Host species differences in the thermal mismatch of host-parasitoid interactions.

Extreme high temperatures associated with climate change can affect species directly, and indirectly through temperature-mediated species interactions. In most host-parasitoid systems, parasitization inevitably kills the host, but differences in heat tolerance between host and parasitoid, and between different hosts, may alter their interactions. Here, we explored the effects of extreme high temperatures on the ecological outcomes - including, in some rare cases, escape from the developmental disruption of parasitism - of the parasitoid wasp, Cotesia congregata, and two co-occurring congeneric larval hosts, Manduca sexta and M. quinquemaculata. Both host species had higher thermal tolerance than C. congregata, resulting in a thermal mismatch characterized by parasitoid (but not host) mortality under extreme high temperatures. Despite parasitoid death at high temperatures, hosts typically remain developmentally disrupted from parasitism. However, high temperatures resulted in a partial developmental recovery from parasitism (reaching the wandering stage at the end of host larval development) in some host individuals, with a significantly higher frequency of this partial developmental recovery in M. quinquemaculata than in M. sexta. Hosts species also differed in their growth and development in the absence of parasitoids, with M. quinquemaculata developing faster and larger at high temperatures relative to M. sexta. Our results demonstrate that co-occurring congeneric species, despite shared environments and phylogenetic histories, can vary in their responses to temperature, parasitism and their interaction, resulting in altered ecological outcomes.

Responses of Manduca sexta larvae to heat waves.

Climate change is increasing the frequency of heat waves and other extreme weather events experienced by organisms. How does the number and developmental timing of heat waves affect survival, growth and development of insects? Do heat waves early in development alter performance later in development? We addressed these questions using experimental heat waves with larvae of the tobacco hornworm, Manduca sexta. The experiments used diurnally fluctuating temperature treatments differing in the number (0-3) and developmental timing (early, middle and/or late in larval development) of heat waves, in which a single heat wave involved three consecutive days with a daily maximum temperature of 42°C. Survival to pupation declined with increasing number of heat waves. Multiple (but not single) heat waves significantly reduced development time and pupal mass; the best models for the data indicated that both the number and developmental timing of heat waves affected performance. In addition, heat waves earlier in development significantly reduced growth and development rates later in larval development. Our results illustrate how the frequency and developmental timing of sublethal heat waves can have important consequences for life history traits in insects.

Responses of Manduca sexta larvae to heat waves.

Climate change is increasing the frequency of heat waves and other extreme weather events experienced by organisms. How does the number and developmental timing of heat waves affect survival, growth and development of insects? Do heat waves early in development alter performance later in development? We addressed these questions using experimental heat waves with larvae of the tobacco hornworm, Manduca sexta. The experiments used diurnally fluctuating temperature treatments differing in the number (0-3) and developmental timing (early, middle and/or late in larval development) of heat waves, in which a single heat wave involved three consecutive days with a daily maximum temperature of 42°C. Survival to pupation declined with increasing number of heat waves. Multiple (but not single) heat waves significantly reduced development time and pupal mass; the best models for the data indicated that both the number and developmental timing of heat waves affected performance. In addition, heat waves earlier in development significantly reduced growth and development rates later in larval development. Our results illustrate how the frequency and developmental timing of sublethal heat waves can have important consequences for life history traits in insects.

Compensating for climate change-induced cue-environment mismatches: evidence for contemporary evolution of a photoperiodic reaction norm in Colias butterflies.

Anthropogenic climate change alters seasonal conditions without altering photoperiod and can thus create a cue-environment mismatch for organisms that use photoperiod as a cue for seasonal plasticity. We investigated whether evolution of the photoperiodic reaction norm has compensated for this mismatch in Colias eurytheme. This butterfly's wing melanization has a thermoregulatory function and changes seasonally. In 1971, Hoffmann quantified how larval photoperiod determines adult wing melanization. We recreated his experiment 47 years later using a contemporary population. Comparing our results to his, we found decreased melanization at short photoperiods but no change in melanization at long photoperiods, which is consistent with the greater increase in spring than summer temperatures recorded for this region. Our study shows that evolution can help correct cue-environment mismatches but not in the same way under all conditions. Studies of contemporary evolution may miss important changes if they focus on only a limited range of conditions.

A Stochastic Model for Predicting Age and Mass at Maturity of Insects.

Variation in age and mass at maturity is commonly observed in populations, even among individuals with the same genetic and environmental backgrounds. Accounting for such individual variation with a stochastic model is important for estimating optimal evolutionary strategies and for understanding potential trade-offs among life-history traits. However, most studies employ stochastic models that are either phenomenological or account for variation in only one life-history trait. We propose a model based on the developmental biology of the moth Manduca sexta that accounts for stochasticity in two key life-history traits, age and mass at maturity. The model is mechanistic, describing feeding behavior and common insect developmental processes, including the degradation of juvenile hormone prior to molting. We derive a joint probability density function for the model and explore how the distribution of age and mass at maturity is affected by different parameter values. We find that the joint distribution is generally nonnormal and highly sensitive to parameter values. In addition, our model predicts previously observed effects of temperature change and nutritional quality on the expected values of insect age and mass. Our results highlight the importance of integrating multiple sources of stochasticity into life-history models.

The ghost of temperature past: interactive effects of previous and current thermal conditions on gene expression in Manduca sexta.

High temperatures can negatively impact the performance and survival of organisms, particularly ectotherms. While an organism's response to high temperature stress clearly depends on current thermal conditions, its response may also be affected by the temporal pattern and duration of past temperature exposures. We used RNA sequencing of Manduca sexta larvae fat body tissue to evaluate how diurnal temperature fluctuations during development affected gene expression both independently and in conjunction with subsequent heat stress. Additionally, we compared gene expression between two M. sexta populations, a lab colony and a genetically related field population that have been separated for >300 generations and differ in their thermal sensitivities. Lab-adapted larvae were predicted to show increased expression responses to both single and repeated thermal stress, whereas recurrent exposure could decrease later stress responses for field individuals. We found large differences in overall gene expression patterns between the two populations across all treatments, as well as population-specific transcriptomic responses to temperature; more differentially expressed genes were upregulated in the field compared with lab larvae. Developmental temperature fluctuations alone had minimal effects on long-term gene expression patterns, with the exception of a somewhat elevated stress response in the lab population. Fluctuating rearing conditions did alter gene expression during exposure to later heat stress, but this effect depended on both the population and the particular temperature conditions. This study contributes to increased knowledge of molecular mechanisms underlying physiological responses of organisms to temperature fluctuations, which is needed for the development of more accurate thermal performance models.

Climate Warming, Resource Availability, and the Metabolic Meltdown of Ectotherms.

Climate warming may lower environmental resource levels, growth, and fitness of many ectotherms. In a classic experiment, Brett and colleagues documented that growth rates of salmon depended strikingly on both temperature and food levels. Here we develop a simple bioenergetic model that explores how fixed temperatures and food jointly alter the thermal sensitivity of net energy gain. The model incorporates differing thermal sensitivities of energy intake and metabolism. In qualitative agreement with Brett's results, it predicts that decreased food intake reduces growth rates, lowers optimal temperatures for growth, and lowers the highest temperatures sustaining growth (upper thermal limit). Consequently, ectotherms facing reduced food intake in warm environments should restrict activity to times when low body temperatures are biophysically feasible, but-in a warming world-that will force ectotherms to shorten activity times and thus further reduce food intake. This "metabolic meltdown" is a consequence of declining energy intake coupled with accelerating metabolic costs at high temperatures and with warming-imposed restrictions on activity. Next, we extend the model to explore how increasing mean environmental temperatures alter the thermal sensitivity of growth: when food intake is reduced, optimal temperatures and upper thermal limits for growth are lowered. We discuss our model's key assumptions and caveats as well as its relationship to a recent model for phytoplankton. Both models illustrate that the deleterious impacts of climate warming on ectotherms will be amplified if food intake is also reduced, either because warming reduces standing food resources or because it restricts foraging time.

No evidence that warmer temperatures are associated with selection for smaller body sizes.

Reductions in animal body size over recent decades are often interpreted as an adaptive evolutionary response to climate warming. However, for reductions in size to reflect adaptive evolution, directional selection on body size within populations must have become negative, or where already negative, to have become more so, as temperatures increased. To test this hypothesis, we performed traditional and phylogenetic meta-analyses of the association between annual estimates of directional selection on body size from wild populations and annual mean temperatures from 39 longitudinal studies. We found no evidence that warmer environments were associated with selection for smaller size. Instead, selection consistently favoured larger individuals, and was invariant to temperature. These patterns were similar in ectotherms and endotherms. An analysis using year rather than temperature revealed similar patterns, suggesting no evidence that selection has changed over time, and also indicating that the lack of association with annual temperature was not an artefact of choosing an erroneous time window for aggregating the temperature data. Although phenotypic trends in size will be driven by a combination of genetic and environmental factors, our results suggest little evidence for a necessary ingredient-negative directional selection-for declines in body size to be considered an adaptive evolutionary response to changing selection pressures.

The analysis and interpretation of critical temperatures.

Critical temperatures are widely used to quantify the upper and lower thermal limits of organisms. But measured critical temperatures often vary with methodological details, leading to spirited discussions about the potential consequences of stress and acclimation during the experiments. We review a model based on the simple assumption that failure rate increases with increasing temperature, independent of previous temperature exposure, water loss or metabolism during the experiment. The model predicts that mean critical thermal maximal temperature (CTmax) increases non-linearly with starting temperature and ramping rate, a pattern frequently observed in empirical studies. We then develop a statistical model that estimates a failure rate function (the relationship between failure rate and current temperature) using maximum likelihood; the best model accounts for 58% of the variation in CTmax in an exemplary dataset for tsetse flies. We then extend the model to incorporate potential effects of stress and acclimation on the failure rate function; the results show how stress accumulation at low ramping rate may increase the failure rate and reduce observed values of CTmax We also applied the model to an acclimation experiment with hornworm larvae that used a single starting temperature and ramping rate; the analyses show that increasing acclimation temperature significantly reduced the slope of the failure rate function, increasing the temperature at which failure occurred. The model directly applies to critical thermal minima, and can utilize data from both ramping and constant-temperature assays. Our model provides a new approach to analyzing and interpreting critical temperatures.

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.

Evolution of plasticity and adaptive responses to climate change along climate gradients.

The relative contributions of phenotypic plasticity and adaptive evolution to the responses of species to recent and future climate change are poorly understood. We combine recent (1960-2010) climate and phenotypic data with microclimate, heat balance, demographic and evolutionary models to address this issue for a montane butterfly, Colias eriphyle, along an elevational gradient. Our focal phenotype, wing solar absorptivity, responds plastically to developmental (pupal) temperatures and plays a central role in thermoregulatory adaptation in adults. Here, we show that both the phenotypic and adaptive consequences of plasticity vary with elevation. Seasonal changes in weather generate seasonal variation in phenotypic selection on mean and plasticity of absorptivity, especially at lower elevations. In response to climate change in the past 60 years, our models predict evolutionary declines in mean absorptivity (but little change in plasticity) at high elevations, and evolutionary increases in plasticity (but little change in mean) at low elevation. The importance of plasticity depends on the magnitude of seasonal variation in climate relative to interannual variation. Our results suggest that selection and evolution of both trait means and plasticity can contribute to adaptive response to climate change in this system. They also illustrate how plasticity can facilitate rather than retard adaptive evolutionary responses to directional climate change in seasonal environments.

Beyond Thermal Performance Curves: Modeling Time-Dependent Effects of Thermal Stress on Ectotherm Growth Rates.

Thermal performance curves have been widely used to model the ecological responses of ectotherms to variable thermal environments and climate change. Such models ignore the effects of time dependence-the temporal pattern and duration of temperature exposure-on performance. We developed and solved a simple mathematical model for growth rate of ectotherms, combining thermal performance curves for ingestion rate with the temporal dynamics of gene expression and protein production in response to high temperatures to predict temporal patterns of growth rate in constant and diurnally fluctuating temperatures. We used the model to explore the effects of heat shock proteins on larval growth rates of Manduca sexta. The model correctly captures two empirical patterns for larval growth rate: first, maximal growth rate and optimal temperature decline with increasing duration of temperature exposure; second, mean growth rates decline with time in diurnally fluctuating temperatures at higher mean temperatures. These qualitative results apply broadly to cases where proteins or other molecules produced in response to high temperatures reduce growth rates. We discuss some of the critical assumptions and predictions of the model and suggest potential extensions and alternatives. Incorporating time-dependent effects will be essential for making more realistic predictions about the physiological and ecological consequences of temperature fluctuations and climate change.

Plasticity of upper thermal limits to acute and chronic temperature variation in Manduca sexta larvae.

In many ectotherms, exposure to high temperatures can improve subsequent tolerance to higher temperatures. However, the differential effects of single, repeated or continuous exposure to high temperatures are less clear. We measured the effects of single heat shocks and of diurnally fluctuating or constant rearing temperatures on the critical thermal maximum (CTmax) for final instar larvae of Manduca sexta Brief (2 h) heat shocks at temperatures of 35°C and above significantly increased CTmax relative to control temperatures (25°C). Increasing mean temperatures (from 25 to 30°C) or greater diurnal fluctuations (from constant to ±10°C) during larval development also significantly increased CTmax Combining these data showed that repeated or continuous temperature exposure during development improved heat tolerance beyond the effects of a single exposure to the same maximum temperature. These results suggest that both acute and chronic temperature exposure can result in adaptive plasticity of upper thermal limits.

Geographic divergence in upper thermal limits across insect life stages: does behavior matter?

Insects with complex life cycles vary in size, mobility, and thermal ecology across life stages. We examine how differences in the capacity for thermoregulatory behavior influence geographic differences in physiological heat tolerance among egg and adult Colias butterflies. Colias adults exhibit differences in morphology (wing melanin and thoracic setal length) along spatial gradients, whereas eggs are morphologically indistinguishable. Here we compare Colias eriphyle eggs and adults from two elevations and Colias meadii from a high elevation. Hatching success and egg development time of C. eriphyle eggs did not differ significantly with the elevation of origin. Egg survival declined in response to heat-shock temperatures above 38-40 °C and egg development time was shortest at intermediate heat-shock temperatures of 33-38 °C. Laboratory experiments with adults showed survival in response to heat shock was significantly greater for Colias from higher than from lower elevation sites. Common-garden experiments at the low-elevation field site showed that C. meadii adults initiated heat-avoidance and over-heating behaviors significantly earlier in the day than C. eriphyle. Our study demonstrates the importance of examining thermal tolerances across life stages. Our findings are inconsistent with the hypothesis that thermoregulatory behavior inhibits the geographic divergence of physiological traits in mobile stages, and suggest that sessile stages may evolve similar heat tolerances in different environments due to microclimatic variability or evolutionary constraints.

Genetic variation, simplicity, and evolutionary constraints for function-valued traits.

Understanding the patterns of genetic variation and constraint for continuous reaction norms, growth trajectories, and other function-valued traits is challenging. We describe and illustrate a recent analytical method, simple basis analysis (SBA), that uses the genetic variance-covariance (G) matrix to identify "simple" directions of genetic variation and genetic constraints that have straightforward biological interpretations. We discuss the parallels between the eigenvectors (principal components) identified by principal components analysis (PCA) and the simple basis (SB) vectors identified by SBA. We apply these methods to estimated G matrices obtained from 10 studies of thermal performance curves and growth curves. Our results suggest that variation in overall size across all ages represented most of the genetic variance in growth curves. In contrast, variation in overall performance across all temperatures represented less than one-third of the genetic variance in thermal performance curves in all cases, and genetic trade-offs between performance at higher versus lower temperatures were often important. The analyses also identify potential genetic constraints on patterns of early and later growth in growth curves. We suggest that SBA can be a useful complement or alternative to PCA for identifying biologically interpretable directions of genetic variation and constraint in function-valued traits.

Climate variability slows evolutionary responses of Colias butterflies to recent climate change.

How does recent climate warming and climate variability alter fitness, phenotypic selection and evolution in natural populations? We combine biophysical, demographic and evolutionary models with recent climate data to address this question for the subalpine and alpine butterfly, Colias meadii, in the southern Rocky Mountains. We focus on predicting patterns of selection and evolution for a key thermoregulatory trait, melanin (solar absorptivity) on the posterior ventral hindwings, which affects patterns of body temperature, flight activity, adult and egg survival, and reproductive success in Colias. Both mean annual summer temperatures and thermal variability within summers have increased during the past 60 years at subalpine and alpine sites. At the subalpine site, predicted directional selection on wing absorptivity has shifted from generally positive (favouring increased wing melanin) to generally negative during the past 60 years, but there is substantial variation among years in the predicted magnitude and direction of selection and the optimal absorptivity. The predicted magnitude of directional selection at the alpine site declined during the past 60 years and varies substantially among years, but selection has generally been positive at this site. Predicted evolutionary responses to mean climate warming at the subalpine site since 1980 is small, because of the variability in selection and asymmetry of the fitness function. At both sites, the predicted effects of adaptive evolution on mean population fitness are much smaller than the fluctuations in mean fitness due to climate variability among years. Our analyses suggest that variation in climate within and among years may strongly limit evolutionary responses of ectotherms to mean climate warming in these habitats.

Elevational differences in developmental plasticity determine phenological responses of grasshoppers to recent climate warming.

Annual species may increase reproduction by increasing adult body size through extended development, but risk being unable to complete development in seasonally limited environments. Synthetic reviews indicate that most, but not all, species have responded to recent climate warming by advancing the seasonal timing of adult emergence or reproduction. Here, we show that 50 years of climate change have delayed development in high-elevation, season-limited grasshopper populations, but advanced development in populations at lower elevations. Developmental delays are most pronounced for early-season species, which might benefit most from delaying development when released from seasonal time constraints. Rearing experiments confirm that population, elevation and temperature interact to determine development time. Population differences in developmental plasticity may account for variability in phenological shifts among adults. An integrated consideration of the full life cycle that considers local adaptation and plasticity may be essential for understanding and predicting responses to climate change.

Fluctuating temperatures and ectotherm growth: distinguishing non-linear and time-dependent effects.

Most terrestrial ectotherms experience diurnal and seasonal variation in temperature. Because thermal performance curves are non-linear, mean performance can differ in fluctuating and constant thermal environments. However, time-dependent effects--effects of the order and duration of exposure to temperature--can also influence mean performance. We quantified the non-linear and time-dependent effects of diurnally fluctuating temperatures for larval growth rates in the tobacco hornworm, Manduca sexta L., with four main results. First, the shape of the thermal performance curve for growth rate depended on the duration of exposure: for example, optimal temperature and thermal breadth were greater for growth rates measured over short (24 h during the last instar) compared with long (the entire period of larval growth) time periods. Second, larvae reared in diurnally fluctuating temperatures had significantly higher optimal temperatures and maximal growth rates than larvae reared in constant temperatures. Third, for larvae maintained at three mean temperatures (20, 25 and 30°C) and three diurnal temperature ranges (±0, ±5 and ±10°C), diurnal fluctuations had opposite effects on mean growth rates at low versus high mean temperature. Fourth, both short- and long-term thermal performance curves yielded poor predictions of the non-linear effects of fluctuating temperature on mean growth rates (compared with our experimental results) at higher mean temperatures. Our results suggest caution in using constant temperature studies to model the consequences of variable thermal environments.

Phenotypic clines, energy balances and ecological responses to climate change.

The Metabolic Theory of Ecology has renewed interest in using energetics to scale across levels of ecological organization. Can scaling from individual phenotypes to population dynamics provides insight into why species have shifted their phenologies, abundances and distributions idiosyncratically in response to recent climate change? We consider how the energetic implications of phenotypes may scale to understand population and species level responses to climate change using four focal grasshopper species along an elevation gradient in Colorado. We use a biophysical model to translate phenotypes and environmental conditions into estimates of body temperatures. We measure thermal tolerances and preferences and metabolic rates to assess rates of energy use and acquisition. Body mass declines along the elevation gradient for all species, but mass-specific metabolic rates increases only modestly. We find interspecific differences in both overall thermal tolerances and preferences and in the variation of these metrics along the elevation gradient. The more dispersive species exhibit significantly higher thermal tolerance and preference consistent with much of their range spanning hot, low elevation areas. When integrating these metrics to consider metabolic constraints, we find that energetic costs decrease along the elevation gradient due to decreasing body size and temperature. Opportunities for energy acquisition, as reflected by the proportion of time that falls within a grasshopper's thermal tolerance range, peak at mid elevations. We discuss methods for translating these energetic metrics into population dynamics. Quantifying energy balances and allocation offers a viable approach for predicting how populations will respond to climate change and the consequences for species composed of populations that may be locally adapted.