The effect of experimental pollinator decline on pollinator-mediated selection on floral traits.

Human-mediated environmental change, by reducing mean fitness, is hypothesized to strengthen selection on traits that mediate interactions among species. For example, human-mediated declines in pollinator populations are hypothesized to reduce mean seed production by increasing the magnitude of pollen limitation and thus strengthen pollinator-mediated selection on floral traits that increase pollinator attraction or pollen transfer efficiency. To test this hypothesis, we measured two female fitness components and six floral traits of Lobelia siphilitica plants exposed to supplemental hand-pollination, ambient open-pollination, or reduced open-pollination treatments. The reduced treatment simulated pollinator decline, while the supplemental treatment was used to estimate pollen limitation and pollinator-mediated selection. We found that plants in the reduced pollination treatment were significantly pollen limited, resulting in pollinator-mediated selection for taller inflorescences and more vibrant petals, both traits that could increase pollinator attraction. This contrasts with plants in the ambient pollination treatment, where reproduction was not pollen limited and there was not significant pollinator-mediated selection on any floral trait. Our results support the hypothesis that human-mediated environmental change can strengthen selection on traits of interacting species and suggest that these traits have the potential to evolve in response to changing environments.

Evolvability and trait function predict phenotypic divergence of plant populations.

Understanding the causes and limits of population divergence in phenotypic traits is a fundamental aim of evolutionary biology, with the potential to yield predictions of adaptation to environmental change. Reciprocal transplant experiments and the evaluation of optimality models suggest that local adaptation is common but not universal, and some studies suggest that trait divergence is highly constrained by genetic variances and covariances of complex phenotypes. We analyze a large database of population divergence in plants and evaluate whether evolutionary divergence scales positively with standing genetic variation within populations (evolvability), as expected if genetic constraints are evolutionarily important. We further evaluate differences in divergence and evolvability-divergence relationships between reproductive and vegetative traits and between selfing, mixed-mating, and outcrossing species, as these factors are expected to influence both patterns of selection and evolutionary potentials. Evolutionary divergence scaled positively with evolvability. Furthermore, trait divergence was greater for vegetative traits than for floral (reproductive) traits, but largely independent of the mating system. Jointly, these factors explained ~40% of the variance in evolutionary divergence. The consistency of the evolvability-divergence relationships across diverse species suggests substantial predictability of trait divergence. The results are also consistent with genetic constraints playing a role in evolutionary divergence.

Simulated pollinator decline has similar effects on seed production of female and hermaphrodite Lobelia siphilitica, but different effects on selection on floral traits.

PREMISE: Pollinator decline, by reducing seed production, is predicted to strengthen natural selection on floral traits. However, the effect of pollinator decline on gender dimorphic species (such as gynodioecious species, where plants produce female or hermaphrodite flowers) may differ between the sex morphs: if pollinator decline reduces the seed production of females more than hermaphrodites, then it should also have a larger effect on selection on floral traits in females than in hermaphrodites. METHODS: To simulate pollinator decline, we experimentally reduced pollinator access to female and hermaphrodite Lobelia siphilitica plants. We compared the seed production of plants in the reduced pollination treatment to plants that were exposed to ambient pollination conditions. Within each treatment, we also measured directional selection on four floral traits of females and hermaphrodites. RESULTS: Experimentally reducing pollination decreased seed production of both females and hermaphrodites by ~21%. Reducing pollination also strengthened selection on floral traits, but this effect was not larger in females than in hermaphrodites. Instead, reducing pollination intensified selection for taller inflorescences in hermaphrodites, but did not intensify selection on any floral trait in females. CONCLUSIONS: Our results suggest that pollinator decline will not have a larger effect on either seed production or selection on floral traits of female plants. As such, any effect of pollinator decline on seed production may be similar for gender dimorphic and monomorphic species. However, the potential for floral traits of females (and thus of gender dimorphic species) to evolve in response to pollinator decline may be limited.

Do Genetic Drift and Gene Flow Affect the Geographic Distribution of Female Plants in Gynodioecious Lobelia siphilitica?

Variation in population sex ratio is particularly pronounced in gynodioecious angiosperms. Extremely high female frequencies in gynodioecious populations cannot be readily explained by selective forces alone. To assess the contributions of drift and gene flow to extreme sex-ratio variation, we documented sex ratio and population size in 92 populations of Lobelia siphilitica across its range and genotyped plants using plastid and nuclear genetic markers. Similarity in spatial patterns of genetic and demographic variables may suggest that drift and/or gene flow have contributed to population sex-ratio variation in L. siphilitica. We found strong spatial structuring of extremely high female frequencies: populations with >50% female plants are restricted to the south−central portion of the range. However, we did not detect any spatial structuring in population size nor metrics of genetic diversity, suggesting that extreme variation in female frequency is not strongly affected by drift or gene flow. Extreme sex-ratio variation is frequently observed in gynodioecious plants, but its causes are difficult to identify. Further investigation into mechanisms that create or maintain the spatial structure of sex ratios in gynodioecious species will provide much needed insight.

Plasticity in floral longevity and sex-phase duration of Lobelia siphilitica in response to simulated pollinator declines.

PREMISE: Pollinator declines can reduce the quantity and quality of pollination services, resulting in less pollen deposited on flowers and lower seed production by plants. In response to these reductions, plant species that cannot autonomously self-pollinate and thus are dependent on pollinators to set seed could plastically adjust their floral traits. Such plasticity could increase the opportunity for outcross pollination directly, as well as indirectly by affecting inflorescence traits. METHODS: To test whether plants can respond to pollinator declines by plastically adjusting their floral traits, we simulated declines by experimentally reducing pollinator access to Lobelia siphilitica plants and measuring two traits of early- and late-season flowers: (1) floral longevity; and (2) sex-phase duration. To test whether plasticity in these floral traits affected inflorescence traits, we measured daily display size and phenotypic gender. RESULTS: We found that experimentally reducing pollination did not affect female-phase duration, but did extend the male-phase duration of early-season flowers by 13% and the longevity of late-season flowers by 12.8%. However, plants with an extended male phase did not have a more male-biased phenotypic gender, and plants with an extended floral longevity did not have a larger daily display. CONCLUSIONS: Our results suggest that plants can respond to pollinator declines by plastically adjusting both the longevity and sex-phase duration of their flowers. If this plasticity increases the opportunity for outcross pollination, then it could be one mechanism by which pollinator-dependent plant species maintain seed production as pollinators decline.

Defence by duplication: The relation between phenotypic glyphosate resistance and EPSPS gene copy number variation in Amaranthus palmeri.

Gene copy number variation (CNV) has been increasingly associated with organismal responses to environmental stress, but we know little about the quantitative relation between CNV and phenotypic variation. In this study we quantify the relation between variation in EPSPS (5-enolpyruvylshikimate-3-phosphate synthase) copy number using digital drop PCR and variation in phenotypic glyphosate resistance in 22 populations of Amaranthus palmeri (Palmer Amaranth), a range-expanding agricultural weed. Overall, we detected a significant positive relation between population mean copy number and resistance. The majority of populations exhibited high glyphosate resistance yet maintained low-resistance individuals, resulting in bimodality in many populations. We also investigated threshold models for the relation between copy number and resistance, and found evidence for a threshold of ~15 EPSPS copies: there was a steep increase in resistance below the threshold, followed by a much shallower increase. Across 924 individuals, as copy number increased the range of variation in resistance decreased, yielding an increasing frequency of high phenotypic resistance individuals. Among populations we detected a decline in variation (s.d.) as mean phenotypic resistance increased from moderate to high, consistent with the prediction that as phenotypic resistance increases in populations, stabilizing selection decreases variation in the trait. Our study demonstrates that populations of A. palmeri can harbour wide variation in EPSPS copy number and phenotypic glyphosate resistance, reflecting the history of, and template for future, resistance evolution.

Phenotypic plasticity masks range-wide genetic differentiation for vegetative but not reproductive traits in a short-lived plant.

Genetic differentiation and phenotypic plasticity jointly shape intraspecific trait variation, but their roles differ among traits. In short-lived plants, reproductive traits may be more genetically determined due to their impact on fitness, whereas vegetative traits may show higher plasticity to buffer short-term perturbations. Combining a multi-treatment greenhouse experiment with observational field data throughout the range of a widespread short-lived herb, Plantago lanceolata, we (1) disentangled genetic and plastic responses of functional traits to a set of environmental drivers and (2) assessed how genetic differentiation and plasticity shape observational trait-environment relationships. Reproductive traits showed distinct genetic differentiation that largely determined observational patterns, but only when correcting traits for differences in biomass. Vegetative traits showed higher plasticity and opposite genetic and plastic responses, masking the genetic component underlying field-observed trait variation. Our study suggests that genetic differentiation may be inferred from observational data only for the traits most closely related to fitness.

A sit-and-wait predator, but not an active-pursuit predator, alters pollinator-mediated selection on floral traits.

Indirect species interactions are ubiquitous in nature, often outnumbering direct species interactions. Yet despite evidence that indirect interactions have strong ecological effects, relatively little is known about whether they can shape adaptive evolution by altering the strength and/or direction of natural selection. We tested whether indirect interactions affect the strength and direction of pollinator-mediated selection on floral traits of the bumble-bee pollinated wildflower Lobelia siphilitica. We estimated the indirect effects of two pollinator predators with contrasting hunting modes: dragonflies (Aeshnidae and Corduliidae) and ambush bugs (Phymata americana, Reduviidae). Because dragonflies are active pursuit predators, we hypothesized that they would strengthen pollinator-mediated selection by weakening plant-pollinator interactions (i.e., a density-mediated indirect effect). In contrast, because ambush bugs are sit-and-wait predators, we hypothesized that they would weaken or reverse the direction of pollinator-mediated selection by altering pollinator foraging behavior (i.e., a trait-mediated indirect effect). Specifically, if ambush bugs hunt from plants with traits that attract pollinators (i.e., prey), then pollinators will spend less time visiting those plants, weakening or reversing the direction of selection on attractive floral traits. We did not find evidence that high dragonfly abundance strengthened selection on floral traits via a density-mediated indirect effect: neither pollen limitation (a proxy for the strength of plant-pollinator interactions) nor directional selection on floral traits of L. siphilitica differed significantly between high- and low-dragonfly abundance treatments. In contrast, we did find evidence that ambush bug presence affected selection on floral traits via a trait-mediated indirect effect: ambush bugs hunted from L. siphilitica plants with larger daily floral displays, reversing the direction of pollinator-mediated selection on daily display size. These results suggest that indirect species interactions have the potential to shape adaptive evolution by altering natural selection.

Global gene flow releases invasive plants from environmental constraints on genetic diversity.

When plants establish outside their native range, their ability to adapt to the new environment is influenced by both demography and dispersal. However, the relative importance of these two factors is poorly understood. To quantify the influence of demography and dispersal on patterns of genetic diversity underlying adaptation, we used data from a globally distributed demographic research network comprising 35 native and 18 nonnative populations of Plantago lanceolata Species-specific simulation experiments showed that dispersal would dilute demographic influences on genetic diversity at local scales. Populations in the native European range had strong spatial genetic structure associated with geographic distance and precipitation seasonality. In contrast, nonnative populations had weaker spatial genetic structure that was not associated with environmental gradients but with higher within-population genetic diversity. Our findings show that dispersal caused by repeated, long-distance, human-mediated introductions has allowed invasive plant populations to overcome environmental constraints on genetic diversity, even without strong demographic changes. The impact of invasive plants may, therefore, increase with repeated introductions, highlighting the need to constrain future introductions of species even if they already exist in an area.

Simulated pollinator declines intensify selection on floral traits that facilitate selfing and outcrossing in Impatiens capensis.

PREMISE: Anthropogenic environmental change is causing pollinator populations to decline. These declines should intensify selection for floral traits that facilitate outcrossing by making plants more attractive to pollinators and/or for floral traits that facilitate selfing in the absence of pollinators. However, the effect of pollinator declines on selection on floral traits could be modified by other environmental factors such as herbivores. METHODS: We studied the effect of simulated pollinator declines on selection on floral traits of Impatiens capensis, a mixed-mating species that produces both obligately selfing cleistogamous flowers and primarily outcrossing chasmogamous flowers. We measured directional selection differentials via seeds per plant on two traits that facilitate outcrossing (chasmogamous flower size and number) and one trait that facilitates selfing (cleistogamous flower number) in ambient, reduced pollinator access, and reduced pollinator access combined with increased foliar herbivory treatments. RESULTS: Reduced pollinator access intensified selection for larger chasmogamous flowers and more cleistogamous flowers. In contrast, increased herbivory did not affect selection on any floral trait. CONCLUSIONS: Reduced pollinator access intensified selection for a trait that facilitates outcrossing, suggesting that even species such as I. capensis that can autonomously self-pollinate have the potential to respond to pollinator declines by evolving floral traits that reinforce interactions between plants and pollinators. However, reduced pollinator access also intensified selection for a trait that facilitates selfing, suggesting that I. capensis could adapt to pollinator declines by evolving floral traits that maintain the production of both selfed and outcrossed seeds.

Beyond balancing selection: frequent mitochondrial recombination contributes to high-female frequencies in gynodioecious Lobelia siphilitica (Campanulaceae).

Gynodioecy is a sexual system in which females and hermaphrodites co-occur. In most gynodioecious angiosperms, sex is determined by an interaction between mitochondrial male-sterility genes (CMS) that arise via recombination and nuclear restorer alleles that evolve to suppress them. In theory, gynodioecy occurs when multiple CMS types are maintained at equilibrium frequencies by balancing selection. However, some gynodioecious populations contain very high frequencies of females. High female frequencies are not expected under balancing selection, but could be explained by the repeated introduction of novel CMS types. To test for balancing selection and/or the repeated introduction of novel CMS, we characterised cytoplasmic haplotypes from 61 populations of Lobelia siphilitica that vary widely in female frequency. We confirmed that mitotype diversity and female frequency were positively correlated across populations, consistent with balancing selection. However, while low-female populations hosted mostly common mitotypes, high-female populations and female plants hosted mostly rare, recombinant mitotypes likely to carry novel CMS types. Our results suggest that balancing selection maintains established CMS types across this species, but extreme female frequencies result from frequent invasion by novel CMS types. We conclude that balancing selection alone cannot account for extreme population sex-ratio variation within a gynodioecious species.

Load Carriage and Physical Exertion Influence Cognitive Control in Military Scenarios.

INTRODUCTION: Physical exertion has both beneficial and detrimental effects on cognitive performance, particularly cognitive control. Research into physical exertion under conditions of load carriage is particularly important given that military personnel and first responders must perform optimally under such combinatorial physical stressors. The present work sought to characterize cognitive control as a function of physical exertion and load carriage in a military operational scenario. METHODS: Thirty-one active-duty soldiers underwent a 4-h operationally relevant and fatiguing scenario that included two 1-h foot marches under load carriage conditions of 8.8, 47.2, 50.7 kg on each of three separate days. During each foot march, they completed five 5-min blocks of an auditory go/no-go task of response inhibition. RESULTS: Results showed that response inhibition declined with increasing load carriage and physical exertion, as evidenced by lower proportion of correct responses, higher proportion of false alarms, and lower response sensitivity between all three load conditions, particularly upon successive foot marches and time blocks within each foot march. CONCLUSIONS: The results support previous laboratory-based work on load carriage and physical exertion and suggest that deteriorations in cognitive control witnessed in laboratory settings are more pronounced within realistic operational scenarios akin to those encountered by military personnel and first responders.

A meta-analysis of the agents of selection on floral traits.

Floral traits are hypothesized to evolve primarily in response to selection by pollinators. However, selection can also be mediated by other environmental factors. To understand the relative importance of pollinator-mediated selection and its variation among trait and pollinator types, we analyzed directional selection gradients on floral traits from experiments that manipulated the environment to identify agents of selection. Pollinator-mediated selection was stronger than selection by other biotic factors (e.g., herbivores), but similar in strength to selection by abiotic factors (e.g., soil water), providing partial support for the hypothesis that floral traits evolve primarily in response to pollinators. Pollinator-mediated selection was stronger on pollination efficiency traits than on other trait types, as expected if efficiency traits affect fitness via interactions with pollinators, but other trait types also affect fitness via other environmental factors. In addition to varying among trait types, pollinator-mediated selection varied among pollinator taxa: selection was stronger when bees, long-tongued flies, or birds were the primary visitors than when the primary visitors were Lepidoptera or multiple animal taxa. Finally, reducing pollinator access to flowers had a relatively small effect on selection on floral traits, suggesting that anthropogenic declines in pollinator populations would initially have modest effects on floral evolution.

Response to Comment on "Precipitation drives global variation in natural selection".

The comment by Myers-Smith and Myers focuses on three main points: (i) the lack of a mechanistic explanation for climate-selection relationships, (ii) the appropriateness of the climate data used in our analysis, and (iii) our focus on estimating climate-selection relationships across (rather than within) taxonomic groups. We address these critiques in our response.

What Are the Environmental Determinants of Phenotypic Selection? A Meta-analysis of Experimental Studies.

Although many selection estimates have been published, the environmental factors that cause selection to vary in space and time have rarely been identified. One way to identify these factors is by experimentally manipulating the environment and measuring selection in each treatment. We compiled and analyzed selection estimates from experimental studies. First, we tested whether the effect of manipulating the environment on selection gradients depends on taxon, trait type, or fitness component. We found that the effect of manipulating the environment was larger when selection was measured on life-history traits or via survival. Second, we tested two predictions about the environmental factors that cause variation in selection. We found support for the prediction that variation in selection is more likely to be caused by environmental factors that have a large effect on mean fitness but not for the prediction that variation is more likely to be caused by biotic factors. Third, we compared selection gradients from experimental and observational studies. We found that selection varied more among treatments in experimental studies than among spatial and temporal replicates in observational studies, suggesting that experimental studies can detect relationships between environmental factors and selection that would not be apparent in observational studies.

Precipitation drives global variation in natural selection.

Climate change has the potential to affect the ecology and evolution of every species on Earth. Although the ecological consequences of climate change are increasingly well documented, the effects of climate on the key evolutionary process driving adaptation-natural selection-are largely unknown. We report that aspects of precipitation and potential evapotranspiration, along with the North Atlantic Oscillation, predicted variation in selection across plant and animal populations throughout many terrestrial biomes, whereas temperature explained little variation. By showing that selection was influenced by climate variation, our results indicate that climate change may cause widespread alterations in selection regimes, potentially shifting evolutionary trajectories at a global scale.

Physiological effects of temperature do not explain prevalence of females in populations of gynodioecious Lobelia siphilitica growing in warmer climates.

PREMISE OF THE STUDY: Gynodioecy is a sexual polymorphism whereby female and hermaphroditic plants co-occur within populations. In many gynodioecious species, stressful abiotic environments are associated with higher frequencies of females. This association suggests that abiotic stress affects the relative fitness of females and hermaphrodites and, thus, the maintenance of gynodioecy. METHODS: To test whether abiotic stress affects the fitness of females and hermaphrodites, we grew open-pollinated Lobelia siphilitica families in temperature regimes characteristic of the southern portion of the species' range (where females are common) and the northern portion of the range (where females are rare). We measured physiological and phenological traits that are indicative of heat stress, and fitness components of females and hermaphrodites that could affect the maintenance of gynodioecy. KEY RESULTS: Contrary to expectations if growth at high temperatures is stressful, we found that the hot treatment increased leaf chlorophyll content, decreased the percentage of plants that delayed flowering initiation, and did not affect the quantum efficiency of photosystem II. Growth at high temperatures did not affect the magnitude of the difference in rosette size (a correlate of flower number) between females and hermaphrodites, or the variance in pollen viability among hermaphrodites. CONCLUSIONS: We found that growing-season temperatures typical of high female L. siphilitica populations were not stressful and did not affect either the fitness of females compared to hermaphrodites or variation in fitness among hermaphrodites. Consequently, further research is necessary to explain correlations between abiotic environmental factors and the frequency of females in this and other gynodioecious species.

Why is gynodioecy a rare but widely distributed sexual system? Lessons from the Lamiaceae.

Gynodioecy, a sexual system where females and hermaphrodites co-occur, is found in << 1% of angiosperm species. To understand why gynodioecy is rare, we need to understand why females are maintained in some lineages, but not in others. We modelled the evolution of gynodioecy in the Lamiaceae, and investigated whether transition rates between gynodioecious and nongynodioecious states varied across the family. We also investigated whether the evolution of gynodioecy was correlated with the evolution of a herbaceous growth form and temperate distribution. Transition rates differed between Lamiaceae subfamilies. In the Nepetoideae, there were many transitions towards gynodioecy (n = 11), but also many reversions to nongynodioecy (n = 29). In addition, a herbaceous growth form, but not a temperate distribution, affected the rate of transitions both towards and away from gynodioecy; transitions towards gynodioecy occurred ˜16 times more frequently and transitions away from gynodioecy occurred ˜11 times more frequently in herbaceous lineages than in woody lineages. Within the Lamiaceae, lineages in which gynodioecy has frequently evolved also have a high rate of reversions to the nongynodioecious state. Consequently, to understand why gynodioecy is rare, we need to understand why sexual systems are more evolutionarily labile in some lineages than in others.

Do Plants Eavesdrop on Floral Scent Signals?

Plants emit a diverse array of volatile organic compounds that can function as cues to other plants. Plants can use volatiles emitted by neighbors to gain information about their environment, and respond by adjusting their phenotype. Less is known about whether the many different volatile signals that plants emit are all equally likely to function as cues to other plants. We review evidence for the function of floral volatile signals and conclude that plants are as likely to perceive and respond to floral volatiles as to other, better-studied volatiles. We propose that eavesdropping on floral volatile cues is particularly likely to be adaptive because plants can respond to these cues by adjusting traits that directly affect pollination and mating.

Frequency-dependent fitness in gynodioecious Lobelia siphilitica.

Selection is frequency dependent when an individual's fitness depends on the frequency of its phenotype. Frequency-dependent selection should be common in gynodioecious plants, where individuals are female or hermaphroditic; if the fitness of females is limited by the availability of pollen to fertilize their ovules, then they should have higher fitness when rare than when common. To test whether the fitness of females is frequency dependent, we manipulated the sex ratio in arrays of gynodioecious Lobelia siphilitica. To test whether fitness was frequency dependent because of variation in pollen availability, we compared open-pollinated and supplemental hand-pollinated plants. Open-pollinated females produced more seeds when they were rare than when they were common, as expected if fitness is negatively frequency dependent. However, hand-pollinated females also produced more seeds when they were rare, indicating that variation in pollen availability was not the cause of frequency-dependent fitness. Instead, fitness was frequency dependent because both hand- and open-pollinated females opened more flowers when they were rare than when they were common. This plasticity in the rate of anthesis could cause fitness to be frequency dependent even when reproduction is not pollen limited, and thus expand the conditions under which frequency-dependent selection operates in gynodioecious species.