level. Nevertheless, our results are mostly consistent with the hypothesis of pollinator‐mediated competition shaping the segregated flowering pattern reported previously for this temperate plant assemblage. The mechanisms likely involve effects on male function, whereby pollen‐transport loss during heterospecific flower visits limit pollen export, and more variable effects on female function through pollen limitation.     

Interspecific Pollen Transfer: Magnitude,Prevalence and Consequences for Plant Fitness

Interspecific pollen transfer (IPT) is one of the mechanisms underlying potential competition among plants for pollinators, and it refers to movement of pollen between different plant species by pollinators that visit their flowers simultaneously. Two components of IPT, related to each other, are distinguished: (a) heterospecific pollen deposition (HPD) on conspecific stigmas, which may interfere with fertilization by conspecific pollen; and (b) conspecific pollen loss (CPL) on heterospecific flowers, which may reduce the amount of pollen transferred between conspecific flowers. Thus, IPT may lead to reciprocal losses for male and female functions of the plant, with potentially important ecological and evolutionary consequences. In this review, we explore the magnitude and prevalence of IPT, examining documented mechanisms and evaluating such potential ecological and evolutionary consequences. We compiled existing evidence of interspecific pollinator sharing and interspecific pollinator switching between flowers of different species in natural communities. We evaluated the relative importance of both HPD and CPL from studies comparing these variables in pure vs. mixed floral neighborhoods, analyzing evidence for the claim that IPT is an evolutionary force promoting character displacement in habitat affinity, flowering times, and floral morphology. We also examined the findings of hand-pollination experiments carried out to reveal different mechanisms by which heterospecific pollen can affect performance of native pollen. Finally, we review evidence for impacts of alien plant species on native species' reproduction, and briefly comment on risks of crop-to-wild gene flow imposed by the release of genetically modified (transgenic) crops through IPT.     

Evaluating the effects of pollinator‐mediated interactions using pollen transfer networks: evidence of widespread facilitation in south Andean plant communities

Information about the relative importance of competitive or facilitative pollinator‐mediated interactions in a multi‐species context is limited. We studied interspecific pollen transfer (IPT) networks to evaluate quantity and quality effects of pollinator sharing among plant species on three high‐Andean communities at 1600, 1800 and 2000 m a.s.l. To estimate the sign of the effects (positive, neutral or negative), the relation between conspecific and heterospecific pollen deposited on stigmas was analysed with GLMMs. Network analyses showed that communities were characterised by the presence of pollen hub‐donors and receptors. We inferred that facilitative and neutral pollinator‐mediated interactions among plants prevailed over competition. Thus, the benefits from pollinator sharing seem to outweigh the costs (i.e. heterospecific deposition and conspecific pollen loss). The largest proportion of facilitated species was found at the highest elevation community, suggesting that under unfavourable conditions for the pollination service and at lower plant densities facilitation can be more common.     

Multi‐year stigmatic pollen‐load sampling reveals temporal stability in interspecific pollination of flowers in a subalpine meadow

Co‐flowering plants may commonly experience interspecific pollination. It remains unknown, however, whether interspecific pollination is a largely stochastic process or consistent enough over years to exert selection for traits that can reduce interspecific pollination or ameliorate its deleterious effects on reproduction. To assess the likelihood of this precondition being met, stigmatic pollen loads on 17–34 insect‐pollinated plant species over three consecutive years were scored in a subalpine meadow in southwestern China. Plant species varied significantly in the amount and proportion of heterospecific pollen (HP) on stigmas. Both the number of HP species and the proportion of the pollen load that was HP for each recipient species correlated positively between years (reflected in pairwise correlations for all year‐by‐year combinations). Although inter‐annual variation was smaller for conspecific pollen (CP) than for HP loads, species tended to experience either consistently high or consistently low HP proportions across years. We found that species with higher stigmatic HP proportions generally experienced lower proportional variation in stigmatic HP, an unexpected result if high HP loads are the result of rare stochastic events. The novel finding of between‐year consistency in stigmatic loads of heterospecific pollen suggests that adaption to stigmatic loads of HP is possible, and two divergent strategies may have evolved: HP avoidance and HP tolerance. The observation of temporally consistent differences among species in levels of HP supports the idea that natural selection may be operating either to increase tolerance or to minimize arrival of heterospecific pollen on stigmas in co‐flowering plants. Such adaptations may be important for the maintenance of high levels of local plant diversity in biodiversity hotspots such as our study area.     

Incomplete partitioning of pollinators by Linum suffruticosum and its coflowering congeners

Premise

Linum suffruticosum shows variations in pollinator fit, pollen pickup, and local pollinators that predict pollen deposition rates. The species often coflowers with other Linum species using the same pollinators. We investigated whether L. suffruticosum trait variation could be explained by local patterns of pollinator sharing and associated evolution to reduce interspecific pollen transfer.

Methods

Pollinator observations were made in different localities (single species, coflowering with other congeners). Floral traits were measured to detect differences across populations and from coflowering species. Reproductive costs were quantified using interspecific hand pollinations and measures of pollen-tube formation, combined with observations of pollen arrival on stigmas and pollen-tube formation after natural pollination in allopatric and sympatric localities.

Results

The size and identity of the most important pollinator of L. suffruticosum and whether there was pollinator sharing with coflowering species appeared to explain floral trait variation related to pollinator fit. The morphological overlap of the flowers of L. suffruticosum with those of coflowering species varied, depending on coflowering species identity. A post-pollination incompatibility system maintains reproductive isolation, but conspecific pollen-tube formation was lower after heterospecific pollination. Under natural pollination at sites of coflowering with congeners, conspecific pollen-tube formation was lower than at single-species localities.

Conclusions

Trait variation in L. suffruticosum appears to respond to the most important local pollinator. Locally, incomplete pollinator partitioning might cause interspecific pollination, imposing reproductive costs. These reproductive costs may generate selection on floral traits for reduced morphological overlap with coflowering congeners, leading to the evolution of pollination ecotypes.     

Pollen transfer in fragmented plant populations: insight from the pollen loads of pollinators and stigmas in a mass‐flowering species

Pollinator and/or mate scarcity affects pollen transfer, with important ecological and evolutionary consequences for plant reproduction. However, the way in which the pollen loads transported by pollinators and deposited on stigmas are affected by pollination context has been little studied. We investigated the impacts of plant mate and visiting insect availabilities on pollen transport and receipt in a mass‐flowering and facultative autogamous shrub (Rhododendron ferrugineum). First, we recorded insect visits to R. ferrugineum in plant patches of diverse densities and sizes. Second, we analyzed the pollen loads transported by R. ferrugineum pollinators and deposited on stigmas of emasculated and intact flowers, in the same patches. Overall, pollinators (bumblebees) transported much larger pollen loads than the ones found on stigmas, and the pollen deposited on stigmas included a high proportion of conspecific pollen. However, comparing pollen loads of emasculated and intact flowers indicated that pollinators contributed only half the conspecific pollen present on the stigma. At low plant density, we found the highest visitation rate and the lowest proportion of conspecific pollen transported and deposited by pollinators. By contrast, at higher plant density and lower visitation rate, pollinators deposited larger proportion of conspecific pollen, although still far from sufficient to ensure that all the ovules were fertilized. Finally, self‐pollen completely buffered the detrimental effects on pollination of patch fragmentation and pollinator failure. Our results indicate that pollen loads from pollinators and emasculated flowers should be quantified for an accurate understanding of the relative impacts of pollinator and mate limitation on pollen transfer in facultative autogamous species.     

Pollinator‐mediated interactions between cultivated papaya and co‐flowering plant species

Many modern crop varieties rely on animal pollination to set fruit and seeds. Intensive crop plantations usually do not provide suitable habitats for pollinators so crop yield may depend on the surrounding vegetation to maintain pollination services. However, little is known about the effect of pollinator‐mediated interactions among co‐flowering plants on crop yield or the underlying mechanisms. Plant reproductive success is complex, involving several pre‐ and post‐pollination events; however, the current literature has mainly focused on pre‐pollination events in natural plant communities. We assessed pollinator sharing and the contribution to pollinator diet in a community of wild and cultivated plants that co‐flower with a focal papaya plantation. In addition, we assessed heterospecific pollen transfer to the stigmatic loads of papaya and its effect on fruit and seed production. We found that papaya shared at least one pollinator species with the majority of the co‐flowering plants. Despite this, heterospecific pollen transfer in cultivated papaya was low in open‐pollinated flowers. Hand‐pollination experiments suggest that heterospecific pollen transfer has no negative effect on fruit production or weight, but does reduce seed production. These results suggest that co‐flowering plants offer valuable floral resources to pollinators that are shared with cultivated papaya with little or no cost in terms of heterospecific pollen transfer. Although HP reduced seed production, a reduced number of seeds per se are not negative, given that from an agronomic perspective the number of seeds does not affect the monetary value of the papaya fruit.     

Interspecific pollinator movements reduce pollen deposition and seed production in Mimulus ringens (Phrymaceae)

Movement of pollinators between coflowering plant species may influence conspecific pollen deposition and seed set. Interspecific pollinator movements between native and showy invasive plants may be particularly detrimental to the pollination and reproductive success of native species. We explored the effects of invasive Lythrum salicaria on the reproductive success of Mimulus ringens, a wetland plant native to eastern North America. Pollinator flights between these species significantly reduced the amount of conspecific pollen deposited on Mimulus stigmas and the number of seeds in Mimulus fruits, suggesting that pollen loss is an important mechanism of competition for pollination. Although pollen loss is often attributed to pollen wastage on heterospecific floral structures, our novel findings suggest that grooming by bees as they forage on a competitor may also significantly reduce outcross pollen export and seed set in Mimulus ringens.     

Intrinsic and extrinsic factors acting on the reproductive process in alpine‐snowbed plants: roles of phenology,biological interaction,and breeding system

Pollinator activity and competition for pollinators lead to quantitative and qualitative pollen limitations on seed production and affect the reproductive success of plant species, depending on their breeding system (e.g., self‐compatibility and heterospecific compatibility) and genetic load (e.g., inbreeding depression and hybrid inviability). In alpine ecosystems, snowmelt regimes determine the distribution and phenology of plant communities. Plant species growing widely along a snowmelt gradient often grow with different species among local populations. Their pollinators also vary in their abundance, activity, and behavior during the season. These variations may modify plant–pollinator and plant–plant interactions. We integrated a series of our studies on the alpine dwarf shrub, Phyllodoce aleutica (Ericaceae), to elucidate the full set of intrinsic (species‐specific breeding system) and extrinsic factors (snow condition, pollinator activity, and interspecific competition) acting on their reproductive process. Seasonality of pollinator activity led to quantitative pollen limitation in the early‐blooming populations, whereas in the late‐blooming populations, high pollinator activity ensured pollination service, but interspecific competition for pollinators led to qualitative and quantitative pollen limitation in less competitive species. However, negative effects of illegitimate pollen receipt on seed‐set success might be reduced when cryptic incompatibility systems (i.e., outcross pollen grains took priority over self‐ and heterospecific pollen grains) could effectively prevent ovule and seed discounting. Our studies highlight the importance of species‐specific responses of plant reproduction to changing pollinator availability along environmental gradients to understand the general features of pollination networks in alpine ecosystems.     

Individual flowering phenology shapes plant–pollinator interactions across ecological scales affecting plant reproduction

The balance of pollination competition and facilitation among co-flowering plants and abiotic resource availability can modify plant species and individual reproduction. Floral resource succession and spatial heterogeneity modulate plant–pollinator interactions across ecological scales (individual plant, local assemblage, and interaction network of agroecological infrastructure across the farm). Intraspecific variation in flowering phenology can modulate the precise level of spatio-temporal heterogeneity in floral resources, pollen donor density, and pollinator interactions that a plant individual is exposed to, thereby affecting reproduction. We tested how abiotic resources and multi-scale plant–pollinator interactions affected individual plant seed set modulated by intraspecific variation in flowering phenology and spatio-temporal floral heterogeneity arising from agroecological infrastructure. We transplanted two focal insect-pollinated plant species (Cyanus segetum and Centaurea jacea, n = 288) into agroecological infrastructure (10 sown wildflower and six legume–grass strips) across a farm-scale experiment (125 ha). We applied an individual-based phenologically explicit approach to match precisely the flowering period of plant individuals to the concomitant level of spatio-temporal heterogeneity in plant–pollinator interactions, potential pollen donors, floral resources, and abiotic conditions (temperature, water, and nitrogen). Individual plant attractiveness, assemblage floral density, and conspecific pollen donor density (C. jacea) improved seed set. Network linkage density increased focal species seed set and modified the effect of local assemblage richness and abundance on C. segetum. Mutual dependence on pollinators in networks increased C. segetum seed set, while C. jacea seed set was greatest where both specialization on pollinators and mutual dependence was high. Abiotic conditions were of little or no importance to seed set. Intra- and interspecific plant–pollinator interactions respond to spatio-temporal heterogeneity arising from agroecological management affecting wild plant species reproduction. The interplay of pollinator interactions within and between ecological scales affecting seed set implies a co-occurrence of pollinator-mediated facilitative and competitive interactions among plant species and individuals.     

Pollen dispersal and fruit production in Vaccinium oxycoccos and comparison with its sympatric congener V. uliginosum

Investigating plant–pollinator interactions and pollen dispersal are particularly relevant for understanding processes ensuring long‐term viability of fragmented plant populations. Pollen dispersal patterns may vary strongly, even between similar congeneric species, depending on the mating system, pollinator assemblages and floral traits. We investigated pollen dispersal and fruit production in a population of Vaccinium oxycoccos, an insect‐pollinated shrub, and compared the pollen dispersal pattern with a co‐flowering, sympatric congener, V. uliginosum. We examined whether they share pollinators (through interspecific fluorescent dye transfers) and may differently attract pollinators, by comparing their floral colour as perceived by insects. Fluorescent dyes were mainly dispersed over short distances (80% within 40.4 m (max. 94.5 m) for V. oxycoccos and 3.0 m (max. 141.3 m) for V. uliginosum). Dye dispersal in V. oxycoccos was not significantly affected by plant area, floral display or the proximity to V. uliginosum plants. Interspecific dye transfers were observed, indicating pollinator sharing. The significantly lower dye deposition on V. oxycoccos stigmas suggests lower visitation rates by pollinators, despite higher flower density and local abundance. The spectral reflectance analysis indicates that bees are unlikely to be able to discriminate between the two species based on floral colour alone. Fruit production increased with increasing floral display, but was not affected by proximity to V. uliginosum plants. Our study highlights that fragmented populations of V. oxycoccos, when sympatric with co‐flowering V. uliginosum, might incur increased competition for the shared pollinators in the case of pollination disruption, which might then reduce outcrossed seed set.     

Secondary pollen presentation and the temporal dynamics of stylar hair retraction and style elongation in Campanula trachelium (Campanulaceae)

To increase the accuracy of pollen capture and transfer by pollinators some plant species have developed secondary pollen presentation structures. Because the presence of secondary pollen presentation structures at the pistil may reduce the spatial separation between the sexual functions and increase the risk of self‐interference and selfing, temporal segregation of the sexual organs, triggered by visiting insects, can be expected to occur. We investigated secondary pollen presentation and the temporal dynamics of the sexual phases in combination with the physiological self‐incompatibility system in Campanula trachelium, a protandrous insect‐pollinated herb. Stylar hair retraction (male function) and curling of the stigmatic lobes (female function) were modelled using Gompertz growth functions. Finally, we performed pollination experiments in the lab and field to assess seed set and pollen limitation under natural conditions. About 68% of the total pollen load was captured by stylar hairs. Manual manipulation of the stylar hairs, mimicking pollinator visitation, significantly shortened the male phase and accelerated the female phase, resulting in a significant decline in temporal overlap between the two sexual functions. Conversely, when pollinators and/or manual manipulations were lacking, the male phase was substantially prolonged and sexual overlap was maximal. This suggests that spreading of the sexual phases and thus the risk of sexual interference are largely determined by the interaction between stylar hairs and visiting pollinators. Natural seed set was high and not pollinator limited. Overall, these results indicate that secondary pollen presentation and partial protandry resulted in efficient pollen capture, transfer and deposition.     

Interspecific pollen transfer and competition between co-occurring plant species

Summary Computer simulations of a pollinator foraging in a mixture of two species were used to explore how plant reproduction can be influenced by interspecific pollination movements. Interspecific pollen transfer led to strong competitive effects when availabilities of pollen, receptive stigma surfaces, or pollinator movements were limited relative to the total number of fertilizations possible in the mixed population. Results from simulations suggest that competition for pollination through interspecific pollen transfer can result in rapid exclusion of one of two species, and that such competition represents a selective force promoting stable divergence of potential competitors in habitat affinity, flowering time, or other characteristics related to pollinator sharing.     

Interspecific pollen transfer as a mechanism of competition: Consequences of foreign pollen contamination for seed set in the alpine wildflower,Polemonium viscosum

Summary Earlier studies showed that under natural conditions foreign pollen receipt is correlated with loss of seed set in populations of P. viscosum. Here, we report on experimental pollinations using foreign pollen from two co-flowering species that share pollinators with P. viscosum. Experiments were designed to ascertain whether improper pollen transfer causes reduced seed set under controlled conditions and, if so, to determine the stage at which interference occurs. In flowers pre-treated by pollination with either Mertensia or Castilleja pollen, adherence of subsequently deposited conspecific pollen grains was unaffected. However, conspecific grains had significantly lower germination and ovule fertilization success in flowers receiving foreign pollen than in controls. These results suggest that pollinator infidelity in high alpine plant communities can reduce female reproductive success of P. viscosum by inhibiting the performance of conspecific pollen grains. Negative effects of Mertensia and Castilleja pollen were comparable despite large interspecific differences in pollen grain morphology. Accordingly, susceptibility to foreign pollen interference appears to be an intrinsic property of P. viscosum that is not dependent on the unique structural properties of pollen produced by competitive species.     

Seasonal changes in pollinator activity influence pollen dispersal and seed production of the alpine shrub Rhododendron aureum (Ericaceae)

In alpine ecosystems, microscale variation in snowmelt timing often causes different flowering phenology of the same plant species and seasonal changes in pollinator activity. We compared the variations in insect visitation, pollen dispersal, mating patterns, and sexual reproduction of Rhododendron aureum early and late in the flowering season using five microsatellites. Insects visiting the flowers were rare early in the flowering season (mid-June), when major pollinators were bumblebee queens and flies. In contrast, frequent visitations by bumblebee workers were observed late in the season (late July). Two-generation analysis of pollen pool structure demonstrated that quality of pollen-mediated gene flow was more diverse late in the season in parallel with the high pollinator activity. The effective number of pollen donors per fruit (N(ep)) increased late in the season (N(ep) = 2.2-2.7 early, 3.4-4.4 late). However, both the outcrossing rate (t(m)) and seed-set ratio per fruit were smaller late in the season (t(m) = 0.89 and 0.71, seed-set ratio = 0.52 and 0.18, early and late in the season, respectively). In addition, biparental inbreeding occurred only late in the season. We conclude that R. aureum shows contrasting patterns of pollen movement and seed production between early and late season: in early season, seed production can be high but genetically less diverse and, during late season, be reduced, possibly due to higher inbreeding and inbreeding depression, but have greater genetic diversity. Thus, more pollinator activity does not always mean more pollen movement.     

Temporal variations in the linkages between plants and flower visitors and the pollination success of Primula modesta along the snowmelt gradient

Although pollination networks between plants and flower visitors are diverse and flexible, seed production of many plant species is restricted by pollen limitation. Obligate outcrossers often suffer from low pollinator activity or severe interspecific competition for pollinator acquisition among co-flowering species. This study focused on seasonal changes in plant–flower visitor linkages in an alpine ecosystem and examined whether and how this seasonality affected the seed-set of Primula modesta, a self-incompatible distylous herb having long-tubed flowers. First, we recorded the linkages between plants and flower visitors along the snowmelt gradient. Then, pollination experiment was conducted to estimate the degree of pollen limitation over the course of flowering season of P. modesta. Flower visitors were classified by their tongue length based on the morphological matching with P. modesta flowers. As the season progressed, plant–visitor linkages became more diverse and generalized, and the visitation frequency to P. modesta flowers increased. In the later part of the season, however, the seed set of P. modesta was significantly reduced due to severe pollen limitation, presumably because of increased competition for long-tongued pollinators among co-flowering species. The present study revealed that pollinator availability for specialist species may be restricted even when plant–visitor linkages are diverse and generalized as a whole. In the case of P. modesta, morphological matching and competition for pollinators might be the main factors explaining this discrepancy.     

Experimental sympatry suggests geographic isolation as an essential reproductive barrier between two sister species of Pedicularis

Although mechanical isolation mediated by shared pollinators has been considered as a classic model of pollinator-mediated floral isolation in Pedicularis, a superdiverse genus in Hengduan Mountains, southwest China, there has been no empirical study of interspecific pollen flow between closely related species. We examined reproductive barriers at six different stages between Pedicularis cranolopha and Pedicularis tricolor, two sister species. The two sister species were geographically isolated from each other based on our field survey and the records of herbarium specimens. Translocation experiments showed that flowering phenology partly overlapped and bumblebee pollinators did not discriminate between flowers of the two species. Bumblebee interspecific moves could mediate interspecific pollination as traced using fluorescent powder, in which pollen analogs placed on one species were transferred to the stigmas of the other species in experimental plots containing both species. Heterospecific pollen tubes grew in the style as well as conspecific pollen in hand-pollination experiments. Reciprocal hybridization between the two species could produce (partially) viable seeds, suggesting weak post-pollination barriers. Our results showed that geographic isolation was an important barrier of two species, and the total reproductive isolation between two species was incomplete when without geographical isolation. The formation of Big Snow Mountains could introduce an important pre-zygotic reproductive barrier between the two sister species of Pedicularis; such geographical isolation could be responsible for allopatric speciation, giving a clue to understanding the rapid radiation on mountain areas.     

Effects of multiple competitors for pollination on bumblebee foraging patterns and Mimulus ringens reproductive success

When co‐occurring plant species overlap in flowering phenology they may compete for the service of shared pollinators. Competition for pollination may lower plant reproductive success by reducing the number of pollinator probes or by decreasing the quality of pollen transport to or from a focal species. Pair‐wise interactions between plants sharing pollinators have been well documented. However, relatively few studies have examined interactions for pollination among three or more plant species, and little is known about how the outcomes and mechanisms of competition for pollination may vary with competitor species composition. To better understand how the dynamics of competition for pollination may be influenced by changes in the number of competitors, we manipulated the presence of two competitors, Lythrum salicaria and Lobelia siphilitica, and quantified reproductive success for a third species, Mimulus ringens. Patterns of pollinator preference and interspecific transitions in mixed‐species arrays were significantly influenced by the species composition of competitor plants present. Both pair‐wise and three‐species competition treatments led to a similar ～ 40% reduction in Mimulus ringens seed set. However, the patterns of pollinator foraging we observed suggest that the relative importance of different mechanisms of competition for pollination may vary with the identity and number of competitors present. This variation in mechanisms of competition for pollination may be especially important in diverse plant communities where many species interact through shared pollinators.