Spring temperature drives phenotypic selection on plasticity of flowering time.

In seasonal environments, a high responsiveness of development to increasing temperatures in spring can infer benefits in terms of a longer growing season, but also costs in terms of an increased risk of facing unfavourable weather conditions. Still, we know little about how climatic conditions influence the optimal plastic response. Using 22 years of field observations for the perennial forest herb Lathyrus vernus, we assessed phenotypic selection on among-individual variation in reaction norms of flowering time to spring temperature, and examined if among-year variation in selection on plasticity was associated with spring temperature conditions. We found significant among-individual variation in mean flowering time and flowering time plasticity, and that plants that flowered earlier also had a more plastic flowering time. Selection favoured individuals with an earlier mean flowering time and a lower thermal plasticity of flowering time. Less plastic individuals were more strongly favoured in colder springs, indicating that spring temperature influenced optimal flowering time plasticity. Our results show how selection on plasticity can be linked to climatic conditions, and illustrate how we can understand and predict evolutionary responses of organisms to changing environmental conditions.

Exposure to ultraviolet-B radiation increases the susceptibility of mosquitoes to infection with dengue virus.

By 2100, greenhouse gases are predicted to reduce ozone and cloud cover over the tropics causing increased exposure of organisms to harmful ultraviolet-B radiation (UVBR). UVBR damages DNA and is an important modulator of immune function and disease susceptibility in humans and other vertebrates. The effect of UVBR on invertebrate immune function is largely unknown, but UVBR together with ultraviolet-A radiation impairs an insect immune response that utilizes melanin, a pigment that also protects against UVBR-induced DNA damage. If UVBR weakens insect immunity, then it may make insect disease vectors more susceptible to infection with pathogens of socioeconomic and public health importance. In the tropics, where UVBR is predicted to increase, the mosquito-borne dengue virus (DENV), is prevalent and a growing threat to humans. We therefore examined the effect of UVBR on the mosquito Aedes aegypti, the primary vector for DENV, to better understand the potential implications of increased tropical UVBR for mosquito-borne disease risk. We found that exposure to a UVBR dose that caused significant larval mortality approximately doubled the probability that surviving females would become infected with DENV, despite this UVBR dose having no effect on the expression of an effector gene involved in antiviral immunity. We also found that females exposed to a lower UVBR dose were more likely to have low fecundity even though this UVBR dose had no effect on larval size or activity, pupal cuticular melanin content, or adult mass, metabolic rate, or flight capacity. We conclude that future increases in tropical UVBR associated with anthropogenic global change may have the benefit of reducing mosquito-borne disease risk for humans by reducing mosquito fitness, but this benefit may be eroded if it also makes mosquitoes more likely to be infected with deadly pathogens.

A novel and high-throughput approach to assess photosynthetic thermal tolerance of kelp using chlorophyll α fluorometry.

Foundation seaweed species are experiencing widespread declines and localized extinctions due to increased instability of sea surface temperature. Characterizing temperature thresholds are useful for predicting patterns of change and identifying species most vulnerable to extremes. Existing methods for characterizing seaweed thermal tolerance produce diverse metrics and are often time-consuming, making comparisons between species and techniques difficult, hindering insight into global patterns of change. Using three kelp species, we adapted a high-throughput method - previously used in terrestrial plant thermal biology - for use on kelps. This method employs temperature-dependent fluorescence (T-F0 ) curves under heating or cooling regimes to determine the critical temperature (Tcrit ) of photosystem II (PSII), i.e., the breakpoint between slow and fast rise fluorescence response to changing temperature, enabling rapid assays of photosynthetic thermal tolerance using a standardized metric. This method enables characterization of Tcrit for up to 48 samples per two-hour assay, demonstrating the capacity of T-F0 curves for high-throughput assays of thermal tolerance. Temperature-dependent fluorescence curves and their derived metric, Tcrit , may offer a timely and powerful new method for the field of phycology, enabling characterization and comparison of photosynthetic thermal tolerance of seaweeds across many populations, species, and biomes.

Phenotypic plasticity and exotic plant invasions: Effects of soil nutrients, species nutrient requirements, and types of traits.

High-phenotypic plasticity has long been considered as a characteristic promoting exotic plant invasions. However, the results of the studies testing this hypothesis are still inconsistent. Overlooking the effects of species resource requirements and environmental resource availability may be the main reasons for the ambiguous conclusions. Here, we compared phenotypic plasticity between five noxious invasive species with different nutrient requirements (evaluated using the soil nutrient status of their natural distribution ranges) and their phylogenetically related natives under five nutrient levels. We found that species with high-nutrient requirements showed greater plasticity of total biomass than species with low-nutrient requirements, regardless of their status (invasive or native). Invasives with high-nutrient requirements had greater growth plasticity than their related natives, which may contribute to their invasiveness under high-nutrient environments. However, compared with the related natives, a higher growth plasticity may not help exotic species with low-nutrient requirements to invade nutrient-rich habitats, and exotic species with high-nutrient requirements to invade nutrient-limited habitats. In contrast, invasives with low-nutrient requirements exhibited lower growth plasticity than their related natives, contributing to their invasiveness under nutrient-limited habitats. Functional traits showed growth-related plasticity in only 10 cases (3.8%), and there was no functional trait whose plastic response to soil nutrients was beneficial to exotic plant invasions. Our study indicates that low-growth plasticity could also promote exotic plant invasions, high plasticity may not necessarily lead to invasiveness. We must test the adaptive significance of plasticity of functional traits when studying its biological roles.

Ecological responses to variation in seasonal snow cover.

Seasonal snow is among the most important factors governing the ecology of many terrestrial ecosystems, but rising global temperatures are changing snow regimes and driving widespread declines in the depth and duration of snow cover. Loss of the insulating snow layer will fundamentally change the environment. Understanding how individuals, populations, and communities respond to different snow conditions is thus essential for predicting and managing future ecosystem change. We synthesized 365 studies that examined ecological responses to variation in winter snow conditions. This research encompasses a broad range of methods (experimental manipulations, measurement of natural snow gradients, and long-term monitoring), locations (35 countries), study organisms (plants, mammals, arthropods, birds, fish, lichen, and fungi), and response measures. Earlier snowmelt was consistently associated with advanced spring phenology in plants, mammals, and arthropods. Reduced snow depth often increased mortality or physical injury in plants, although there were few clear effects on animals. Neither snow depth nor snowmelt timing had clear or consistent directional effects on body size of animals or biomass of plants. However, because 96% of studies were from the northern hemisphere, the generality of these trends across ecosystems and localities is also unclear. We identified substantial research gaps for several taxonomic groups and response types; research on wintertime responses was notably scarce. Future research should prioritize examination of the mechanisms underlying responses to changing snow conditions and the consequences of those responses for seasonally snow-covered ecosystems.

Respuestas Ecológicas a la Variación de la Cobertura Estacional de Nieve Resumen La nieve estacional se encuentra entre los factores más importantes que determinan la ecología de muchos ecosistemas terrestres, pero las crecientes temperaturas mundiales están cambiando los sistemas de nieve y causando declinaciones generalizadas en la profundidad y la duración de la capa de nieve. La pérdida de la capa de nieve aislante cambiará fundamentalmente el ambiente. El entendimiento de cómo los individuos, las poblaciones y las comunidades responden a las diferentes condiciones de nieve es esencial para predecir y manejar los cambios del ecosistema en el futuro. Sintetizamos 365 estudios que examinaron las respuestas ecológicas a la variación en las condiciones invernales de nieve. Esta investigación engloba una gama amplia de métodos (manipulaciones experimentales, medida de los gradientes naturales de nieve y monitoreo a largo plazo), localidades (35 países), organismos de estudio (plantas, mamíferos, artrópodos, aves, peces, líquenes y hongos) y medidas de respuesta. El deshielo temprano estuvo asociado continuamente con el adelanto de la fenología de plantas, mamíferos y artrópodos en primavera. La reducción de la profundidad de la nieve con frecuencia incrementó la mortalidad o las lesiones físicas en las plantas, aunque tuvo pocos efectos visibles sobre los animales. Ni la profundidad de la nieve ni la temporalidad del deshielo tuvieron efectos direccionales claros o consistentes sobre el tamaño corporal de los animales o la biomasa de las plantas. Sin embargo, ya que el 96% de los estudios se realizó en el hemisferio norte, la generalidad de estas tendencias en todos los ecosistemas y localidades tampoco está clara. Identificamos vacíos importantes en la investigación en torno a varios grupos taxonómicos y los tipos de respuesta; la información sobre las respuestas invernales estaba particularmente reducida. Las futuras investigaciones deberían priorizar el análisis de los mecanismos subyacentes a las respuestas ante las condiciones cambiantes de nieve y las consecuencias de aquellas respuestas para los ecosistemas cubiertos de nieve.

Inherent conflicts between reaction norm slope and plasticity indices when comparing plasticity: a conceptual framework and empirical test.

Phenotypic plasticity index (PI), the slope of reaction norm (K) and relative distances plasticity index (RDPI), the most commonly used estimators, have occasionally been found to generate different plasticity rankings between groups (species, populations, cultivars or genotypes). However, no effort has been made to determine how frequent this incongruence is, and the factors that influence the occurrence of the incongruence. To address these problems, we first proposed a conceptual framework and then tested the framework (its predictions) by reanalyzing 1248 sets of published data. Our framework reveals inherent conflicts between K and PI or RDPI when comparing plasticity between two groups, and the frequency of these conflicts increases with increasing inter-group initial trait difference and/or K values of the groups compared. More importantly, the estimators also affect the magnitude of the inter-group plasticity differences even when they do not change groups' plasticity rankings. The above-mentioned effects of plasticity estimators were confirmed by our empirical test using data from the literature, and the conflicts occur in 203 (16%) of the 1248 comparisons between K and indices, indicating that a considerable proportion of the comparative conclusions on plasticity in literature are estimator-dependent. The frequency of the conflicts is influenced by phylogenetic relatedness of the groups compared, being lower when comparing within relative to between species, but not by specific types of environments, traits and species. Our study indicates that care is needed to select estimator when comparing groups' plasticity, and that the conclusions in relevant literature should be treated with great caution.

Ensuring Prevention Science Research is Synthesis-Ready for Immediate and Lasting Scientific Impact.

When seeking to inform and improve prevention efforts and policy, it is important to be able to robustly synthesize all available evidence. But evidence sources are often large and heterogeneous, so understanding what works, for whom, and in what contexts can only be achieved through a systematic and comprehensive synthesis of evidence. Many barriers impede comprehensive evidence synthesis, which leads to uncertainty about the generalizability of intervention effectiveness, including inaccurate titles/abstracts/keywords terminology (hampering literature search efforts), ambiguous reporting of study methods (resulting in inaccurate assessments of study rigor), and poorly reported participant characteristics, outcomes, and key variables (obstructing the calculation of an overall effect or the examination of effect modifiers). To address these issues and improve the reach of primary studies through their inclusion in evidence syntheses, we provide a set of practical guidelines to help prevention scientists prepare synthesis-ready research. We use a recent mindfulness trial as an empirical example to ground the discussion and demonstrate ways to ensure the following: (1) primary studies are discoverable; (2) the types of data needed for synthesis are present; and (3) these data are readily synthesizable. We highlight several tools and practices that can aid authors in these efforts, such as using a data-driven approach for crafting titles, abstracts, and keywords or by creating a repository for each project to host all study-related data files. We also provide step-by-step guidance and software suggestions for standardizing data design and public archiving to facilitate synthesis-ready research.

The thermal tolerance of photosynthetic tissues: a global systematic review and agenda for future research.

Understanding plant thermal tolerance is fundamental to predicting impacts of extreme temperature events that are increasing in frequency and intensity across the globe. Extremes, not averages, drive species evolution, determine survival and increase crop performance. To better prioritize agricultural and natural systems research, it is crucial to evaluate how researchers are assessing the capacity of plants to tolerate extreme events. We conducted a systematic review to determine how plant thermal tolerance research is distributed across wild and domesticated plants, growth forms and biomes, and to identify crucial knowledge gaps. Our review shows that most thermal tolerance research examines cold tolerance of cultivated species; c. 5% of articles consider both heat and cold tolerance. Plants of extreme environments are understudied, and techniques widely applied in cultivated systems are largely unused in natural systems. Lastly, we find that lack of standardized methods and metrics compromises the potential for mechanistic insight. Our review provides an entry point for those new to the methods used in plant thermal tolerance research and bridges often disparate ecological and agricultural perspectives for the more experienced. We present a considered agenda of thermal tolerance research priorities to stimulate efficient, reliable and repeatable research across the spectrum of plant thermal tolerance.

How to analyse plant phenotypic plasticity in response to a changing climate.

Contents Summary 1235 I. Introduction 1235 II. The many shapes of phenotypic plasticity 1236 III. Random regression mixed model framework 1237 IV. Conclusions 1240 Acknowledgements 1240 References 1240 SUMMARY: Plant biology is experiencing a renewed interest in the mechanistic underpinnings and evolution of phenotypic plasticity that calls for a re-evaluation of how we analyse phenotypic responses to a rapidly changing climate. We suggest that dissecting plant plasticity in response to increasing temperature needs an approach that can represent plasticity over multiple environments, and considers both population-level responses and the variation between genotypes in their response. Here, we outline how a random regression mixed model framework can be applied to plastic traits that show linear or nonlinear responses to temperature. Random regressions provide a powerful and efficient means of characterising plasticity and its variation. Although they have been used widely in other fields, they have only recently been implemented in plant evolutionary ecology. We outline their structure and provide an example tutorial of their implementation.

Drosophila melanogaster does not exhibit a behavioural fever response when infected with Drosophila C virus.

Behavioural fever is a widely conserved response to infection. The host increases body temperature (Tb) by altering their preferred temperature (Tp), generating a fever and delaying or avoiding pathogen-induced mortality. This response is not ubiquitous in insects, however, although few studies have investigated this response to viral infection. Here, we examined the change in Tp of Drosophila in response to virus infection using a thermal gradient. No difference in Tp was observed. We suggest that the lack of behavioural fever could be due to the increased energy cost of maintaining a higher Tb whilst the immune response is active. To the best of our knowledge, this is the first study to assay for changes in Tp of infected Drosophila.

Wolbachia-mediated antiviral protection in Drosophila larvae and adults following oral infection.

Understanding viral dynamics in arthropods is of great importance when designing models to describe how viral spread can influence arthropod populations. The endosymbiotic bacterium Wolbachia spp., which is present in up to 40% of all insect species, has the ability to alter viral dynamics in both Drosophila spp. and mosquitoes, a feature that in mosquitoes may be utilized to limit spread of important arboviruses. To understand the potential effect of Wolbachia on viral dynamics in nature, it is important to consider the impact of natural routes of virus infection on Wolbachia antiviral effects. Using adult Drosophila strains, we show here that Drosophila-Wolbachia associations that have previously been shown to confer antiviral protection following systemic viral infection also confer protection against virus-induced mortality following oral exposure to Drosophila C virus in adults. Interestingly, a different pattern was observed when the same fly lines were challenged with the virus when still larvae. Analysis of the four Drosophila-Wolbachia associations that were protective in adults indicated that only the w1118-wMelPop association conferred protection in larvae following oral delivery of the virus. Analysis of Wolbachia density using quantitative PCR (qPCR) showed that a high Wolbachia density was congruent with antiviral protection in both adults and larvae. This study indicates that Wolbachia-mediated protection may vary between larval and adult stages of a given Wolbachia-host combination and that the variations in susceptibility by life stage correspond with Wolbachia density. The differences in the outcome of virus infection are likely to influence viral dynamics in Wolbachia-infected insect populations in nature and could also have important implications for the transmission of arboviruses in mosquito populations.

Temperature and nutrition do not interact to shape the evolution of metabolic rate.

Metabolic cold adaptation, or Krogh's rule, is the controversial hypothesis that predicts a monotonically negative relationship between metabolic rate and environmental temperature for ectotherms living along thermal clines measured at a common temperature. Macrophysiological patterns consistent with Krogh's rule are not always evident in nature, and experimentally evolved responses to temperature have failed to replicate such patterns. Hence, temperature may not be the sole driver of observed variation in metabolic rate. We tested the hypothesis that temperature, as a driver of energy demand, interacts with nutrition, a driver of energy supply, to shape the evolution of metabolic rate to produce a pattern resembling Krogh's rule. To do this, we evolved replicate lines of Drosophila melanogaster at 18, 25 or 28°C on control, low-calorie or low-protein diets. Contrary to our prediction, we observed no effect of nutrition, alone or interacting with temperature, on adult female and male metabolic rates. Moreover, support for Krogh's rule was only in females at lower temperatures. We, therefore, hypothesize that observed variation in metabolic rate along environmental clines arises from the metabolic consequences of environment-specific life-history optimization, rather than because of the direct effect of temperature on metabolic rate. This article is part of the theme issue 'The evolutionary significance of variation in metabolic rates'.

Physiological and metabolic consequences of viral infection in Drosophila melanogaster.

An extensively used model system for investigating anti-pathogen defence and innate immunity involves Drosophila C virus (DCV) and Drosophila melanogaster. While there has been a significant effort to understand infection consequences at molecular and genetic levels, an understanding of fundamental higher-level physiology of this system is lacking. Here, we investigate the metabolic rate, locomotory activity, dry mass and water content of adult male flies injected with DCV, measured over the 4 days prior to virus-induced mortality. DCV infection resulted in multiple pathologies, notably the depression of metabolic rate beginning 2 days post-infection as a response to physiological stress. Even in this depressed metabolic state, infected flies did not decrease their activity until 1 day prior to mortality, which further suggests that cellular processes and synthesis are disrupted because of viral infection. Growth rate was also reduced, indicating that energy partitioning is altered as infection progresses. Microbial infection in insects typically results in an increase in excretion; however, water appeared to be retained in DCV-infected flies. We hypothesise that this is due to a fluid intake-output imbalance due to disrupted transport signalling and a reduced rate of metabolic processing. Furthermore, infected flies had a reduced rate of respiration as a consequence of metabolic depression, which minimised water loss, and the excess mass as a result of water retention is concurrent with impaired locomotory ability. These findings contribute to developing a mechanistic understanding of how pathologies accumulate and lead to mortality in infected flies.

Consistently high heat tolerance acclimation in response to a simulated heatwave across species from the broadly distributed Acacia genus.

When leaves exceed their thermal threshold during heatwaves, irreversible damage to the leaf can accumulate. However, few studies have explored short-term acclimation of leaves to heatwaves that could help plants to prevent heat damage with increasing heatwave intensity. Here, we studied the heat tolerance of PSII (PHT) in response to a heatwave in Acacia species from across a strong environmental gradient in Australia. We compared PHT metrics derived from temperature-dependent chlorophyll fluorescence response curves (T-F 0 ) before and during a 4-day 38°C heatwave in a controlled glasshouse experiment. We found that the 15 Acacia species displayed surprisingly large and consistent PHT acclimation responses with a mean tolerance increase of 12°C (range, 7.7-19.1°C). Despite species originating from diverse climatic regions, neither maximum temperature of the warmest month nor mean annual precipitation at origin were clear predictors of PHT. To our knowledge, these are some of the largest measured acclimation responses of PHT from a controlled heatwave experiment. This remarkable capacity could partially explain why this genus has become more diverse and common as the Australian continent became more arid and suggests that the presence of Acacia in Australian ecosystems will remain ubiquitous with climate change.

Meta-analysis reveals that resting metabolic rate is not consistently related to fitness and performance in animals.

Explaining variation in the fitness of organisms is a fundamental goal in evolutionary ecology. Maintenance energy metabolism is the minimum energy required to sustain biological processes at rest (resting metabolic rate: RMR) and is proposed to drive or constrain fitness of animals; however, this remains debated. Hypotheses have been proposed as to why fitness might increase with RMR (the 'increased intake' or 'performance' hypothesis), decrease with RMR (the 'compensation' or 'allocation' hypothesis), or vary among species and environmental contexts (the 'context dependent' hypothesis). Here, we conduct a systematic review and meta-analysis of the literature, finding 114 studies with 355 relationships between RMR and traits that may be related to fitness. We show that individuals with relatively high RMR generally have high fitness overall, which might be supported by an increased energy intake. However, fitness proxies are not interchangeable: the nature of the RMR-fitness relationship varied substantially depending on the specific trait in question, and we found no consistent relationship between RMR and those traits most closely linked with actual fitness (i.e., lifetime reproductive success). We hypothesise that maintaining high RMR is not costly when resources are unlimited, and we propose ideas for future studies to identify mechanisms underlying RMR-fitness relationships.

A high-throughput method for measuring critical thermal limits of leaves by chlorophyll imaging fluorescence.

Plant thermal tolerance is a crucial research area as the climate warms and extreme weather events become more frequent. Leaves exposed to temperature extremes have inhibited photosynthesis and will accumulate damage to PSII if tolerance thresholds are exceeded. Temperature-dependent changes in basal chlorophyll fluorescence (T-F0) can be used to identify the critical temperature at which PSII is inhibited. We developed and tested a high-throughput method for measuring the critical temperatures for PSII at low (CTMIN) and high (CTMAX) temperatures using a Maxi-Imaging fluorimeter and a thermoelectric Peltier plate heating/cooling system. We examined how experimental conditions of wet vs dry surfaces for leaves and heating/cooling rate, affect CTMIN and CTMAX across four species. CTMAX estimates were not different whether measured on wet or dry surfaces, but leaves were apparently less cold tolerant when on wet surfaces. Heating/cooling rate had a strong effect on both CTMAX and CTMIN that was species-specific. We discuss potential mechanisms for these results and recommend settings for researchers to use when measuring T-F0. The approach that we demonstrated here allows the high-throughput measurement of a valuable ecophysiological parameter that estimates the critical temperature thresholds of leaf photosynthetic performance in response to thermal extremes.

Sparse evidence for selection on phenotypic plasticity in response to temperature.

Phenotypic plasticity is frequently assumed to be an adaptive mechanism by which organisms cope with rapid changes in their environment, such as shifts in temperature regimes owing to climate change. However, despite this adaptive assumption, the nature of selection on plasticity within populations is still poorly documented. Here, we performed a systematic review and meta-analysis of estimates of selection on thermal plasticity. Although there is a large literature on thermal plasticity, we found very few studies that estimated coefficients of selection on measures of plasticity. Those that did do not provide strong support for selection on plasticity, with the majority of estimates of directional selection on plasticity being weak and non-significant, and no evidence for selection on plasticity overall. Although further estimates are clearly needed before general conclusions can be drawn, at present there is not clear empirical support for any assumption that plasticity in response to temperature is under selection. We present a multivariate mixed model approach for robust estimation of selection on plasticity and demonstrate how it can be implemented. Finally, we highlight the need to consider the environments, traits and conditions under which plasticity is (or is not) likely to be under selection, if we are to understand phenotypic responses to rapid environmental change. This article is part of the theme issue 'The role of plasticity in phenotypic adaptation to rapid environmental change'.

The origin and maintenance of metabolic allometry in animals.

Organisms vary widely in size, from microbes weighing 0.1 pg to trees weighing thousands of megagrams - a 1021-fold range similar to the difference in mass between an elephant and the Earth. Mass has a pervasive influence on biological processes, but the effect is usually non-proportional; for example, a tenfold increase in mass is typically accompanied by just a four- to sevenfold increase in metabolic rate. Understanding the cause of allometric scaling has been a long-standing problem in biology. Here, we examine the evolution of metabolic allometry in animals by linking microevolutionary processes to macroevolutionary patterns. We show that the genetic correlation between mass and metabolic rate is strong and positive in insects, birds and mammals. We then use these data to simulate the macroevolution of mass and metabolic rate, and show that the interspecific relationship between these traits in animals is consistent with evolution under persistent multivariate selection on mass and metabolic rate over long periods of time.