Local adaptation to hummingbirds and bees in Salvia stachydifolia: insights into pollinator shifts in a Southern Andean sage.

BACKGROUND AND AIMS: Differences among populations in pollinator assemblages can lead to local adaptation mosaics in which plants evolve different floral morphologies and attractive traits. Mountain habitats may promote local adaptation because of differences in environmental conditions with altitude, causing changes in pollinators, and because mountaintops can act as isolated habitats. We studied if the differences in floral shape, size and nectar traits in Salvia stachydifolia can be attributed to variations in the relative contribution of hummingbirds and insects. METHODS: We studied eight populations of S. stachydifolia in natural and under common garden conditions, to assess whether population differences have a genetic component. We recorded pollinators, their behaviour and visitation rates, and characterized pollinator assemblages. In addition, we measured nectar volume and concentration, and collected flowers to describe floral shape and size variation using geometric morphometric methods. We then applied an unsupervised learning algorithm to identify ecotypes based on morphometric traits. Finally, we explored whether populations with different pollinator assemblages had different climatic and/or elevation preferences. KEY RESULTS: We found that variation in the identity of the main pollinators was associated with differences among populations in all traits, as expected under a local adaptation scenario. These differences persisted in the common garden, suggesting that they were not due to phenotypic plasticity. We found S. stachydifolia populations were pollinated either by bees, by hummingbirds or had mixed pollination. We identified two ecotypes that correspond to the identity of the main pollinator guilds, irrespective of climate or altitude. CONCLUSIONS: Variation in S. stachydifolia floral traits did not follow any evident association with bioclimatic factors, suggesting that populations may have diverged as the product of historical isolation on mountaintops. We suggest that differences among populations point to incipient speciation and an ongoing pollinator shift.

Annual molt period and seasonal color variation in the Eared Dove´s crown.

Molting is an important process in which old and worn feathers are exchanged for new ones. Plumage color is determined by pigments such as carotenes, melanin and by the ultrastructure of the feather. The importance of plumage coloration has been widely studied in different groups of birds, generally at a particular time of the year. However, plumage coloration is not static and few studies have addressed the change in plumage color over time and its relationship to reproductive tasks. The Eared Dove (Zenaida auriculata, Des Murs, 1847) has a melanistic coloration with sexual dichromatism in different body regions. The Eared Dove´s crown is the most exposed body region during the bowing display. Our objective was therefore to accurately determine the molting period of the crown feathers and study the seasonal variation in their coloration in females and males. Our findings indicate a molting period of 6 months (January to June). The new feathers are undergoing changes in their coloration from July to December. During that period we apply an avian vision model then enabled us to reveal a seasonal variation in the coloration of the crown feathers in both sexes, as given by a change in the chromatic distances. The highest values in the chromatic distances towards the reproductive period are given by a change in the UV-violet component of the spectrum, indicating changes in the microstructure of the feather. This change in crown coloration towards the breeding season could be linked to reproductive behaviors.

Did early shifts to bird pollination impose constraints on Salvia flower evolution?

A recent article by Kriebel et al. (2020) examines the relationship between floral shape evolution and pollination shifts in Salvia, a plant genus almost worldwide distributed, but particularly diverse in the New World. Kriebel et al. (2020) argue that a major shift to bird pollination at the origin of the subg. Calosphace (~20 million years ago [Mya]) imposed a legacy of constraints, resulting in significant differences in flower morphology between New World and Old World Salvia. However, reanalyses of the data using hidden states to account for the heterogeneity in evolutionary rates do not support an early origin of bird pollination in this group. Instead, bird pollination may have appeared after the arrival of modern hummingbirds to North America (15.5-12 Mya), as in other North American plant clades. The use of more complex models of ancestral state reconstruction into comparative analyses provides a different perspective to explain morphological differences within Salvia. Our results indicate that bird pollination did not impose constraints on corolla shape evolution. Evolutionary constraints in anther connective and style shapes may have arisen at the origin of Calosphace but they were not associated with shifts to hummingbird pollination, being more likely the product of contingent evolution.

Phenotypic decanalization driven by social determinants could explain variance patterns for glycemia in adult urban Argentinian population.

Type 2 diabetes, one of the major causes of death and disability worldwide, is characterized by problems in the homeostasis of blood glucose. Current preventive policies focus mainly on individual behaviors (diet, exercise, salt and alcohol consumption). Recent hypotheses state that the higher incidence of metabolic disease in some human populations may be related to phenotypic decanalization causing a heightened phenotypic variance in response to unusual or stressful environmental conditions, although the nature of these conditions is under debate. Our aim was to explore variability patterns of fasting blood glucose to test phenotypic decanalization as a possible explanation of heightened prevalence for type 2 diabetes in some groups and to detect variables associated with its variance using a nation-wide survey of Argentinian adult population. We found patterns of higher local variance for fasting glycemia associated with lower income and educational attainment. We detected no meaningful association of glycemia or its variability with covariates related to individual behaviors (diet, physical activity, salt or alcohol consumption). Our results were consistent with the decanalization hypothesis for fasting glycemia, which appears associated to socioeconomic disadvantage. We therefore propose changes in public policy and discuss the implications for data gathering and further analyses.

The role of ontogenetic allometry and nonallometric flower shape variation in species-level adaptive diversification - Calceolaria polyrhiza (Calceolariaceae) as a case study.

Organism shape changes predictably during ontogeny, resulting in specific patterns of ontogenetic allometry. In several plant and animal lineages, among-species variation in the shape of mature organisms mirrors variation along their growth trajectories. Hence, ontogenetic allometry is an important bias in evolution. This bias should be stronger at reduced evolutionary time scales, in which among-trait correlations had less time to evolve. Nevertheless, it was shown that adaptation of organism shape frequently involved departures from the ancestral ontogenetic allometry. Moreover, only a moderate fraction of shape variation is correlated with size during ontogeny. Hence, nonallometric variation in shape (NAVSh) is likely to contribute to adaptation, even at reduced evolutionary time scales. We explored the contributions of allometric variation in shape (AVSh), NAVSh, and size variation to adaptive evolution in the angiosperm species Calceolaria polyrhiza. This strongly relies on oil-collecting bees for pollination and experienced transitions in the size of pollinators during the last 2 Ma. Using geometric morphometrics, we described corolla morphology in several populations across its distribution range. Variation in corolla shape was decomposed into an allometric and a nonallometric component, and corolla size was estimated. We then looked for the correlation between these aspects of morphology and the pollinator. Our results suggest that adaptation to pollinators with different sizes relied on NAVSh, which resulted from shifts in the allometric slope and from shape changes that occurred early in flower development. We conclude that NAVSh can contribute to adaptation in flowering plants, even at the species-level.

Daily fluctuations in pollination effectiveness explain higher efficiency of native over exotic bees in Lepechinia floribunda (Lamiaceae).

BACKGROUND: Despite Stebbins' principle of the most efficient pollinator being proposed decades ago, the most important pollinators are still mainly identified using the frequency of visits to flowers. This shortcoming results in a gap between the characterization of the flower visitors of a plant species and a reliable estimation of the plant fitness consequences of the mutualistic interaction. The performance of a mutualistic visitor depends on its abundance, behaviour, effectiveness (pollen removal and deposition per unit time) and efficiency (seed set per unit time) conditioned by the temporal matching between pollinator activity and temporal patterns of maturation of the sexual functions of flowers. Although there have been recent attempts to provide a conceptual and methodological framework to characterize pollinators' performance, few have combined all key elements of visitors and plants to provide an accurate estimation of pollinators' performance under natural conditions. METHODS: We complement information on the flower biology and mating system of the sub-shrub Lepechinia floribunda (Lamiaceae) to provide a daily quantitative estimation of performance (effectiveness and efficiency) of the more abundant pollinators, i.e. native bumble-bees (Bombus spp.) and leafcutter bees (Megachile sp.), and the exotic honey-bee (Apis mellifera). KEY RESULTS: Unlike honey-bees or leafcutter bees, native bumble-bees matched the daily pattern of nectar production and stigma receptivity, and showed higher effectiveness and efficiency. Despite the overabundance of honey-bees, visits occurred mainly when stigmas were not receptive, thus reducing the honey-bees' overall performance. CONCLUSIONS: Bumble-bees appear to be the most important pollinators and potential historical mediators of reproductive trait evolution in L. floribunda. Because the production of seeds by bumble-bees involved fewer pollen grains for plants and less investment in floral display than honey-bees, contemporary and expected changes in pollinator abundance may affect future L. floribunda floral evolution. If bumble-bees were to be further displaced by anthropogenic disturbance or by competition with honey-bees, their lower efficiency will select for a larger floral display increasing reproductive costs. This scenario may also impose selection to reduce dichogamy to match honey-bee foraging activity.

Evolutionary transition between bee pollination and hummingbird pollination in Salvia: Comparing means, variances and covariances of corolla traits.

Covariation among traits can modify the evolutionary trajectory of complex structures. This process is thought to operate at a microevolutionary scale, but its long-term effects remain controversial because trait covariation can itself evolve. Flower morphology, and particularly floral trait (co)variation, has been envisioned as the product of pollinator-mediated selection. Available evidence suggests that major changes in pollinator assemblages may affect the joint expression of floral traits and their phenotypic integration. We expect species within a monophyletic lineage sharing the same pollinator type will show not only similarity in trait means but also similar phenotypic variance-covariance structures. Here, we tested this expectation using eighteen Salvia species pollinated either by bees or by hummingbirds. Our findings indicated a nonsignificant multivariate phylogenetic signal and a decoupling between means and variance-covariance phenotypic matrices of floral traits during the evolution to hummingbird pollination. Mean trait value analyses revealed significant differences between bee- and hummingbird-pollinated Salvia species although fewer differences were detected in the covariance structure between groups. Variance-covariance matrices were much more similar among bee- than hummingbird-pollinated species. This pattern is consistent with the expectation that, unlike hummingbirds, bees physically manipulate the flower, presumably exerting stronger selection pressures favouring morphological convergence among species. Overall, we conclude that the evolution of hummingbird pollination proceeded through different independent transitions. Thus, although the evolution of hummingbird pollination led to a new phenotypic optimum, the process involved the diversification of the covariance structure.

The evolution of floral ontogenetic allometry in the Andean genus Caiophora (Loasaceae, subfam. Loasoideae).

The astounding variety of angiosperm flower morphologies has evolved in response to many selective forces. Flower development is highly coordinated and involves developmental associations between size and shape, ontogenetic allometry, which in turn affect the morphology of mature flowers. Although ontogenetic allometries can act as a developmental constraint and may influence adaptive evolution, allometries can evolve themselves and may change rapidly in response to selection. We explored the evolution of ontogenetic allometry in the flowers of 11 species of Loasoideae. Seven species belong to Caiophora, which radiated recently in the central Andes, and contains species that are pollinated by bees, hummingbirds, and small rodents. According to a previous study, the diversification of Caiophora involved departures from simple allometric scaling, but the changes to allometry that enabled flower diversification have not been explored yet. We characterized the ontogenetic allometry of each species with the methods of geometric morphometrics. We studied the evolution of allometries by constructing allometric spaces, in which the allometry of each species is represented by a point and the arrangement of points indicates the relations among allometric trajectories. To examine the history of changes of ontogenetic allometries, we projected the phylogeny into the allometric spaces. Inspection of allometric spaces suggests that ontogenetic variation is limited to a few dominant features. The allometries of the two main functional flower parts under study differ in their evolutionary labilities, and patterns of variation reflect pollination systems, differences in structural organization, and abiotic environmental factors.

Pericarpial nectary-visiting ants do not provide fruit protection against pre-dispersal seed predators regardless of ant species composition and resource availability.

Extrafloral nectaries can occur in both vegetative and reproductive plant structures. In many Rubiaceae species in the Brazilian Cerrado, after corolla abscission, the floral nectary continues to secret nectar throughout fruit development originating post-floral pericarpial nectaries which commonly attract many ant species. The occurrence of such nectar secreting structures might be strategic for fruit protection against seed predators, as plants are expected to invest higher on more valuable and vulnerable parts. Here, we performed ant exclusion experiments to investigate whether the interaction with ants mediated by the pericarpial nectaries of Tocoyena formosa affects plant reproductive success by reducing the number of pre-dispersal seed predators. We also assessed whether ant protection was dependent on ant species composition and resource availability. Although most of the plants were visited by large and aggressive ant species, such as Ectatomma tuberculatum and species of the genus Camponotus, ants did not protect fruits against seed predators. Furthermore, the result of the interaction was neither related to ant species composition nor to the availability of resources. We suggest that these results may be related to the nature and behavior of the most important seed predators, like Hemicolpus abdominalis weevil which the exoskeleton toughness prevent it from being predated by most ant species. On the other hand, not explored factors, such as reward quality, local ant abundance, ant colony characteristics and/or the presence of alternative energetic sources could also account for variations in ant frequency, composition, and finally ant protective effects, highlighting the conditionality of facultative plant-ant mutualisms.

A Spectrophotometric Study of Plumage Color in the Eared Dove (Zenaida auriculata), the Most Abundant South American Columbiforme.

For birds, plumage color perception is critical in social interactions such as courtship, in both monochromatic and dichromatic species. In the Eared Dove (Zenaida auriculata), perhaps the most abundant South American Columbiforme, the plumage of males and females looks alike and both sexes share the same melanistic coloration with gray and pink tones. The aim of this study was therefore to determine whether evident sexual dichromatism exists in the plumage of the Eared Dove using a spectrophotometry technique in the avian-visible range (300-700 nm). The results of the classic colorimetric variables analysis (hue, chroma and brightness) show that males are in general brighter and have higher UV chroma values than females. The avian visual model points to differences in achromatic and chromatic levels between males and females in body regions possibly involved in sexual selection (e.g. the crown). The model also indicates chromatic or achromatic differences in body regions not subject to sexual selection such as the black spots on the wing coverts and white tail bands, both of which may be involved in intra- or inter-gender-specific communication.

Exploring the ontogenetic scaling hypothesis during the diversification of pollination syndromes in Caiophora (Loasaceae, subfam. Loasoideae).

BACKGROUND AND AIMS: Phenotypic diversification of flowers is frequently attributed to selection by different functional groups of pollinators. During optimization of floral phenotype, developmental robustness to genetic and non-genetic perturbations is expected to limit the phenotypic space available for future evolutionary changes. Although adaptive divergence can occur without altering the basic developmental programme of the flower (ontogenetic scaling hypothesis), the rarity of reversion to ancestral states following adaptive radiations of pollination syndromes suggests that changes in the ancestral developmental programme of the flower are common during such evolutionary transitions. Evidence suggests that flower diversification into different pollination syndromes in the Loasoideae genus Caiophora took place during a recent adaptive radiation in the central Andes. This involved transitions from bee to hummingbird and small rodent pollination. The aim of this work was to examine if the adaptive radiation of pollination syndromes in Caiophora occurred through ontogenetic scaling or involved a departure from the ontogenetic pattern basal to this genus. METHODS: We used geometric morphometric variables to describe the shape and size of floral structures taking part in the pollination mechanism of Loasoideae. This approach was used to characterize the developmental trajectories of three species basal to the genus Caiophora through shape-size relationships (ontogenetic allometry). We then tested if the shape-size combinations of these structures in mature flowers of derived Caiophora species fall within the phenotypic space predicted by the development of basal species. KEY RESULTS: Variation in the size and shape of Caiophora flowers does not overlap with the pattern of ontogenetic allometry of basal species. Derived bee-, hummingbird- and rodent-pollinated species had divergent ontogenetic patterns of floral development from that observed for basal bee-pollinated species. CONCLUSIONS: The adaptive radiation of Caiophora involved significant changes in the developmental pattern of the flowers, rejecting the ontogenetic scaling hypothesis.

Beyond neutral and forbidden links: morphological matches and the assembly of mutualistic hawkmoth-plant networks.

A major challenge in evolutionary ecology is to understand how co-evolutionary processes shape patterns of interactions between species at community level. Pollination of flowers with long corolla tubes by long-tongued hawkmoths has been invoked as a showcase model of co-evolution. Recently, optimal foraging models have predicted that there might be a close association between mouthparts' length and the corolla depth of the visited flowers, thus favouring trait convergence and specialization at community level. Here, we assessed whether hawkmoths more frequently pollinate plants with floral tube lengths similar to their proboscis lengths (morphological match hypothesis) against abundance-based processes (neutral hypothesis) and ecological trait mismatches constraints (forbidden links hypothesis), and how these processes structure hawkmoth-plant mutualistic networks from five communities in four biogeographical regions of South America. We found convergence in morphological traits across the five communities and that the distribution of morphological differences between hawkmoths and plants is consistent with expectations under the morphological match hypothesis in three of the five communities. In the two remaining communities, which are ecotones between two distinct biogeographical areas, interactions are better predicted by the neutral hypothesis. Our findings are consistent with the idea that diffuse co-evolution drives the evolution of extremely long proboscises and flower tubes, and highlight the importance of morphological traits, beyond the forbidden links hypothesis, in structuring interactions between mutualistic partners, revealing that the role of niche-based processes can be much more complex than previously known.

The evolution of signal-reward correlations in bee- and hummingbird-pollinated species of Salvia.

Within-individual variation in floral advertising and reward traits is a feature experienced by pollinators that visit different flowers of the same plant. Pollinators can use advertising traits to gather information about the quality and amount of rewards, leading to the evolution of signal-reward correlations. As long as plants differ in the reliability of their signals and pollinators base their foraging decisions on this information, natural selection should act on within-individual correlations between signals and rewards. Because birds and bees differ in their cognitive capabilities, and use different floral traits as signals, we tested the occurrence of adaptive divergence of the within-individual signal-reward correlations among Salvia species that are pollinated either by bees or by hummingbirds. They are expected to use different floral advertising traits: frontal traits in the case of bees and side traits in the case of hummingbirds. We confirmed this expectation as bee- and hummingbird-pollinated species differed in which specific traits are predominantly associated with nectar reward at the within-individual level. Our findings highlight the adaptive value of within-individual variation and covariation patterns, commonly disregarded as 'environmental noise', and are consistent with the hypothesis that pollinator-mediated selection affects the correlation pattern among floral traits.

Armament imbalances: match and mismatch in plant-pollinator traits of highly specialized long-spurred orchids.

BACKGROUND: Some species of long-spurred orchids achieve pollination by a close association with long-tongued hawkmoths. Among them, several Habenaria species present specialized mechanisms, where pollination success depends on the attachment of pollinaria onto the heads of hawkmoths with very long proboscises. However, in the Neotropical region such moths are less abundant than their shorter-tongued relatives and are also prone to population fluctuations. Both factors may give rise to differences in pollinator-mediated selection on floral traits through time and space. METHODOLOGY/PRINCIPAL FINDINGS: We characterized hawkmoth assemblages and estimated phenotypic selection gradients on orchid spur lengths in populations of three South American Habenaria species. We examined the match between hawkmoth proboscis and flower spur lengths to determine whether pollinators may act as selective agents on flower morphology. We found significant directional selection on spur length only in Habenaria gourlieana, where most pollinators had proboscises longer than the mean of orchid spur length. CONCLUSIONS/SIGNIFICANCE: Phenotypic selection is dependent on the mutual match between pollinator and flower morphologies. However, our findings indicate that pollinator-mediated selection may vary through time and space according to local variations in pollinator assemblages.

How to look like a mallow: evidence of floral mimicry between Turneraceae and Malvaceae.

Abundant, many-flowered plants represent reliable and rich food sources for animal pollinators, and may even sustain guilds of specialized pollinators. Contrastingly, rare plants need alternative strategies to ensure pollinators' visitation and faithfulness. Flower mimicry, i.e. the sharing of a similar flower colour and display pattern by different plant species, is a means by which a rare species can exploit a successful model and increase its pollination services. The relationship between two or more rewarding flower mimic species, or Müllerian mimicry, has been proposed as mutualistic, in contrast to the unilaterally beneficial Batesian floral mimicry. In this work, we show that two different geographical colour phenotypes of Turnera sidoides ssp. pinnatifida resemble co-flowering Malvaceae in colour as seen by bees' eyes, and that these pollinators do not distinguish between them when approaching flowers in choice tests. Main pollinators of T. sidoides are bees specialized for collecting pollen in Malvaceae. We demonstrate that the similarity between at least one of the geographical colour phenotypes of T. sidoides and co-flowering Malvaceae is adaptive, since the former obtains more pollination services when growing together with its model than when growing alone. Instead of the convergent evolution pattern attributed to Müllerian mimicry, our data rather suggest an advergent evolution pattern, because only T. sidoides seems to have evolved to be more similar to its malvaceous models.