Pollination success increases with plant diversity in high-Andean communities

BackgroundIn flowering communities, plant species commonly share pollinators and therefore plant individuals receive heterospecific pollen (HP). However, the patterns of HP transfers can deviate from patterns of plant-pollinator visitations. Although flower-visitor interactions are known to be mediated by floral traits, e.g. floral size or nectar tube depth, the explanatory power of these traits for HP transfer patterns remains elusive. Here, we have explored pollen transfer patterns at three sites in Southern Germany on three dates (early, mid and late summer). At the plant level, we tested whether flower abundance and floral traits are correlated with HP reception and donation. At the community level, we determined whether flower and bee diversity are correlated with network modularity and whether floral traits explain the module affiliation of plant species. We collected the stigmas of flowering plant species, analysed HP and conspecific pollen (CP) loads and measured floral traits, flower and bee diversity.ResultsOur results show that the degree and intensity of HP reception or donation at the plant level do not correlate with floral traits, whereas at the community level, the module affiliation of who is sharing pollen with whom is well-explained by floral traits. Additionally, variation in network modularity between communities is better explained by plant diversity and abundance than by bee diversity and abundance.ConclusionsOverall, our results indicate that floral traits that are known to mediate flower-visitor interactions can improve our understanding of qualitative HP transfer but only provide limited information about the quantity of HP transfer, which more probably depends on other floral traits, flower-visitor identity or community properties.

Heterospecific pollen (HP) deposition varies widely among species in communities, which has been explicated by two adaptation strategies: HP avoidance and HP tolerance. Studies of the plant-pollinator network have uncovered that oceanic island communities are highly generalized and strongly connected. It remains unclear, however, which strategy prevails in such communities. We examined stigma pollen deposition on 29 plant species, and assessed patterns of HP load size and diversity in the Yongxing Island community. We assessed the effects of phenotypic specialization and species-level network structural properties of plant species on pollen deposition among species. The hypothesis of three accrual patterns of HP within species was tested by illustrating the relationship between conspecific pollen (CP) and HP receipt. Extensive variation occurred among species in HP receipt, while 75.9% of species received less than 10% HP and one species received more than 40% HP throughout the community. Flower size strongly drives the variation of HP receipt, while network structural properties had no effect on the pollen receipt. Nineteen species showed no relationship between the number of HP and CP loads, and they received smaller HP load sizes and lower HP proportions. Most plant species evolved HP avoidance strategy, and HP receipt was an occasional event for most plant species in the generalized community. HP and CP receipts are independent of each other in plant species with the HP avoidance mechanism. Our results highlight that plants in the generalized pollination system may preferentially select to minimize the HP load on stigmas.

The study of plant responses to touch, particularly the responses of leaves, stems, and roots, has a long history. By contrast, floral responses are relatively unexplored. Stigma closure is common in the Bignoniaceae, but the factors influencing it are not well understood. We investigated factors influencing stigma closure and reopening and its effects on pollen receipt in seven Oroxylum indicum trees near Hat Yai, Thailand. The effects of pressure, conspecific and heterospecific pollen, and pollen load (the amount of pollen deposited) on stigma behavior were examined in 270 flowers (of the total 430 flowers evaluated in the entire study). Pressure alone resulted in faster closure than did conspecific pollination and faster reopening than did heterospecific pollination. Stigmas never reopened after conspecific pollination. Pollen load had no effect on stigma behavior. Stigmas discriminated between conspecific and foreign pollen; they reopened only after pollination with the latter. A manipulative experiment revealed that stigma closure did not affect the number of conspecific pollen grains received. We also counted pollen tubes in styles that were either hand-supplemented with outcross conspecific pollen or open-pollinated. Pollen tube numbers were highest after light pollination (∼900 grains), indicating that interference among pollen grains may occur after pollination with very heavy loads (>6000 grains). Possible fitness consequences of these responses are discussed.

Co-flowering species may have evolved strategies to avoid or tolerate the adverse effects of heterospecific pollen deposition. However, the precondition for this evolutionary response is spatial–temporal stability, an aspect currently understudied. Here, we examined the spatial–temporal stability in conspecific and heterospecific pollen loads on stigmas across 19 co-flowering species in six sub-alpine meadow communities over four consecutive years. We found that, although conspecific and heterospecific pollen loads, as well as proportions of heterospecific pollen, differed significantly among species, with heterospecific pollen proportion ranging from 0.1% to 41.8%, variation in heterospecific pollen proportion among species was stable across different years and communities. The most important predictor of variation in both conspecific and heterospecific pollen loads, as well as heterospecific pollen proportions, was species identity; furthermore, this factor was independent of phylogenetic relationship. The proportion of heterospecific pollen varied less within species that had high proportions of heterospecific pollen. Furthermore, both the proportion of heterospecific pollen and its coefficient of variation were more strongly driven by heterospecific pollen than by conspecific pollen. Our study suggests that variation in stigmatic pollen load among co-flowering species is spatially and temporally consistent, a precondition for the tolerance-avoidance strategy. This study provides new insights into how different plant species respond to heterospecific pollen deposition.

Pollinator sharing often leads to receipt of heterospecific pollen (HP) along with conspecific pollen. As a result, flowering plants can accumulate diverse communities of HP on stigmas. While variation in HP diversity is an important selective force contributing to flowering plant fitness, evolution and community assembly, our understanding of the extent and drivers of heterogeneity of HP diversity is limited. In this study, we examined the species compositions and abundances of ~1000 HP communities across 59 co‐flowering plant species in three serpentine seep communities in California, USA. We evaluated the variation in HP diversity (γ diversity) across plant species in each seep and asked whether the variation in HP γ diversity was caused by variation in HP diversity within stigmas (α diversity) or HP compositional variation among stigmas (β diversity) due to the replacement of HP species (turnover) or their loss (nestedness) from one stigma to another. We further evaluated the potential drivers of variation in HP α and β diversity using phylogenetic structural equation models. We found that variation in HP γ diversity across plant species was driven strongly by differences among species in HP α diversity and to a lesser extent by HP β diversity. HP community turnover contributed more to HP β diversity than nestedness consistently across plant species and seeps, suggesting a general pattern of HP compositional heterogeneity from stigma to stigma. The phylogenetic structural equation models further revealed that floral traits (e.g., stigma area, stigma‐anther distance, stigma exposure) and floral abundance were key in determining HP α diversity by influencing HP abundance (load size), while floral traits and abundance showed variable impact on HP β diversity (turnover and nestedness). Pollination generalism contributed relatively less to HP‐α and β diversity. These findings disentangle the heterogeneity in HP diversity at different levels, which is essential for understanding the process underlying patterns of HP receipt in plant communities. That floral traits drive the heterogeneity in HP diversity points to additional avenues by which HP receipt may contribute to plant evolution. Read the free Plain Language Summary for this article on the Journal blog.

Plants may experience reduced reproductive success at low densities, due to lower numbers of pollinator visits or reduced visit quality. Co-occurring plant species that share pollinators have the potential to facilitate pollination by either increasing numbers of pollinator visits or increasing the quality of visits, but also have the potential to reduce plant reproductive success through competition for pollination. I used a field experiment with a common distylous perennial (Piriqueta caroliniana) in the presence and absence of a co-flowering species (Coreopsis leavenworthii) in plots with one of four different distances between conspecific plants. I found strong negative effects of increasing interplant distance (related to conspecific density) on several components of P. caroliniana reproductive success: pollinator visits to plants per plot visit, visits received by individual plants, conspecific pollen grains on stigmas, outcross pollen grains on stigmas, and probability of fruit production. Although P. caroliniana and C. leavenworthii share pollinators, the co-flowering species did not affect visitation, pollen receipt or reproductive effort in P. caroliniana. Pollinators moved very infrequently between species in this experiment, so floral constancy might explain the lack of effect of the co-flowering species on P. caroliniana reproductive success at low densities. In co-occurring self-incompatible plants with floral rewards, reproductive success at low density may depend more on conspecific densities than on the presence of other species.

Negative effects of heterospecific pollen receipt vary with abiotic conditions: ecological and evolutionary implications

Co-flowering species that have not evolved an avoidance mechanism may have tolerance to heterospecific pollen (HP) deposition as an adaptive strategy to minimize any deleterious effects of HP transfer, but empirical evidence for the tolerance hypothesis remains scarce. To estimate the potential effects of heterospecific pollen deposition (HPD) on female reproductive success, we counted conspecific (CP) and HP pollen grains deposited on stigmas and assessed subsequent seed set of both open- and hand-pollinated flowers in three co-flowering Silene species with exposed stigmas that usually received numerous HP grains on the elongated receptive area. The percentage of HP grains per flower (HP%) varied from 16.6% to 43.0% among three species. Silene chungtienensis had lower HP%, and the CP-HP relationship was neutral; S. gracilicaulis and S. yunnanensis had a relatively higher HP% with a positive CP-HP relationship. The effects of CP and HP number on natural seed set were positive for all three species, but HP% had stronger negative effects in S. chungtienensis and S. gracilicaulis. In hand-pollinated flowers of the three Silene species, seed set did not decrease with HP whether CP was in excess or insufficient, indicating no negative effects of HPD on seed production. Consistent with the tolerance hypothesis, our results indicated that species with higher HP interference are likely to be tolerant to an increase in HP%. These species with generalist-pollinated flowers and exposed large stigmas may benefit from an increase of conspecific pollen deposition, despite the associated increase in heterospecific pollen deposition.

The extragynoecial compitum formed by the incomplete fusion of carpel margins, while allowing intercarpellary growth of pollen tubes in apocarpous angiosperms, may also increase the risk of reproductive interference caused by heterospecific pollen (HP) deposition. In Sagittaria, congeneric HP tubes grow via different paths and enter the ovules later than conspecific pollen (CP) tubes. However, it is unclear how the growth advantage of the CP tube helps ensure reproductive success when HP is deposited on the stigmas. We performed molecular characterization of interspecies-pollinated seeds to examine the consequences of interspecific pollen deposition between Sagittaria pygmaea and S. trifolia. We also conducted CP–HP (1:1) mixed pollination and delayed CP pollination treatments to explore the seed-siring abilities of CP and HP. Our results showed that although HP could trigger the development of fruits, the interspecies-pollinated seeds contained partially developed embryos and could not germinate. More than 70% of the embryos in these seeds were molecularly identified as hybrids of both species, suggesting that HP tubes could enter the ovules and fertilize the egg cells. Moreover, CP could sire more offspring (≥70%) after the CP–HP (1:1) mixed pollination treatment, even when HP reached the stigma 0.5–1 h earlier than CP (≥50%). Following adequate CP vs. HP (1:1) pollination on carpels on two sides of the apocarpous gynoecium, both species produced > 70% conspecific seeds, indicating that the CP tubes could occupy ovules that should be occupied by HP via the extragynoecial compitum. Our results reveal that in Sagittaria, pollen deposition from co-existing congeneric heterospecies leads to interspecific seed discounting. However, the CP advantage mediated by the extragynoecial compitum is an effective strategy to mitigate the effects of interspecific pollen deposition. This study improves our understanding of how apocarpous angiosperms with an extragynoecial compitum can maintain species stability and mitigate the negative reproductive interference effect from sympatrically distributed related species.

ABSTRACTPollinator sharing among coflowering plants can reduce plant fitness through heterospecific pollen (HP) deposition. Compatible HP is expected to cause stronger reproductive interference, but the factors determining HP–pistil compatibility and its reproductive consequences remain poorly understood. Moreover, in some apocarpous taxa, the extragynoecial compitum, a basal pore formed by the incomplete fusion of carpel margins, can enhance reproductive assurance by facilitating conspecific pollen (CP) tube reallocation across free pistils. What remains unclear is how these taxa balance the trade‐off between this benefit and the potential risk of reproductive interference from intercarpellary heterospecific pollen (HP) tube growth. In this study, widely distributed apocarpous Sagittaria trifolia with typical extragynoecial compitum was selected as a pollen recipient. Hand‐pollination experiments were performed with 42 distributed and coflowering HP donors to assess pollen–pistil interaction and HP effects on seed quantity. Results showed that HP–pistil compatibility was independent of species origin (native/alien), pollen size, and aperture number of the HP donor. Instead, it was negatively correlated with phylogenetic distance: closely related HP exhibited higher compatibility with the pistil and caused greater reductions in seed set. Nevertheless, CP consistently sired over 78% of seeds whether applied simultaneously with or after HP. In the half‐and‐half pollination treatments with CP and compatible HP, CP sired significantly more than 50% of seeds. These indicated that CP advantage and intercarpellary CP tube growth collectively mitigate interspecific pollen interference. However, these compensatory mechanisms were less effective when faced with alien congeners. Our results provide a good basis for understanding the variation in HP‐mediated fitness costs and shed light on how apocarpous lineages with extragynoecial compitum adaptively tolerate HP interference and maintain reproductive success.

Generalized pollinators visit multiple co‐flowering plant species and may transfer heterospecific pollen grains. Recent studies have indicated that the effect of heterospecific pollen (HP) on reproduction success is variable and depends on the identity of donor and recipient species. However, few studies have documented variation in HP receipt and evaluated the reproductive effects of HP receipt across geographic locations under natural conditions. We investigated the spatial variation of pollen deposition across eight sites and how the pollen receipt related to the seed set of Salvia przewalskii, a subalpine perennial herb in Hengduan Mountain in southwest China. We found that stigmatic pollen loads substantially varied among sites for several metrics, including quantities of conspecific and heterospecific pollen, the proportion of HP, and species composition of HP donors. Five different plant families were the most common HP source at one or two sites, and the proportion of HP ranged from 3.4% to 51.3% across sites. The association of conspecific pollen with seed set was positive and variable among sites, whereas the association of HP receipt and seed set was negative and not significantly different among sites. Our results demonstrate variation in the quantity and fitness effect of pollen receipt across sites, which is a precondition for evolution of local adaptation. Further study of variation in patterns and effects of HP receipt for the same recipient species across natural communities would allow better understanding of the ecological and evolutionary consequences of HP receipt.

Coflowering plants are at risk for receiving pollen from heterospecifics as well as conspecifics, yet evidence shows wide variation in the degree that heterospecific pollen transfer occurs. Evaluation of patterns and correlates of among- and within-species variation in heterospecific pollen (HP) receipt is key to understanding its importance for floral evolution and species coexistence; however, the rarity of deeply sampled multispecies comparisons has precluded such an evaluation. We evaluated patterns of among- and within-species variation in HP load size and diversity in 19 species across three distinct plant communities. We assessed the importance of phenotypic specialization (floral phenotype), ecological specialization (contemporary visitor assemblage), and conspecific flower density as determinants of among-species variation. We present hypotheses for different accrual patterns of HP within species based on the evenness and quality of floral visitors and evaluated these by characterizing the relationship between conspecific pollen (CP) and HP receipt. We found that within-species variation in HP receipt was greater than among-species and among-communities variation. Among species, ecological generalization emerged as the strongest driver of variation in HP receipt irrespective of phenotypic specialization. Within-species variation in HP load size and diversity was predicted most often from two CP-HP relationships (linear or exponentially decreasing), suggesting that two distinct types of plant-pollinator interactions prevail. Our results give important insights into the potential drivers of among- and within-species variation in HP receipt. They also highlight the value of explorations of patterns at the intraspecific level, which can ultimately shed light on plant-pollinator-mediated selection in diverse plant communities.

PremiseHeterospecific pollen transfer, the transfer of pollen between species, is common among co‐flowering plants, yet the amount of pollen received is extremely variable among species. Intraspecific variation in heterospecific pollen receipt can be even greater, but we lack an understanding of its causes and fitness consequences in wild populations.MethodsWe examined potential drivers of variation in heterospecific pollen receipt in Oenothera fruticosa. We evaluated the relationship between heterospecific and conspecific pollen receipt and considered how visitation by different pollinator groups, local floral neighborhood composition, and flowering phenology affect the total amount and proportion of heterospecific pollen received. Finally, we tested whether variation in heterospecific pollen receipt translated into lower seed production.ResultsHeterospecific pollen was ubiquitous on O. fruticosa stigmas, but the amount received was highly variable and unrelated to conspecific pollen receipt. Heterospecific pollen receipt depended on pollinator type, the proportion of nearby conspecific flowers, and flowering date. Significant interactions revealed that the effects of pollinator type and neighborhood were not independent, further contributing to variation in heterospecific pollen. Naturally occurring levels of heterospecific pollen were sufficient to negatively impact seed set, but large amounts of conspecific pollen counteracted this detrimental effect.ConclusionsAlthough selection could act on floral traits that attract quality pollinators and promote synchronous flowering in O. fruticosa, the risk of heterospecific pollen is equally dependent on local floral context. This work highlights how extrinsic and intrinsic factors contribute to intraspecific variation in heterospecific pollen receipt in wild plants, with significant fitness consequences.

Plant species vary greatly in the degree to which floral morphology restricts access to the flower interior. Restrictiveness of flower corollas may influence heterospecific pollen receipt, but the impact of floral morphology on heterospecific pollen transfer has received little attention. We characterized patterns of pollinator visitation and quantities of conspecific and heterospecific pollen receipt for 29 species with a range of floral morphologies in a prairie community dominated by the introduced plant Euphorbia esula (leafy spurge) which has an unrestrictive morphology. Pollinator overlap was significantly greater between Euphorbia and other unrestrictive flowers than restrictive flowers. Compared to flowers with restrictive morphologies, unrestrictive flowers received significantly more Euphorbia pollen, more heterospecific pollen from other sources, and a greater diversity of pollen species, but not more conspecific pollen. However, stigmatic surface area was significantly larger for flowers with unrestrictive morphologies, and the density of Euphorbia and other heterospecific pollen per stigmatic area did not significantly differ between flower types. These findings suggest that the smaller stigma size in restrictive flowers partly accounts for their decreased heterospecific pollen receipt, but that restrictiveness also allows species to increase the purity of pollen loads they receive. Given that restrictive flowers receive fewer heterospecific pollen grains but at a higher density, the effect of restrictiveness on fecundity depends on whether absolute quantity or density of heterospecific pollen affects fecundity more. Our results also indicate that abundant neighbors are not necessarily important heterospecific pollen sources since Euphorbia pollen was rarely abundant on heterospecifics.

Reproductive interference is a negative interspecific interaction that can drive mutually exclusive distributions of closely related species. While recent research indicates that reproductive interference in plants frequently occurs during pollen-pistil interactions, comprehensive descriptions of these interactions are scarce. Understanding the mechanisms underlying reproductive interference requires studies integrating empirical observations with interaction analyses. This study investigates pollen-pistil interactions between three Veronica species recently observed to exhibit asymmetric reproductive interference.Our experiments revealed similar pollen tube behavior in V. polita var. lilacina and V. cymbalaria pistils, irrespective of whether they were pollinated with conspecific or V. persica pollen. Conversely, in V. persica pistils, the number of pollen tubes significantly decreased following heterospecific pollination compared to conspecific pollination. Furthermore, half of the pollen grains on V. persica stigmas, presumably heterospecific pollen grains, occasionally appeared non-luminous under the fluorescence microscope after mixed pollination. Conspecific pollen tubes appeared to grow faster within V. persica pistils; however, statistical analysis did not support this trend.These results suggest that V. polita var. lilacina and V. cymbalaria pistils exhibit limited discrimination against V. persica pollen, resulting in ovule discounting by the heterospecific pollen. This mechanism explains the empirical observation that these species experience reduced seed set after mixed pollination, even when conspecific pollen is applied first. In contrast, V. persica pistils demonstrate a degree of discrimination between conspecific and heterospecific pollen, particularly when conspecific pollen arrives on the stigma prior to heterospecific pollen. This selectivity accounts for the empirical findings that V. persica exhibits reduced seed set only when surrounded by V. polita var lilacina or after mixed pollination with V. cymbalaria pollen preceding conspecific pollen.