The timing of visits by large and small bees differentially affects pollination success in Mimulus ringens.

PREMISE: Cross-fertilization in most flowering plants is facilitated by mobile animals that transport pollen while foraging for floral rewards. The contributions of different visitors can vary widely, depending on the amount of pollen transferred during a single visit and on the frequency and timing of the visits of each pollinator taxon. METHODS: We used three approaches to measure the pollination value of bees that visit Mimulus ringens: pollinator interviews, field population observations, and caging studies. RESULTS: The single-visit effectiveness of small bees (primarily Halictidae) was only half that of larger bees (primarily Bombus) for pollen delivery and removal. In five field populations, we found substantial temporal and spatial variation in visitation and pollination. In most sites big bees were active before 08:00 hours, and by 10:00-11:00 hours, stigmas were usually fully pollinated and closed, and little pollen remained in anthers. Small bees seldom visited before 10:00 hours. Excluding big bees from plants confirmed that pollination is reduced and delayed in this ecological context. CONCLUSIONS: Big bees are the primary pollinators of M. ringens, accounting for at least 75% of seed production. Not only are they more effective per visit, in most situations they also visit before small bees become active. Although small bees are not usually important pollinators of M. ringens, they have the potential to partially replace them as a "fail-safe" pollinator in contexts where big bees are not abundant. In a world where pollinator abundance is declining, such backup pollinators may be important for maintaining plant reproduction.

Among-individual variation in flowering phenology affects flowering synchrony and mating opportunity.

PREMISE: The timing and pattern of a plant's flowering can have important consequences for reproductive success. Variation in flowering phenology may influence the number of prospective mates, the risk of mating with lower quality individuals, and the likelihood of self-pollination. Here we use a common garden experiment to explore within- and among-population variation in phenology. Our work provides new insights into how flowering phenology shapes mating opportunity and flowering synchrony in a self-compatible perennial. METHODS: To quantify variation in flowering phenology we raised progeny from nine populations of Mimulus ringens in a common garden. For each individual, we measured phenological traits including age at flowering onset, daily floral display size, total flower number, and flowering synchrony with other members of the population, and related these traits to mating opportunity. We also tested how individual flowering schedules influence the magnitude of synchrony. RESULTS: Flowering phenology and synchrony varied substantially within and among populations. From day to day, plants often oscillated between large and small daily floral displays. Additionally, flowering schedules of individual plants strongly influenced flowering synchrony and, along with the number of flowering days, markedly affected plants' mating opportunity. CONCLUSIONS: Phenological traits such as flowering synchrony can affect the quantity of mating opportunities and may be important targets of natural selection. Our results highlight the need for studies that quantify flowering patterns of individuals as well as populations.

Selfing rates vary with floral display, pollinator visitation and plant density in natural populations of Mimulus ringens.

Variation in selfing rates within and among populations of hermaphroditic flowering plants can strongly influence the evolution of reproductive strategies and the genetic structure of populations. This intraspecific variation in mating patterns may reflect both genetic and ecological factors, but the relative importance of these factors remains poorly understood. Here, we explore how selfing in 13 natural populations of the perennial wildflower Mimulus ringens is influenced by (a) pollinator visitation, an ecological factor, and (b) floral display, a trait with a genetic component that also responds to environmental variation. We also explore whether genetically based floral traits, including herkogamy, affect selfing. We found substantial variation among populations in selfing rate (0.13-0.55). Selfing increased strongly and significantly with floral display, among as well as within populations. Selfing also increased at sites with lower pollinator visitation and low plant density. However, selfing was not correlated with floral morphology. Overall, these results suggest that pollinator visitation and floral display, two factors that interact to affect geitonogamous pollinator movements, can influence the selfing rate. This study identifies mechanisms that may play a role in maintaining selfing rate variation among populations.

Pollination intensity and paternity in flowering plants.

BACKGROUND: Siring success plays a key role in plant evolution and reproductive ecology, and variation among individuals creates an opportunity for selection to act. Differences in male reproductive success can be caused by processes that occur during two stages, the pollination and post-pollination phases of reproduction. In the pollination phase, heritable variation in floral traits and floral display affect pollinator visitation patterns, which in turn affect variation among plants in the amount of pollen exported and deposited on recipient stigmas. In the post-pollination phase, differences among individuals in pollen grain germination success and pollen tube growth may cause realized paternity to differ from patterns of pollen receipt. The maternal plant can also preferentially provision some developing seeds or fruits to further alter variation in siring success. SCOPE: In this review, we describe studies that advance our understanding of the dynamics of the pollination and post-pollination phases, focusing on how variation in male fitness changes in response to pollen limitation. We then explore the interplay between pollination and post-pollination success, and how these processes respond to ecological factors such as pollination intensity. We also identify pressing questions at the intersection of pollination and paternity and describe novel experimental approaches to elucidate the relative importance of pollination and post-pollination factors in determining male reproductive success. CONCLUSIONS: The relative contribution of pollination and post-pollination processes to variation in male reproductive success may not be constant, but rather may vary with pollination intensity. Studies that quantify the effects of pollination and post-pollination phases in concert will be especially valuable as they will enable researchers to more fully understand the ecological conditions influencing male reproductive success.

Hermaphroditism promotes mate diversity in flowering plants.

PREMISE: Genetically diverse sibships are thought to increase parental fitness through a reduction in the intensity of sib competition, and through increased opportunities for seedling establishment in spatially or temporally heterogeneous environments. Nearly all research on mate diversity in flowering plants has focused on the number of fathers siring seeds within a fruit or on a maternal plant. Yet as hermaphrodites, plants can also accrue mate diversity by siring offspring on several pollen recipients in a population. Here we explore whether mate composition overlaps between the dual sex functions, and discuss the implications for plant reproductive success. METHODS: We established an experimental population of 49 Mimulus ringens (monkeyflower) plants, each trimmed to a single flower. Following pollination by wild bees, we quantified mate composition for each flower through both paternal and maternal function. Parentage was successfully assigned to 240 progeny, 98% of the sampled seeds. RESULTS: Comparison of mate composition between male and female function revealed high mate diversity, with almost no outcross mates shared between the two sexual functions of the same flower. CONCLUSIONS: Dual sex roles contribute to a near doubling of mate diversity in our experimental population of Mimulus ringens. This finding may help explain the maintenance of hermaphroditism under conditions that would otherwise favor the evolution of separate sexes.

Effects of pollination and postpollination processes on selfing rate in Mimulus ringens.

PREMISE OF THE STUDY: Selfing rates vary widely within and among populations of self-compatible flowering plants. This variation is often attributed to differences in the amount and timing of self and outcross pollen deposition on stigmas, as well as to the influence of postpollination mechanisms that control fertilization success. This study explores the relative importance of pollination and postpollination processes in determining selfing rates in monkeyflower, Mimulus ringens. METHODS: We hand-pollinated flowers on 17 unrelated mothers with pollen from one of three experimental treatments intended to replicate field conditions: (1) simultaneous deposition of 50% self pollen and 50% outcross pollen from 5 unrelated donors; (2) self pollen followed 15 min later by application of an equal amount of outcross pollen from five unrelated donors; and (3) outcross pollen from 5 unrelated donors followed 15 min later by application of an equal amount of self pollen. We genotyped 757 progeny at 8 polymorphic microsatellite loci and used paternity exclusion to determine whether each seedling was selfed or outcrossed. KEY RESULTS: When self and outcross pollen arrived simultaneously, and when self pollen arrived 15 min prior to outcross pollen, the observed proportions of self and outcross progeny did not deviate from the expected 1:1 ratio. However, when outcross pollen was applied 15 min prior to self pollen, there was a significant excess of outcross progeny. CONCLUSIONS: Selfing rate in Mimulus ringens is influenced by small differences in the timing of pollen arrival, but not by nonrandom postpollination sorting.

Forecasting climate change impacts on the distribution of wetland habitat in the Midwestern United states.

Shifting precipitation patterns brought on by climate change threaten to alter the future distribution of wetlands. We developed a set of models to understand the role climate plays in determining wetland formation on a landscape scale and to forecast changes in wetland distribution for the Midwestern United States. These models combined 35 climate variables with 21 geographic and anthropogenic factors thought to encapsulate other major drivers of wetland distribution for the Midwest. All models successfully recreated a majority of the variation in current wetland area within the Midwest, and showed that wetland area was significantly associated with climate, even when controlling for landscape context. Inferential (linear) models identified a consistent negative association between wetland area and isothermality. This is likely the result of regular inundation in areas where precipitation accumulates as snow, then melts faster than drainage capacity. Moisture index seasonality was identified as a key factor distinguishing between emergent and forested wetland types, where forested wetland area at the landscape scale is associated with a greater seasonal variation in water table depth. Forecasting models (neural networks) predicted an increase in potential wetland area in the coming century, with areas conducive to forested wetland formation expanding more rapidly than areas conducive to emergent wetlands. Local cluster analyses identified Iowa and Northeastern Missouri as areas of anticipated wetland expansion, indicating both a risk to crop production within the Midwest Corn Belt and an opportunity for wetland conservation, while Northern Minnesota and Michigan are potentially at risk of wetland losses under a future climate.

Effects of floral display size on male and female reproductive success in Mimulus ringens.

BACKGROUND AND AIMS: The number of flowers blooming simultaneously on a plant may have profound consequences for reproductive success. Large floral displays often attract more pollinator visits, increasing outcross pollen receipt. However, pollinators frequently probe more flowers in sequence on large displays, potentially increasing self-pollination and reducing pollen export per flower. To better understand how floral display size influences male and female fitness, we manipulated display phenotypes and then used paternity analysis to quantify siring success and selfing rates. METHODS: To facilitate unambiguous assignment of paternity, we established four replicate (cloned) arrays of Mimulus ringens, each consisting of genets with unique combinations of homozygous marker genotypes. In each array, we trimmed displays to two, four, eight or 16 flowers. When fruits ripened, we counted the number of seeds per fruit and assigned paternity to 1935 progeny. KEY RESULTS: Siring success per flower declined sharply with increasing display size, while female success per flower did not vary with display. The rate of self-fertilization increased for large floral displays, but siring losses due to geitonogamous pollen discounting were much greater than siring gains through increased self-fertilization. As display size increased, each additional seed sired through geitonogamous self-pollination was associated with a loss of 9·7 seeds sired through outcrossing. CONCLUSIONS: Although total fitness increased with floral display size, the marginal return on each additional flower declined steadily as display size increased. Therefore, a plant could maximize fitness by producing small displays over a long flowering period, rather than large displays over a brief flowering period.

New perspectives on the evolution of plant mating systems.

BACKGROUND: The remarkable diversity of mating patterns and sexual systems in flowering plants has fascinated evolutionary biologists for more than a century. Enduring questions about this topic include why sexual polymorphisms have evolved independently in over 100 plant families, and why proportions of self- and cross-fertilization often vary dramatically within and among populations. Important new insights concerning the evolutionary dynamics of plant mating systems have built upon a strong foundation of theoretical models and innovative field and laboratory experiments. However, as the pace of advancement in this field has accelerated, it has become increasingly difficult for researchers to follow developments outside their primary area of research expertise. SCOPE: In this Viewpoint paper we highlight three important themes that span and integrate different subdisciplines: the changes in morphology, phenology, and physiology that accompany the transition to selfing; the evolutionary consequences of pollen pool diversity in flowering plants; and the evolutionary dynamics of sexual polymorphisms. We also highlight recent developments in molecular techniques that will facilitate more efficient and cost-effective study of mating patterns in large natural populations, research on the dynamics of pollen transport, and investigations on the genetic basis of sexual polymorphisms. This Viewpoint also serves as the introduction to a Special Issue on the Evolution of Plant Mating Systems. The 15 papers in this special issue provide inspiring examples of recent discoveries, and glimpses of exciting developments yet to come.

Characterization of 42 polymorphic microsatellite loci in Mimulus ringens (Phrymaceae) using Illumina sequencing.

PREMISE OF THE STUDY: Microsatellite markers were isolated and characterized in Mimulus ringens (Phrymaceae), a herbaceous wetland perennial, to facilitate studies of mating patterns and population genetic structure. • METHODS AND RESULTS: A total of 42 polymorphic loci were identified from a sample of 24 individuals from a single population in Ohio, USA. The number of alleles per locus ranged from two to nine, and median observed heterozygosity was 0.435. • CONCLUSIONS: This large number of polymorphic loci will enable researchers to quantify male fitness, patterns of multiple paternity, selfing, and biparental inbreeding in large natural populations of this species. These markers will also permit detailed study of fine-scale patterns of genetic structure.

Increased relative abundance of an invasive competitor for pollination, Lythrum salicaria, reduces seed number in Mimulus ringens.

When exotic plant species share pollinators with native species, competition for pollination may lower the reproductive success of natives by reducing the frequency and/or quality of visits they receive. Exotic species often become numerically dominant in plant communities, and the relative abundance of these potential competitors for pollination may be an important determinant of their effects on the pollination and reproductive success of co-occurring native species. Our study experimentally tests whether the presence and abundance of an invasive exotic, Lythrum salicaria L. (Lythraceae), influences reproductive success of a co-flowering native species, Mimulus ringens L. (Phrymaceae). We also examine the mechanisms of competition for pollination and how they may be altered by changes in competitor abundance. We found that the presence of Lythrum salicaria lowered mean seed number in Mimulus ringens fruits. This effect was most pronounced when the invasive competitor was highly abundant, decreasing the number of seeds per fruit by 40% in 2006 and 33% in 2007. Reductions in the number of seeds per fruit were likely due to reduced visit quality resulting from Mimulus pollen loss when bees foraged on neighboring Lythrum plants. This study suggests that visit quality to natives may be influenced by the presence and abundance of invasive flowering plants.

Edge effects and mating patterns in a bumblebee-pollinated plant.

Researchers have long assumed that plant spatial location influences plant reproductive success and pollinator foraging behaviour. For example, many flowering plant populations have small, linear or irregular shapes that increase the proportion of plants on the edge, which may reduce mating opportunities through both male and female function. Additionally, plants that rely on pollinators may be particularly vulnerable to edge effects if those pollinators exhibit restricted foraging and pollen carryover is limited. To explore the effects of spatial location (edge vs. interior) on siring success, seed production, pollinator foraging patterns and pollen-mediated gene dispersal, we established a square experimental array of 49 Mimulus ringens (monkeyflower) plants. We observed foraging patterns of pollinating bumblebees and used paternity analysis to quantify male and female reproductive success and mate diversity for plants on the edge versus interior. We found no significant differences between edge and interior plants in the number of seeds sired, mothered or the number of sires per fruit. However, we found strong differences in pollinator behaviour based on plant location, including 15 % lower per flower visitation rates and substantially longer interplant moves for edge plants. This translated into 40 % greater pollen-mediated gene dispersal for edge than for interior plants. Overall, our results suggest that edge effects are not as strong as is commonly assumed, and that different plant reproduction parameters respond to spatial location independently.

Ecology and evolution of plant-pollinator interactions.

BACKGROUND: Some of the most exciting advances in pollination biology have resulted from interdisciplinary research combining ecological and evolutionary perspectives. For example, these two approaches have been essential for understanding the functional ecology of floral traits, the dynamics of pollen transport, competition for pollinator services, and patterns of specialization and generalization in plant-pollinator interactions. However, as research in these and other areas has progressed, many pollination biologists have become more specialized in their research interests, focusing their attention on either evolutionary or ecological questions. We believe that the continuing vigour of a synthetic and interdisciplinary field like pollination biology depends on renewed connections between ecological and evolutionary approaches. SCOPE: In this Viewpoint paper we highlight the application of ecological and evolutionary approaches to two themes in pollination biology: (1) links between pollinator behaviour and plant mating systems, and (2) generalization and specialization in pollination systems. We also describe how mathematical models and synthetic analyses have broadened our understanding of pollination biology, especially in human-modified landscapes. We conclude with several suggestions that we hope will stimulate future research. This Viewpoint also serves as the introduction to this Special Issue on the Ecology and Evolution of Plant-Pollinator Interactions. These papers provide inspiring examples of the synergy between evolutionary and ecological approaches, and offer glimpses of great accomplishments yet to come.

Pollinator visitation patterns strongly influence among-flower variation in selfing rate.

BACKGROUND AND AIMS: Adjacent flowers on Mimulus ringens floral displays often vary markedly in selfing rate. We hypothesized that this fine-scale variation in mating system reflects the tendency of bumble-bee pollinators to probe several flowers consecutively on multiflower displays. When a pollinator approaches a display, the first flower probed is likely to receive substantial outcross pollen. However, since pollen carryover in this species is limited, receipt of self pollen should increase rapidly for later flowers. Here the first direct experimental test of this hypothesis is described. METHODS: In order to link floral visitation sequences with selfing rates of individual flowers, replicate linear arrays were established, each composed of plants with unique genetic markers. This facilitated unambiguous assignment of paternity to all sampled progeny. A single wild bumble-bee was permitted to forage on each linear array, recording the order of floral visits on each display. Once fruits had matured, 120 fruits were harvested (four flowers from each of five floral displays in each of six arrays). Twenty-five seedlings from each fruit were genotyped and paternity was unambiguously assigned to all 3000 genotyped progeny. KEY RESULTS: The order of pollinator probes on Mimulus floral displays strongly and significantly influenced selfing rates of individual fruits. Mean selfing rates increased from 21 % for initial probes to 78 % for the fourth flower probed on each display. CONCLUSIONS: Striking among-flower differences in selfing rate result from increased deposition of geitonogamous (among-flower, within-display) self pollen as bumble-bees probe consecutive flowers on each floral display. The resulting heterogeneity in the genetic composition of sibships may influence seedling competition and the expression of inbreeding depression.

New frontiers in competition for pollination.

BACKGROUND: Co-flowering plant species frequently share pollinators. Pollinator sharing is often detrimental to one or more of these species, leading to competition for pollination. Perhaps because it offers an intriguing juxtaposition of ecological opposites - mutualism and competition - within one relatively tractable system, competition for pollination has captured the interest of ecologists for over a century. SCOPE: Our intent is to contemplate exciting areas for further work on competition for pollination, rather than to exhaustively review past studies. After a brief historical summary, we present a conceptual framework that incorporates many aspects of competition for pollination, involving both the quantity and quality of pollination services, and both female and male sex functions of flowers. Using this framework, we contemplate a relatively subtle mechanism of competition involving pollen loss, and consider how competition might affect plant mating systems, overall reproductive success and multi-species interactions. We next consider how competition for pollination might be altered by several emerging consequences of a changing planet, including the spread of alien species, climate change and pollinator declines. Most of these topics represent new frontiers whose exploration has just begun. CONCLUSIONS: Competition for pollination has served as a model for the integration of ecological and evolutionary perspectives in the study of species interactions. Its study has elucidated both obvious and more subtle mechanisms, and has documented a range of outcomes. However, the potential for this interaction to inform our understanding of both pure and applied aspects of pollination biology has only begun to be realized.

Pollination success following loss of a frequent pollinator: the role of compensatory visitation by other effective pollinators.

Pollinator abundance is declining worldwide and may lower the quantity and quality of pollination services to flowering plant populations. Loss of an important pollinator is often assumed to reduce the amount of pollen received by stigmas of a focal species (pollination success), yet this assumption has rarely been tested experimentally. The magnitude of the effect, if any, may depend on the relative efficiency of the remaining pollinators, and on whether the loss of one pollinator leads to changes in visitation patterns by other pollinators. To explore how a change in pollinator composition influences pollination of Asclepias verticillata, we excluded bumble bees from plots in large and small populations of this milkweed species. We then quantified pollinator visitation rates, pollen export and pollen receipt for control plots and for plots where bumble bees were experimentally excluded. We found that exclusion of bumble bees did not reduce pollen receipt by A. verticillata flowers. Visitation by Polistes wasps increased markedly following bumble bee exclusion, both in small populations (186 % increase) and in large populations (400 % increase). Because Polistes wasps were as efficient as bumble bees at pollen transfer, increased wasp visitation offset lost bumble bee pollination services. Thus, loss of a frequent pollinator will not necessarily lead to a decline in pollination success. When pollinator loss is followed by a shift in the composition and abundance of remaining pollinators, pollination success will depend on the net change in the quantity and quality of pollination services.

Response to Comment on "Worldwide evidence of a unimodal relationship between productivity and plant species richness".

Tredennick et al. criticize one of our statistical analyses and emphasize the low explanatory power of models relating productivity to diversity. These criticisms do not detract from our key findings, including evidence consistent with the unimodal constraint relationship predicted by the humped-back model and evidence of scale sensitivities in the form and strength of the relationship.

Effects of pollination intensity on Lesquerella fendleri seed set: variation among plants.

The amount of pollen arriving on a flower can be an important determinant of seed production. I investigated the effect of varying pollen loads on seed set of the perennial desert mustard Lesquerella fendleri. To do this, I quantified the dose response relationship between stigmatic pollen load and seed set per fruit using over 400 flowers from 13 greenhouse-grown plants. Seed set per fruit generally increased with pollen up to about 100 pollen grains, then reached a plateau. A negative exponential regression of seed set on pollen load for the pooled data explained less than 10% of the observed variation in seeds per fruit. However, accounting for variation among individual plants in the dose-response relationship increased explained variation to 40%, indicating that plants responded differently to the same amount of available pollen. Plants varied little in the initial slope of the dose-response curve, but differed substantially in the asymptote, which ranged from 3 to 16 seeds. This limit is not imposed by ovule number, and may instead result from variation among plants in vigor, propensity to abort seeds, or in gender specialization. Such variation among plants in dose-response relationships has important consequences for understanding pollination limitation and pollen competition.

Competition for pollination: effects of pollen of an invasive plant on seed set of a native congener.

Competition for pollination can be an important factor in plant reproduction, but little attention has been given to the effect of the growing number of invasive plant species on pollination of native species. As a first step in understanding this threat, we used hand pollination to investigate the effects of pollen from an invasive species (Lythrum salicaria) on seed set in a sympatric and co-flowering native congener (L. alatum). Dispersal of fluorescent dyes in the field confirms that pollinators (bumble bees and honey bees) transfer pollen between species. To determine the potential effect of such interspecific pollen transfer on seed set of the native, we pollinated 773 flowers on 20 plants with one of three treatments: legitimate conspecific pollen, a mixture of conspecific and foreign pollen, and foreign pollen. The mixed-pollen treatment resulted in 28.8% lower seed set relative to conspecific pollination. Foreign crosses resulted in extremely low seed set. Observations of pollen germination indicate that events at the stigmatic surface contribute to the reduction in seed set for mixed pollination. Our results indicate that the impacts of invasive species may extend beyond vegetative competition to include competition for pollination.

Influence of pollen transport dynamics on sire profiles and multiple paternity in flowering plants.

In many flowering plants individual fruits contain a mixture of half- and full- siblings, reflecting pollination by several fathers. To better understand the mechanisms generating multiple paternity within fruits we present a theoretical framework linking pollen carryover with patterns of pollinator movement. This 'sire profile' model predicts that species with more extensive pollen carryover will have a greater number of mates. It also predicts that flowers on large displays, which are often probed consecutively during a single pollinator visitation sequence, will have a lower effective number of mates. We compared these predictions with observed values for bumble bee-pollinated Mimulus ringens, which has restricted carryover, and hummingbird-pollinated Ipomopsis aggregata, which has extensive carryover. The model correctly predicted that the effective number of mates is much higher in the species with more extensive carryover. This work extends our knowledge of plant mating systems by highlighting mechanisms influencing the genetic composition of sibships.