Evolution of floral traits and mating systems under drought: a range-wide study of Mimulus cardinalis

PremiseWidespread associations between selfing rate and floral size within and among taxa suggest that these traits may evolve in concert. Does this association develop immediately because of shared genetic and/or developmental control, or stepwise with selection shaping the evolution of one trait following the other? If the former, then association ought to appear within and across selfing populations. We explore this fundamental question in three populations of the mixed‐mater Collinsia verna where autonomous selfing (AS) ability has been shown to be under selection by the pollination environment.MethodsWe grew clonal replicates of C. verna in a controlled environment to characterize broad‐sense genetic correlations among traits within populations and to assess whether divergence in mating system and floral traits among these populations is consistent with their previously observed selection pressures.ResultsAs predicted by their respective pollination environments, we demonstrate significant genetic divergence among populations in AS ability. However, patterns of divergence in floral traits (petal, stamen, and style size, stigmatic receptivity, and stigma‐anther distance) were not as expected. Within populations, genetic variation in AS appeared largely independent from floral traits, except for a single weak negative association in one population between flower size and AS rate.ConclusionsTogether, these results suggest that associations between selfing rate and floral traits across Collinsia species are not reflected at microevolutionary scales. If C. verna were to continue evolving toward the selfing syndrome, floral trait evolution would likely follow stepwise from mating system evolution.

PremiseTo avoid inbreeding depression, plants have evolved diverse breeding systems to favor outcrossing, such as self‐incompatibility. However, changes in biotic and abiotic conditions can result in selective pressures that lead to a breakdown in self‐incompatibility. The shift to increased selfing is commonly associated with reduced floral features, lower attractiveness to pollinators, and increased inbreeding. We tested the hypothesis that the loss of self‐incompatibility, a shift to self‐fertilization (autogamy), and concomitant evolution of the selfing syndrome (reduction in floral traits associated with cross‐fertilization) will lead to increased inbreeding and population differentiation in Oenothera primiveris. Across its range, this species exhibits a shift in its breeding system and floral traits from a self‐incompatible population with large flowers to self‐compatible populations with smaller flowers.MethodsWe conducted a breeding system assessment, evaluated floral traits in the field and under controlled conditions, and measured population genetic parameters using RADseq data.ResultsOur results reveal a bimodal transition to the selfing syndrome from the west to the east of the range of O. primiveris. This shift includes variation in the breeding system and the mating system, a reduction in floral traits (flower diameter, herkogamy, and scent production), a shift to greater autogamy, reduced genetic diversity, and increased inbreeding.ConclusionsThe observed variation highlights the importance of range‐wide studies to understand breeding system variation and the evolution of the selfing syndrome within populations and species.

Understanding the biology of floral traits underlying plant‐pollinator relationships in a community context is important to predict the occurrence and frequency of pairwise mutualistic interactions. Flower nectar is a sugar‐rich solution that is key in plant–pollinator interactions. Nectar sugar concentration directly determines viscosity, regulating nectar ingestion rates by pollinators and, together with nectar volume, shapes foraging decisions. Furthermore, the link between nectar ingestion efficiency and sugar concentration in bee pollinators is mediated by their proboscis length and the spatial positioning of nectar within flowers. We therefore hypothesize that nectar concentration can be related to the structure of bee–flower interactions in a community. We tested the hypothesis in a species‐rich alpine meadow in the Hengduan Mountains, China, a global biodiversity hotspot. The study meadow sustains an extremely diverse bumblebee community. We constructed the bee–flower network in the 2020 flowering season. Visitation frequency of the bumblebees to each flowering plant was recorded while nectar sugar concentration and initial volume of those plants were quantified in addition to other pollination‐related floral traits. We conducted phylogenetically comprehensive analyses to assess factors influencing the network structure, bees' visitation frequency and flower preference, and to detect the correlations of bees' proboscis length with nectar sugar concentration and corolla tube length and the relationships among the pollination‐related floral traits. Modularity analysis indicated that the community bee–flower network was structured by nectar sugar concentration and corolla tube length, which was associated with bees' proboscis length. However, the overall bees' visitation frequency to flowers was correlated to nectar sugar concentration but not to corolla tube length. Flowers visited by long‐proboscid bee species had comparatively dilute nectar. Moreover, the bumblebees' flower preferences were positively influenced by floral abundance. Additionally, variation in nectar sugar concentration was linked with pollination‐related floral traits and pollinator composition, suggesting a diversity of pollination strategies among the plants in the community. Synthesis: To our knowledge, this is the first field investigation uncovering how bee‐flower interactions and nectar sugar concentration are linked at the community level, which broadens the niche dimensions for resource partitioning among co‐occurring species and thus fosters biodiversity.

In flowering plants, the shift from outcrossing to selfing is associated with a set of correlated changes in morphological and reproductive features known as the ‘selfing syndrome’. Species of the sub-endemic Mediterranean genus Centaurium Hill (Gentianaceae) exhibit a wide array of flower traits related to pollination biology and different ploidy levels. We explored if the evolutionary transitions of seven flower traits and life cycle, typically associated with the selfing syndrome, are related to polyploidy, diversification patterns, divergence times and the geological and climatic history of the Mediterranean Basin. Using 26 species of Centaurium we reconstructed a phylogenetic tree, inferred the ancestral states of the selected traits, estimated their phylogenetic signal and tested the correlative evolution among them. We found a significant increase in diversification rates during the Quaternary. Anther length, flower size, herkogamy and polyploidy undergo rapid state transitions without phylogenetic signal that could be the result of adaptation for selfing. Changes in character states do not show evidence of correlative evolution among them, as would be predicted during the evolution of selfing syndrome. The evolution of reproductive traits in Centaurium has probably relied on a more diverse array of drivers than just reproductive assurance or polyploidy events.

How and why plants evolve to become selfing is a long-standing evolutionary puzzle. The transition from outcrossing to highly selfing is less well understood in self-compatible (SC) mixed-mating (MM) species where potentially subtle interactions between floral phenotypes and the environment are at play. We examined floral morphological and developmental traits across an entire SC MM genus, Collinsia, to determine which, if any, predict potential autonomous selfing ability when pollinators are absent (AS) and actual selfing rates in the wild, s(m), and to best define the selfing syndrome for this clade. Using polymorphic microsatellite markers, we obtained 30 population-level estimates of s(m) across 19 Collinsia taxa. Species grand means for the timing of herkogamy (stigma-anther contact) and dichogamy (stigmatic receptivity, SR), AS, floral size, longevity and their genetic correlations were quantified for 22 taxa. Species fell into discrete selfing and outcrossing groups based on floral traits. Loss of dichogamy defines Collinsia's selfing syndrome. Floral size, longevity and herkogamy also differ significantly between these groups. Most taxa have high AS rates (>80 %), but AS is uncorrelated with any measured trait. In contrast, s(m) is significantly correlated only with SR. High variance in s(m) was observed in the two groups. Collinsia species exhibit clear morphological and developmental traits diagnostic of 'selfing' or 'outcrossing' groups. However, many species in both the 'selfing' and the 'outcrossing' groups were MM, pointing to the critical influence of the pollination environment, the timing of AS and outcross pollen prepotency on s(m). Flower size is a poor predictor of Collinsia species' field selfing rates and this result may apply to many SC species. Assessment of the variation in the pollination environment, which can increase selfing rates in more 'outcrossing' species but can also decrease selfing rates in more 'selfing' species, is critical to understanding mating system evolution of SC MM taxa.

In a controlled environment, we artificially induced drought during flowering of Epilobium angustifolium, an animal-pollinated plant. Leaf water potential (ψ(l)) and floral traits were monitored over a 12-d period of soil moisture depletion. Soil moisture depletion induced drought stress over time, as revealed by significant treatment × day interactions for predawn and midday ψ(l). Nectar volume and flower size showed significant negative responses to drought stress, but nectar sugar concentration did not vary between treatments. Floral traits were more buffered from drought than leaf water potentials. We used path analysis to examine direct and indirect effects of ψ(l) on floral traits for plants in well-watered (control) vs. drought treatments. According to the best-fit path models, midday ψ(l) has significant positive effects on flower size and nectar volume in both environments. However, for controls midday ψ(l) also had a significant negative effect on nectar sugar concentration. Results indicate that traits influencing floral attractiveness to pollinators in E. angustifolium vary with plant water status, such that pollinator-mediated selection could indirectly target physiological or biochemical controls on ψ(l). Moreover, under mesic conditions selection for greater nectar sugar reward may be constrained by the antagonistic effects of plant water status on nectar volume and sugar concentration.

ABSTRACTPlant domestication primarily targets traits of direct human interest, such as fruit and seed characteristics; however, its indirect effects on other traits, including floral morphology and rewards (nectar and pollen), remain less understood. In this study, we investigated how domestication has influenced floral traits and rewards in domesticated and wild species of the genus Cucurbita. We compared three domesticated and three wild Cucurbita species in an experimental plot. We measured floral morphological traits, nectar volume, sugar (fructose, glucose, and sucrose), and amino acid concentrations in staminate and pistillate flowers. In addition, we evaluated pollen production and size, as well as protein and lipid concentrations, and the protein: lipid ratio in staminate Cucurbita flowers. Our results show that domesticated Cucurbita species exhibit larger floral morphological traits in both pistillate and staminate flowers compared to their wild relatives. While nectar volume increased in domesticated species, sugar and amino acid concentrations remained unchanged. In contrast, domestication had no significant effect on pollen traits, including production, size, and protein and lipid content. These findings highlight that domestication differentially affects floral traits: while floral morphology is significantly altered, most of the traits of floral rewards remain largely unaffected. This conservation may arise from the recent evolutionary history of these species and their close coevolutionary relationship with Eucera bees, emphasizing pollinator nutritional needs over artificial selection. These results underscore the complex interplay between domestication, resource allocation, and plant–pollinator interactions in shaping floral traits.

We tested for an association between nectar and various floral traits and investigated their roles as primary and secondary pollinator attractants in hummingbird-pollinated Silene virginica. Our goal was to gain insight into the mechanisms of pollinator-mediated selection that underlies floral trait divergence within the genus. In a field population of S. virginica, we measured five floral and eight vegetative traits and quantified nectar volume, nectar sugar concentration, and total sugar reward (nectar volume × nectar sugar concentration). All three components of nectar reward were positively correlated to flower size, and nectar volume varied significantly among individuals within the population. To ascertain whether the correlation of specific floral traits with nectar reward influences the behavior of the primary pollinator of S. virginica, the ruby-throated hummingbird, Archilochus colubris, we investigated whether A. colubris preferred the expression of floral traits associated with high nectar volume and total sugar reward. We accomplished this by constructing floral arrays consisting of artificial flowers that had equal nectar quantity and total sugar reward but that differed in petal area and corolla tube diameter, which were positively correlated with nectar quantity and total sugar reward in our field study. In observations of visitation frequencies to the various floral-trait combinations, hummingbirds preferentially visited artificial floral phenotypes with larger petal displays, with the greatest preference for floral phenotypes with both larger petals and wider corolla-tube diameters. This association between primary and secondary floral attractants and hummingbird discrimination of floral features supports the concept that the floral traits of S. virginica reflect pollinator-mediated selection by the principal pollinator.

The mating systems and floral traits often among relatives of hermaphroditic plants can exhibit considerable diversity. This diversity can be influenced by the evolution of selfing and associated floral traits as a form of reproductive assurance (RA) when pollen limitation (PL) results from insufficient pollinator availability. To explore whether the degree of PL primarily drives differences in mating systems and floral traits, we conducted a comprehensive study involving two closely related species, Halenia elliptica and Halenia grandiflora, in three sympatric sites. We investigated floral characteristics, pollinator visitation, PL, autonomous selfing ability, RA and mating system in studied populations. Our findings show that H. elliptica produces smaller flowers and less nectar production than H. grandiflora, making it less attractive to pollinators. Compared with H. grandiflora, H. elliptica experienced more severe outcross pollen limitation (OPL), but compensates with a higher capacity for autonomous selfing, ensuring seed production under natural conditions. Moreover, significant differences in mating systems were detected between them, with H. elliptica exhibiting a higher selfing rate than H. grandiflora across all studied sympatric populations. These differences are also reflected in variations in herkogamy and dichogamy. Our study suggested that the degree of OPL impacts the divergence in mating systems and floral traits between sympatric closely related species, offering valuable insights into the evolution of plant mating systems and floral traits.

Although the evolution of the selfing syndrome often involves reductions in floral size, pollen and nectar, few studies of selfing syndrome divergence have examined nectar. We investigate whether nectar traits have evolved independently of other floral size traits in the selfing syndrome, whether nectar traits diverged due to drift or selection, and the extent to which quantitative trait locus (QTL) analyses predict genetic correlations. We use F5 recombinant inbred lines (RILs) generated from a cross between Ipomoea cordatotriloba and Ipomoea lacunosa. We calculate genetic correlations to identify evolutionary modules, test whether trait divergence was due to selection, identify QTLs and perform correlation analyses to evaluate how well QTL properties reflect genetic correlations. Nectar and floral size traits form separate evolutionary modules. Selection has acted to reduce nectar traits in the selfing I. lacunosa. Genetic correlations predicted from QTL properties are consistent with observed genetic correlations. Changes in floral traits associated with the selfing syndrome reflect independent evolution of at least two evolutionary modules: nectar and floral size traits. We also demonstrate directional selection on nectar traits, which is likely to be independent of selection on floral size traits. Our study also supports the expected mechanistic link between QTL properties and genetic correlations.

Plant-pollinator interactions are thought to be major drivers of floral trait diversity. However, the relative importance of divergent pollinator-mediated selection vs. neutral processes in floral character evolution has rarely been explored. We tested for adaptive floral trait evolution by comparing differentiation at neutral genetic loci to differentiation at quantitative floral traits in a putative Ipomopsis aggregata hybrid zone. Typical I.aggregata subsp. candida displays slender white tubular flowers that are typical of flowers pollinated by hawkmoths, and subsp. collina displays robust red tubular flowers typical of flowers pollinated by hummingbirds; yet, hybrid flower morphs are abundant across the East Slope of the Colorado Rockies. We estimated genetic differentiation (FST ) for nuclear and chloroplast microsatellite loci and used a half-sib design to calculate quantitative trait divergence (QST ) from collection sites across the morphological hybrid zone. We found little evidence for population structure and estimated mean FST to be 0.032. QST values for several floral traits including corolla tube length and width, colour, and nectar volume were large and significantly greater than mean FST . We performed multivariate comparisons of neutral loci to genetic correlations within and between populations and found a strong signal for divergent selection, suggesting that specific combinations of floral display and reward traits may be the targets of selection. Our results show little support for historical subspecies categories, yet floral traits are more diverged than expected due to drift alone. Non-neutral divergence for multivariate quantitative traits suggests that selection by pollinators is maintaining a correlation between display and reward traits.

The diversity of floral forms has long been considered a prime example of radiation through natural selection. However, little is still known about the evolution of floral traits, a critical piece of evidence for the understanding of the processes that may have driven flower evolution. We studied the pattern of evolution of quantitative floral traits in a group of Neotropical lianas (Bignonieae, Bignoniaceae) and used a time-calibrated phylogeny as basis to: (1) test for phylogenetic signal in 16 continuous floral traits; (2) evaluate the rate of evolution in those traits; and (3) reconstruct the ancestral state of the individual traits. Variation in floral traits among extant species of Bignonieae was highly explained by their phylogenetic history. However, opposite signals were found in floral traits associated with the attraction of pollinators (calyx and corolla) and pollen transfer (androecium and gynoecium), suggesting a differential role of selection in different floral whorls. Phylogenetic independent contrasts indicate that traits evolved at different rates, whereas ancestral character state reconstructions indicate that the ancestral size of most flower traits was larger than the mean observed sizes of the same traits in extant species. The implications of these patterns for the reproductive biology of Bignonieae are discussed. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 102, 378‐390. ADDITIONAL KEYWORDS: ancestral character states ‐ Bignoniaceae ‐ K-statistic ‐ morphological evolution ‐ Pagel’s ancestral character state reconstruction ‐ phylogenetically independent contrasts ‐ quantitative traits.

Floral traits are commonly thought to be more canalized than vegetative ones. In addition, floral and vegetative traits are hypothesized to be genetically decoupled, enabling vegetative structures to respond plastically to environmental heterogeneity, and to evolve in response to selection without disrupting the reproductive function of flowers. To test these hypotheses, we evaluate the genetic architecture of floral and vegetative traits in natural populations of Arabidopsis thaliana raised under variable light-quality environments. Plants were grown either under high or low ratios of red to far-red (R:FR) light, an aspect of light quality that varies with neighbor proximity and regulates competitive shade-avoidance responses. Across environments, we detected significant genetic variation for the average expression of all measured floral traits (petal length and width, stamen length, pistil length, stigma-anther separation, and exsertion of both the stamen and pistil beyond the corolla). Light quality significantly influenced the absolute size of several floral traits as well as the allometry (i.e., relative scaling) of all floral traits, and genotypes differed in the plasticity of floral traits to the light treatments. Exposure to low relative to high R:FR resulted in significantly greater elongation in the vegetative trait, petiole length, and genotypes again differed in the plasticity of this trait to R:FR. Consistent with prior studies, most floral traits were less plastic than the vegetative trait; herkogamy (i.e., stigma-anther separation) was the exception and expressed more variable trait values across environments than petiole length, apparently as a consequence of the independent responses of stamens and pistils. Flowers also showed strong phenotypic integration; genotypic correlations were significantly positive among floral traits within each light treatment. Although floral-vegetative correlations were not significant in the high R:FR light treatment, significant correlations were detected between petal traits, pistil length, and petiole length under low R:FR, in contrast to the widely held hypothesis that floral and vegetative traits are genetically independent. Finally, we detected selection for reduced herkogamy in the low R:FR light treatment. The observed correlation between functional trait groups suggest that vegetative plasticity may affect the expression of floral traits in some environments, and that environment-specific constraints may exist on the evolution of floral and vegetative traits.

Floral traits and rewards are important in mediating interactions between plants and pollinators. Agricultural management practices can affect abiotic factors known to influence floral traits; however, our understanding of the links between agricultural practices and floral trait expression is still poorly understood. Variation in floral morphological, nectar, and pollen traits of two important agricultural species, Coffea arabica and C. canephora, was assessed under different agricultural practices (sun and shade). Corolla diameter and corolla tube length were larger and pollen total nitrogen content greater in shade plantations of C. canephora than sun plantations. Corolla tube length and anther filament length were larger in shade plantations of C. arabica. No effect of agricultural practice was found on nectar volume, sugar or caffeine concentrations, or pollen production. Pollen total nitrogen content was lower in sun than shade plantations of C. canephora, but no difference was found between sun and shade for C. arabica. This study provides baseline data on the influence of agronomic practices on C. arabica and C. canephora floral traits and also helps fill a gap in knowledge about the effects of shade trees on floral traits, which can be pertinent to other agroforestry systems.

The evolution and expression of floral traits are responsive to selection pressure from biotic and abiotic factors. Although floral traits significantly vary among environments, the flower remains unchanged. We aimed to understand the adaptation of Epimedium chlorandrum of floral traits to a frequently nocturnally rainy and wet environments and the roles of floral traits in pollination and reproduction. We observed flowering phenology, measured floral characteristics including the number of pollen grains and ovules per flower, measured pollen viability and stigma receptivity, tested the volume and sugar concentration of nectar and conducted flower-visit observations in this species. Different pollination treatments were performed to characterize the breeding system. The inner sepals and highly curved longer spurs of E. chlorandrum jointly formed an umbrella that shielded the anthers and stigma from rain wash and prevented nectar dilution. Epimedium chlorandrum was visited by six species, while Bombus trifasciatus was the only effective pollinator and fed on the nectar. One flower secreted approximately 17.06 µL of nectar with a 29.19 g/100 mL sugar concentration, and the pollination efficiency of B. trifasciatus was positively associated with the nectar sugar concentration. The self- and open-pollination treatments resulted in fewer fertile seeds than the cross-pollination treatment. In contrast, the autonomous self-pollination treatment failed to yield fertile seeds. In summary, pollen limitation caused by harsh weather and pollinator shortage occurred during the pollination process of E. chlorandrum, which was partially alleviated by self-compatibility.