Background and aims – Variation in flower color among individuals within plant populations has long fascinated horticulturalists and evolutionary biologists alike. Nevertheless, the diversity of genetic factors and ecological interactions that act to maintain flower-color polymorphisms over time are understood for very few species. Here, we suggest that Solanum carolinense (horsenettle) makes an excellent model system for studying the evolutionary ecology of a flower-color polymorphism, and we begin to shed insight into factors that might affect the stability of the polymorphism. Material and methods – We transplanted 24 horsenettle ramets of each of 40 field-collected genets (10 genets per each of four source fields) into a common-garden plot (in an oldfield with an existing horsenettle population). For each ramet, we visually characterized petal color and quantified production of flowers, fruits, seeds, and damage by 11 species of herbivores. Key results – Rather than varying continuously within ramets and among genets, petal color fell into three relatively discrete categories that are common in most horsenettle populations: purple, white, and mauve. In the common-garden experiment, purple-petalled morphs initiated significantly more flower buds, opened more flowers, and produced more fruits and seeds than did the mauve-petalled or white-petalled morphs. Petal-color morphs differed only slightly in resistance to a minority of the herbivore community, with no consistent advantage for color morphs across species of herbivores or types of tissues fed upon. Conclusion – Horsenettle has numerous characteristics that make it an excellent system for studying the genetics and ecology that drive the maintenance of flower-color polymorphisms. Although pleiotropic effects of flower-color on resistance to herbivory appear to be minimal in horsenettle, potential pleiotropies that differentially affect maternal and paternal reproduction, or sexual and vegetative reproduction, are worthy of further study.

The mechanisms underlying trait conservation over long evolutionary time scales are poorly known. These mechanisms fall into the two broad and nonmutually exclusive categories of constraint and selection. A variety of factors have been hypothesized to constrain trait evolution. Alternatively, selection can maintain similar trait values across many species if the causes of selection are also relatively conserved, while many sources of constraint may be overcome over longer periods of evolutionary divergence. An example of deep trait conservation is tetradynamy in the large family Brassicaceae, where the four medial stamens are longer than the two lateral stamens. Previous work has found selection to maintain this difference in lengths, which we call anther separation, in wild radish, Raphanus raphanistrum. Here, we test the constraint hypothesis using five generations of artificial selection to reduce anther separation in wild radish. We found a rapid linear response to this selection, with no evidence for depletion of genetic variation and correlated responses to this selection in only four of 15 other traits, suggesting a lack of strong constraint. Taken together, available evidence suggests that tetradynamy is likely to be conserved due to selection, but the function of this trait remains unclear.

The study of adaptation helps explain biodiversity and predict future evolution. Yet the process of adaptation can be difficult to observe due to limited phenotypic variation in contemporary populations. Furthermore, the scarcity of male fitness estimates has made it difficult to both understand adaptation and evaluate sexual conflict hypotheses. We addressed both issues in our study of two anther position traits in wild radish (Raphanus raphanistrum): anther exsertion (long filament - corolla tube lengths) and anther separation (long - short filament lengths). These traits affect pollination efficiency and are particularly interesting due to the unusually high correlations among their component traits. We measured selection through male and female fitness on wild radish plants from populations artificially selected to recreate ancestral variation in each anther trait. We found little evidence for conflicts between male and female function. We found strong evidence for stabilizing selection on anther exsertion and disruptive selection on anther separation, indicating positive and negative correlational selection on the component traits. Intermediate levels of exsertion are likely an adaptation to best contact small bees. The function of anther separation is less clear, but future studies might investigate pollen placement on pollinators and compare species possessing multiple stamen types.

Iris verna L. var. smalliana Fernald ex M.E. Edwards (Family Iridaceae, hereafter referred to as Iris verna) is currently listed as endangered in Pennsylvania. The six extant populations in Pennsylvania are all located in one state forest and represent the northern end of the range for the species. There is little known about the basic biology of I. verna throughout its range, making preservation of threatened populations difficult. Iris verna was tested for self-compatibility and the ability to autonomously self-pollinate in 2016. Pollinator limitation studies were conducted on three of the Pennsylvania populations in 2015 and 2016 by comparing hand-pollinated and naturally pollinated flowers. Potential animal pollinators or seed dispersers were identified through in-person observation and video camera use. Iris verna was found to be self-compatible, with no significant reduction in seed production in selfed flowers, but the species does require a pollinator for pollen transfer. Seed production in I. verna was limited by pollinators during both years of the study and seed production in both control and hand-pollinated flowers was variable across years, suggesting resource availability also limits sexual reproduction. Ants were the only observed animals potentially coming in contact with reproductive organs while visiting flowers, and were also identified as seed dispersers. Our findings suggest that pollinators can be an important limiting factor to sexual reproduction in these endangered populations of I. verna in Pennsylvania.

Background and AimsPlant-pollinator interactions shape the evolution of flowers. Floral attraction and reward traits have often been shown to affect pollinator behaviour, but the possible effect of efficiency traits on visitation behaviour has rarely been addressed. Anther position, usually considered a trait that influences efficiency of pollen deposition on pollinators, was tested here for its effect on pollinator visitation rates and visit duration in flowers of wild radish, Raphanus raphanistrum .MethodsArtificial selection lines from two experiments that expanded the naturally occurring phenotypic variation in anther position were used. In one experiment, plant lines were selected either to increase or to decrease anther exsertion. The other experiment decreased anther dimorphism, which resulted in increased short stamen exsertion. The hypothesis was that increased exsertion would increase visitation of pollen foragers due to increased visual attraction. Another hypothesis was that exsertion of anthers above the corolla would interfere with nectar foragers and increase the duration of visit per flower.Key ResultsIn the exsertion selection experiment, increased exsertion of both short and long stamens resulted in an increased number of fly visits per plant, and in the dimorphism experiment bee visits increased with increased short stamen exsertion. The duration of visits of nectar feeders declined significantly with increasing long stamen exsertion, which was opposite to the hypothesis.ConclusionsUntil now, anther position was considered to be an efficiency trait to enhance pollen uptake and deposition. Anther position in wild radish is shown here also to have an ecological significance in attracting pollen foragers. This study suggests an additional adaptive role for anther position beyond efficiency, and highlights the multiple ecological functions of floral traits in plant-pollinator interactions.

Abstract: Invasive ants can influence abundance and diversity of insect pollinators, and this effect may be expected to be especially strong on oceanic islands. We examined abundance of native Hylaeus bees and patterns of flower visitation to three native Hawaiian plant species (Leptecophylla tameiameiae, Santalum haleakalae, and Geranium cuneatum) in presence and absence of invasive Argentine ants (Linepithema humile) in the subalpine shrubland of Haleakalā volcano. Native bees, nonnative honeybees, and microlepidoptera were responsible for the majority of visits to flowers of the three plant species. Ants visited flowers of S. haleakalae and G. cuneatum relatively commonly in ant-invaded zones but were less common or absent on flowers of L. tameiameiae in the same zones. Zones with ants had fewer Hylaeus bees in yellow pan traps than the zone without ants, and overall flower visitation rates to one plant species, G. cuneatum, were six times lower in sites containing ants. In one of two observation years, flower visitation to L. tameiameiae was also reduced in sites containing ants. Results suggest that Argentine ants may cause a reduction of native Hylaeus bee abundances; however, other factors correlated with elevation cannot be ruled out as the cause of differences in Hylaeus abundances across our sites.

One explanation for the success of exotic plants in their introduced habitats is that, upon arriving to a new continent, plants escaped their native herbivores or pathogens, resulting in less damage and lower abundance of enemies than closely related native species (enemy release hypothesis). We tested whether the three exotic plant species, Rubus phoenicolasius (wineberry), Fallopia japonica (Japanese knotweed), and Persicaria perfoliata (mile-a-minute weed), suffered less herbivory or pathogen attack than native species by comparing leaf damage and invertebrate herbivore abundance and diversity on the invasive species and their native congeners. Fallopia japonica and R. phoenicolasius received less leaf damage than their native congeners, and F. japonica also contained a lower diversity and abundance of invertebrate herbivores. If the observed decrease in damage experienced by these two plant species contributes to increased fitness, then escape from enemies may provide at least a partial explanation for their invasiveness. However, P. perfoliata actually received greater leaf damage than its native congener. Rhinoncomimus latipes, a weevil previously introduced in the United States as a biological control for P. perfoliata, accounted for the greatest abundance of insects collected from P. perfoliata. Therefore, it is likely that the biocontrol R. latipes was responsible for the greater damage on P. perfoliata, suggesting this insect may be effective at controlling P. perfoliata populations if its growth and reproduction is affected by the increased herbivore damage.

Little is known regarding pollination webs involving island coastal plants and pollinators, and roles that nonnative flower visitors may play in these interaction networks. Plant-pollinator observations made in March 2008 and 2009 were used to describe the pollination network for Ka'ena Point, one of Hawai'i's best-conserved coastal communities. The network includes 15 native plant species, two native bee species, and 26 nonnative insect taxa, forming 119 interactions. Network connectance is 29.4% and weighted nestedness is 17.9, which are similar to values of other dry-habitat, island networks. The network's structure has a core of generalized pollinators plus several more-specialized pollinators. Nearly all plant species interact with two or more generalist pollinators and a variable number of specialists. Small, nonnative bees (Lasioglossum, Ceratina), wasps (Proconura), and flies (mostly Tachinidae) were responsible for 72.7% of flower visits, and they visit five plant species not visited by native bees. The two native visitors were the bees Hylaeus anthracinus and H. longiceps (both proposed as endangered). Hylaeus spp. (especially females) provided 19.8% of flower visits, foraging at high visitation rates and on many species, including the endangered Scaevola coriacea and Sesbania tomentosa. In Hawai'i's coastal habitat, nonnative insects form novel interactions with native species and may maintain an ecosystem's function following loss of most of the original native pollinators. However, their high visitation rates suggest that the two remaining native Hylaeus species are potentially important pollinators for many of the native plants on which they rely for nectar and pollen resources.

Numerous studies have examined foraging strategies of pollinators in continental landscapes, but relatively little is known about island pollinators. We investigated four native plant communities and their potential pollinators on Mauna Loa Volcano on Hawai'i Island across two environmental gradients: elevation (middle and high) and succession (early and late). The effects of elevation and successional stage on visitation rates and degree of plant specialization were compared. Elevation had a significant effect on visitation rates with plants at mid elevation receiving the most visits, whereas succession had little effect on visitation rates. Both succession and elevation impacted plant specialization, with plants at late-succession and high-elevation sites being more specialized in terms of their diversity of flower visitors. Metrosideros polymorpha (Myrtaceae) received the most forager visits, even when it was not the most abundant plant flowering. Endemic Hylaeus bees (Hymenoptera: Colletidae) were the dominant visitors across all sites (64%-91% of visits), and endemic birds were observed only at mid elevation, primarily in the late-succession site. Considering the paucity of natural history studies on Hawaiian Hylaeus, further investigation of their potential role as pollinators of native Hawaiian plants is necessary.

Genetic correlations are the most commonly studied of all potential constraints on adaptive evolution. We present a comprehensive test of constraints caused by genetic correlation, comparing empirical results to predictions from theory. The additive genetic correlation between the filament and the corolla tube in wild radish flowers is very high in magnitude, is estimated with good precision (0.85 ± 0.06), and is caused by pleiotropy. Thus, evolutionary changes in the relative lengths of these two traits should be constrained. Still, artificial selection produced rapid evolution of these traits in opposite directions, so that in one replicate relative to controls, the difference between them increased by six standard deviations in only nine generations. This would result in a 54% increase in relative fitness on the basis of a previous estimate of natural selection in this population, and it would produce the phenotypes found in the most extreme species in the family Brassicaceae in less than 100 generations. These responses were within theoretical expectations and were much slower than if the genetic correlation was zero; thus, there was evidence for constraint. These results, coupled with comparable results from other species, show that evolution can be rapid despite the constraints caused by genetic correlations.

Although conflicting selection from different resources is thought to play a critical role in the evolution of specialized species, the prevalence of conflicting selection in generalists is poorly understood. Plants may experience conflicting selection on floral traits by different pollinators and between genders. Using artificial selection to increase phenotypic variation, we tested for conflicting and nonadditive selection on wild radish (Raphanus raphanistrum) flowers. To do this, we measured selection by each of the major pollinator taxa through both male and female fitness, and tested for a single-generation response to selection by a subset of these pollinators. We found some evidence for conflicting selection on anther exertion--sweat bees exerted stabilizing selection and larger bees selected for increased exertion. Stamen dimorphism was only under selection by honey bees, causing a response to selection in the next generation, and flower size was under similar selection by multiple pollinators. Selection differed significantly between genders for two traits, but there was no evidence for stronger selection through male fitness or for conflicting selection between genders. Our results suggest wild radish flowers can adapt to multiple pollinators, as we found little evidence for conflicting selection and no evidence for nonadditive selection among pollinators.

BACKGROUNDThere are a number of difficulties associated with the study of adaptation. One is a lack of variation in the trait, which is common in adaptations because past selection has removed unfit variants. This lack of variation makes it difficult to determine the relationship between trait variation and fitness. Another difficulty is proving causation in this trait-fitness relationship, because a correlated trait might be the actual adaptation. These difficulties can be ameliorated at least partially by combining studies of natural variation with studies of experimentally manipulated traits and traits whose variance has been augmented by artificial selection.SCOPEWe review here a number of our studies on the adaptive value of two aspects of anther position in wild radish (Raphanus raphanistrum, Brassicaceae): anther exsertion, i.e. the degree to which anthers protrude from the mouth of the corolla tube, and anther height dimorphism, i.e. the difference in lengths of the filaments between the two short and four long stamens. We have used both functional analyses, in which the response variable is pollen removal, and measurements of selection, in which the response variable is lifetime male fitness estimated by molecular genetic paternity analyses. In these studies we use both the natural variation in populations as well as manipulated variation, the latter through both stamen removal and artificial selection, to re-create the ancestral trait conditions.CONCLUSIONSOur work provides convincing evidence that intermediate anther exsertion values are adaptive, and that this is probably an adaptation to a subset of the pollinator fauna, small bees. The picture for anther height dimorphism is much less clear, as the weight of current evidence suggests that current values of this trait might actually be maladaptive; however, if this is true it is difficult to understand how the dimorphism is maintained across the family Brassicaceae.

Weedy species with wide geographical distributions may face strong selection to adapt to new environments, which can lead to adaptive genetic differentiation among populations. However, genetic drift, particularly due to founder effects, will also commonly result in differentiation in colonizing species. To test whether selection has contributed to trait divergence, we compared differentiation at eight microsatellite loci (measured as FST) to differentiation of quantitative floral and phenological traits (measured as QST) of wild radish (Raphanus raphanistrum) across populations from three continents. We sampled eight populations: seven naturalized populations and one from its native range. By comparing estimates of QST and FST, we found that petal size was the only floral trait that may have diverged more than expected due to drift alone, but inflorescence height, flowering time, and rosette formation have greatly diverged between the native and nonnative populations. Our results suggest the loss of a rosette and the evolution of early flowering time may have been the key adaptations enabling wild radish to become a major agricultural weed. Floral adaptation to different pollinators does not seem to have been as necessary for the success of wild radish in new environments.

Plant-pollinator interactions are one of the most important and variable mutualisms in nature. Multiple pollinators often visit plants and can vary in visitation rates, pollen removal and deposition, and spatial and temporal distribution, altering plant reproduction and patterns of pollinator-mediated selection. Although some visitors may not be effective pollinators, pollinator effectiveness is rarely estimated directly as seed set resulting from a single visit by each taxon visiting generalist plants. For two years, effectiveness of visitors to wild radish, Raphanus raphanistrum, was quantified by counting seeds set and pollen grains removed as a result of a single visit. We calculated a pollinator's importance to plant reproduction as the product of visitation rate and single-visit seed set, and regressed pollinator body size on pollen-removal and on seed set effectiveness. Although pollinators differed in effectiveness and visitation rates, pollinator importance was primarily determined by visitation rates. In contrast to similar 2-yr studies, pollinator assemblage composition varied little, suggesting pollinator-mediated selection can be consistent across years for this generalist. Larger pollinators were more effective than smaller at effecting seed set, but body size was a poor predictor of pollen removal ability. Instead, pollen-removal effectiveness may be more influenced by foraging behavior than size.

We developed 15 polymorphic microsatellite loci from the Hawaiian tree Metrosideros polymorpha. These loci were screened against two varieties from several populations and from 23 individuals from one mid-elevation population on Hawaii Island. Loci were variable with the number of alleles per locus ranging from three to 24. Polymorphic information content ranged from 0.222 to 0.941, and observed heterozygosity ranged from 0.261 to 0.955.

Despite the development of diversity indices in community ecology that incorporate both richness and evenness, pollination biologists commonly use only pollinator richness to estimate generalization. Similarly, while pollination biologists have stressed the utility of pollinator importance, incorporating both pollinator abundance and effectiveness, importance values have not been included in estimates of generalization in pollination systems. In this study, we estimated pollinator generalization for 17 plant species using Simpson’s diversity index, which includes richness and evenness. We compared these estimates with estimates based on only pollinator richness, and compared diversity estimates calculated using importance data with those using only visitation data. We found that pollinator richness explains only 57–65% of the variation in diversity, and that, for most plant species, pollinator importance was determined primarily by differences in visitation rather than by differences in effectiveness. While simple richness may suffice for broad comparisons of pollinator generalization, measures that incorporate evenness will provide a much more accurate understanding of generalization. Although incorporating labor-intensive measurements of pollinator effectiveness are less necessary for broad surveys, effectiveness estimates will be important for detailed studies of some plant species. Unfortunately, at this point it is impossible to predict a priori which species these are.

How species evolve depends on the communities in which they are embedded. Here, we briefly review the ideas underlying concepts of diffuse coevolution, evolution, and selection. We discuss criteria to identify when evolution will be diffuse. We advocate a more explicitly trait-oriented approach to diffuse (co)evolution, and discuss how considering effects of interacting species on fitness alone tells us little about evolution. We endorse the view that diffuse evolution occurs whenever the response to selection by one interacting species on a given trait is altered by the presence of a second interacting species. Building on the work of others, we clarify and expand the criteria for diffuse evolution and present a simple experimental design that will allow the detection of diffuse selection. We argue that a greater focus on selection on specific traits and the evolutionary response to that selection will improve our conceptual understanding of how communities affect the evolution of species embedded within them.