Incorporating plant phenological responses into species distribution models reduces estimates of future species loss and turnover.

Anthropogenetic climate change has caused range shifts among many species. Species distribution models (SDMs) are used to predict how species ranges may change in the future. However, most SDMs rarely consider how climate-sensitive traits, such as phenology, which affect individuals' demography and fitness, may influence species' ranges. Using > 120 000 herbarium specimens representing 360 plant species distributed across the eastern United States, we developed a novel 'phenology-informed' SDM that integrates phenological responses to changing climates. We compared the ranges of each species forecast by the phenology-informed SDM with those from conventional SDMs. We further validated the modeling approach using hindcasting. When examining the range changes of all species, our phenology-informed SDMs forecast less species loss and turnover under climate change than conventional SDMs. These results suggest that dynamic phenological responses of species may help them adjust their ecological niches and persist in their habitats as the climate changes. Plant phenology can modulate species' responses to climate change, mitigating its negative effects on species persistence. Further application of our framework will contribute to a generalized understanding of how traits affect species distributions along environmental gradients and facilitate the use of trait-based SDMs across spatial and taxonomic scales.

Heritability and variance components of seed size in wild species: influences of breeding design and the number of genotypes tested.

Seed size affects individual fitness in wild plant populations, but its ability to evolve may be limited by low narrow-sense heritability (h2). h2 is estimated as the proportion of total phenotypic variance (σ2P) attributable to additive genetic variance (σ2A), so low values of h2 may be due to low σ2A (potentially eroded by natural selection) or to high values of the other factors that contribute to σ2P, such as extranuclear maternal effects (m2) and environmental variance effects (e2). Here, we reviewed the published literature and performed a meta-analysis to determine whether h2 of seed size is routinely low in wild populations and, if so, which components of σ2P contribute most strongly to total phenotypic variance. We analyzed available estimates of narrow-sense heritability (h2) of seed size, as well as the variance components contributing to these parameters. Maternal and environmental components of σ2P were significantly greater than σ2A, dominance, paternal, and epistatic components. These results suggest that low h2 of seed size in wild populations (the mean value observed in this study was 0.13) is due to both high values of maternally derived and environmental (residual) σ2, and low values of σ2A in seed size. The type of breeding design used to estimate h2 and m2 also influenced their values, with studies using diallel designs generating lower variance ratios than nested and other designs. e2 was not influenced by breeding design. For some breeding designs, the number of genotypes included in a study also influenced the resulting h2 and e2 estimates, but not m2. Our data support the view that a diallel design is better suited than the alternatives for the accurate estimation of σ2A in seed size due to its factorial design and the inclusion of reciprocal crosses, which allows the independent estimation of both additive and non-additive components of variance.

Genotype × environment interaction obscures genetic sources of variation in seed size in Dithyrea californica but provides the opportunity for selection on phenotypic plasticity.

PREMISE: In many species, seed size influences individual fitness, but its heritability is low, impeding its evolution. In heterogeneous environments, even if heritability of seed size is low, genetic variation in phenotypic plasticity for seed size may provide the opportunity for selection, but this possibility has rarely been investigated in wild species. The evolutionary trajectory of seed size depends on whether additive, maternal, or non-additive genetic variance dominates; moreover, the expression of any of these sources of variance may be environment-dependent, reflecting genetic variation in plasticity. In this study, we examined these sources of variation in seed size and their response to drought in Dithyrea californica. METHODS: We used a diallel design to estimate variance components for seed size in three greenhouse-raised populations sampled from California and northern Mexico. We replicated diallels in two watering treatments to examine genetic parameters and genotype × environment interactions affecting seed size. We estimated general (GCA) and specific (SCA) combining ability, reciprocal effects (RGCA and RSCA), and their interactions with water availability, and we sought evidence that sexual conflict influences seed size. RESULTS: Norms of reaction revealed genetic variation in plasticity for seed size in each population. Seed size in D. californica is determined by the combination of watering treatment, GCA and RGCA; parental identity and water availability do not consistently affect seed size, and we detected no evidence for sexual conflict. CONCLUSIONS: Multiple sources of genetic variation in phenotypic plasticity for seed size have the potential to influence its evolutionary trajectory in heterogenous environments.

Context-dependent concordance between physiological divergence and phenotypic selection in sister taxa with contrasting phenology and mating systems.

PREMISE: The study of phenotypic divergence of, and selection on, functional traits in closely related taxa provides the opportunity to detect the role of natural selection in driving diversification. If the strength or direction of selection in field populations differs between taxa in a pattern that is consistent with the phenotypic difference between them, then natural selection reinforces the divergence. Few studies have sought evidence for such concordance for physiological traits. METHODS: Herbarium specimen records were used to detect phenological differences between sister taxa independent of the effects on flowering time of long-term variation in the climate across collection sites. In the field, physiological divergence in photosynthetic rate, transpiration rate, and instantaneous water-use efficiency were recorded during vegetative growth and flowering in 13 field populations of two taxon pairs of Clarkia, each comprising a self-pollinating and a outcrossing taxon. RESULTS: Historically, each selfing taxon flowered earlier than its outcrossing sister taxon, independent of the effects of local long-term climatic conditions. Sister taxa differed in all focal traits, but the degree and (in one case) the direction of divergence depended on life stage. In general, self-pollinating taxa had higher gas exchange rates, consistent with their earlier maturation. In 6 of 18 comparisons, patterns of selection were concordant with the phenotypic divergence (or lack thereof) between sister taxa. CONCLUSIONS: Patterns of selection on physiological traits measured in heterogeneous conditions do not reliably reflect divergence between sister taxa, underscoring the need for replicated studies of the direction of selection within and among taxa.

What determines the evolutionary trajectories of wild plant species? Approaches to the study of quantitative fitness-related traits.

Wild plant species provide excellent examples of qualitative traits that evolve in response to environmental challenges (e.g., flower color, heavy metal tolerance, cyanogenesis, and male sterility). In addition to such discrete characters, a dazzling array of continuously distributed, quantitative traits are expressed at every phase of the life cycle. These traits are known or suspected to have evolved by natural selection because they are heritable, differ among populations or closely related taxa occupying distinct habitats, and have individual phenotypes associated with survival and reproductive success. This special issue [American Journal of Botany 109(11)] focuses on the tools and approaches for detecting or inferring the ecological and genetic factors contributing to changes in genetically based variation of quantitative traits within or among populations, or causing their divergence among taxa. The assembled articles use one or more of three primary approaches to detect the process or outcome of natural selection on morphological, life history, reproductive, chemical, and physiological quantitative traits: the analysis of phenotypic or artificially imposed selection to detect direct and indirect selection on traits whose function is well-understood; common garden experiments, including reciprocal transplants and "resurrection" experiments; and quantitative genetic analyses designed to detect and to estimate the environmental and genetic sources of phenotypic variation or to forecast short-term evolutionary change. Together, these articles examine and reveal the adaptive capacity of quantitative traits and the genetically based constraints that may limit their directional evolutionary change, thereby informing and testing inferences, hypotheses, and predictions concerning the evolutionary trajectories of wild plant species.

Advancing frost dates have reduced frost risk among most North American angiosperms since 1980.

In recent decades, the final frost dates of winter have advanced throughout North America, and many angiosperm taxa have simultaneously advanced their flowering times as the climate has warmed. Phenological advancement may reduce plant fitness, as flowering prior to the final frost date of the winter/spring transition may damage flower buds or open flowers, limiting fruit and seed production. The risk of floral exposure to frost in the recent past and in the future, however, also depends on whether the last day of winter frost is advancing more rapidly, or less rapidly, than the date of onset of flowering in response to climate warming. This study presents the first continental-scale assessment of recent changes in frost risk to floral tissues, using digital records of 475,694 herbarium specimens representing 1,653 angiosperm species collected across North America from 1920 to 2015. For most species, among sites from which they have been collected, dates of last frost have advanced much more rapidly than flowering dates. As a result, frost risk has declined in 66% of sampled species. Moreover, exotic species consistently exhibit lower frost risk than native species, primarily because the former occupy warmer habitats where the annual frost-free period begins earlier. While reducing the probability of exposure to frost has clear benefits for the survival of flower buds and flowers, such phenological advancement may disrupt other ecological processes across North America, including pollination, herbivory, and disease transmission.

Region-specific phenological sensitivities and rates of climate warming generate divergent temporal shifts in flowering date across a species' range.

PREMISE: Forecasting how species will respond phenologically to future changes in climate is a major challenge. Many studies have focused on estimating species- and community-wide phenological sensitivities to climate to make such predictions, but sensitivities may vary within species, which could result in divergent phenological responses to climate change. METHODS: We used 743 herbarium specimens of the mountain jewelflower (Streptanthus tortuosus, Brassicaceae) collected over 112 years to investigate whether individuals sampled from relatively warm vs. cool regions differ in their sensitivity to climate and whether this difference has resulted in divergent phenological shifts in response to climate warming. RESULTS: During the past century, individuals sampled from warm regions exhibited a 20-day advancement in flowering date; individuals in cool regions showed no evidence of a shift. We evaluated two potential drivers of these divergent responses: differences between regions in (1) the degree of phenological sensitivity to climate and (2) the magnitude of climate change experienced by plants, or (3) both. Plants sampled from warm regions were more sensitive to temperature-related variables and were subjected to a greater degree of climate warming than those from cool regions; thus our results suggest that the greater temporal shift in flowering date in warm regions is driven by both of these factors. CONCLUSIONS: Our results are among the first to demonstrate that species exhibited intraspecific variation in sensitivity to climate and that this variation can contribute to divergent responses to climate change. Future studies attempting to forecast temporal shifts in phenology should consider intraspecific variation.

Machine Learning Using Digitized Herbarium Specimens to Advance Phenological Research.

Machine learning (ML) has great potential to drive scientific discovery by harvesting data from images of herbarium specimens-preserved plant material curated in natural history collections-but ML techniques have only recently been applied to this rich resource. ML has particularly strong prospects for the study of plant phenological events such as growth and reproduction. As a major indicator of climate change, driver of ecological processes, and critical determinant of plant fitness, plant phenology is an important frontier for the application of ML techniques for science and society. In the present article, we describe a generalized, modular ML workflow for extracting phenological data from images of herbarium specimens, and we discuss the advantages, limitations, and potential future improvements of this workflow. Strategic research and investment in specimen-based ML methods, along with the aggregation of herbarium specimen data, may give rise to a better understanding of life on Earth.

Heteranthery in Clarkia: pollen performance of dimorphic anthers contradicts expectations.

PREMISE OF THE STUDY: Wild plant species that require the services of pollen-feeding insects for reliable pollination may evolve features that attract and reward their mutualistic partners. Heterantherous species have been proposed to exhibit a "division of labor" whereby "feeding anthers" (which produce pollen that may be consumed by an insect) are distinguished from "reproductive anthers" (which produce pollen more likely to contribute to reproduction). In some heterantherous species, including Clarkia unguiculata (Onagraceae), these two anther types differ with respect to stamen length, anther size, pollen production, and pollen color. METHODS: The primary goal of this study was to test one component of the "division of labor" hypothesis by comparing the performance of the pollen produced by each type of anther in C. unguiculata. To achieve this goal, under greenhouse conditions, we hand pollinated and assessed pollen performance (using epifluorescence microscopy) within ~228 flowers. KEY RESULTS: The pollen produced by the two anther types differed significantly with respect to both stigma and style penetration. The inner series of anthers produce pollen with higher performance than the outer series of longer, dark red anthers. CONCLUSIONS: These findings contradict previous descriptions of the genus, reporting that the inner diminutive series of anthers in Clarkia produce "abortive and nonfunctional" pollen. We outline the future research required to demonstrate the ecological function of heteranthery in this iconic wildflower group.

Floral traits influence the opportunity for selection among male gametophytes: independent and combined effects of style length and petal area.

PREMISE: Strong correlations between traits can obscure their independent effects on components of reproduction. Style length (SL) and petal area (PA) vary within species, for example, but their independent effects on the opportunity for selection among pollen genotypes are poorly understood. Previous work in Clarkia detected a positive effect of SL on pollen receipt, potentially intensifying selection. However, this apparent effect of SL may be influenced by a correlated trait, such as PA. Here, we examine the independent effects of these two traits on pollen receipt and performance. METHODS: We collected petals and styles from wild populations of two insect-pollinated Clarkia taxa and estimated the independent and combined effects of SL and PA on pollen receipt and performance. RESULTS: In both taxa, SL and PA are positively correlated. In C. unguiculata, both traits positively and independently affect pollen receipt, but in C. xantiana ssp. xantiana, the two traits act only in combination to affect pollen receipt. In both taxa, pollen receipt positively affects the numbers of pollen tubes entering and penetrating the style, as well as pollen tube attrition. CONCLUSIONS: The effects of SL and PA on pollen receipt and performance are taxon specific. In C. unguiculata, both traits may be independent targets of selection due to their effects on pollen receipt. In C. xantiana ssp. xantiana, by contrast, the combined (but not independent) effects of SL and PA influence pollen receipt. Ecological differences between these taxa require exploration to understand the mechanisms by which these traits affect pollinator behavior.

Overlooked climate parameters best predict flowering onset: Assessing phenological models using the elastic net.

Determining the manner in which plant species shift their flowering times in response to climatic conditions is essential to understanding and forecasting the impacts of climate change on the world's flora. The limited taxonomic diversity and duration of most phenological datasets, however, have impeded a comprehensive, systematic determination of the best predictors of flowering phenology. Additionally, many studies of the relationship between climate conditions and plant phenology have included only a limited set of climate parameters that are often chosen a priori and may therefore overlook those parameters to which plants are most phenologically sensitive. This study harnesses 894,392 digital herbarium records and 1,959 in situ observations to produce the first assessment of the effects of a large number (25) of climate parameters on the flowering time of a very large number (2,468) of angiosperm taxa throughout North America. In addition, we compare the predictive capacity of phenological models constructed from the collection dates of herbarium specimens vs. repeated in situ observations of individual plants using a regression approach-elastic net regularization-that has not previously been used in phenological modeling, but exhibits several advantages over ordinary least squares and stepwise regression. When herbarium-derived data and in situ phenological observations were used to predict flowering onset, the multivariate models based on each of these data sources had similar predictive capacity (R2  = 0.27). Further, apart from mean maximum temperature (TMAX), the two best predictors of flowering time have not commonly been included in phenological models: the number of frost-free days (NFFD) and the quantity of precipitation as snow (PAS) in the seasons preceding flowering. By vetting these models across an unprecedented number of taxa, this work demonstrates a new approach to phenological modeling.

Could seasonally deteriorating environments favour the evolution of autogamous selfing and a drought escape physiology through indirect selection? A test of the time limitation hypothesis using artificial selection in Clarkia.

Background and Aims: The evolution of selfing from outcrossing may be the most common transition in plant reproductive systems and is associated with a variety of ecological circumstances and life history strategies. The most widely discussed explanation for these associations is the reproductive assurance hypothesis - the proposition that selfing is favoured because it increases female fitness when outcross pollen receipt is limited. Here an alternative explanation, the time limitation hypothesis, is addressed, one scenario of which proposes that selfing may evolve as a correlated response to selection for a faster life cycle in seasonally deteriorating environments. Methods: Artificial selection for faster maturation (early flowering) or for low herkogamy was performed on Clarkia unguiculata (Onagraceae), a largely outcrossing species whose closest relative, C. exilis, has evolved higher levels of autogamous selfing. Direct responses to selection and correlated evolutionary changes in these traits were measured under greenhouse conditions. Direct responses to selection on early flowering and correlated evolutionary changes in the node of the first flower, herkogamy, dichogamy, gas exchange rates and water use efficiency (WUE) were measured under field conditions. Key Results: Lines selected for early flowering and for low herkogamy showed consistent, statistically significant responses to direct selection. However, there was little or no evidence of correlated evolutionary changes in flowering date, floral traits, gas exchange rates or WUE. Conclusions: These results suggest that the maturation rate and mating system have evolved independently in Clarkia and that the time limitation hypothesis does not explain the repeated evolution of selfing in this genus, at least through its indirect selection scenario. They also suggest that the life history and physiological components of drought escape are not genetically correlated in Clarkia, and that differences in gas exchange physiology between C. unguiculata and C. exilis have evolved independently of differences in mating system and life history.

Historical changes in flowering phenology are governed by temperature × precipitation interactions in a widespread perennial herb in western North America.

For most species, a precise understanding of how climatic parameters determine the timing of seasonal life cycle stages is constrained by limited long-term data. Further, most long-term studies of plant phenology that have examined relationships between phenological timing and climate have been local in scale or have focused on single climatic parameters. Herbarium specimens, however, can expand the temporal and spatial coverage of phenological datasets. Using Trillium ovatum specimens collected over > 100 yr across its native range, we analyzed how seasonal climatic conditions (mean minimum temperature (Tmin ), mean maximum temperature and total precipitation (PPT)) affect flowering phenology. We then examined long-term changes in climatic conditions and in the timing of flowering across T. ovatum's range. Warmer Tmin advanced flowering, whereas higher PPT delayed flowering. However, Tmin and PPT were shown to interact: the advancing effect of warmer Tmin was strongest where PPT was highest, and the delaying effect of higher PPT was strongest where Tmin was coldest. The direction of temporal change in climatic parameters and in the timing of flowering was dependent on geographic location. Tmin , for example, decreased across the observation period in coastal regions, but increased in inland areas. Our results highlight the complex effects of climate and geographic location on phenology.

Outcrossing and photosynthetic rates vary independently within two Clarkia species: implications for the joint evolution of drought escape physiology and mating system.

Background and Aims Mating systems of plants are diverse and evolutionarily labile. Abiotic environmental factors, such as seasonal drought, may impose selection on physiological traits that could lead to transitions in mating system if physiological traits are genetically correlated with traits that influence mating system. Within Clarkia, self-fertilizing taxa have higher photosynthetic rates, earlier flowering phenology, faster individual floral development and more compressed flowering periods than their outcrossing sister taxa, potentially reducing the selfing taxa's exposure to drought. In theory, this contrast in trait combinations between sister taxa could have arisen via correlated evolution due to pleiotropy or genetic linkage. Alternatively, each trait may evolve independently as part of a life history that is adaptive in seasonally dry environments. Methods To evaluate these hypotheses, we examined relationships between photosynthetic rates (adjusted for plant height and leaf node position) and outcrossing rates (estimated by allozyme variation in progeny arrays) during two consecutive years in multiple wild populations of two mixed-mating Clarkia taxa, each of which is sister to a derived selfing taxon. If the negative association between photosynthetic rate and outcrossing previously observed between sister taxa reflects correlated evolution due to a strong negative genetic correlation between these traits, then a similarly negative relationship would be observed within populations of each taxon. By contrast, if the combination of elevated photosynthetic rates and reduced outcrossing evolved independently within taxa, we predicted no consistent relationship between photosynthetic rate and outcrossing rate. Key Results We found no significant difference in outcrossing rates within populations between groups of plants with high versus low photosynthetic rates. Conclusions Overall, these results provide support for the hypothesis that the joint divergence in photosynthetic rate and mating system observed between Clarkia sister taxa is the result of independent evolutionary transitions.

Pollen--tiny and ephemeral but not forgotten: New ideas on their ecology and evolution.

Ecologists and evolutionary biologists have been interested in the functional biology of pollen since the discovery in the 1800s that pollen grains encompass tiny plants (male gametophytes) that develop and produce sperm cells. After the discovery of double fertilization in flowering plants, botanists in the early 1900s were quick to explore the effects of temperature and maternal nutrients on pollen performance, while evolutionary biologists began studying the nature of haploid selection and pollen competition. A series of technical and theoretic developments have subsequently, but usually separately, expanded our knowledge of the nature of pollen performance and how it evolves. Today, there is a tremendous diversity of interests that touch on pollen performance, ranging from the ecological setting on the stigma, structural and physiological aspects of pollen germination and tube growth, the form of pollen competition and its role in sexual selection in plants, virus transmission, mating system evolution, and inbreeding depression. Given the explosion of technical knowledge of pollen cell biology, computer modeling, and new methods to deal with diversity in a phylogenetic context, we are now more than ever poised for a new era of research that includes complex functional traits that limit or enhance the evolution of these deceptively simple organisms.

Geographic variation in climate as a proxy for climate change: Forecasting evolutionary trajectories from species differentiation and genetic correlations.

PREMISE OF THE STUDY: Climate change models for California predict a warmer, drier future, potentially resulting in shorter growing seasons. If phenotypic differences between closely related species currently distributed across a moisture and temperature gradient represent adaptations to their abiotic environment, then as conditions become warmer and drier, populations presently adapted to cooler and wetter conditions may evolve to become more similar to those adapted to warmer and drier conditions. Two sister species, Clarkia unguiculata and C. exilis, are distributed across a moisture and temperature gradient in the southern Sierra Nevada, providing an opportunity to predict how this process may occur. METHODS: In a greenhouse experiment using wild-collected seeds from 11 populations in the southern Sierra Nevada, we examined relationships among elevation, climatic conditions, and population means for each trait, then evaluated bivariate relationships among maternal family means, using raw values and controlling for population and seed mass effects on phenotype. KEY RESULTS: Clarkia exilis occupied warmer, drier conditions, typically at lower elevations, than C. unguiculata did and flowered earlier and faster, producing smaller flowers with lower herkogamy. In C. unguiculata, petal area, herkogamy, and the rate of flower production were positively correlated with days to first flower. CONCLUSIONS: If selection favors earlier flowering, smaller petals, or faster flower production in C. unguiculata, then the genetic correlations among these traits should reinforce their joint evolution. Moreover, the correlations between these traits and herkogamy may promote the evolution of self-fertilization as an indirect response to selection, a previously unrecognized potential outcome of climate change.

Winning in style: Longer styles receive more pollen, but style length does not affect pollen attrition in wild Clarkia populations.

PREMISE OF THE STUDY: One proposed function of long styles is to intensify selection among male gametophytes relative to short styles. If so, given sufficient competition, longer styles will have higher rates of pollen tube attrition (failure to reach the style base) within the style than shorter ones. Alternatively, style length may influence pollen receipt, which itself may affect attrition rates. METHODS: We tested these predictions by collecting senescing styles from wild populations of two insect-pollinated Clarkia species. We examined the number of pollen grains adhering to the stigma, length of styles, and rates of attrition from the stigma surface to the stigma-style junction (SSJ), from the SSJ to the style base, and from the stigma surface to the style base. Multivariate analyses estimated the independent effects of pollen grains per stigma, the number of pollen tubes at the SSJ, and style length on attrition. KEY RESULTS: Style length was generally positively correlated with pollen receipt, and the number of pollen grains per stigma was positively correlated with all three attrition rates. In neither species was any attrition rate affected by style length independent of the number of pollen grains per stigma. CONCLUSIONS: Pollen attrition was mediated by style length, but the function of style length was primarily to increase the number of germinating pollen grains, which affected attrition rates either through stigma clogging or pollen-pollen interactions. Style length may have a direct effect on pollen receipt due to the stigma's position relative to pollinator body parts, but traits correlated with style length may also directly affect pollen receipt.

Project Baseline: An unprecedented resource to study plant evolution across space and time.

PREMISE OF THE STUDY: Project Baseline is a seed bank that offers an unprecedented opportunity to examine spatial and temporal dimensions of microevolution during an era of rapid environmental change. Over the upcoming 50 years, biologists will withdraw genetically representative samples of past populations from this time capsule of seeds and grow them contemporaneously with modern samples to detect any phenotypic and molecular evolution that has occurred during the intervening time. METHODS: We carefully developed this living genome bank using protocols to enhance its experimental value by collecting from multiple populations and species across a broad geographical range in sites that are likely to be preserved into the future. Seeds are accessioned with site and population data and are stored by maternal line under conditions that maximize seed longevity. This open-access resource will be available to researchers at regular intervals to evaluate contemporary evolution. KEY RESULTS: To date, the Project Baseline collection includes 100-200 maternal lines of each of 61 species collected from over 831 populations on sites that are likely to be preserved into the future across the United States (∼78,000 maternal lines). Our strategically designed collection circumvents some problems that can cloud the results of "resurrection" studies involving naturally preserved or existing seed collections that are available fortuitously. CONCLUSIONS: The resurrection approach can be coupled with long-established and newer techniques over the next five decades to elucidate genetic change and thereby vastly improve our understanding of temporal and spatial changes in phenotype and the evolutionary processes underlying it.

Seasonal changes in physiological performance in wild Clarkia xantiana populations: Implications for the evolution of a compressed life cycle and self-fertilization.

PREMISE OF THE STUDY: One explanation for the evolution of selfing, the drought escape hypothesis, proposes that self-fertilization may evolve under conditions of intensifying seasonal drought as part of a suite of traits that enable plants to accelerate the completion of their life cycle, thereby escaping late-season drought. Here, we test two fundamental assumptions of this hypothesis in Clarkia xantiana: (1) that a seasonal decline in precipitation causes an increase in drought stress and (2) that this results in changes in physiological performance, reflecting these deteriorating conditions. METHODS: We examined seasonal and interannual variation in abiotic environmental conditions (estimated by ambient temperature, relative humidity, predawn leaf water potentials, and carbon isotope ratios) and physiological traits (photosynthesis, conductance, transpiration, instantaneous water-use efficiency, ascorbate peroxidase and glutathione reductase activities, quantum yield of photosystem II, PSII potential efficiency) in field populations of C. xantiana in 2009 and 2010. KEY RESULTS: In both years, plants experienced intensifying drought across the growing season. Gas exchange rates decreased over the growing season and were lower in 2009 (a relatively dry year) than in 2010, suggesting that the temporal changes from early to late spring were directly linked to the deteriorating environmental conditions. CONCLUSIONS: Seasonal declines in transpiration rate may have increased survival by protecting plants from desiccation. Concomitant declines in photosynthetic rate likely reduced the availability of resources for seed production late in the season. Thus, the physiological patterns observed are consistent with the conditions required for the drought escape hypothesis.

Plasticity and not adaptation is the primary source of temperature-mediated variation in flowering phenology in North America.

Phenology varies widely over space and time because of its sensitivity to climate. However, whether phenological variation is primarily generated by rapid organismal responses (plasticity) or local adaptation remains unresolved. Here we used 1,038,027 herbarium specimens representing 1,605 species from the continental United States to measure flowering-time sensitivity to temperature over time (Stime) and space (Sspace). By comparing these estimates, we inferred how adaptation and plasticity historically influenced phenology along temperature gradients and how their contributions vary among species with different phenology and native climates and among ecoregions differing in species composition. Parameters Sspace and Stime were positively correlated (r = 0.87), of similar magnitude and more frequently consistent with plasticity than adaptation. Apparent plasticity and adaptation generated earlier flowering in spring, limited responsiveness in late summer and delayed flowering in autumn in response to temperature increases. Nonetheless, ecoregions differed in the relative contributions of adaptation and plasticity, from consistently greater importance of plasticity (for example, southeastern United States plains) to their nearly equal importance throughout the season (for example, Western Sierra Madre Piedmont). Our results support the hypothesis that plasticity is the primary driver of flowering-time variation along temperature gradients, with local adaptation having a widespread but comparatively limited role.