Interspecific Pollen Transfer Research Articles

Post-pollination barriers contribute to coexistence of partially pollinator-sharing Arisaema species (Araceae).

Reproductive isolation plays an important role in maintaining the species integrity of sympatric close relatives. For sympatric Arisaema species, interspecific gene flow is expected to be effectively prevented by pre-pollination barriers, particularly strong pollinator isolation mediated by fungus gnats. However, due to the lack of quantitative studies combining multiple pre- and post-pollination barriers, it is not known whether pre-pollination isolation is complete, and whether post-pollination barriers also contribute to reproductive isolation among some Arisaema species. In this study, we quantified the individual strengths and absolute contributions of four pre- and post-pollination barriers (phenological isolation, pollinator isolation, hybrid fruit formation, and hybrid seed formation) among three sympatric Arisaema species (A. bockii, A. lobatum, and A. erubescens). Although phenological isolation and pollinator isolation reduced the frequencies of interspecific pollen transfer among these species, the partial overlap of flowering times and pollinator assemblages resulted in incomplete pre-pollination isolation. Post-pollination barriers also contributed to reproductive isolation at the hybrid fruit and seed formation stages. We propose that, although pre-pollination barriers are expected to contribute more to total isolation than post-pollination barriers in Arisaema, pre-pollination barriers may not completely prevent interspecific pollen transfer among some Arisaema species. Post-pollination barriers, which are generally ignored, may also have contributed significantly to reproductive isolation in Arisaema.

Incomplete partitioning of pollinators by Linum suffruticosum and its coflowering congeners.

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. 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. 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. 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.

Interspecific variation in plant abundance can contribute to the diversity and evolutionary assembly of floral communities

Angiosperms show remarkable floral diversity in pollinator use. However, how this diversity evolves in a community context remains poorly understood. In this article I propose that different plant species abundances promote adaptation to different pollinators and different degrees of specialization. In this view, interspecific variation in species abundance can foster floral diversity. I develop a mathematical model of pollen transfer considering the interaction of several pollination processes – pollen removal and carryover, intra‐ and interspecific competition for pollinator visitation, and interspecific pollen transfer – that are linked to floral abundance. I integrate the model to an adaptive dynamic framework to simulate the evolutionary assembly of pollination networks. The model demonstrates that different pollinator attributes and degrees of generalization are favored at different plant relative abundances. Within communities, interspecific variation in plant abundance increased diversity in pollinator use and degree of generalization while also promoting the evolution of pollination networks with higher nestedness and lower connectance, leading to networks more consistent with natural pollination networks. The model helps understand the evolutionary assembly of flower communities and suggests a new mechanism by which floral diversity can be generated, contributing to our understanding of floral evolution and diversification.

Pollinator sharing between reproductively isolated genetic lineages of Silene nutans.

High reciprocal pollination specialization leading to pollinator isolation can prevent interspecific pollen transfer and competition for pollinators. Sharing pollinators may induce mating costs, but it may also increase pollination services and pollen dispersal and offer more resources to pollinators, which may be important in case of habitat fragmentation leading to pollination disruption. We estimated pollen dispersal and pollinator isolation or sharing between two reproductively isolated genetic lineages of Silene nutans (Caryophyllaceae), which are rare and occur in parapatry in southern Belgium, forming two edaphic ecotypes. As inter-ecotypic crosses may lead to pollen wastage and inviable progeny, pollinator isolation might have evolved between ecotypes. Silene nutans is mainly pollinated by nocturnal moths, including nursery pollinators, which pollinate and lay their eggs in flowers, and whose caterpillars feed on flowers and seeds. Pollinator assemblages of the two ecotypes are largely unknown and inter-ecotypic pollen flows have never been investigated. Fluorescent powdered dyes were used as pollen analogues to quantify intra- and inter-ecotypic pollen transfers and seeds were germinated to detect chlorotic seedlings resulting from inter-ecotypic pollination. Nocturnal pollinators were observed using infrared cameras on the field, and seed-eating caterpillars were collected and reared to identify nursery pollinator species. No pollinator isolation was found: we detected long-distance (up to 5km) inter-ecotypic dye transfers and chlorotic seedlings, indicating inter-ecotypic fertilization events. The rare moth Hadena albimacula, a nursery pollinator specialized on S. nutans, was found on both ecotypes, as well as adults visiting flowers (cameras recordings) as seed-eating caterpillars. However, S. nutans populations harbor different abundance and diversity of seed predator communities, including other rare nursery pollinators, suggesting a need for distinct conservation strategies. Our findings demonstrate the efficiency of moths, especially of nursery pollinators, to disperse pollen over long distances in natural landscapes, so to ensure gene flow and population sustainability of the host plant. Seed-predator specificities between the two reproductively isolated genetic lineages of S. nutans, and pollinator sharing instead of pollinator isolation when plants occur in parapatry, suggest that conservation of the host plant is also essential for sustaining (rare) pollinator and seed predator communities.

The selfing syndrome and beyond: diverse evolutionary consequences of mating system transitions in plants.

The shift from outcrossing to self-fertilization (selfing) is considered one of the most prevalent evolutionary transitions in flowering plants. Selfing species tend to share similar reproductive traits in morphology and function, and such a set of traits is called the 'selfing syndrome'. Although the genetic basis of the selfing syndrome has been of great interest to evolutionary biologists, knowledge of the causative genes or mutations was limited until recently. Thanks to advances in population genomic methodologies combined with high-throughput sequencing technologies, several studies have successfully unravelled the molecular and genetic basis for evolution of the selfing syndrome in Capsella, Arabidopsis, Solanum and other genera. Here we first introduce recent research examples that have explored the loci, genes and mutations responsible for the selfing syndrome traits, such as reductions in petal size or in pollen production, that are mainly relevant to pre-pollination processes. Second, we review the relationship between the evolution of selfing and interspecific pollen transfer, highlighting the findings of post-pollination reproductive barriers at the molecular level. We then discuss the emerging view of patterns in evolution of the selfing syndrome, such as the pervasive involvement of loss-of-function mutations and the relative importance of selection versus neutral degradation. This article is part of the theme issue 'Genetic basis of adaptation and speciation: from loci to causative mutations'.

Effects of heterospecific pollen on stigma behavior in Campsis radicans: Causes and consequences.

Pollinator sharing of co-flowering plants may result in interspecific pollen receipt with a fitness cost. However, the underlying factors that determine the effects of heterospecific pollen (HP) are not fully understood. Moreover, the cost of stigma closure induced by HP may be more severe for plants with special touch-sensitive stigmas than for plants with non-touch-sensitive stigmas. Very few studies have assessed HP effects on stigma behavior. We conducted hand-pollination experiments with 10 HP donors to estimate HP effects on stigma behavior and stigmatic pollen germination in Campsis radicans (Bignoniaceae) at low and high pollen loads. We assessed the role of phylogenetic distance between donor and recipient, pollen size, and pollen aperture number in mediating HP effects. Additionally, we observed pollen tube growth to determine the conspecific pollen-tube-growth advantage. Stigma behavior differed significantly with HP of different species. Pollen load increased, while pollen size decreased, the percentage of permanent closure and stigmatic germination of HP. Stigmatic HP germination increased with increasing aperture number. However, HP effects did not depend on phylogenetic distance. In addition, conspecific pollen had a pollen-tube-growth advantage over HP. Our results provide a good basis for understanding the stigma-pollen recognition process of plant taxa with touch-sensitive stigmas. We concluded that certain flowering traits drive the HP effects on the post-pollination period. To better understand the impact of pollinator sharing and interspecific pollen transfer on plant evolution, we highlight the importance of evaluating more factors that determine HP effects at the community level.

Pollinators contribute to the maintenance of flowering plant diversity

Mechanisms that favour rare species are key to the maintenance of diverse communities1-3. One of the most critical tasks for conservation of flowering plant biodiversity is to understand how plant-pollinator interactions contribute to the maintenance of rare species4-7. Here we show that niche partitioning in pollinator use and asymmetric facilitation confer fitness advantage of rarer species in a biodiversity hotspot using phylogenetic structural equation modelling that integrates plant-pollinator and interspecific pollen transfer networks with floral functional traits. Co-flowering species filtered pollinators via floral traits, and rarer species showed greater pollinator specialization leading to higher pollination-mediated male and female fitness than more abundant species. When plants shared pollinator resources, asymmetric facilitation via pollen transport dynamics benefitted the rarer species at the cost of more abundant species, serving as an alternative diversity-promoting mechanism. Our results emphasize the importance of community-wide plant-pollinator interactions that affect reproduction for biodiversity maintenance.

Relationship between interspecific pollen transfer and pistil length in sympatric congeners, Clerodendrum trichotomum and C. izuinsulare

AbstractInterspecific pollen transfer (IPT) often leads to reproductive interference. Although character displacement of reproductive traits in plants is often considered a consequence of reproductive interference, few studies have tested whether intraspecific variation in floral morphology changes the intensity of reproductive interference among pollinator‐sharing plants. We investigated whether intraspecific variation in pistil length changes the proportion of IPT (interspecific pollen transfer) in sympatric populations of Clerodendrum trichotomum and C. izuinsulare on the two islands. On Toshima Island, C. izuinsulare flowers with shorter pistils were significantly less likely to receive C. trichotomum pollen, and there was a slight tendency for longer pistiles of C. trichotomum flowers to receive less C. izuinsulare pollen, suggesting that IPT has caused character displacement in pistil length in these congeneric plants. In contrast, we did not detect any relationship between pistil length and the proportion of IPT for the two species on Niijima Island. The discrepancy between the islands may reflect differences in pollinator assemblages.

Moth versus fly: a preliminary study of the pollination mode of two species of endemic Asteraceae from St Helena (Commidendrum robustum and C. rugosum) and its conservation implications.

Commidendrum robustum (Roxb.) DC. (St Helena gumwood) and C. rugosum (Dryand.) DC. (St Helena scrubwood) are ecologically important, endemic woody Asteraceae from the isolated South Atlantic island of St Helena. Once very abundant, they now exist in sparse fragmented populations due to 500 years of environmental destruction. They are sister taxa that evolved on the island and are reported to hybridise. Commidendrum rugosum has a saucer-like erect capitulum, whereas C. robustum has a somewhat globular hanging capitulum. Using daytime timelapse photography to follow capitula through their life cycle, we found that C. rugosum appears to be myophilous, visited largely by flies (including the endemic syrphid, Sphaerophoria beattiei Doesburg & Doesburg) and occasionally by Lepidoptera. Commidendrum robustum, on the other hand, although visited by flies, strongly attracts moths (especially noted at the Millennium Forest site). Our data suggest that moth visits may reduce visits from flies due to the sensitivity of flies to interference by other insects. We conclude that C. robustum may have a mixed syndrome of myophily/phalaenophily and that there is apparently some divergence of the pollination niche between the two species. Its potential in attracting moths, coupled with its former abundance, suggests that it may have been a major food source for adults of the numerous endemic moths. Pollinator activity was measured by insect visitation rates (mean visits per capitulum per day, V) and insect residence time (mean pollinator kiloseconds per capitulum per day, R). Both are higher for C. robustum (C. rugosum, V = 16.4, R = 3.101; C. robustum, V = 34.0, R = 8.274), reflecting the abundance of moths on the capitula at the Millennium Forest site. The conservation implications of the pollination mode are that: (1) there is considerable pollinator activity on the capitula and pollination is not currently a limiting factor for plant reproduction; (2) gene exchange between geographically-isolated populations of C. rugosum is likely to be minimal due to the apparent reliance of the species for pollination on small flies (especially Sphaerophoria beattiei), which are believed to be not effective as pollinators over long distances (> 1 km). A possible exception is the strong-flying drone-fly, Eristalis tenax Linn. which, although not as abundant as Sphaerophoria, does visit the flowers; (3) there is considerable overlap between the two species in flower visitors and interspecific pollen transfer is possible where the two species grow intermixed (which has potential positive and negative implications for species survival).

Character displacement drives floral variation in Pelargonium (Geraniaceae) communities.

Interactions between plant community members are an underexplored driver of angiosperm floral variation. We investigate character displacement as a potential contributor to floral variation in Pelargonium communities. Pelargoniums all place pollen on the ventral sides of their pollinators, potentially leading to interspecific pollen transfer (IPT) in sympatry. We show that the positions of pollen placement and receipt are determined by anther and style exsertion lengths. Using field experiments, we demonstrate that heterospecific species experience higher IPT if they have similar style lengths than when they have greater style length differences. Using crosses, we show that IPT has negative consequences on seed set. In combination, these results suggest that character displacement in style length is likely to reduce IPT and increase female fitness in sympatry. Patterns of style length variation across 29 different Pelargonium communities suggest that character displacement has occurred in multiple communities. Furthermore, analyses using a wide-ranging species pair show that style lengths are more different between sympatric populations than they are between allopatric populations. In addition to pollinators as agents of floral divergence, this study suggests that variation in Pelargonium community structure has driven style length variation through character displacement.

Importance of Pollinator-Mediated Interspecific Pollen Transfer for Angiosperm Evolution

Understanding how pollen moves between species is critical to understanding speciation, diversification, and evolution of flowering plants. For co-flowering species that share pollinators, competition through interspecific pollen transfer (IPT) can profoundly impact floral evolution, decreasing female fitness via heterospecific pollen deposition on stigmas and male fitness via pollen misplacement during visits to heterospecific flowers. The pollination literature demonstrates that such reproductive interference frequently selects for reproductive character displacement in floral traits linked to pollinator attraction, pollen placement, and mating systems and has also revealed that IPT between given pairs of species is typically asymmetric. More recent work is starting to elucidate its importance to the speciation process, clarifying the link between IPT and current and historical patterns of hybridization, the evolution of phenotypic novelty through adaptive introgression, and the rise of reproductive isolation. Our review aims to stimulate further research on IPT as a ubiquitous mechanism that plays a central role in angiosperm diversification.

Reproductive isolation among three sympatric Achimenes species: pre- and post-pollination components.

Closely related species occurring in sympatry may experience the negative consequences of interspecific pollen transfer if reproductive isolation (RI) barriers are not in place. We evaluated the importance of pre- and post-pollination RI barriers in three sympatric species of Achimenes (Gesneriaceae), including ecogeographic, phenological, floral isolation, self-pollination, and hybrid viability (fruit and seed set). We recorded geographic distribution throughout species ranges and assessed flowering phenology and pollinator visitation at one site in central Mexico. In the greenhouse, we measured floral traits involved in RI and quantified fruit and seed set for from self, intraspecific, and interspecific crosses. Ecogeographic barriers were important in RI, but under sympatry, phenological and floral barriers contributed more to total RI. Phenological RI varied between species and years, while floral RI was 100% effective at preventing interspecific visitation. Species showed differences in floral morphology, color, and scents associated with specialized pollination systems (A. antirrhina-hummingbirds, A. flava-bees, A. patens-butterflies); heterospecific visitation events were restricted to rare secondary pollinators. Hybrid crosses consistently yielded progeny in lower numbers than intraspecific crosses. This study indicated that neither autogamy nor early post-pollination barriers prevent interspecific pollen flow between Achimenes species. However, floral isolation, acting through a combination of attraction and reward traits, consistently ensures specificity of the pollination system. These results suggest that selection on floral traits to reduce the costs of hybrid progeny production may have played a role in evolution or maintenance of specialized pollination systems in Achimenes.

Prior selfing can mitigate the negative effects of mutual reproductive interference between coexisting congeners

Abstract When more than one closely related plant species share the same pollination niche, reproductive interference via interspecific pollen transfer should limit their coexistence. However, some studies have reported the sympatric coexistence of two native close relatives pollinated by the same pollinators under reproductive interference, even without niche partitioning. We examined the frequency dependency of reproductive interference between close relatives in natural conditions and the potential roles of autonomous selfing in mitigating the negative reproductive interference effects between congeneric species. We investigated sympatrically growing Commelina communis (Cc) and C. c. f. ciliata (Ccfc) populations. These species exhibit very large overlaps in habitat preference, flowering phenology and pollination niche, but seldom produce hybrids. First, we conducted a hand‐pollination experiment to examine the negative effects of heterospecific pollen deposition on seed production and the potential of self‐pollination to mitigate the effects in both species. Then, we examined the effects of reproductive interference on reproductive success and the potential for autonomous selfing in the field. We found significant negative effects of heterospecific pollen deposition on seed production and the mitigation effects of prior and competing self‐pollination, in both Cc and Ccfc. For both species in the field, intraspecific pollinator movements and reproductive success significantly decreased with an increase in the relative floral abundance of competing species, although the negative reproductive interference effect on reproductive success was lower in Cc than in Ccfc. We also found greater potential for prior autonomous selfing in Cc than in Ccfc. Our findings suggest that Cc flowers were less affected by reproductive interference from competing species, which was likely due to a higher prior selfing ability compared to Ccfc flowers. The asymmetry in susceptibility to reproductive interference may explain the Cc‐biased distribution in the study area. The study improves our understanding of how prior autonomous selfing can reduce the negative reproductive interference effect from competing species in mixed‐mating species with frequent pollinator visits. A plain language summary is available for this article.

A gradient of pollination specialization in three species of Bolivian Centropogon.

Closely related plant species with overlapping ranges often experience competition for pollination services. Such competition can select for divergence in floral traits that attract pollinators or determine pollen placement. While most species in Centropogon (Campanulaceae: Lobelioideae) have flowers that suggest adaptation to bat or hummingbird pollination, actual pollinators are rarely documented, and a few species have a mix of traits from both pollination syndromes. We studied the pollination biology of a "mixed-syndrome" species and its co-occurring congeners to examine the relationship between floral traits and visitation patterns for Centropogon. Fieldwork at two sites in Bolivian cloud forests involved filming floral visitors, quantifying pollen transfer, and measuring floral traits. Stamen exsertion, which determines pollen placement, was measured from herbarium specimens across the geographic range of these species to test for character displacement. Results show a generalization gradient, from primarily bat pollination in white-flowered Centropogon incanus, to bat pollination with secondary hummingbird pollination in the cream-flowered C. brittonianus, to equal reliance on both pollinators in the red-flowered, mixed-syndrome C. mandonis. Pollen transfer between these species is further reduced by differences in stamen exsertion that are accentuated in zones of sympatry, a pattern consistent with character displacement. Our results demonstrate that key differences in floral color and shape mediate a gradient of specialization in Bolivian Centropogon. Interspecific pollen transfer is further reduced by potential character displacement of a key trait. Broadly, our results have implications for understanding the hyper-diversity of Andean cloud forests, in which multiple species of the same genus frequently co-occur.

Evidence of temporal niche separation via low flowering time overlap in an old-field plant community.

Flowering time is a trait that reflects the timing of specific resource requirements by plants. Consequently, several predictions have been made related to how species are assembled within communities according to flowering time. Strong overlap in flowering time among coexisting species may result from clustered abiotic resources, or contribute to improved pollination success. Conversely, low flowering time overlap (asynchrony) among coexisting species may reduce competition for soil, light, or pollinator resources and alleviate interspecific pollen transfer. Here, we present evidence that coexisting species in an old-field community generally overlap less in flowering time than expected under a commonly used and statistically validated null model. Flowering time asynchrony was more pronounced when abundance data were used (compared to presence-absence data), and when analyses focused on species that share bees as pollinators. Control and herbivore-exclusion plots did not differ in flowering time overlap, providing no evidence of the reduction in overlap expected to result from increased competition. Our results varied with the randomization algorithm used, emphasizing that the choice of algorithm can influence the outcome of null models. Our results varied between 2years, with patterns being less clear in the second year, when both growing season and flowering times were contracted. Finally, we found evidence that further supports a previous finding that higher plot-level flowering time overlap was associated with higher proportions of introduced species. Reduced flowering time overlap among species in our focal community may promote coexistence via temporal niche differentiation and reduced competition for pollinators and other abiotic resources.

Variation in sampling effort affects the observed richness of plant-plant interactions via heterospecific pollen transfer: implications for interpretation of pollen transfer networks.

There is growing interest in understanding plant-plant interactions via pollen transfer at the community level. Studies on the structure and spatial variability of pollen transfer networks have been valuable to this understanding. However, there is high variability in the intensity of sampling used to characterize pollen transfer interactions, which could influence network structure. To date, there is no knowledge of how sampling effort influences the richness of pollen on stigmas and thereby transfer interactions observed, nor how this may vary across species and study sites. We use rarefaction curves on 16 species to characterize the relationship between sampling effort (number of stigmas analyzed) and the richness of pollen transfer interactions recorded. We further assess variability in this relationship among species, plant community types, and sites within a single plant community. We show high among-species variation in the amount of sampling required to sufficiently characterize interspecific pollen transfer. We further reveal variability in the sampling effort-interaction richness relationship among different plant communities and even for the same species growing in different sites. The wide heterogeneity in the sampling effort required to accurately characterize pollen transfer interactions observed has the potential to influence the characterization of pollen transfer dynamics. Thus, sampling completeness should be considered in future studies to avoid overestimation of modularity and specialization in pollen transfer networks that may bias the predicted causes and expected consequences of such processes for plant-plant interactions.

Impact of pre- and post-pollination barriers on pollen transfer and reproductive isolation among three sympatric Pedicularis (Orobanchaceae) species.

Pedicularis is the largest genus in the Orobanchaceae (>300) with many species co-occurring and co-blooming in subalpine to alpine meadows in the Himalayas. Although it is well known that different Pedicularis species place pollen on different parts of the same bumblebee's body, thus reducing interspecific pollen transfer, it is not known whether post-pollination components also contribute to reproductive isolation (RI). In this study, we quantified the individual strengths and absolute contributions of six pre- and post-pollination components of RI between three sympatric species in two pairs; Pedicularis gruina×Pedicularis tenuisecta (gru×ten) and Pedicularis comptoniifolia×Pedicularis tenuisecta (com×ten). All three Pedicularis species shared the same Bombus species. Individual foragers showed a high, but incomplete, floral constancy for each species. Therefore, pre-pollination barriers were potentially 'leaky' as Bombus species showed a low but consistent frequency of interspecific visitation. The RI strength of pre-pollination was lower in com×ten than in gru×ten. In contrast, post-pollination barriers completely blocked gene flow between both sets of species pairs. Two post-pollination recognition sites were identified. Late acting rejection of interspecific pollen tube growth occurred in com♀×ten♂, while seeds produced in bi-directional crosses of gru×ten failed to germinate. We propose that, although floral isolation based on pollen placement on pollinators in the genus Pedicularis is crucial to avoid interspecific pollen transfer, the importance of this mode of interspecific isolation may be exaggerated. Post-pollination barriers may play even larger roles for currently established populations of co-blooming and sympatric species in this huge genus in the Himalayas.

The effect of the dispersion of rewarding and rewardless flowers on visitation and constancy by bumblebees (<i>Bombus impatiens</i>)

Deceptive pollination strategies, in which plants rely on animal pollinators but provide no benefits to their pollinators, have evolved many times in angiosperms. However, the conditions that favour deceptive pollination strategies over mutualistic strategies are poorly understood. One factor that may be important for the success of deceptive strategies is the plants’ dispersion in relation to co-flowering, rewarding species. We compared pollinator behaviour across two artificial environments, one in which a rewardless species was mixed with a rewarding species and one in which the two species occurred in contiguous patches. Bumblebees both encountered and visited rewardless flowers more often when they were mixed with rewarding species. However, the rate of switching was also higher under those conditions, which could result in higher rates of interspecific pollen transfer. The environmental conditions most favourable to deceptive pollination strategies may vary depending on the vulnerability of the plant species to interspecific pollen transfer.

Mutualism between co-occurring plant species in South Africa's Mediterranean climate heathland is mediated by birds.

Interactions among plant species via pollinators vary from competitive to mutualistic and can influence the probability of stable coexistence of plant species. We aimed to determine the nature of the interaction via flower visitors between Leucospermum conocarpodendron and Mimetes fimbriifolius, two shrubs in the Proteaceae that share many ecological traits and coexist on the Cape Peninsula, South Africa. To assess the extent of pollinator sharing we analysed nectar properties and recorded the pollinator fauna, their behaviour and contribution to seed set. To test for competition via interspecific pollen transfer, we recorded the movement patterns of pollinators and quantified pollen loads. To determine the effect of co-flowering on visitation rates we recorded visits in stands that varied in the density of the two species. We found that the species produce similar rewards and share pollinating Cape Sugarbirds (Promerops cafer). Interspecific pollen transfer is avoided by placing pollen on different parts of the bird. Both species are visited by nectar-thieving Orange-breasted Sunbirds (Anthobaphes violacea). Insects and autonomous self-pollination contributed little to seed set. Pollinator visits increased with conspecific density in both species, and the slope of the increase was steepest in the presence of high densities of the co-occurring plant species. Nectar thief visits also increased with conspecific density in both species, but the slope declined with increasing density of the co-occurring species. Co-occurrence enhanced pollinator visits and alleviated nectar robbing in both plant species, consistent with mutualisms. Mutualism within a trophic level is unusual, but may help to explain the stable coexistence of ecologically similar species.

Low interspecific pollen transfer between invasive aquatic Ludwigia grandiflora and native co-flowering plants

Showy invasive alien plants are often integrated in the diet of generalist pollinators and because of the lack of co-evolvement with the native plant community, a high amount of interspecific pollen transfer (IPT) can be expected. We investigated pollinator switching and magnitude plus distance of IPT between the alien aquatic Ludwigia grandiflora and the native Lythrum salicaria in both directions in uninvaded and invaded sites with a different relative abundance of L. grandiflora (% cover of the alien plant: no cover; low cover: <5%; high cover: 50–75%). A field experiment was conducted to include both pollinator interspecific movements and tracking of IPT, using fluorescent dye as a pollen analogue. Despite a substantial overlap in pollinators between L. grandiflora and the native L. salicaria, less than 10% of the observed flights were interspecific. Similar results were found in dye transfer patterns. The proportions of stigmas with conspecific dye were always higher than the proportions of stigmas with heterospecific dye for L grandiflora and L. salicaria. There were no differences in conspecific dye loads for L. salicaria between uninvaded and invaded sites. Conspecific pollen loss (native CPL) and heterospecific pollen deposition (alien HPD) were in general low and species-specific. The distance of HPD ranged respectively from 1.7 to 39 m and from 0.3 to 54.8 m in the low cover and high cover sites while CPL ranged respectively from 6.40 to 68.02 m and from 0.60 to 40.18 m in the low cover and high cover sites. We can conclude that, in this system, CPL and HPD will play a minor role in pollinator-mediated interaction. Furthermore, interspecific competition for pollinators will cover a larger distance than just neighboring individuals. Our results suggest the necessity to consider the combined effect of insect visitation, pollen deposition, relative alien abundance, distance and seed set when investigating pollinator-mediated interactions of invasive plants.