Simulation analyses of the evolution of intra-inflorescence flowering patterns assuming selection on anthesis interval among individual flowers.

What conditions select flowering patterns within inflorescences, or variation in the anthesis interval within inflorescences among plants? Under what conditions are gradual blooming and simultaneous blooming, both traits related to floral display size, advantageous? We constructed a simulation model in which the opening times and longevities of individual flowers within inflorescences, the sizes of attractive structures of individual flowers, and the numbers of ovules and pollen grains produced by individual flowers evolve. Individual plants in the population compete for pollinators, and plants are selected by pollinators according to their floral display sizes and amounts of resources allocated to attractive structures. We found that, if the proportion of pollen on a pollinator deposited on a stigma was low, gradual blooming did not evolve even if inbreeding depression was greater than 0.5. This is because the amount of outcross-pollen on pollinators decreased at a low rate during flower visits within a single inflorescence, and the selfing rate was suppressed to a low level even if the floral display size was large. On the other hand, if the proportion of pollen deposition was high, gradual blooming evolved even if inbreeding depression was smaller than 0.5. This may be because gradual blooming can enhance pollen delivery to other plants by reducing the loss of self-pollen by geitonogamy. On the other hand, allocation ratios among floral organs (female and male organs and attractive structures) were independent of the degree of simultaneous and gradual blooming within inflorescences. We concluded that the evolution of gradual blooming is more strongly affected by the proportion of pollen on a pollinator deposited on a stigma than by inbreeding depression.

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.

In Which Way Do the Flower Properties of the Specialist Orchid Goodyera repens Meet the Requirements of Its Generalist Pollinators?

This article is the next part of a series of studies documenting the influence of flower traits on the reproductive success (RS) of orchids. Knowledge of factors influencing RS helps to understand the mechanisms and processes crucial for shaping plant-pollinator interactions. The aim of the present study was to determine the role of flower structure and nectar composition in shaping the RS of the specialist orchid Goodyea repens, which is pollinated by generalist bumblebees. We found a high level of pollinaria removal (PR) and female reproductive success (fruiting, FRS) as well as a high level of variation between populations, although in certain populations pollination efficiency was low. Floral display traits, mainly inflorescence length, influenced FRS in certain populations. Among the flower traits, only the height of flowers was correlated with FRS in one population, suggesting that the flower structure of this orchid is well adapted to pollination by bumblebees. The nectar of G. repens is diluted and dominated by hexoses. Sugars were less important in shaping RS than amino acids. At the species level, twenty proteogenic and six non-proteogenic AAs were noted, along with their differentiated amounts and participation in particular populations. We found that distinct AAs or their groups mainly shaped PR, especially when correlations were considered at the species level. Our results suggest that both the individual nectar components and the ratios between them have an impact on G. repens RS. Because different nectar components influence the RS parameters in different ways (i.e., negatively or positively), we suggest that different Bombus species play the role of main pollinators in distinct populations.

Does Reproductive Success in Natural and Anthropogenic Populations of Generalist Epipactis helleborine Depend on Flower Morphology and Nectar Composition?

The purpose of our study was to determine the role of flower structure and nectar composition in shaping the reproductive success (RS) of the generalist orchid Epipactis helleborine in natural and anthropogenic populations. We supposed that the distinct character of two groups of habitats creates different conditions for plant-pollinator relationships, thus influencing reproductive success in E. helleborine populations. Both pollinaria removal (PR) and fruiting (FRS) were differentiated between the populations. On average, FRS was almost two times higher in the anthropogenic than in the natural populations. The difference between the two population groups in PR was smaller but still statistically significant. RS parameters were correlated with some floral display and flower traits. Floral display influenced RS only in three anthropogenic populations. Flower traits had a weak influence on RS (10 of the 192 cases analyzed). The more important trait in shaping RS was nectar chemistry. The nectar of E. helleborine is relatively diluted with a lower sugar concentration in the anthropogenic than in the natural populations. In the natural populations, domination of sucrose over hexoses was found, while in the anthropogenic populations, hexoses were more abundant and the participation of sugars was balanced. In some populations, sugars influenced RS. In E. helleborine nectar, 20 proteogenic and 7 non-proteogenic amino acids (AAs) were found with a clear domination of glutamic acid. We noted relationships between some AAs and RS, but distinct AAs shaped RS in different populations, and their impact was independent of their participation. Our results indicate that the flower structure and nectar composition of E. helleborine reflect its generalistic character and meet the requirements of a wide range of pollinators. Simultaneously, the differentiation of flower traits suggests a variation in pollinator assemblages in particular populations. Knowledge about the factors influencing RS in distinct habitats helps to understand the evolutionary potential of species and to understand mechanisms and processes crucial for shaping interactions between plants and pollinators.

Effect of grazing disturbance on floral display, pollen limitation and plant pollination efficiency in the desert steppe.

BACKGROUND: Grazing disturbance usually affects floral display and pollination efficiency in the desert steppe, which may cause pollen limitation in insect-pollinated plants. Effective pollination is essential for the reproductive success of insect-pollinated plants and insufficient pollen transfer may result in pollen limitation. Caragana microphylla Lam is an arid region shrub with ecological importance. Few studies have been conducted on how grazing disturbance influences pollen limitation and pollination efficiency of C. microphylla. Here, we quantify the effect of different grazing intensities on floral display, pollinator visitation frequency and seed production in the Urat desert steppe. RESULTS: In C. microphylla, supplemental hand pollination increased the seed set, and pollen limitation was the predominant limiting factor. As the heavy grazing significantly reduced the seed set in plants that underwent open-pollination, but there was no significant difference in the seed set between plants in the control plots and plants in the moderate grazing plots. Furthermore, there was a higher pollinator visitation frequency in plants in the control plots than in plants in the heavy grazing plots. CONCLUSIONS: We found that pollinator visitation frequency was significantly associated with the number of open flowers. Our findings also demonstrated that seed production is associated with pollinator visitation frequency, as indicated by increased seed production in flowers with higher pollinator visitation frequency. Therefore, this study provides insight into the effect of different grazing intensities on floral display that are important for influencing pollinator visitation frequency and pollination efficiency in desert steppes.

Disruptive selection via pollinators and seed predators on the height of flowers on a wind-dispersed alpine herb.

PREMISE: Floral stalk height is known to affect seed dispersal of wind-dispersed grassland species, but it may also affect the attractiveness of flowers and fruits of animal-pollinated and animal-dispersed plants. Stalk height may thus be responsive to selection via interactions with both mutualist pollinators and seed dispersers, but also antagonist florivores and seed predators. In this study, we aimed to determine the effect of pollinators and seed predators on selection on floral stalk height in the insect-pollinated and wind-dispersed, alpine, andromonoecious herb Pulsatilla alpina, whose flowers also vary in their sex allocation and thus in the resources available to both mutualists and antagonists. METHODS: We measured the resource status of individuals in terms of their size and the height of the vegetation surrounding plants of P. alpina at 11 sites. In one population, we recorded floral stalk height over an entire growing season and investigated its association with floral morphology and floral sex allocation (pistil and stamen number) and used leaf-removal manipulations to assess the effect of herbivory on floral stalk height. Finally, in four populations, we quantified phenotypic selection on floral stalk height in four female components of reproductive success before seed dispersal. RESULTS: Stalk height was positively associated with female allocation of the respective flower, the resource status of the individual, and the height of the surrounding vegetation, and negatively affected by leaf removal. Our results point to disruptive selection on stalk height in terms of both selection differentials and selection gradients for fertilization, seed predation, and seed maturation rates and to positive selection on stalk height in terms of a selection differential for mature seed number. CONCLUSIONS: Stalk height of P. alpina is a costly trait that affects female reproductive success via interactions with both mutualists and antagonists. We discuss the interplay between the resource status and selection imposed on female reproductive success and its likely role in the evolution of sex-allocation strategies, especially andromonoecy.

Plasticity in floral longevity and sex-phase duration of Lobelia siphilitica in response to simulated pollinator declines.

PREMISE: Pollinator declines can reduce the quantity and quality of pollination services, resulting in less pollen deposited on flowers and lower seed production by plants. In response to these reductions, plant species that cannot autonomously self-pollinate and thus are dependent on pollinators to set seed could plastically adjust their floral traits. Such plasticity could increase the opportunity for outcross pollination directly, as well as indirectly by affecting inflorescence traits. METHODS: To test whether plants can respond to pollinator declines by plastically adjusting their floral traits, we simulated declines by experimentally reducing pollinator access to Lobelia siphilitica plants and measuring two traits of early- and late-season flowers: (1) floral longevity; and (2) sex-phase duration. To test whether plasticity in these floral traits affected inflorescence traits, we measured daily display size and phenotypic gender. RESULTS: We found that experimentally reducing pollination did not affect female-phase duration, but did extend the male-phase duration of early-season flowers by 13% and the longevity of late-season flowers by 12.8%. However, plants with an extended male phase did not have a more male-biased phenotypic gender, and plants with an extended floral longevity did not have a larger daily display. CONCLUSIONS: Our results suggest that plants can respond to pollinator declines by plastically adjusting both the longevity and sex-phase duration of their flowers. If this plasticity increases the opportunity for outcross pollination, then it could be one mechanism by which pollinator-dependent plant species maintain seed production as pollinators decline.

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.

Bumblebees can detect floral humidity.

Floral humidity, a region of elevated humidity in the headspace of the flower, occurs in many plant species and may add to their multimodal floral displays. So far, the ability to detect and respond to floral humidity cues has been only established for hawkmoths when they locate and extract nectar while hovering in front of some moth-pollinated flowers. To test whether floral humidity can be used by other more widespread generalist pollinators, we designed artificial flowers that presented biologically relevant levels of humidity similar to those shown by flowering plants. Bumblebees showed a spontaneous preference for flowers that produced higher floral humidity. Furthermore, learning experiments showed that bumblebees are able to use differences in floral humidity to distinguish between rewarding and non-rewarding flowers. Our results indicate that bumblebees are sensitive to different levels of floral humidity. In this way floral humidity can add to the information provided by flowers and could impact pollinator behaviour more significantly than previously thought.

Asymmetry in fitness-related traits of later-generation hybrids between two invasive species.

PREMISE: The importance of hybridization to invasion has been frequently discussed, with most studies focusing on the comparison of fitness-related traits between F1 hybrids and their parents and the consequences of such fitness differences. However, relatively little attention has been given to late-generation hybrids. Different fitness landscapes could emerge in later generations after hybrids cross with each other or backcross with their parents, which may play an important role in plant invasion and subsequent speciation. METHODS: In this study, artificial crosses were conducted to generate multiple generations, including F1, F2, and backcrosses between two invasive species: Cakile edentula (self-compatible) and C. maritima (self-incompatible). Putative hybrids were also collected in the sympatric zone and compared with their co-occurring parents for phenotypic and genetic differences. RESULTS: Genetic data provided evidence of hybridization happening in the wild, and phenotypic comparisons showed that natural hybrids had intermediate traits between the two species but showed more similarity to C. maritima than to C. edentula. The asymmetry was further identified in artificial generations for several phenotypic characters. Furthermore, backcrosses exhibited different patterns of variation, with backcrosses to C. maritima having higher reproductive output than their counterparts. CONCLUSIONS: Our results suggest that hybridization and introgression (backcrossing) in Cakile species is asymmetric and most likely to favor the proliferation of C. maritima genes in the mixed population and thus help its establishment, a finding that could not be predicted by characterizing F1 hybrids.

Nectar Chemistry or Flower Morphology-What Is More Important for the Reproductive Success of Generalist Orchid Epipactis palustris in Natural and Anthropogenic Populations?

The aim of this study was to determine the level of reproductive success (RS) in natural and anthropogenic populations of generalist orchid Epipactis palustris and its dependence on flower structure and nectar composition, i.e., amino acids and sugars. We found that both pollinaria removal and female reproductive success were high and similar in all populations, despite differences in flower traits and nectar chemistry. Flower structures were weakly correlated with parameters of RS. Nectar traits were more important in shaping RS; although, we noted differentiated selection on nectar components in distinct populations. Individuals in natural populations produced nectar with a larger amount of sugars and amino acids. The sucrose to (fructose and glucose) ratio in natural populations was close to 1, while in anthropogenic ones, a clear domination of fructose and glucose was noted. Our results indicate that the flower traits and nectar composition of E. palustris reflect its generalist character and meet the requirements of a wide range of pollinators, differing according to body sizes, mouth apparatus, and dietary needs. Simultaneously, differentiation of nectar chemistry suggests a variation of pollinator assemblages in particular populations or domination of their some groups. To our knowledge, a comparison of nectar chemistry between natural and anthropogenic populations of orchids is reported for the first time in this paper.

Pollination outcomes reveal negative density-dependence coupled with interspecific facilitation among plants.

Pollination is thought to be under positive density-dependence, destabilising plant coexistence by conferring fitness disadvantages to rare species. Such disadvantage is exacerbated by interspecific competition but can be mitigated by facilitation and intraspecific competition. However, pollinator scarcity should enhance intraspecific plant competition and impose disadvantage on common over rare species (negative density-dependence, NDD). We assessed pollination proxies (visitation rate, pollen receipt, pollen tubes) in a generalised plant community and related them to conspecific and heterospecific density, expecting NDD and interspecific facilitation due to the natural pollinator scarcity. Contrary to usual expectations, all proxies indicated strong intraspecific competition for common plants. Moreover interspecific facilitation prevailed and was stronger for rare than for common plants. Both NDD and interspecific facilitation were modulated by specialisation, floral display and pollinator group. The combination of intraspecific competition and interspecific facilitation fosters plant coexistence, suggesting that pollination can be a niche axis maintaining plant diversity.

Floral Trait Evolution of Angiosperms on Pacific Islands.

Animals frequently evolve unique suites of traits on islands, but whether plants evolve comparable island syndromes remains unresolved. Here, we test the prediction on the basis of natural history observations that insect-pollinated plants evolve smaller flowers on islands than on mainland communities. We examined 556 plant species representing 136 phylogenetically independent contrasts between island and mainland sister taxa. We focused on endemic taxa originating from the Americas associated with seven tropical and subtropical islands of the Pacific Ocean. Contrary to conventional wisdom, flowers were not on average smaller on islands than on the mainland. On specific archipelagos (the Galápagos Islands and Revillagigedo Islands), however, island taxa did evolve smaller flowers. Divergence in flower size between island and mainland taxa also varied among taxonomic families, such that some plant families evolved smaller flowers on islands, other families evolved larger flowers on islands, and some families exhibited no divergence in flower size between island and mainland taxa. Overall, our results show that there is no general island syndrome for flower size, but instead that the evolution of floral morphology is complex and context dependent, depending on variation among islands and plant families. Our results also suggest that if island floras are typically dominated by small flowered species, as suggested by natural history observations, then ecological filtering is a more likely explanation of this pattern than evolutionary divergence postcolonization. We propose future studies that could disentangle the relative roles of ecological filtering and evolution in the distribution of floral traits on islands.

Temporal dynamics of sexual dimorphism in a dioecious species.

BACKGROUND AND AIMS: Sexual dimorphism for floral traits is common in dioecious plant species. Beyond its significance for understanding how selection acts on plant traits through male vs. female reproductive function, sexual dimorphism has also been proposed as a possible risky characteristic for insect-pollinated plants, as it could drive pollinators to forage mostly on male plants. However, even though most flowering plant species spread their flowering across several weeks or months, the temporal variation of floral phenotypes and sexual dimorphism have rarely been investigated. METHODS: We performed a survey of male and female plants from the dioecious generalist-pollinated Silene dioica (Caryophyllaceae) in a common garden experiment, over two consecutive flowering seasons. Flower number and floral size were measured each week, as well as pollen quantity and viability in male plants. KEY RESULTS: Sexual dimorphism was found for all investigated floral traits, with males showing an overall higher investment in flower production and flower size. Males and females showed a similar temporal decline in flower size. The temporal dynamics of daily flower number differed between sexes, with males showing a peak in the middle of their flowering season, whereas flower production by females was quite stable over time. At the scale of the experimental population, both individual and floral sex ratios appeared to vary across the flowering season. Moreover, because the onset of flowering varied among plants, the magnitude of sexual dimorphism in floral size also fluctuated strongly through time. CONCLUSIONS: Capturing male/female differences with only one temporal measurement per population may not be informative. This opens stimulating questions about how pollinator behaviour and resulting pollination efficiency may vary across the flowering season.

Local adaptation mediates floral responses to water limitation in an annual wildflower.

PREMISE: Understanding how environmental stress affects the strength of mutualisms is critically important given observed and projected environmental changes. In particular, the frequency and duration of drought have been increasing worldwide. We investigated how water availability affects plant traits that mediate a pollination mutualism. METHODS: For butterfly-pollinated Phlox drummondii, we determined how moisture availability affects flower size, nectar volume, and nectar sugar amount. Furthermore, we explored the role that local adaptation may play in responses to moisture availability by including plants collected from regions that differ in aridity. Finally, we determined whether responses of plant populations to selection may differ under drought by calculating heritability of traits under control and dry conditions. RESULTS: Flower size was generally smaller in dry plants than in control plants. Early in the treatment period, nectar volume and sugar were higher in dry plants than in control plants for plants from both arid and wetter regions. With prolonged dry treatment, nectar volume and sugar remained higher only in plants from the arid region. Heritability of floral traits was lower for water-limited plants than for control plants. CONCLUSIONS: Plant investment into pollination mutualisms under environmental stress may depend on the extent to which populations are already locally adapted to such conditions, suggesting that mutualism may remain strong, at least in arid regions. However, decreases in heritability under water-limitation suggest that responses to selection imposed by pollinators may be low, even if drought-adapted plants maintain production of rewards to pollinators.

The Influence of Arbuscular Mycorrhizal Fungi on Plant Reproduction.

Arbuscular mycorrhizal (AM) fungi can influence all components of plant reproduction including pollen delivery, pollen germination, pollen tube growth, fertilization, and seed germination. AM fungi associate with plant roots, uptake nutrients, and prime plants for faster defense responses. Our literature review first identified four testable hypotheses describing how AM fungi could alter pollen delivery: (1) We hypothesize AM fungi promote floral display size. The influence of AM fungi on flower size and number is supported by literature, however there are no studies on floral color. (2) We hypothesize AM fungi promote pollen and nectar quality and quantity, and, as reported before, AM fungi promote male fitness over female fitness. (3) We hypothesize AM fungi promote both earlier and longer flowering times, but we found no consistent trend in the data for earlier or later or longer flowering times. (4) We hypothesize AM fungi alter floral secondary chemistry and VOCs, and find there is clear evidence for the alteration of floral chemistry but little data on VOCs. Second, we focus on how AM fungi could alter pollen germination, pollen tube growth, and fertilization, and present three testable hypotheses. We found evidence that AM fungi influence pollen germination and pollen tube growth, production of seeds, and seed germination. However, while most of these influences are positive they are not conclusive, because studies have been conducted in small numbers of systems and groups. Therefore, we conclude that the majority of research to date may not be measuring the influence of AM fungi on the most important components of plant reproduction: pollen germination, pollen tube growth, fertilization, and seed germination.

Fitness benefits and costs of floral advertising traits: insights from rayed and rayless phenotypes of Anacyclus (Asteraceae).

PREMISE OF THE STUDY: Ray flowers commonly observed in daisies' flowering heads are a well-known example of advertising structures for enhancing pollinator attraction. Despite this, ray loss has occurred in multiple lineages, which still rely on pollinators, suggesting that rayless phenotypes could also be adaptive for animal-pollination. Here, we investigate the benefits and costs of these specialized floral advertising structures by comparing rayed and rayless phenotypes in two hybridizing closely related species. METHODS: We assessed the advantages and costs of ray production in terms of floral visitor's attraction, pollen limitation, and female reproductive success using the broad natural variation on ray size and number at the contact zone of A. clavatus (rayed) and A. valentinus (rayless). In addition, we experimentally explored the effect of rays under controlled neighborhoods and the effect of ray removal on fruit production. KEY RESULTS: In sympatry, rayed phenotypes attracted significantly more visitors than rayless plants, in which seed production was pollen limited. However, rayed phenotypes did not show higher fruit set or seed production than rayless phenotypes. Fruit set and seed production benefited from denser neighborhood displays and larger individual floral displays, respectively. The removal of ray florets did not appear to enable resource reallocation to fruit production. CONCLUSIONS: Rayless heads compensated their lower visitation rate by means of a higher number of flowers per head achieving similar fecundity levels to rayed plants. The larger size of rayless heads might thus indicate an inflorescence-level trade-off between attraction and fertility.

A meta-analysis of whole genome duplication and the effects on flowering traits in plants.

PREMISE OF THE STUDY: Polyploidy, or whole genome duplication (WGD), is common in plants despite theory suggesting that polyploid establishment is challenging and polyploids should be evolutionarily transitory. There is renewed interest in understanding the mechanisms that could facilitate polyploid establishment and explain their pervasiveness in nature. In particular, premating isolation from their diploid progenitors is suggested to be a crucial factor. To evaluate how changes in assortative mating occur, we need to understand the phenotypic effects of WGD on reproductive traits. METHODS: We used literature surveys and a meta-analysis to assess how WGD affects floral morphology, flowering phenology, and reproductive output in plants. We focused specifically on comparisons of newly generated polyploids (neopolyploids) and their parents to mitigate potential confounding effects of adaptation and drift that may be present in ancient polyploids. KEY RESULTS: The results indicated that across a broad representation of angiosperms, floral morphology traits increased in size, reproductive output decreased, and flowering phenology was unaffected by WGD. Additionally, we found that increased trait variation after WGD was uncommon for the phenotypic traits examined. CONCLUSIONS: Our results suggest that the phenotypic effects on traits important to premating isolation of neopolyploids are small, in general. Changes in flowering phenology, reproductive output, and phenotypic variation resulting from WGD may be less critical in facilitating premating isolation and neopolyploid establishment. However, floral traits for which size is an important component of function (e.g., pollen transfer) could be strongly influenced by WGD.

Phenotypic selection on floral traits in an urban landscape.

Native species are increasingly living in urban landscapes associated with abiotic and biotic changes that may influence patterns of phenotypic selection. However, measures of selection in urban and non-urban environments, and exploration of the mechanisms associated with such changes, are uncommon. Plant-animal interactions have played a central role in the evolution of flowering plants and are sensitive to changes in the urban landscape, and thus provide opportunities to explore how urban environments modify selection. We evaluated patterns of phenotypic selection on the floral and resistance traits of Gelsemium sempervirens in urban and non-urban sites. The urban landscape had increased florivory and decreased pollen receipt, but showed only modest differences in patterns of selection. Directional selection for one trait, larger floral display size, was stronger in urban compared to non-urban sites. Neither quadratic nor correlational selection significantly differed between urban and non-urban sites. Pollination was associated with selection for larger floral display size in urban compared to non-urban sites, due to the differences in the translation of pollination into seeds rather than pollinator selectivity. Thus, our data suggest that urban landscapes may not result in sweeping differences in phenotypic selection but rather modest differences for some traits, potentially mediated by species interactions.

Scale-dependent effects of conspecific flower availability on pollination quantity and quality in an invasive shrub.

Pollen limitation can strongly influence reproduction of pollinator-dependent plants. Flower abundance can affect pollination 'quantity' and 'quality' due to its influence on pollen availability and foraging patterns of pollinators, ultimately impacting on seed production. We complemented individual-based measurements with landscape-level metrics to assess the influence of conspecific flower availability at different spatial scales on the quantity and quality components of pollination, and their impact on seed production in the invasive shrub Cytisus scoparius. In 2013-2014, we sampled 40 C. scoparius populations in Nahuel Huapi National Park, Patagonia (Argentina). In each population, we estimated the proportion of tripped flowers, fruit- and seed-set in five randomly selected individuals. The proportion of tripped flowers and the proportion of them setting fruit were used as proxies of pollination quantity and quality, respectively. Conspecific flower availability at distinct spatial scales (5-1000 m) was estimated as the area covered by flowering C. scoparius from color aerial photographs. Flower availability influenced seed output due to contrasting scale-dependent effects on pollination quantity and quality. Increasing flower availability at the landscape-scale reduced pollination quantity, whereas at the neighborhood-scale it increased pollination quality. The overall positive effect of flower availability on seed output at the neighborhood scale was slightly higher than the overall negative effect at the landscape scale. Moreover, pollination quality had a higher positive effect on seed output than pollination quantity. Our results demonstrate that pollination quality may severely limit plant reproduction. Pollination quality limitation can act independently of pollination quantity limitation because these factors operate at different spatial scales.