Direct and indirect selection on floral pigmentation by pollinators and seed predators in a color polymorphic South African shrub

The coexistence of different color morphs is often attributed to variable selection pressures across space, time, morph frequencies, or selection agents, but the routes by which each morph is favored are rarely identified. In this study we investigated factors that influence floral color polymorphisms on a local scale in Protea, within which approximately 40% of species are polymorphic. Previous work shows that seed predators and reproductive differences likely contribute to maintaining polymorphism in four Protea species. We explored whether selection acts directly or indirectly on floral color in two populations of Protea aurea, using path analysis of pollinator behavior, nectar production, seed predation, color, morphology, and maternal fecundity fitness components. We found that avian pollinators spent more time on white morphs, likely due to nectar differences, but that this had no apparent consequences for fecundity. Instead, the number of flowers per inflorescence underpinned many of the reproductively important differences between color morphs. White morphs had more flowers per inflorescence, which itself was positively correlated with nectar production, seed predator occurrence, and total long-term seed production. The number of seeds per plant to survive predation, in contrast, was not directly associated with color or any other floral trait. Thus, although color differences may be associated with conflicting selection pressures, the selection appears to be associated with the number of flowers per inflorescence and its unmeasured correlates, rather than with inflorescence color itself.     

Discovery of gynoecium color polymorphism in an aquatic plant

Flower color polymorphlsm exhibited by natural populations provides an opportunity for understanding the evolutionary mechanisms contributing to the diversity of floral morphology.However,little is known about the color polymorphism of female organs in flowering plants.Here we report gynoecium color polymorphism in Butomus umbellatus (Butomaceae),an emergent,aquatic monocot.Populations from Mishan,northeastern China comprised two morphs; gynoecia are either pink,as observed in other areas,or white.We measured floral traits and female fecundity in the two gynoecium color morphs in the field.There was no significant difference in plant height,pedicel length,and flower size including petal,sepal and gynoecium between the two morphs,but plants with pink gynoecia had wider inflorescence stalks,larger inner whorl anthers and produced more pollen and ovules than those with white gynoecia.Correspondingly,we found that seed production was significantly higher in the pink than in the white morph.This new finding suggested selection against white gynoecia in part because of low fecundity,consistent with the rarity of the white gynoecium morph in this species.     

Flower Color Polymorphism in Rhododendron cyanocarpum (Ericaceae), an Endangered Alpine Species Endemic to NW Yunnan,China

Rhododendron cyanocarpum is a narrow endemic species with pink and white floral color. In the present study, to investigate the significance of petal color morphs, we examined color morph frequencies, petal color reflectance and other associated floral characters, effective pollinators, visitation frequencies, and fruit production in the field. In all surveyed known populations, plants with pink color morph dominanted and comprised 77%-100% of individuals. Two peaks at 430 nm and 650 nm were found in the petal color reflectance of pink flowers, and only one peak at 430 nm was found in the reflectance spectrum of white flowers. In addition, color morphs were also associated with colors of style and stigma, lengths of corolla, calyx and pedicel, the closest distance between stigma and stamen, but not with style length and nectar production. Moreover, higher visitation frequency of their shared pollinators (bumblebees) and fruit production were observed of pink flowers than white flowers. Despite a briefly temporal and spacial study, we suggest that color morph frequencies, visit frequencies of bumblebees and fruit production, all favor to be stablilizing selection for the pink color morph.     

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.     

滇西北特有植物蓝果杜鹃的花色多态性研究(英文)

蓝果杜鹃(Rhododendron cyanocarpum)为大理苍山特有的濒危植物,有粉色和白色两种花冠类型。为了探讨该物种花色多态性的意义,本研究调查了粉色花和白色花植株在已知的各居群的分布频率、花冠的反射光谱及其它的花部特征、有效传粉者及其访花频率与结实情况。结果表明:粉色花植株在所有调查的居群中占优势(77%~100%)。粉色花的花冠反射光谱在430 nm和650 nm有两个峰,而白色花只在430 nm有一个反射峰。同时,花特征如:花柱与柱头颜色、花冠长度、花萼长度、花梗长度以及雌雄蕊最短距离,两种花冠存在显著差异。另外,尽管熊蜂作为这两种花冠的主要传粉者,但粉红花的访花频率以及自然条件下的结实情况显著高于白色花。本研究结果推测粉红色花可能受到了稳定性选择的作用。     

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.     

Florivores prefer white versus pink petal color morphs in wild radish, Raphanus sativus

Many hypotheses suggest that pollinators act to maintain or change floral color morph frequencies in nature, although pollinator preferences do not always match color morph frequencies in the field. Therefore, non-pollinating agents may also be responsible for color morph frequencies. To test this hypothesis, we examined whether Raphanus sativus plants with white flowers received different amounts of florivory than plants with pink flowers, and whether florivores preferred one floral color over the other. We found that white-flowered plants received significantly more floral damage than pink-flowered plants in eight populations over 4 years in northern California. Both generalists and specialists on Brassicaceae preferred white petals in choice and short-term no choice tests. In performance tests, generalists gained more weight on white versus pink petals whereas specialists gained similar amounts of weight on pink and white morphs. Because our results suggest that florivores prefer and perform better on white versus pink flowers, these insects may have the opportunity to affect the frequency of color morphs in the field.     

Herbivore pressure by weevils associated with flower color polymorphism in Geranium thunbergii (Geraniaceae)

Although floral herbivory has recently received increased attention as an important factor influencing plant reproduction, relatively little is known about how its frequency and intensity vary depending on traits of host plants. Here we report that herbivore pressure by a weevil, Zacladus geranii, is associated with a flower color polymorphism of Geranium thunbergii (Geraniaceae). Pink and white flower color morphs have been reported in G. thunbergii, and we found in a three-year field survey in multiple populations that, generally, adult weevils more preferentially visited white flowers than pink flowers. Consistently, we found more severe damage by weevil larvae in white flowers. Overall herbivore pressure for G. thunbergii varied strongly between populations, and the difference seems to be partly explained by the co-occurrence of a related plant species, Geranium yezoense, in a population, as weevils preferred it to both color morphs of G. thunbergii, thereby relaxing overall herbivore pressure for G. thunbergii. Nonetheless, despite such high variability, the preference of weevils for white morphs over pink morphs of G. thunbergii was found across multiple populations. We discuss possible mechanisms causing the association between flower color and herbivore preference as well as its evolutionary consequences.     

Floral Color Polymorphism and Reproductive Success in Annatto (Bixa orellana L.)

Floral color polymorphism of annatto (Bixa orellana L.) offers a wide range of colors that are maintained in the population by either pollinators or non pollinator agents of selection. In the present study, maintenance of different floral colors was analyzed in relation to reproductive success of Bixa orellana. The different floral petal colors (white, amaranth rose, petunia purple or cobalt violet) were determined from selected plants with reflectance spectrophotometry. Phenotypic measures of other floral traits, female reproductive success, seed set, seed output and seed weight also revealed variation between different floral morphs. Records on seed set varied significantly for different floral color morphs. Maximum fruit maturation (58 %) was observed in amaranth rose and least fruit maturation (25 %) in the white morph. Seed set data indicates pollinators’ preference for more intensely colored flowers. This preference may be due to ability of the pollinators to distinguish the morphs through differentially reduced sensitivity at the green wavelengths. In flowers which received fewer insect visits, polymorphism might be maintained by self fertilization. The color morphs showed differences in Random Amplified Polymorphic DNA (RAPD) profile indicating a genetic basis for floral color variation and consequent differences in seed set. Out of 88 bands generated with nine operon primers, 70 were polymorphic. The present study provides valuable information on the influence of petal color on maternal fitness in B. orellana.     

Effects of a Locus Affecting Floral Pigmentation in Ipomoea Purpurea on Female Fitness Components

A locus influencing floral pigment intensity in the morning glory, Ipomoea purpurea, is polymorphic throughout the southeastern United States. Previous work has suggested that the white allele at this locus has a transmission advantage during mating because of the effect of flower color on pollinator behavior. The experiment described here was designed to determine whether other effects of the W locus may contribute an opposing selective advantage to the dark allele. Dark homozygotes were vegetatively smaller and produced fewer flowers, seed capsules and seeds than either light heterozygotes or white homozygotes. In addition, dark homozygotes produced smaller seeds than heterozygotes, and there is some indication that white homozygotes also produced smaller seeds than heterozygotes. Pleiotropic effects on seed number thus do not seem to contribute to selection opposing the mating advantage associated with the white allele. However, pleiotropic effects on seed size might contribute to overdominance that could stabilize the W locus polymorphism.     

Direct and indirect effects of pollinators and seed predators to selection on plant and floral traits

Although flowering traits are often assumed to be under strong selection by pollinators, significant variation in such traits remains the norm for most plant species. Thus, it is likely that the interactions among plants, mutualists, and other selective agents, such as antagonists, ultimately shape the evolution of floral and flowering traits. We examined the importance of pollination vs pre-dispersal seed predation to selection on plant and floral characters via female plant-reproductive success in Castilleja linariaefolia (Scrophulariaceae). C. linariaefolia is pollinated by hummingbirds and experiences high levels of pre-dispersal seed predation by plume moth and fly larvae in the Rocky Mountains of Colorado, USA, where this work was conducted. We first examined whether female reproduction in C. linariaefolia was limited by pollination. Supplemental pollination only marginally increased components of female reproduction, likely because seed predation masked, in part, the beneficial effects of pollen addition. In unmanipulated populations, we measured calyx length, flower production, and plant height and used path analysis combined with structural equation modeling to quantify their importance to relative seed set through pathways involving pollination vs seed predation. We found that the strength of selection on calyx length, flower production, and plant height was greater for seed predation pathways than for pollination pathways, and one character, calyx length, experienced opposing selection via pollination vs seed predation. These results suggest that the remarkable intraspecific variation in plant and floral characters exhibited by some flowering plants is likely the result of selection driven, at least in part, by pollinators in concert with antagonists, such as pre-dispersal seed predators. This work highlights the subtle but complex interactions that shape floral and vegetative design in natural ecosystems.     

Flower color microevolution in wild radish: evolutionary response to pollinator-mediated selection

Evolutionary ecologists are fundamentally interested in how species interactions affect evolutionary change. We tested the degree to which plant-pollinator interactions affect the frequency of flower color morphs of Raphanus sativus. Petal color in R. sativus is determined by two independently assorting loci, producing four petal colors (yellow, white, pink, and bronze). We assessed the impact of pollinator discrimination on changes in flower color variation by comparing the frequency of colors produced in the presence (open pollination) versus absence (null pollination) of pollinator discrimination. We also assessed the impact of postpollination and developmental effects on progeny colors using equal pollinations with all four color morphs. Our results from open pollinations found an overrepresentation of yellow progeny in the next generation, when compared with both null pollinations and cumulative ratios based on Hardy-Weinberg and linkage equilibria assumptions. When these results were combined with those from equal pollinations, the overrepresentation of yellow could be attributed to selection from pollinators. Yet, surveys in the field the following year found no flower color frequency changes in the next generation. These results illustrate that flower color microevolution can be driven by both pollinator discrimination and other nonpollinator selective forces acting during the seed-to-adult transition, countering selection imposed by pollinators.     

Pollen limitation, seed predation and scape length in Primula farinosa

Floral display and reward production may affect the attractiveness of a plant to a range of interacting animals including pollinators, herbivores, and vectors of pathogenic fungi. The optimal floral phenotype should therefore depend on the relative importance of selection exerted by both mutualistic and antagonistic animals. The perennial, rosette herb Primula farinosa is polymorphic for scape length. Natural populations may include both plants with flowers displayed well above the ground (the long-scaped morph) and those with flowers positioned very close to the ground (the short-scaped morph). In this study, we conducted a field experiment to examine how the relative fitness of the two scape morphs is affected by interactions with pollinators and fruit predators in two different microhabitats (high and low vegetation). As predicted based on the difference in floral display, supplemental hand-pollination showed that fruit initiation was more strongly pollen-limited in the short-scaped than in the long-scaped morph, and that this difference was significantly larger in high than in low vegetation. Moreover, plants with a short scape experienced lower levels of fruit predation than plants with a long scape. Among open-pollinated controls, there was no significant difference in seed output between the two scape morphs. However, among plants receiving supplemental hand-pollination, short-scaped plants produced significantly more seeds than long-scaped plants. The results suggest that the positive and negative effects of a prominent floral display (increased pollination and seed predation, respectively) balance in the study population, but also that the short-scaped morph would have an advantage at higher pollination intensities. Spatial and temporal variation in pollinator activity and seed predation should result in associated variation in the relative fecundity of the two scape morphs.     

Comparison of floral properties and breeding system in dimorphic Buddleja delavayi (Scrophulariaceae)

In this study, floral color, scent composition and emission rate, nectar property, pollinators, and breeding system of dimorphic Buddleja delavayi Gagnep. were investigated. Flower color of B. delavayi was determined using a standard color chart and spectrophotometer, and two distinct color polymorphisms were observed having purple or white flowers. Floral scents of B. delavayi were collected using dynamic headspace adsorption and identified with coupled gas chromatography and mass spectrometry. In total, 28 compounds were identified from the flowers of B. delavayi. The identified scents were divided into three chemical classes based on their biosynthetic origin: terpenes, fatty acid derivatives, and benzenoids. The scent profiles in all individuals were dominated by a few components, such as lilac aldehyde and alcohol, 4-oxoisophorone, benaldehyde, and oxoisophorone oxide. Floral scent composition (benzenoids and terpenes) showed a significant difference between white and purple flower morphs. Flower color–flower scent associations in B. delavayi were identified with two distinct scent profiles in the two color phenotypes. The studies of other floral characteristics (nectar, floral visitors, breeding system, and fruit set) indicated that floral scent emission rate, nectar volume, visitor visitation frequency, and natural fruit set were not significantly different between the two flower color morphs. Bagging experiments revealed that seed production of B. delavayi is dependent mainly on honeybee Apis cerana. Lastly, this study implies that dimorphic floral color in B. delavayi may have been maintained by floral visitors and nectar guide color.     

Linking frugivores to the dynamics of a fruit color polymorphism

Although fruit color polymorphisms are a widespread phenomenon, the role of frugivores in their maintenance is unknown. Selection would require that frugivores interact differentially with fruit color morphs to alter their relative fitnesses, but such a pattern has yet to be demonstrated. In a 3-yr field study, the interactions of ants and birds with Acacia ligulata, an Australian shrub with a red/yellow/ orange aril color polymorphism, were examined. Bird species fell into three feeding guilds: seed dispersers, seed predators, and aril thieves; ant species acted either as seed dispersers or aril thieves. While there was no evidence of morph bias in ants, in some years birds fed more frequently on the yellow and orange morphs. Based on patterns of seedling survival and juvenile recruitment in seed deposition sites, bird seed dispersers increased the fitness of yellow and orange morphs (relative to red) in some populations, but decreased their relative fitness in others. Bird seed predators uniformly reduced relative fitness of yellow and orange morphs, while bird aril thieves had unknown effects. Altogether, consumer biases produced spatiotemporal variability in the relative fitness of A. ligulata color morphs, a pattern qualitatively consistent with maintenance of the polymorphism.     

Pollination biology and the impact of floral display, pollen donors, and distyly on seed production in Arcytophyllum lavarum (Rubiaceae)

In animal-pollinated plants, two factors affecting pollen flow and seed production are changes in floral display and the availability of compatible mates. Changes in floral display may affect the number of pollinator visits and the availability of compatible mates will affect the probability of legitimate pollination and seed production. Distyly is a floral polymorphism where long-styled (pin) and short-styled (thrum) floral morphs occur among different individuals. Distylous plants frequently exhibit self and intra-morph incompatibility. Therefore changes in morph abundance directly affect the arrival of compatible pollen to the stigmas. Floral morph by itself may also affect female reproductive success because floral morphs may display differences in seed production. We explored the effects of floral display, availability of neighboring compatible mates, and floral morph on seed production in the distylous herb ARCYTOPHYLLUM LAVARUM. We found that floral display does not affect the mean number of seeds produced per flower. There is also no effect of the proportion of neighboring legitimate pollen donors on seed production in pin or thrum flowers. However, floral morphs differed in their female reproductive success and the thrum morph produced more seeds. Hand pollination experiments suggest that differences in seed production between morphs are the result of pollen limitation. Future research will elucidate if the higher seed production in thrum flowers is a consequence of higher availability of pollen donors in the population, or higher efficiency of the pin morph as pollen donor.     

Sex‐biased seed predation in gynodioecious Dianthus superbus var. longicalycinus (Capryophyllaceae) and differential influence of two seed predator species on the floral traits

Gynodioecy, the co‐occurrence of hermaphrodite and female individuals within a species, is maintained by differential reproductive success between sexes. Recently, researchers recognized that not only pollinators but also herbivores are important agents in the evolution and maintenance of gynodioecy, when herbivory is hermaphrodite biased. In this study, we investigated whether there is hermaphrodite‐biased herbivory in a gynodioecious plant, Dianthus superbus var. longicalycinus, and if so, what floral traits influenced hermaphrodite‐biased herbivory. We measured flower morphology (flower diameter, calyx tube length, corolla height and petal width) and phenology of flowers of female individuals, hermaphrodites and gynomonoecious individuals in a natural population. We also investigated seed predation and predator species. At the study site, Sibinia weevils (Curculionidae; Coleoptera) and Coleophora moths (Coleophoridae; Lepidoptera) were common pre‐dispersal seed predators in this species. The weevil appeared early in the flowering season, and weevil predation correlated with flower phenology. Because female individuals did not flower early in the season, weevil predation was less frequent in female individuals. Moth predation correlated with calyx length. The calyx length of flowers of female individuals was smaller than those of hermaphrodites, but a direct comparison of moth predation rates failed to find a significant difference among sex morphs. We found that the two seed predators had different effects on floral traits in D. superbus var. longicalycinus. We suggest that weevil predation contributes to the maintenance of gynodioecy because female individuals successfully escaped weevil predation by flowering late. It remains unclear why flower phenology is different among sex morphs.     

Flower color-flower scent associations in polymorphic Hesperis matronalis (Brassicaceae)

Floral scent emission rate and composition of purple and white flower color morphs of Hesperis matronalis (Brassicaceae) were determined for two populations and, for each, at two times of day using dynamic headspace collection and GC-MS. The floral volatile compounds identified for this species fell into two main categories, terpenoids and aromatics. Principal component analysis of 30 compounds demonstrated that both color morphs emitted more scent at dusk than at dawn. Color morphs varied in chemical composition of scent, but this differed between populations. The white morphs exhibited significant differences between populations, while the purple morphs did not. In the white morphs, one population contains color-scent associations that match expectations from classical pollination syndrome theory, where the flowers have aromatic scents, which are expected to maximize night-flying moth pollinator attraction; in the second population, white morphs were strongly associated with terpenoid compounds. The potential impact that pollinators, conserved biosynthetic pathways, and the genetics of small colonizing populations may have in determining population-specific associations between floral color and floral scent are discussed.     

Equal and Opposite Effects of Floral Offer and Spatial Distribution on Fruit Production and Predispersal Seed Predation in Xanthosoma daguense (Araceae)1

Inflorescences of the terrestrial aroid Xanthosoma daguense in the Andes of Colombia are visited by Dinastinae and Nitidulidae beetles. Plants produce one inflorescence at a time, which is pollinated during the first night of opening. Dynastine beetles act as pollinators, whereas Nitidulids lay eggs in the inflorescence and the larvae damage the seeds. We explored the effects of floral offer and distance among inflorescences on the number of pollinator visits, fruit production, and predispersal seed predation. Number of Dynastine visits per inflorescence tended to increase with increasing distances among inflorescences, but fruit predation increased when inflorescences were more clumped. Both pollinator visitation rates and predispersal seed predation were low at high floral offer. Fruit set increased when inflorescences were visited by two or more Dynastines, but the proportion of fruits damaged by Nitidulid larvae was equivalent to the increase in fruit production due to more Dinastine visits. The net result was a similar number of undamaged fruits in all infructescences produced, independent of the number of Dinastine visits. Our results revealed that both pollinators and predators responded to the number of available inflorescences and their spatial distribution, but they had opposing effects on the infructescences. Thus, our study suggests that the interaction of two ecological processes, pollination and predispersal seed predation, may cancel each other's effects under natural conditions.     

Perspective: Evolution of flower color in the desert annual Linanthus parryae: Wright revisited

Linanthus parryae, a diminutive desert annual with white or blue flowers, has been the focus of a long-standing debate among evolutionary biologists. At issue is whether the flower color polymorphism in this species is the product of random genetic drift, as Sewall Wright argued, or of natural selection, as proposed by Carl Epling and his colleagues. Our long-term studies of three polymorphic populations in the Mojave Desert demonstrate that flower color is subject to selection that varies in both time and space in its direction and magnitude. For all sites taken together, blue-flowered plants produced more seeds than white-flowered plants in years of relatively low seed production, whereas white-flowered plants had higher fitness in years of high seed production. Evidence of selection on flower color was found in two of the three study sites. Differences in fitness between the color morphs were sometimes large, with selection coefficients as high as 0.60 in some years. Our longest period of observations was at Pearblossom site 1, where plants reached appreciable densities in seven of the 11 years of study. Here we found significant differences in the seed production of the color morphs in six years, with three years of blue advantage and three years of white advantage. For all sites taken together, total spring precipitation (March and April) was positively correlated with both absolute and relative seed production of the color morphs. At Pearblossom site 1, blue-flowered plants typically had a fitness advantage in years of low spring precipitation, whereas white-flowered plants had a fitness advantage in years of high spring precipitation. This temporal variation in selection may contribute to the maintenance of the flower-color polymorphism at Pearblossom site 1, whereas gene flow from neighboring populations is proposed as the principal factor maintaining the polymorphism at the other study sites. We found no significant differences between the color morphs in pollinator visitation rate or in their carbon isotope ratio, a measure of water-use efficiency. Although the mechanism of selection remains elusive, our results refute Wright's conclusion that the flower color polymorphism in L. parryae is an example of isolation by distance, a key component of his shifting balance theory of evolution.