Developmental plasticity in thermal tolerance: Ontogenetic variation, persistence, and future directions.

Understanding the factors affecting thermal tolerance is crucial for predicting the impact climate change will have on ectotherms. However, the role developmental plasticity plays in allowing populations to cope with thermal extremes is poorly understood. Here, we meta-analyse how thermal tolerance is initially and persistently impacted by early (embryonic and juvenile) thermal environments by using data from 150 experimental studies on 138 ectothermic species. Thermal tolerance only increased by 0.13°C per 1°C change in developmental temperature and substantial variation in plasticity (~36%) was the result of shared evolutionary history and species ecology. Aquatic ectotherms were more than three times as plastic as terrestrial ectotherms. Notably, embryos expressed weaker but more heterogenous plasticity than older life stages, with numerous responses appearing as non-adaptive. While developmental temperatures did not have persistent effects on thermal tolerance overall, persistent effects were vastly under-studied, and their direction and magnitude varied with ontogeny. Embryonic stages may represent a critical window of vulnerability to changing environments and we urge researchers to consider early life stages when assessing the climate vulnerability of ectotherms. Overall, our synthesis suggests that developmental changes in thermal tolerance rarely reach levels of perfect compensation and may provide limited benefit in changing environments.

Genetic and environmental drivers of colour and pattern in the Australian jacky dragon (Amphibolurus muricatus).

The underlying drivers of variation in the colouration (colour and pattern) of animals can be genetic, non-genetic, or more likely, a combination of both. Understanding the role of heritable genetic elements, as well as non-genetic factors such as age, habitat or temperature, in shaping colouration can provide insight into the evolution and function of these traits, as well as the speed of response to changing environments. This project examined the genetic and non-genetic drivers of continuous variation in colouration in a lizard, the jacky dragon (Amphibolurus muricatus). We leveraged a large captive experiment that manipulated parental and offspring thermal environment to simultaneously estimate the genetic and non-genetic drivers of variation in colouration. We found that the overall brightness, the elongation of the longitudinal stripes on the dorsum and the contrast between light and dark patches of the pattern were all heritable. Colouration varied according to the age of the hatchling; however, the thermal environment of neither the parents nor offspring contributed significantly to colouration. It appears that developmental plasticity and maternal effects associated with temperature are not important drivers of variation in our measures of colouration.

Impact of fluctuating developmental temperatures on phenotypic traits in reptiles: a meta-analysis.

During the vulnerable stages of early life, most ectothermic animals experience hourly and diel fluctuations in temperature as air temperatures change. While we know a great deal about how different constant temperatures impact the phenotypes of developing ectotherms, we know remarkably little about the impacts of temperature fluctuations on the development of ectotherms. In this study, we used a meta-analytic approach to compare the mean and variance of phenotypic outcomes from constant and fluctuating incubation temperatures across reptile species. We found that fluctuating temperatures provided a small benefit (higher hatching success and shorter incubation durations) at cool mean temperatures compared with constant temperatures, but had a negative effect at warm mean temperatures. In addition, more extreme temperature fluctuations led to greater reductions in embryonic survival compared with moderate temperature fluctuations. Within the limited data available from species with temperature-dependent sex determination, embryos had a higher chance of developing as female when developing in fluctuating temperatures compared with those developing in constant temperatures. With our meta-analytic approach, we identified average mean nest temperatures across all taxa where reptiles switch from receiving benefits to incurring costs when incubation temperatures fluctuate. More broadly, our study indicates that the impact of fluctuating developmental temperature on some phenotypes in ectothermic taxa are likely to be predictable via integration of developmental temperature profiles with thermal performance curves.

Best practices for building and curating databases for comparative analyses.

Comparative analyses have a long history of macro-ecological and -evolutionary approaches to understand structure, function, mechanism and constraint. As the pace of science accelerates, there is ever-increasing access to diverse types of data and open access databases that are enabling and inspiring new research. Whether conducting a species-level trait-based analysis or a formal meta-analysis of study effect sizes, comparative approaches share a common reliance on reliable, carefully curated databases. Unlike many scientific endeavors, building a database is a process that many researchers undertake infrequently and in which we are not formally trained. This Commentary provides an introduction to building databases for comparative analyses and highlights challenges and solutions that the authors of this Commentary have faced in their own experiences. We focus on four major tips: (1) carefully strategizing the literature search; (2) structuring databases for multiple use; (3) establishing version control within (and beyond) your study; and (4) the importance of making databases accessible. We highlight how one's approach to these tasks often depends on the goal of the study and the nature of the data. Finally, we assert that the curation of single-question databases has several disadvantages: it limits the possibility of using databases for multiple purposes and decreases efficiency due to independent researchers repeatedly sifting through large volumes of raw information. We argue that curating databases that are broader than one research question can provide a large return on investment, and that research fields could increase efficiency if community curation of databases was established.

Ovotestes suggest cryptic genetic influence in a reptile model for temperature-dependent sex determination.

Sex determination and differentiation in reptiles is complex. Temperature-dependent sex determination (TSD), genetic sex determination (GSD) and the interaction of both environmental and genetic cues (sex reversal) can drive the development of sexual phenotypes. The jacky dragon (Amphibolurus muricatus) is an attractive model species for the study of gene-environment interactions because it displays a form of Type II TSD, where female-biased sex ratios are observed at extreme incubation temperatures and approximately 50 : 50 sex ratios occur at intermediate temperatures. This response to temperature has been proposed to occur due to underlying sex determining loci, the influence of which is overridden at extreme temperatures. Thus, sex reversal at extreme temperatures is predicted to produce the female-biased sex ratios observed in A. muricatus. The occurrence of ovotestes during development is a cellular marker of temperature sex reversal in a closely related species Pogona vitticeps. Here, we present the first developmental data for A. muricatus, and show that ovotestes occur at frequencies consistent with a mode of sex determination that is intermediate between GSD and TSD. This is the first evidence suggestive of underlying unidentified sex determining loci in a species that has long been used as a model for TSD.

Heightened among-individual variation in life history but not morphology is related to developmental temperature in reptiles.

Increases in phenotypic variation under extreme (e.g. novel or stressful) environmental conditions are emerging as a crucial process through which evolutionary adaptation can occur. Lack of prior stabilizing selection, as well as potential instability of developmental processes in these environments, may lead to a release of phenotypic variation that can have important evolutionary consequences. Although such patterns have been shown in model study organisms, we know little about the generality of trait variance across environments for non-model organisms. Here, we test whether extreme developmental temperatures increase the phenotypic variation across diverse reptile taxa. We find that the among-individual variation in a key life-history trait (post-hatching growth) increases at extreme cold and hot temperatures. However, variations in two measures of hatchling morphology and in hatchling performance were not related to developmental temperature. Although extreme developmental temperatures may increase the variation in growth, our results suggest that plastic responses to stressful incubation conditions do not generally make more extreme phenotypes available to selection. We discuss the reasons for the general lack of increased variability at extreme incubation temperatures and the implications this has for local adaptation in hatchling morphology and physiology.

Climate change, sex reversal and lability of sex-determining systems.

Sex reversal at high temperatures during embryonic development (e.g., ZZ females) provides the opportunity for new genotypic crosses (e.g., ZZ male × ZZ female). This raises the alarming possibility that climatic warming could lead to the loss of an entire chromosome-one member of the sex chromosome pair (the Y or W)-and the transition of populations to environmental sex determination (ESD). Here we examine the evolutionary dynamics of sex-determining systems exposed to climatic warming using theoretical models. We found that the loss of sex chromosomes is not an inevitable consequence of sex reversal. A large frequency of ZZ sex reversal (50% reversal from male to female) typically divides the outcome between loss of the ZW genotype and the stable persistence of ZZ males, ZW females and ZZ females. The amount of warming associated with sex chromosome loss depended on several features of wild populations-environmental fluctuation, immigration, heritable variation in temperature sensitivity and differential fecundity of sex-reversed individuals. Chromosome loss was partially or completely buffered when sex-reversed individuals suffered a reproductive fitness cost, when immigration occurred or when heritable variation for temperature sensitivity existed. Thus, under certain circumstances, sex chromosomes may persist cryptically in systems where the environment is the predominant influence on sex.

Context dependence of transgenerational plasticity: the influence of parental temperature depends on offspring environment and sex.

Under environmental change, the relationship between phenotype and fitness can change rapidly, leaving populations vulnerable. Plasticity within and between generations could provide the fastest mitigation to environmental change. However, plasticity may depend on interactions among parental environment, offspring environment and offspring sex, and we know little of how these interactions manifest. We examine the importance of parental and offspring thermal environment in the context of a terrestrial ectotherm (the jacky lizard, Amphibolurus muricatus), where the thermal environment depends on complex thermoregulatory behaviours. By manipulating both parental and offspring thermoregulatory (basking) opportunities in a full factorial design, we found that transgenerational plasticity was highly context dependent. Overall, longer parental thermoregulatory opportunities led to increased growth in offspring, providing a clear fitness benefit to daughters but inducing a cost of increased oxidative stress in sons. Daughters, but not sons, received the greatest advantage when their thermal environment matched that of their parents. The offspring thermal environment had little independent effect on offspring phenotype. Together, the results suggest that both directional selection on offspring size (leading to growth being dependent on thermal conditions) and selection for anticipatory parental effects operate, but with a different balance in males and females. More broadly, restrictions in thermoregulatory opportunities under climatic warming will have negative consequences both within and between generations, but the exact nature will depend on the evolved functional form of plasticity.

Drop it like it's hot: Interpopulation variation in thermal phenotypes shows counter-gradient pattern.

Ectotherms utilise a complex array of behavioural and physiological mechanisms to cope with variation in suboptimal thermal environments. However, these mechanisms may be insufficient for population persistence under contemporary climate change, resulting in a greater need to understand how local populations respond to geographic variation in climate. In this study, we explored the potential for local adaptation and acclimation in thermal traits and behaviours using wild and captive populations of a small agamid lizard (the jacky lizard, Amphibolurus muricatus). We predicted that wild lizards from a high elevation site would have cooler thermal preferences compared to those at low elevation sites to match the more restricted thermal resources at higher, cooler elevations. We additionally explored whether variation in thermal traits was due to recent acclimation or fixed population differences, such as due to developmental plasticity or local adaptation. In contrast to our predictions, we found high-elevation lizards began panting at higher temperatures and had higher thermal preferences relative to lower elevation lizards. When allowed to bask freely, there was no difference in the intensity of basking or daily duration of time spent basking between lizards from different elevations. Although the high-elevation lizards appeared to show stronger acclimation to recent air temperatures compared to low-elevation lizards, this difference was not significant. Similarly, captive lizards acclimated under long and short basking regimes showed no major differences in thermal traits or basking behaviour. Our results are consistent with the presence of counter-gradient variation in thermal phenotypes of lizards, and suggest that these are driven by local adaptive responses or developmental effects rather than recent acclimation.

Troubleshooting public data archiving: suggestions to increase participation.

An increasing number of publishers and funding agencies require public data archiving (PDA) in open-access databases. PDA has obvious group benefits for the scientific community, but many researchers are reluctant to share their data publicly because of real or perceived individual costs. Improving participation in PDA will require lowering costs and/or increasing benefits for primary data collectors. Small, simple changes can enhance existing measures to ensure that more scientific data are properly archived and made publicly available: (1) facilitate more flexible embargoes on archived data, (2) encourage communication between data generators and re-users, (3) disclose data re-use ethics, and (4) encourage increased recognition of publicly archived data.

Parental thermal environment alters offspring sex ratio and fitness in an oviparous lizard.

The environment experienced by parents can impact the phenotype of their offspring (parental effects), a critical component of organismal ecology and evolution in variable or changing environments. Although temperature is a central feature of the environment for ectotherms, its role in parental effects has been little explored until recently. Here, parental basking opportunity was manipulated in an oviparous lizard with temperature-dependent sex determination, the jacky dragon (Amphibolurus muricatus). Eggs were incubated at a temperature that typically produces a 50:50 sex ratio, and hatchlings were reared in a standard thermal environment. Offspring of parents in short bask conditions appeared to have better fitness outcomes in captive conditions than those of parents in long bask conditions - they had greater growth and survival as a function of their mass. In addition, the sex of offspring (male or female) depended on the interaction between parental treatment and egg mass, and treatment impacted whether sons or daughters grew larger in their first season. The interactive effects of treatment on offspring sex and growth are consistent with adaptive explanations for the existence of temperature-dependent sex determination in this species. Moreover, the greater performance recorded in short bask offspring may represent an anticipatory parental effect to aid offspring in predicted conditions of restricted thermal opportunity. Together, these responses constitute a crucial component of the population response to spatial or temporal variation in temperature.

Demography can favour female-advantageous alleles.

When female fecundity is relatively independent of male abundance, while male reproduction is proportional to female abundance, females have a larger effect on population dynamics than males (i.e. female demographic dominance). This population dynamic phenomenon might not appear to influence evolution, because male and female genomes still contribute equally much to the next generation. However, here we examine two evolutionary scenarios to provide a proof of principle that spatial structure can make female demographic dominance matter. Our two simulation models combine dispersal evolution with local adaptation subjected to intralocus sexual conflict and environmentally driven sex ratio biases, respectively. Both models have equilibria where one environment (without being intrinsically poorer) has so few reproductive females that trait evolution becomes disproportionately determined by those environments where females survive better (intralocus sexual conflict model), or where daughters are overproduced (environmental sex determination model). Surprisingly, however, the two facts that selection favours alleles that benefit females, and population growth is improved when female fitness is high, together do not imply that all measures of population performance are improved. The sex-specificity of the source-sink dynamics predicts that populations can evolve to fail to persist in habitats where alleles do poorly when expressed in females.

How do climate-linked sex ratios and dispersal limit range boundaries?

BACKGROUND: Geographic ranges of ectotherms such as reptiles may be determined strongly by abiotic factors owing to causal links between ambient temperature, juvenile survival and individual sex (male or female). Unfortunately, we know little of how these factors interact with dispersal among populations across a species range. We used a simulation model to examine the effects of dispersal, temperature-dependent juvenile survival and sex determining mechanism (temperature-dependent sex determination (TSD) and genotypic sex determination (GSD)) and their interactions, on range limits in populations extending across a continuous range of air temperatures. In particular, we examined the relative importance of these parameters for population persistence to recommend targets for future empirical research. RESULTS: Dispersal influenced the range limits of species with TSD to a greater extent than in GSD species. Whereas male dispersal led to expanded species ranges across warm (female-producing) climates, female dispersal led to expanded ranges across cool (male-producing) climates. Two-sex dispersal eliminated the influence of biased sex ratios on ranges. CONCLUSION: The results highlight the importance of the demographic parameter of sex ratio in determining population persistence and species range limits.

Direct effects of incubation temperature on morphology, thermoregulatory behaviour and locomotor performance in jacky dragons (Amphibolurus muricatus).

Incubation temperature is one of the most studied factors driving phenotypic plasticity in oviparous reptiles. We examined how incubation temperature influenced hatchling morphology, thermal preference and temperature-dependent running speed in the small Australian agamid lizard Amphibolurus muricatus. Hatchlings incubated at 32 °C grew more slowly than those incubated at 25 and 28 °C during their first month after hatching, and tended to be smaller at one month. These differences were no longer significant by three months of age due to selective mortality of the smallest hatchlings. The cooler incubation treatments (25 °C and 28 °C) produced lizards that had deeper and wider heads. Hatchlings from 28 °C had cooler and more stable temperature preferences, and also had lower body temperatures during a 2-h thermoregulatory behaviour trial. Locomotor performance was enhanced at higher body temperatures, but incubation temperature had no measurable effect either independently or in interaction with body temperature. Our study demonstrates that incubation temperature has direct effects on morphology and thermoregulatory behaviour that appears to be independent of any size-dependent effects. We postulate a mechanistic link between these two effects.

Plasticity and the adaptive evolution of switchlike reaction norms under environmental change.

Phenotypic plasticity is often posited as an avenue for adaptation to environmental change, whereby environmental influences on phenotypes could shift trait expression toward new optimal values. Conversely, plastic trait expression may inhibit adaptation to environmental change by reducing selective pressure on ill-adapted traits. While plastic responses are often assumed to be linear, nonlinear phenotype-environment relationships are common, especially in thermally sensitive traits. Here we examine nonlinear plasticity in a trait with great ecological and evolutionary significance: sexual phenotype in species with environmental sex determination (ESD). In species with ESD, development switches between male and female at an environmental threshold (the inflection point). The inflection point is a key trait for adaptive responses to changing environments and should evolve toward the new optimum in order to maintain evolutionarily stable sex ratios. We used an individual-based theoretical model to investigate how two forms of plasticity in the ESD reaction norm-the nonlinear slope of the reaction norm and a linear shift in the inflection point-influence the evolution of the inflection point under climate warming. We found that steeper reaction norm slopes (high nonlinear plasticity) promoted evolution toward new optimal phenotypes (higher inflection points). In contrast, increased linear plasticity in the inflection point (shift) hindered adaptive evolution. Additionally, populations in moderate warming scenarios showed greater adaptive evolution of the inflection point compared with populations in extreme warming scenarios, suggesting that the proximity of existing phenotypes to new optimal phenotypes influences evolutionary outcomes. Unexpectedly, we found greater population persistence under high climate variability, due to the increased production of rare-sex individuals in unusually cold years. Our results demonstrate that different forms of phenotypic plasticity have crucially different effects on adaptive evolution. Plasticity that prevented sex ratio bias hindered the evolution of the inflection point, while plasticity that exacerbated sex ratio bias promoted adaptation to environmental change.

New horizons for comparative studies and meta-analyses.

Comparative analyses and meta-analyses are key tools to elucidate broad biological principles, yet the two approaches often appear different in purpose. We propose an integrated approach that can generate deeper insights into ecoevolutionary processes. Marrying comparative and meta-analytic approaches will allow for (i) a more accurate investigation of drivers of biological variation, (ii) a greater ability to account for sources of non-independence in experimental data, (iii) more effective control of publication bias, and (iv) improved transparency and reproducibility. Stronger integration of meta-analytic and comparative studies can also broaden the scope from species-centric investigations to community-level responses and function-valued traits (e.g., reaction norms). We illuminate commonalities, differences, and the transformative potential of combining these methodologies for advancing ecology and evolutionary biology.

Linking disease and community ecology through behavioural indicators: immunochallenge of white-footed mice and its ecological impacts.

1. Pathogens and immune challenges can induce changes in host phenotype in ways that indirectly impact important community interactions, including those that affect host-pathogen interactions. 2. To explore host behavioural response to immune challenge, we exposed wild white-footed mice (Peromyscus leucopus) to an immunogen from an endemic, zoonotic pathogen, the spirochete Borrelia burgdorferi. White-footed mice are a major reservoir host of Lyme disease (LD) spirochetes in northeastern USA and an abundant member of forest communities. The activity patterns, foraging behaviour, and space use of white-footed mice have implications for population growth rates of community members upon which mice incidentally prey (i.e. gypsy moths and native thrushes), as well as potentially determining host-vector encounter rates and human risk of LD. 3. Immunochallenge led to specific humoral (antibody) and cellular (i.e. elevated neutrophils and eosinophils) immune responses, supporting use of the immunogen as a surrogate for pathogenic infection. 4. Immunochallenged mice had reduced wheel-running activity early in the night when measured in the lab. However, mouse activity, as measured by track plates in natural field experiments, did not differ between mice exposed to the immunogen and unexposed mice. 5. Foraging behaviour of wild mice in the field - assessed with giving-up densities of seed at artificial feeding stations - was affected by exposure to the immunogen. Whereas immunochallenge did not influence whether foraging mice gained information on patch quality while foraging, it led to reductions in predator avoidance during foraging, suggesting that the proportion of space used by foraging mice may be greater as a result of immunochallenge. This increased space use is predicted to increase encounter rates with patchily distributed LD vectors (ticks) and with incidental prey items. 6. Thus, immunochallenge in white-footed mice, and potentially pathogenic infection, have the potential to indirectly impact community interactions, including those important for pathogen transmission.

Sexual Development and the Environment: Conclusions from 40 Years of Theory.

In this review, we consider the insight that has been gained through theoretical examination of environmental sex determination (ESD) and thermolability - how theory has progressed our understanding of the ecological and evolutionary dynamics associated with ESD, the transitional pathways between different modes of sex determination, and the underlying mechanisms. Following decades of theory on the adaptive benefits of ESD, several hypotheses seem promising. These hypotheses focus on the importance of differential fitness (sex-specific effects of temperature on fitness) in generating selection for ESD, but highlight alternative ways differential fitness arises: seasonal impacts on growth, sex-specific ages of maturation, and sex-biased dispersal. ESD has the potential to generate biased sex ratios quite easily, leading to complex feedbacks between the ecology and evolution of ESD. Frequency-dependent selection on sex acts on ESD-related traits, driving local adaptation or plasticity to restore equilibrium sex ratio. However, migration and overlapping generations ("mixing") diminish local adaptation and leave each cohort/population with the potential for biased sex ratios. Incorporating mechanism into ecology and evolution models reveals similarities between different sex-determining systems. Dosage and gene regulatory network models of sexual development are beginning to shed light on how temperature sensitivity and thresholds may arise. The unavoidable temperature sensitivity in sex-determining systems inherent to these models suggests that evolutionary transitions between genotypic sex determination (GSD) and temperature-dependent sex determination, and between different forms of GSD, are simple and elegant. Theoretical models are often best-served by considering a single piece of a puzzle; however, there is much to gain from reflecting on all of the pieces together in one integrative picture.

Inheritance of nesting behaviour across natural environmental variation in a turtle with temperature-dependent sex determination.

Nesting behaviour is critical for reproductive success in oviparous organisms with no parental care. In organisms where sex is determined by incubation temperature, nesting behaviour may be a prime target of selection in response to unbalanced sex ratios. To produce an evolutionary change in response to sex-ratio selection, components of nesting behaviour must be heritable. We estimated the field heritability of two key components of nesting behaviour in a population of painted turtles (Chrysemys picta) with temperature-dependent sex determination by applying the 'animal model' to a pedigree reconstructed from genotype data. We obtained estimates of low to non-detectable heritability using repeated records across all environments. We then determined environment-specific heritability by grouping records with similar temperatures for the winter preceding the nesting season, a variable known to be highly associated with our two traits of interest, nest vegetation cover and Julian date of nesting. The heritability estimates of nest vegetation cover and Julian date of nesting were qualitatively highest and significant, or nearly so, after hot winters. Additive genetic variance for these traits was not detectable after cold winters. Our analysis suggests that the potential for evolutionary change of nesting behaviour may be dependent on the thermal conditions of the preceding winter, a season that is predicted to be especially subject to climate change.

Climate and predation dominate juvenile and adult recruitment in a turtle with temperature-dependent sex determination.

Conditions experienced early in life can influence phenotypes in ecologically important ways, as exemplified by organisms with environmental sex determination. For organisms with temperature-dependent sex determination (TSD), variation in nest temperatures induces phenotypic variation that could impact population growth rates. In environments that vary over space and time, how does this variation influence key demographic parameters (cohort sex ratio and hatchling recruitment) in early life stages of populations exhibiting TSD? We leverage a 17-year data set on a population of painted turtles, Chrysemys picta, to investigate how spatial variation in nest vegetation cover and temporal variation in climate influence early life-history demography. We found that spatial variation in nest cover strongly influenced nest temperature and sex ratio, but was not correlated with clutch size, nest predation, total nest failure, or hatching success. Temporal variation in climate influenced percentage of total nest failure and cohort sex ratio, but not depredation rate, mean clutch size, or mean hatching success. Total hatchling recruitment in a year was influenced primarily by temporal variation in climate-independent factors, number of nests constructed, and depredation rate. Recruitment of female hatchlings was determined by stochastic variation in nest depredation and annual climate and also by the total nest production. Overall population demography depends more strongly on annual variation in climate and predation than it does on the intricacies of nest-specific biology. Finally, we demonstrate that recruitment of female hatchlings translates into recruitment of breeding females into the population, thus linking climate (and other) effects on early life stages to adult demographics.