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.

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.

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.

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.

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.

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.

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.

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.

Sexual competition and the evolution of condition-dependent ageing.

Increased individual resources (condition) can be correlated with either increased or decreased longevity. While variation in resource acquisition and allocation can account for some of this variation, the general conditions that select for either pattern remain unclear. Previous models suggest that nonlinearity of payoffs from investment in reproduction (e.g., male secondary sexual traits) can select for high-condition individuals that sacrifice longevity to increase reproductive opportunity. However, it remains unclear what mating systems or patterns of sexual competition might select for such life-history strategies. We used a model of condition-dependent investment to explore how expected payoffs from increased expression of secondary sexual traits affect optimal investment in lifespan. We find that nonlinearity of these payoffs results in a negative relationship between condition and lifespan under two general conditions: first, when there are accelerating marginal benefits from increasing investment; second, when individuals that invest minimally in secondary sexual trait expression can still achieve matings. In the second scenario, the negative relationship occurs due to selection on low-condition individuals to extend lifespan at the cost of secondary sexual trait expression. Our findings clarify the potential role of sexual selection in shaping patterns of condition-dependent ageing, and highlight the importance of considering the strategies of both low- and high-condition individuals when investigating patterns of condition-dependent ageing.

A comprehensive database of thermal developmental plasticity in reptiles.

How temperature influences development has direct relevance to ascertaining the impact of climate change on natural populations. Reptiles have served as empirical models for understanding how the environment experienced by embryos can influence phenotypic variation, including sex ratio, phenology and survival. Such an understanding has important implications for basic eco-evolutionary theory and conservation efforts worldwide. While there is a burgeoning empirical literature of experimental manipulations of embryonic thermal environments, addressing widespread patterns at a comparative level has been hampered by the lack of accessible data in a format that is amendable to updates as new studies emerge. Here, we describe a database with nearly 10, 000 phenotypic estimates from 155 species of reptile, collected from 300 studies manipulating incubation temperature (published between 1974-2016). The data encompass various morphological, physiological, behavioural and performance traits along with growth rates, developmental timing, sex ratio and survival (e.g., hatching success). This resource will serve as an important data repository for addressing overarching questions about thermal plasticity of reptile embryos.

Thermal plasticity due to parental and early-life environments in the jacky dragon (Amphibolurus muricatus).

Developmental plasticity creates marked variation in individual phenotypes when the environment is patchy, such as when the thermal environment varies. Plasticity may occur in response to the environment experienced during an individual's lifetime or to the environment experienced by parents (transgenerational plasticity), and may be adaptive if it enhances fitness. In particular, plasticity in thermal traits, such as preferred temperatures and thermal limits, may improve performance and fitness based on temperatures in the local environment. This study examined the influence of parental and offspring thermal environments (duration of access to a basking lamp) on offspring thermal traits (preferred temperatures and panting threshold) in jacky dragons (Agamidae: Amphibolurus muricatus). Long-bask parental environments led, indirectly, to higher preferred temperatures of offspring due to increased offspring body mass compared to offspring of short-bask parents. The increase in median temperature preference was associated with a higher voluntary minimum body temperature and a narrower preference range, suggesting tradeoffs in thermal behavior and a matching of offspring preferences to the parental environment. Parental thermal treatment did not influence offspring panting threshold. Instead, the panting threshold tended to be higher in offspring that were reared in the long-bask treatment compared to those in the short-bask treatment, suggesting longer basking environments increased thermal tolerance. Parental and offspring thermal environment did not exhibit any interactive effect on thermal traits. The results indicate that thermal environments experienced by lizards can have both transgenerational and within-generation impacts on thermal traits, thus influencing how populations respond to fluctuating or changing climates.

Patterns of developmental plasticity in response to incubation temperature in reptiles.

Early life environments shape phenotypic development in important ways that can lead to long-lasting effects on phenotype and fitness. In reptiles, one aspect of the early environment that impacts development is temperature (termed 'thermal developmental plasticity'). Indeed, the thermal environment during incubation is known to influence morphological, physiological, and behavioral traits, some of which have important consequences for many ecological and evolutionary processes. Despite this, few studies have attempted to synthesize and collate data from this expansive and important body of research. Here, we systematically review research into thermal developmental plasticity across reptiles, structured around the key papers and findings that have shaped the field over the past 50 years. From these papers, we introduce a large database (the 'Reptile Development Database') consisting of 9,773 trait means across 300 studies examining thermal developmental plasticity. This dataset encompasses data on a range of phenotypes, including morphological, physiological, behavioral, and performance traits along with growth rate, incubation duration, sex ratio, and survival (e.g., hatching success) across all major reptile clades. Finally, from our literature synthesis and data exploration, we identify key research themes associated with thermal developmental plasticity, important gaps in empirical research, and demonstrate how future progress can be made through targeted empirical, meta-analytic, and comparative work.

Mate limitation and sex ratio evolution.

Sex ratio evolution has been one of the most successful areas of evolutionary theory. Pioneered by Düsing and Fisher under panmixia, and later extended by Hamilton to cover local mate competition (LMC), these models often assume, either implicitly or explicitly, that all females are fertilized. Here, we examine the effects of relaxing this assumption, under both panmictic and LMC models with diploid genetics. We revisit the question of the mathematical relationship between sex ratio and probability of fertilization, and use these results to model sex ratio evolution under risk of incomplete fertilization. We find that (i) under panmixia, mate limitation has no effect on the evolutionarily stable strategy (ESS) sex allocation; (ii) under LMC, mate limitation can make sex allocation less female-biased than under complete fertilization; (iii) contrary to what is occasionally stated, a significant fraction of daughters can remain unfertilized at the ESS in LMC with mate limitation; (iv) with a commonly used mating function, the fraction of unfertilized daughters can be quite large, and (v) with more realistic fertilization functions, the deviation becomes smaller. The models are presented in three equivalent forms: individual selection, kin selection and group selection. This serves as an example of the equivalence of the methods, while each approach has their own advantages. We discuss possible extensions of the model to haplodiploidy.

Developmental temperatures and phenotypic plasticity in reptiles: a systematic review and meta-analysis.

Early environments can profoundly influence an organism in ways that persist over its life. In reptiles, early thermal environments (nest temperatures) can impact offspring phenotype and survival in important ways, yet we still lack an understanding of whether general trends exist and the magnitude of impact. Understanding these patterns is important in predicting how climate change will affect reptile populations and the role of phenotypic plasticity in buffering populations. We compiled data from 175 reptile studies to examine, and quantify, the effect of incubation temperature on phenotype and survival. Using meta-analytic approaches (standardized mean difference between incubation treatments, Hedges' g), we show that across all trait types examined there is, on average, a moderate to large magnitude of effect of incubation temperatures (absolute effect: |g| = 0.75). Unsurprisingly, this influence was extremely large for incubation duration, as predicted, with warmer temperatures decreasing incubation time overall (g = -8.42). Other trait types, including behaviour, physiology, morphology, performance, and survival experienced reduced, but still mostly moderate to large effects, with particularly strong effects on survival. Moreover, the impact of incubation temperature persisted at least one-year post-hatching, suggesting that these effects have the potential to impact fitness in the long term. The magnitude of effect increased as the change in temperature increased (e.g. 6°C versus 2°C) in almost all cases, and tended to decrease when temperatures of the treatments fluctuated around a mean temperature compared to when they were constant. The effect also depended on the mid-temperature of the comparison, but not in consistent ways, with some traits experiencing the greatest effects at extreme temperatures, while others did not. The highly heterogeneous nature of the effects we observe, along with a large amount of unexplained variability, indicates that the shape of reaction norms between phenotype and temperature, along with ecological and/or experimental factors, are important when considering general patterns. Our analyses provide new insights into the effects of incubation environments on reptile phenotype and survival and allow general, albeit coarse, predictions for taxa experiencing warming nest temperatures under climatic change.

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.

Costs of Rearing the Wrong Sex: Cross-Fostering to Manipulate Offspring Sex in Tammar Wallabies.

Sex allocation theory assumes that offspring sex (son vs. daughter) has consequences for maternal fitness. The most compelling experiment to test this theory would involve manipulating offspring sex and measuring the fitness consequences of having the "wrong" sex. Unfortunately, the logistical challenges of such an experiment limit its application. In tammar wallabies (Macropus eugenii), previous evidence suggests that mothers in good body condition are more likely to produce sons compared to mothers in poor condition, in support of the Trivers-Willard Hypothesis (TW) of condition-dependent sex allocation. More recently, we have found in our population of tammar wallabies that females with seemingly poor access to resources (based on condition loss over the dry summer) are more likely to produce sons, consistent with predictions from the Local Resource Competition (LRC) hypothesis, which proposes that production of sons or daughters is driven by the level of potential competition between mothers and philopatric daughters. We conducted a cross-fostering experiment in free-ranging tammar wallabies to disassociate the effects of rearing and birthing offspring of each sex. This allowed us to test the prediction of the LRC hypothesis that rearing daughters reduces the future direct fitness of mothers post-weaning and the prediction of the TW hypothesis that rearing sons requires more energy during lactation. Overall, we found limited costs to the mother of rearing the "wrong" sex, with switching of offspring sex only reducing the likelihood of a mother having a pouch young the following year. Thus, we found some support for both hypotheses in that rearing an unexpected son or an unexpected daughter both lead to reduced future maternal fitness. The study suggests that there may be context-specific costs associated with rearing the "wrong" sex.