POLLINATION AND SEED PRODUCTION IN IPOMOPSIS AGGREGATA: DIFFERENCES AMONG AND WITHIN FLOWER COLOR MORPHS

The pollination of red, pink, and white color morphs of Ipomopsis aggregata was evaluated to assess whether ethological isolation based on pollinator color discrimination may occur. We observed animal visitors, assessed pollen delivery, seed set per fruit, percentage of flowers setting fruit, nectar production, and timing of flower opening for different color morphs in the Front Range of Colorado. Based on traditional zoophilous flower classifications, we expected hummingbirds to pollinate red-flowered I. aggregata subsp. collina and hawkmoths to pollinate white-flowered I. aggregata subsp. Candida. However, ethological isolation does not appear to occur among color morphs of I. aggregata in the Front Range. Hummingbirds visited red-flowered plants in excess overall, and, to a lesser extent, so did hawkmoths. Both hummingbirds and hawkmoths visited all color morphs and probably transferred pollen among them. Pollen delivery data and a day-night bagging experiment also suggest that pollinators do not necessarily behave as predicted by flower classifications. In addition, there is little evidence for major differences between red, white, and pink flowers in any aspects of reproductive biology. Indeed, most variation occurs within a given color morph.     

Pollinator response to flower color polymorphism and floral display in a plant with a single-locus floral color polymorphism: consequences for plant reproduction

Variation in flower color, particularly polymorphism, in which two or more different flower color phenotypes occur in the same population or species, may be affected or maintained by mechanisms that depend on pollinators. Furthermore, variation in floral display may affect pollinator response and plant reproductive success through changes in pollinator visitation and availability of compatible pollen. To asses if flower color polymorphism and floral display influences pollinator preferences and movements within and among plants and fitness-related variables we used the self-incompatible species Cosmos bipinnatus Cav. (Asteraceae), a model system with single-locus flower color polymorphism that comprises three morphs: white (recessive homozygous), pink (heterozygous co-dominate), and purple (dominant homozygous) flowers. We measured the preferences of pollinators for each morph and constancy index for each pollinator species, pollination visitation rate, floral traits, and female fitness measures. Flower color morphs differed in floral trait measures and seed production. Pollinators foraged nonrandomly with respect to flower color. The most frequent morph, the pink morph, was the most visited and pollinators exhibited the highest constancy for this morph. Moreover, this morph exhibited the highest female fitness. Pollinators responded strongly to floral display size, while probed more capitulums from plants with large total display sizes, they left a great proportion of them unvisited. Furthermore, total pollinator visitation showed a positive relation with female fitness. Results suggest that although pollinators preferred the heterozygous morph, they alternate indiscriminately among morphs making this polymorphism stable.     

VARIATION AND COVARIATION AMONG FLORAL TRAITS WITHIN AND AMONG FOUR SPECIES OF NORTHERN EUROPEAN PRIMULA (PRIMULACEAE)

Phenotypic and genetic variation and correlations among floral traits within and among four Primula species were measured to seek evidence for potential constraints on the independent evolution of floral characters, to examine the relationship between mating system, ploidy level, and sex allocation, and to determine whether some traits are more conservative than others within and across these congeners. We measured mean flower diameter, corolla depth, pollen production, modal pollen grain volume, ovule number per flower, and pollen: ovule ratios for 64 field-collected genotypes from northern Europe. These represented one heterostylous (P. farinosa: 2n = 18) and three homostylous (P. scotica: 2n = 54, P. scahdinavica: 2n = 74, and P. stricta: 2n ~ 126) species. All traits differed significantly among species and among the six taxon/morph categories identified (including three morphs of P. farinosa: pin, thrum, and homostylous). Pollen production per flower was significantly higher (and individual pollen grain volume lower) in the outcrossing P. farinosa than in any of the homostylous species; also, pin morphs produced significantly more pollen per flower than thrums in P. farinosa. Among the homostylous species, there were significant differences in all traits except modal pollen grain volume. Ovule number per flower and flower size were less variable among taxa than pollen production and pollen volume. Within species, there were several strong negative correlations among genets between pairs of traits, but each species exhibited a unique set of inverse relationships. We detected only one significant positive genetic correlation; in P. stricta, ovule number and pollen production per flower were positively correlated among genets. Among species means, two pairs of traits were negatively correlated: mean ovule number per flower vs. flower diameter (but P = 0.0587), and mean pollen production per flower vs. modal pollen grain volume. These negative correlations within and among taxa suggest that there may be intrinsic genetic constraints on the independent evolution of these floral characters, but that these constraints differ among species.     

THE INFLUENCE OF FLOWER COLOR ON OUTCROSSING RATE AND MALE REPRODUCTIVE SUCCESS IN IPOMOEA PURPUREA

Experimental populations of the annual plant, Ipomoea purpurea, composed of individuals belonging to two flower color morphs were studied to determine the effect of flower color on outcrossing rate and reproductive success as a male parent. Analyses of parent and offspring genotypes show that the pigmented and white morphs outcross at similar rates, but that the white morph is favored as a pollen donor. The result suggests that the dynamics of selection occurring at the locus coding for white versus pigmented flowers are more complex than previously believed. Factors such as frequency-dependent outcrossing rates and epistatic effects of the white allele may be operating. The results also suggest that pollinator observations are unreliable indicators of the actual mating system.     

Strong pollinator‐mediated selection for increased flower brightness and contrast in a deceptive orchid

Contrasting flower color patterns that putatively attract or direct pollinators toward a reward are common among angiosperms. In the deceptive orchid Anacamptis morio, the lower petal, which makes up most of the floral display, has a light central patch with dark markings. Within populations, there is pronounced variation in petal brightness, patch size, amount of dark markings, and contrast between patch and petal margin. We tested whether pollinators mediate selection on these color traits and on morphology (plant height, number of flowers, corolla size, spur length), and whether selection is consistent with facilitated or negative frequency‐dependent pollination. Pollinators mediated strong selection for increased petal brightness (Δβ_poll = 0.42) and contrast (Δβ_poll = 0.51). Pollinators also tended to mediate stabilizing selection on brightness (Δγ_poll = –0.27, n.s.) favoring the most common phenotype in the population. Selection for reduced petal brightness among hand‐pollinated plants indicated a fitness cost associated with brightness. The results demonstrate that flower color traits influence pollination success and seed production in A. morio, indicating that they affect attractiveness to pollinators, efficiency of pollen transfer, or both. The documented selection is consistent with facilitated pollination and selection for color convergence toward cooccurring rewarding species.     

Reproductive conflicts ofPalicourea padifolia (Rubiaceae) a distylous shrub of a tropical cloud forest in Mexico

We studied a population of the distylousPalicourea padifolia (Rubiaceae) in a cloud forest remnant near Xalapa City, Veracruz, México to explore possible asymmetries between floral morphs in the attractiveness to pollinators, seed dispersers, nectar robbers, floral parasites, and herbivores. We first assessed heterostyly and reciprocal herkogamy by measuring floral attributes such as corolla length (buds and open flowers), style and anther heights, stigma and stamen lengths and the distance between the anther tip to the stigma lobe. We then estimated floral and fruit attributes such as flower size, anther height, number and size of pollen grains, fruit size, seed size, nectar production, and flower and fruit standing crops to assess differences between floral morphs in attracting and effectively using mutualistic pollinators and seed dispersers. Also, floral parasitism and nectar robbing were assessed in this study as a measure of flower attractiveness to antagonists. The system seems to conform well to classical heterostyly (e.g. reciprocal stamen/style lengths, pollen and anther dimorphism, intramorph incompatibility) yet, there were several tantalizing differences observed between pin and thrum morphs. Thrum flowers have longer corollas and larger but fewer pollen grains than pin flowers. Both morphs produced the same total number of inflorescences, developed the same number of buds, and opened the same number of flowers per inflorescence during the flowering season. Nectar production and sugar concentration were similar between floral morphs but the reward was not offered symmetrically to floral visitors throughout the day. Nectar concentration was higher in pin flowers in the afternoon. The numbers of developing, fully developed, and ripe fruits were the same between floral morphs, however, fruits and seeds were larger than those of thrums. The incidence of fly larvae was higher among thrum flowers and damage by nectar robbing was the same between floral morphs. Fruit abortion patterns of flowers manually pollinated suggest intra-morph sterility (self and intramorph incompatibility). There were no differences between morphs in fruit and seed set per flower following legitimate pollination although thrums were more leaky than the pins (intramorph compatibility).     

HETEROSTYLY,POPULATION COMPOSITION,AND POLLEN FLOW IN HEDYOTIS CAERULEA

Hedyotis caerulea is a perennial, spring-flowering herb native to eastern North America with distylous flowers that differ in a number of morphological and physiological traits. However, the pin and thrum morphs produce the same numbers of buds, flowers, and fruits per plant, although it is possible that differences in these may occur in some populations at certain times of the flowering period. The two morphs are self-incompatible and cross-compatible. Most populations contained an excess of pins over thrums (anisoplethy); less commonly pins and thrums were equally represented (isoplethy). Populations change from anisoplethy to isoplethy and in the reverse direction. The spatial distribution of pin and thrum flowers in populations was random in some populations but non-random in others. There is some indication that the two morphs in some populations have somewhat different flowering periods. Pollinators seem to be chiefly bombyliid flies and perhaps thrips, but insects were rarely observed visiting flowers. In some populations, the two morphs produced equal numbers of pollen grains per flower; in others they did not. The average pollen viability varied, but on the average there was a moderate level of pollen sterility. High numbers of pollen grains remained in dehisced anthers, probably as a result of low pollinator activity. However, between 5% and 9% of the pollen produced participated in pollination. Stigmas of most pin flowers received more pin pollen grains than thrum pollen grains; on stigmas of thrum flowers pin pollen grains outnumbered thrum pollen grains. Thus, compatible pollen flow from pins to thrums was greater than in the reverse direction. Anisoplethic and isoplethic populations had the same pollen flow patterns. A plant-by-plant examination of stigmas indicated that many stigmas bore few or no pollen grains. Seed production of the two morphs was equal. Despite the inequities in pollen flow patterns, the widespread and occasionally weedy nature of H. caerulea suggests that its breeding system must be viewed as a successful one.     

Variation in expression of trimorphic incompatibility in Pontederia cordata L. (Pontederiaceae)

Summary Pontederia cordata L. (Pontederiaceae), a perennial diploid, possesses the rare genetic polymorphism tristyly. A controlled pollination programme was conducted over a three year period, under glasshouse conditions, on 36 clones of P. cordata var. cordata to examine the nature of the self-incompatibility system. The three major findings of the pollination study were: (1) the three floral morphs display different levels of self-incompatibility, (2) pollen from the two anther levels within a flower exhibits different compatibility behaviour in self-pollinations, (3) considerable individual genetic variation in the expression of self-incompatibility is evident among clones within floral morphs. Similar results were also obtained from a smaller study on 15 clones of P. cordata var. lancifolia conducted over a 6 month period. In common with other Pontederia species the mid-styled morph (M) of P. cordata produces large amounts of seed when self-pollinated with pollen from long-level anthers. A developmental model is proposed to explain the high level of self-compatibility of the M morph in Pontederia species. Self-pollination of segregating progenies from M and S morphs of known incompatibility status demonstrated that the expression of incompatibility is closely associated with style length. It is suggested that overall differences in incompatibility behaviour among the floral morphs may be due to the pleiotropic effects of major genes controlling sub-characters of the tristylous syndrome, rather than linked modifier genes. However, the variable expression of trimorphic incompatibility within floral morphs suggests that this variation may be polygenic in origin.     

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

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

THE IMPACT OF A FLOWER-COLOR POLYMORPHISM ON MATING PATTERNS IN EXPERIMENTAL POPULATIONS OF WILD RADISH (RAPHANUS RAPHANISTRUM L.)

We conducted field experiments to determine how a naturally occurring petal-color polymorphism influences mating patterns in wild radish (Raphanus raphanistrum). The polymorphism is controlled at a single genetic locus, with white petal color being completely dominant to yellow. In experimental populations with equal numbers of yellow- and white-flowered homozygous individuals, insect visitors strongly discriminated against white flowers. Pieris rapae, the most frequent pollinator, was almost 50% more likely to visit yellow than white flowers. Maternal fecundity did not differ between the morphs and was not significantly influenced by a plant's compatibility with potential donors, suggesting that seed production was not limited by receipt of compatible pollen. In contrast, the yellow-flowered morph sired approximately 75% of all seeds produced during the study. This paternity proportion was consistently greater than that expected on the basis of postpollination compatibility measures and was indistinguishable from that expected on the basis of pollinator-visitation frequency. We conclude that the male-fitness advantage of the yellow morph resulted from enhanced pollen export due to the greater attractiveness of its flowers to insect pollinators. With color morphs evenly distributed in experimental arrays, insects did not move assortatively on the basis of petal color, and we found no evidence for assortative pollen flow due to the floral polymorphism. Once postpollination compatibility relationships within populations were taken into account, paternal success of yellow donors did not differ between yellow- and white-flowered maternal plants.     

Pollen quality limitation in the Iberian critically endangered genus <Emphasis Type="Italic">Pseudomisopates</Emphasis> (Antirrhinaceae)

Pollen limitation occurs when sexual reproduction is decreased due to inadequate pollen receipt. Limitation is usually associated with the quality and the quantity of pollen; still, most studies do not discriminate between the two. We used hand-pollination experiments and observations of floral visitors to determine the mating system and limitations to seed production of the Spanish endangered species Pseudomisopates rivas-martinezii. We tested for agamospermy, autogamy, obligated autogamy, cross-pollination and supplementation. The response variables considered were fruit set, seed set and viable seed set. Previous studies indicated that (i) the species exhibits extensive clonal growth, (ii) plants flower profusely in summer, (iii) seeds showed to be highly unviable, and (iv) no seedlings were observed in the field. We found that P. rivas-martinezii is predominantly self-incompatible, has an unexpected generalized pollination system considering its occluded corolla, and is limited by qualitative rather than quantitative pollen limitation. These results indicate that, in addition to other presumed environmental factors, the low seed viability of this self-incompatible species is due to pollen quality limitation as a result of reduced mate availability.     

SPATIAL DIFFERENTIATION FOR FLOWER COLOR IN THE DESERT ANNUAL LINANTHUS PARRYAE: WAS WRIGHT RIGHT?

Understanding the evolutionary mechanisms that contribute to the local genetic differentiation of populations is a major goal of evolutionary biology, and debate continues regarding the relative importance of natural selection and random genetic drift to population differentiation. The desert plant Linanthus parryae has played a prominent role in these debates, with nearly six decades of empirical and theoretical work into the causes of spatial differentiation for flower color. Plants produce either blue or white flowers, and local populations often differ greatly in the frequencies of the two color morphs. Sewall Wright first applied his model of "isolation by distance" to investigate spatial patterns of flower color in Linanthus. He concluded that the distribution of flower color morphs was due to random genetic drift, and that Linanthus provided an example of his shifting balance theory of evolution. Our results from comprehensive field studies do not support this view. We studied an area in which flower color changed abruptly from all-blue to all-white across a shallow ravine. Allozyme markers sampled across these regions showed no evidence of spatial differentiation, reciprocal transplant experiments revealed natural selection favoring the resident morph, and soils and the dominant members of the plant community differed between regions. These results support the hypothesis that local differences in flower color are due to natural selection, not due to genetic drift.     

Pollinator‐mediated selection on floral size and tube color in Linum pubescens: Can differential behavior and preference in different times of the day maintain dimorphism?

Diversity of flower traits is often proposed as the outcome of selection exerted by pollinators. Positive directional pollinator‐mediated selection on floral size has been widely shown to reduce phenotypic variance. However, the underlying mechanism of maintaining within‐population floral color polymorphism is poorly understood. Divergent selection, mediated by different pollinators or by both mutualists and antagonists, may create and maintain such polymorphism, but it has rarely been shown to result from differential behavior of one pollinator. We tested whether different behaviors of the same pollinators in morning and evening are associated with dimorphic floral trait in Linum pubescens, a Mediterranean annual plant that exhibits variable within‐population frequencies of dark‐ and light‐colored flower tubes. Usia bicolor bee‐flies, the major pollinators of L. pubescens, are mostly feeding in the flower in the morning, while in the evening they are mostly visiting the flowers for mating. In 2 years of studying L. pubescens in a single large population in the Carmel, Israel, we found in one year that dark‐centered flowers received significantly higher fraction of visits in the morning. Fitness was positively affected by number of visits, but no fitness differences were found between tube‐color morphs, suggesting that both morphs have similar pollination success. Using mediation analysis, we found that flower size was under positive directional pollinator‐mediated selection in both years, but pollinator behavior did not explain entirely this selection, which was possibly mediated also by other agents, such as florivores or a‐biotic stresses. While most pollinator‐mediated selection studies show that flower size signals food reward, in L. pubescens, it may also signal for mating place, which may drive positive selection. While flower size found to be under pollinator‐mediated selection in L. pubescens, differential behavior of the pollinators in morning and evening did not seem to explain flower color polymorphism.     

Pollinator visitation in populations of tristylous Eichhornia paniculata in northeastern Brazil

Summary The frequencies of floral morphs in populations of tristylous Eichhornia paniculata often deviate from the theoretical expectation of equality. This variation is associated with the breakdown of tristyly and the evolution of self-fertilization. Differences in morph frequencies could result from selection pressures due to variable levels of insect visitation to populations and contrasting foraging behavior among the floral morphs. We estimated pollinator densities in 16 populations and quantified visitation sequences to morphs in five populations of E. paniculata in northeastern Brazil. Foraging behavior among floral morphs was measured as the frequency of visits to morphs relative to their frequency in the population (preference) and number of flights between inflorescences of the same versus different morphs (constancy). Pollinator density (number/m²/minute) was not correlated with population size, plant density or morph diversity. Pollinator densities varied most among populations of less than 200 plants. Whether pollinators discriminated among the morphs, depended on whether they primarily collected nectar or pollen. In four populations, nectar-feeding bees (Ancyloscelis and Florilegus spp.) and butterflies showed no consistent preference or constancy among the morphs. In contrast, pollen-collecting bees (Trigona sp.) visited a lower proportion of longstyled inflorescences than expected and tended to visit more mid-and short-styled inflorescences in succession, once they were encountered. Pollinator constancy for morphs did not result from differences in inflorescence production or spatial patchiness among the morphs. Although non-random pollinator visitation to morphs in heterostylous populations could potentially affect mating and hence morph frequencies, the observed visitation patterns in this study do not provide evidence that pollinators play a major role in influencing floral morph frequencies.     

Floral color variation and associations with fitness‐related traits in Malva moschata (Malvaceae)

Genetic polymorphisms for floral color are interesting phenomena to study because they are likely to be maintained by opposing selective forces. Pollinator preferences may exert direct selection on floral color; however, floral color might also be the indirect target of selection through genetic associations with other traits under selection. Malva moschata (Malvaceae) is a North American species that produces either red or white flowers. In the present study, we present reflectance spectrophotometry data that characterize the nature of floral color variation in this species and show that honey bees and bumble bees should be able to distinguish between the morphs through differential sensitivity at the green (long‐wavelength) photoreceptor. Second, we use a series of phenotypic measures to investigate whether the color morphs differ with respect to other floral traits, vegetative traits or female reproductive success, and use a series of correlation analyses to infer the relative independence of color from these other traits. We found that red‐flowered morphs produced more anthers per flower and had greater leaf area, and that white‐flowered morphs had greater percentage fruit set; however, there were no reproductive success differences between the morphs. The relationship between flower size and anther number was the only correlation that differed between the morphs. Finally, a series of pollinator‐choice experiments showed that bumble bees strongly prefer red morphs in terms of visit frequency and duration, but honey bees have no preference. Taken together, our results suggest that color is rather independent of other phenotypic traits, and that honey bee abundance is likely to play a role in maintaining color variation in this system.     

Flower colour change in Banksia ilicifolia: A signal for pollinators

Both birds and insects visit yellow flower heads of Banksia ilicifolia rather than those in the pink or red phases. Birds carry most pollen. Substantial nectar and pollen rewards are present only in the yellow phase. The timing of flower colour change also corresponds to a decline in viability of presented pollen and stigma receptivity. Colour change is age-dependent rather than pollinator-induced. Bird visits to yellow or red heads are essentially determined by the availability of nectar in each rather than differences in their visibility. Fruit set is negligible in the absence of pollinators but still < 1% in their presence. Banksia ilicifolia has the smallest heads and is the most localized of five co-occurring and partly co-flowering Banksia species. It is hypothesized that the restriction of flower colour change to B. ilicifolia increases the competitiveness of this species: bird visitors are directed to flower heads with abundant nectar, viable pollen and receptive stigmas, foraging and pollination efficiency thereby being enhanced without a marked reduction in long-distance attractiveness of the tree to potential pollinators.     

POPULATION STRUCTURE AND MORPH-SPECIFIC FITNESS DIFFERENCES IN TRISTYLOUS LYTHRUM SALICARIA

In tristylous plant populations, style-morph frequencies are governed by an interaction between frequency-dependent selection due to disassortative mating and stochastic processes. Provided that there are no inherent fitness differences among morphs, frequency-dependent selection should result in equal morph frequencies at equilibrium. Stochastic models indicate that the short-styled morph has the highest and the long-styled morph the lowest probability of being lost from local populations as a result of random processes. We surveyed the morph composition of 82 populations of the tristylous, self-incompatible herb Lythrum salicaria in two archipelagos, one in central and one in northern Sweden, located close to the range-margin of the species. To examine whether deviations from even morph frequencies can be explained by among-morph differences in reproductive success, we quantified flower and seed production in six and three populations in the northern and southern archipelago, respectively, and we recorded segregation ratios in offspring produced in six trimorphic populations in the northern area. Seed germination and offspring growth were studied in the greenhouse. Ninety percent of the populations in the southern archipelago (N = 31) and 69% of the populations in the northern archipelago (N = 35) were trimorphic; the remaining populations were dimorphic (only populations consisting of at least three flowering plants considered). Dimorphic populations were smaller than trimorphic populations, as predicted by stochastic models. There was a striking difference in the morph composition of L. salicaria populations between the two archipelagos. In the southern archipelago, there was a slight excess of the long-styled morph and a corresponding deficiency of the short-styled morph. In contrast, the northern populations were characterized by a marked deficiency of the mid-styled morph: the average frequency of the mid-styled morph in trimorphic populations was 0.21, and nine of eleven dimorphic populations lacked the mid-styled morph. In both archipelagos, the long-styled morph (the most common morph) produced about 20% fewer seeds per fruit than the other morphs. The long-styled morph also tended to produce fewer seeds per plant. A hand-pollination experiment performed in two of the northern populations indicated that seed production per flower was pollen-limited in the long-styled morph but not in the other two morphs. Seed germination and offspring size after 24 weeks of growth did not differ among morphs. The mid-styled morph tended to have a higher representation in the offspring than in the parental generation in all six trimorphic populations studied further indicating that the deficiency of the mid-styled morph in the northern archipelago does not represent an equilibrium. Taken together, the results do not support the hypothesis that morph-specific differences in reproductive success can account for deviations from even morph frequencies in L. salicaria. It is suggested that among-morph differences in other components of fitness and historical factors may contribute to the current morph structure.     

Effects of local density and flower colour polymorphism on pollination and reproduction in the rewardless orchid Dactylorhiza sambucina (L.) Soò

The rewardless orchid Dactylorhiza sambucina shows a stable flower colour polymorphism, with both yellow- and red-flowered morphs growing sympatrically. Pollination biology and breeding system were investigated to examine the effects of density of plants, colour polymorphism, inflorescence dimension, and flower position within inflorescence on male and female reproductive success in three natural populations of D. sambucina. There were significant differences among sites in the number of pollinia removed and in fruit set per inflorescence. Number of removed pollinia and capsule production in D. sambucina were independent from flower and inflorescence size or flower position. As a whole, the red morphs showed the highest number of capsules produced, while the yellow morphs had the greatest male success. The relative male and female reproductive success were independent from plant density but were significantly correlated with the yellow morph frequency at the population level. Overall, our findings show that the contribution to the total reproductive success deriving from the two colour morphs does not conform with the predictions of negative frequency-dependent selection.     

Identification of effective pollinators of Primula sieboldii E. Morren in a wild habitat in Hiroshima,Japan

Primula sieboldii E. Morren is a heterostylous clonal herb that is widely distributed in Japan but in danger of extinction in the wild. The existence of pollinators in each habitat is imperative for its long-term survival, because seeds can be produced only by insect cross-pollination between different flower morphs. In this study, we identified the pollinators of P. sieboldii in a wild habitat in Hiroshima as those insects that we observed to (a) put the proboscis into a corolla tube, (b) deposit pollen grains on the proboscis, and (c) have a proboscis of appropriate length and width. Effective pollinators were identified from their contribution to pollination. In 2015 and 2016, the flower visitations of 232 and 558 insects, respectively, were recorded and 85 and 13 insects were captured. Two Bombylius species, B. major L. and B. shibakawae Matsumura, accounted for 90% of flower-visiting insects in both years. All 14 species that we captured were considered pollinators of P. sieboldii, because they had proboscises that were long enough to reach pollen and that had pollen grains deposited on them. The total visitation rate of “Bombyliidae” was the highest among all pollinator categories. The results of potential pollen transport per flower per hour, which was based on total pollen number and total visitation rate of each pollinator category, indicated that “Bombyliidae” species were the most effective pollinators of P. sieboldii in this habitat.