Evolution Of Phenotypic Plasticity Research Articles

Antagonistic Effects of Assortative Mating on the Evolution of Phenotypic Plasticity along Environmental Gradients.

AbstractPrevious theory has shown that assortative mating for plastic traits can maintain genetic divergence across environmental gradients despite high gene flow. Yet these models did not examine how assortative mating affects the evolution of plasticity. We here describe patterns of genetic variation across elevation for plasticity in a trait under assortative mating, using multiple-year observations of budburst date in a common garden of sessile oaks. Despite high gene flow, we found significant spatial genetic divergence for the intercept, but not for the slope, of reaction norms to temperature. We then used individual-based simulations, where both the slope and the intercept of the reaction norm evolve, to examine how assortative mating affects the evolution of plasticity, varying the intensity and distance of gene flow. Our model predicts the evolution of either suboptimal plasticity (reaction norms with a slope shallower than optimal) or hyperplasticity (slopes steeper than optimal) in the presence of assortative mating when optimal plasticity would evolve under random mating. Furthermore, a cogradient pattern of genetic divergence for the intercept of the reaction norm (where plastic and genetic effects are in the same direction) always evolves in simulations with assortative mating, consistent with our observations in the studied oak populations.

Phenotypic plasticity evolves at multiple biological levels in response to environmental predictability in a long-term experiment with a halotolerant microalga

Phenotypic plasticity, the change in the phenotype of a given genotype in response to its environment of development, is a ubiquitous feature of life, enabling organisms to cope with variation in their environment. Theoretical studies predict that, under stationary environmental variation, the level of plasticity should evolve to match the predictability of selection at the timing of development. However, the extent to which patterns of evolution of plasticity for more integrated traits are mirrored by their underlying molecular mechanisms remains unclear, especially in response to well-characterized selective pressures exerted by environmental predictability. Here, we used experimental evolution with the microalgae Dunaliella salina under controlled environmental fluctuations, to test whether the evolution of phenotypic plasticity in responses to environmental predictability (as measured by the squared autocorrelation ρ2) occurred across biological levels, going from DNA methylation to gene expression to cell morphology. Transcriptomic analysis indicates clear effects of salinity and ρ2 × salinity interaction on gene expression, thus identifying sets of genes involved in plasticity and its evolution. These transcriptomic effects were independent of DNA methylation changes in cis. However, we did find ρ2-specific responses of DNA methylation to salinity change, albeit weaker than for gene expression. Overall, we found consistent evolution of reduced plasticity in less predictable environments for DNA methylation, gene expression, and cell morphology. Our results provide the first clear empirical signature of plasticity evolution at multiple levels in response to environmental predictability, and highlight the importance of experimental evolution to address predictions from evolutionary theory, as well as investigate the molecular basis of plasticity evolution.

Sexual repurposing of juvenile aposematism in locusts

Adaptive plasticity requires an integrated suite of functional responses to environmental variation, which can include social communication across life stages. Desert locusts (Schistocerca gregaria) exhibit an extreme example of phenotypic plasticity called phase polyphenism, in which a suite of behavioral and morphological traits differ according to local population density. Male and female juveniles developing at low population densities exhibit green- or sand-colored background-matching camouflage, while at high densities they show contrasting yellow and black aposematic patterning that deters predators. The predominant background colors of these phenotypes (green/sand/yellow) all depend on expression of the carotenoid-binding "Yellow Protein" (YP). Gregarious (high-density) adults of both sexes are initially pinkish, before a YP-mediated yellowing reoccurs upon sexual maturation. Yellow color is especially prominent in gregarious males, but the reason for this difference has been unknown since phase polyphenism was first described in 1921. Here, we use RNA interference to show that gregarious male yellowing acts as an intrasexual warning signal, which forms a multimodal signal with the antiaphrodisiac pheromone phenylacetonitrile (PAN) to prevent mistaken sexual harassment from other males during scramble mating in a swarm. Socially mediated reexpression of YP thus adaptively repurposes a juvenile signal that deters predators into an adult signal that deters undesirable mates. These findings reveal a previously underappreciated sexual dimension to locust phase polyphenism, and promote locusts as a model for investigating the relative contributions of natural versus sexual selection in the evolution of phenotypic plasticity.

The evolution of neurosensation provides opportunities and constraints for phenotypic plasticity

Phenotypic plasticity is widely regarded as important for enabling species resilience to environmental change and for species evolution. However, insight into the complex mechanisms by which phenotypic plasticity evolves in nature is limited by our ability to reconstruct evolutionary histories of plasticity. By using part of the molecular mechanism, we were able to trace the evolution of pre-feeding phenotypic plasticity across the class Echinoidea and identify the origin of plasticity at the base of the regular urchins. The neurosensory foundation for plasticity was ancestral within the echinoids. However, coincident development of the plastic trait and the neurosensory system was not achieved until the regular urchins, likely due to pleiotropic effects and linkages between the two colocalized systems. Plasticity continues to evolve within the urchins with numerous instances of losses associated with loss of sensory abilities and neurons, consistent with a cost of maintaining these capabilities. Thus, evidence was found for the neurosensory system providing opportunities and constraints to the evolution of phenotypic plasticity.

The seeds of invasion: enhanced germination in invasive European populations of black locust (Robinia pseudoacacia L.) compared to native American populations.

Local adaptation and the evolution of phenotypic plasticity may facilitate biological invasions. Both processes can enhance germination and seedling recruitment, which are crucial life-history traits for plants. The rate, timing and speed of germination have recently been documented as playing a major role during the invasion process. Black locust (Robinia pseudoacacia L.) is a North American tree, which has spread widely throughout Europe. A recent study demonstrated that a few populations are the source of European black locust. Thus, invasive populations can be compared to native ones in order to identify genetic-based phenotypic differentiation and the role of phenotypic plasticity can thereby be assessed. A quantitative genetics experiment was performed to evaluate 13 juvenile traits of both native and invasive black locust populations (3000 seeds, 20 populations) subjected to three different thermal treatments (18°C, 22°C and 31°C). The results revealed European populations to have a higher germination rate than the native American populations (88% versus 60%), and even when genetic distance between populations was considered. Moreover, this trait showed lower plasticity to temperature in the invasive range than in the native one. Conversely, other studied traits showed high plasticity to temperature, but they responded in a similar way to temperature increase: the warmer the temperature, the higher the growth rate or germination traits values. The demonstrated genetic differentiation between native and invasive populations testifies to a shift between ranges for the maximum germination percentage. This pattern could be due to human-mediated introduction of black locust.

Plasticity is a locally adapted trait with consequences for ecological dynamics in novel environments.

Phenotypic plasticity is predicted to evolve in more variable environments, conferring an advantage on individual lifetime fitness. It is less clear what the potential consequences of that plasticity will have on ecological population dynamics. Here, we use an invertebrate model system to examine the effects of environmental variation (resource availability) on the evolution of phenotypic plasticity in two life history traits—age and size at maturation—in long‐running, experimental density‐dependent environments. Specifically, we then explore the feedback from evolution of life history plasticity to subsequent ecological dynamics in novel conditions. Plasticity in both traits initially declined in all microcosm environments, but then evolved increased plasticity for age‐at‐maturation, significantly so in more environmentally variable environments. We also demonstrate how plasticity affects ecological dynamics by creating founder populations of different plastic phenotypes into new microcosms that had either familiar or novel environments. Populations originating from periodically variable environments that had evolved greatest plasticity had lowest variability in population size when introduced to novel environments than those from constant or random environments. This suggests that while plasticity may be costly it can confer benefits by reducing the likelihood that offspring will experience low survival through competitive bottlenecks in variable environments. In this study, we demonstrate how plasticity evolves in response to environmental variation and can alter population dynamics—demonstrating an eco‐evolutionary feedback loop in a complex animal moderated by plasticity in growth.

Reacquisition of light-harvesting genes in a marine cyanobacterium confers a broader solar niche

The evolution of phenotypic plasticity, i.e., the environmental induction of alternative phenotypes by the same genotype, can be an important mechanism of biological diversification.1,2 For example, an evolved increase in plasticity may promote ecological niche expansion as well as the innovation of novel traits;3 however, both the role of phenotypic plasticity in adaptive evolution and its underlying mechanisms are still poorly understood.4,5 Here, we report that the Chlorophyll d-producing marine cyanobacterium Acaryochloris marina strain MBIC11017 has evolved greater photosynthetic plasticity by reacquiring light-harvesting genes via horizontal gene transfer. The genes, which had been lost by the A.marina ancestor, are involved in the production and degradation of the light-harvesting phycobiliprotein phycocyanin. A.marina MBIC11017 exhibits a high degree of wavelength-dependence in phycocyanin production, and this ability enables it to grow with yellow and green light wavelengths that are inaccessible to other A.marina. Consequently, this strain has a broader solar niche than its close relatives. We discuss the role of horizontal gene transfer for regaining a lost phenotype in light of Dollo's Law6 that the loss of a complex trait is irreversible.

Variation in regrowth ability in relation to land-use intensity in three common grassland herbs

Abstract Aims Plant populations in managed grasslands are subject to strong selection exerted by grazing, mowing and fertilization. Many previous studies showed that this can cause evolutionary changes in mean trait values, but little is known about the evolution of phenotypic plasticity in response to land use. In this study, we aimed to elucidate the relationships between phenotypic plasticity—specifically, regrowth ability after biomass removal—and the intensity of grassland management and levels of temporal variation therein. Methods We conducted an outdoor common garden experiment to test if plants from more intensively mown and grazed sites showed an increased ability to regrow after biomass removal. We used three common plant species from temperate European grasslands, with seed material from 58 to 68 populations along gradients of land-use intensity, ranging from extensive (only light grazing) to very intensive management (up to four cuts per year). Important Findings In two out of three species, we found significant population differentiation in regrowth ability after clipping. While variation in regrowth ability was unrelated to the mean land-use intensity of populations of origin, we found a relationship with its temporal variation in Plantago lanceolata, where plants experiencing less variable environmental conditions over the last 11 years showed stronger regrowth in reproductive biomass after clipping. Therefore, while mean grazing and mowing intensity may not select for regrowth ability, the temporal stability of the environmental heterogeneity created by land use may have caused its evolution in some species.

The Evolution of Phenotypic Plasticity in Response to Temperature Stress.

Phenotypic plasticity is the ability of a single genotype to produce different phenotypes in response to environmental variation. The importance of phenotypic plasticity in natural populations and its contribution to phenotypic evolution during rapid environmental change is widely debated. Here, we show that thermal plasticity of gene expression in natural populations is a key component of its adaptation: evolution to novel thermal environments increases ancestral plasticity rather than mean genetic expression. We determined the evolution of plasticity in gene expression by conducting laboratory natural selection on a Drosophila simulans population in hot and cold environments. After more than 60 generations in the hot environment, 325 genes evolved a change in plasticity relative to the natural ancestral population. Plasticity increased in 75% of these genes, which were strongly enriched for several well-defined functional categories (e.g., chitin metabolism, glycolysis, and oxidative phosphorylation). Furthermore, we show that plasticity in gene expression of populations exposed to different temperatures is rather similar across species. We conclude that most of the ancestral plasticity can evolve further in more extreme environments.

Transgenerational Plasticity in the Context of Predator-Prey Interactions

Almost all animal species are engaged in predator-prey interactions. These interactions, variable in time and space, favor the emergence and evolution of phenotypic plasticity, which allows prey to fine-tune their phenotype to the current risk of predation. A famous example is the induction of defensive neck-teeth, spines or helmets in some water fleas when they detect cues of predator presence. In general, the response may involve different types of traits (behavioral, morphological, physiological and life-history traits), alone or in combination. The induced traits may be adaptive anti-predator defenses or reflect more general stress-based responses. Recently, it has been found that predator-induced plasticity occurs not only within but also across generations (transgenerational plasticity), i.e. the phenotype of a generation is influenced by the detection of predator-cues in previous generation(s), even if the current generation is not itself exposed to these cues. In this paper, we aim to review this accumulating literature and propose a current state of key aspects of predator-induced transgenerational plasticity in metazoans. In particular, we review whether patterns of predator-induced transgenerational plasticity depend on the type of traits. We analyze the adaptive value of predator-induced transgenerational plasticity and explored the evidence for its evolution and underlying mechanisms. We also consider its temporal dynamics: What are the time windows during which predator-cues must be detected to be transmitted across generations? Are transgenerational responses in offspring stage-dependent? How many generations does transgenerational plasticity persist? Finally, we discuss other factors highlighted in the literature that influence predator-induced transgenerational plasticity: What are the relative contributions of maternal and paternal exposure to predator-cues in generating transgenerational plasticity? Do transgenerational responses depend on offspring sex? Do they scale with the perceived level of predation risk? This review shows that we are only at the beginning of understanding the processes of predator-induced transgenerational plasticity, and it encourages future research to fill the lack of knowledge highlighted here.

Measuring phenotypes in fluctuating environments

Abstract Despite considerable theoretical interest in how the evolution of phenotypic plasticity should be shaped by environmental variability and stochasticity, how individuals actually respond to these aspects of the environment within their own lifetimes remains unclear. We propose that this understanding has been hampered by experimental approaches that expose organisms to fluctuating environments (typically treatments where fluctuations in the environment are cyclical vs. erratic) for a pre‐determined duration while ensuring that the mean environment over that the entire exposure period is invariable. This approach implicitly assumes that responses to the mean and variance/predictability in the environment occur over the same time‐scale. If this assumption is false, one potential outcome is that phenotypic differences among the treatment groups might arise in response to differences in the mean environment that are present over shorter time periods among those same treatment groups. We illustrate an experimental design that (a) creates variation in the level of environmental predictability, (b) allows for estimation of the time‐scale over which the phenotypic response to the mean environment occurs and (c) permits statistical estimation of the effect of predictability in the environmental variable of interest while controlling for any effect of the mean environment over the relevant temporal scale. Using the clonally reproducing zooplankton species Daphnia magna, we test for within‐generation plasticity in the ability to tolerate high temperature following exposure to multiple temperature treatments with the same overall mean, but where the pattern of fluctuations differed among them. This approach revealed that heat tolerance in Daphnia was not influenced by variability in temperature per se nor the predictability of fluctuations in temperature but adjusted in response to the mean temperature they experienced 24 hr prior to measurement. Our results suggest that conclusions arising from studies that employ a single manipulation of environmental predictability and which cannot consider such potentially confounding effects may be premature. A free Plain Language Summary can be found within the Supporting Information of this article.

The impact of multiple climatic and geographic factors on the chemical defences of Asian toads (Bufo gargarizans Cantor)

Chemical defences are widespread in nature, yet we know little about whether and how climatic and geographic factors affect their evolution. In this study, we investigated the natural variation in the concentration and composition of the main bufogenin toxin in adult Asian toads (Bufo gargarizans Cantor) captured in twenty-two regions. Moreover, we explored the relative importance of eight climatic factors (average temperature, maximum temperature, minimum temperature, average relative humidity, 20–20 time precipitation, maximum continuous precipitation, maximum ground temperature, and minimum ground temperature) in regulating toxin production. We found that compared to toads captured from central and southwestern China, toads from eastern China secreted higher concentrations of cinobufagin (CBG) and resibufogenin (RBG) but lower concentrations of telocinobufagin (TBG) and cinobufotalin (CFL). All 8 climatic variables had significant effects on bufogenin production (ri>0.5), while the plastic response of bufogenin toxin to various climate factors was highly variable. The most important climatic driver of total bufogenin production was precipitation: the bufogenin concentration increased with increasing precipitation. This study indicated that the evolution of phenotypic plasticity in chemical defences may depend at least partly on the geographic variation of defensive toxins and their climatic context.

The evolution of phenotypic plasticity when environments fluctuate in time and space.

Most theoretical studies have explored the evolution of plasticity when the environment, and therefore the optimal trait value, varies in time or space. When the environment varies in time and space, we show that genetic adaptation to Markovian temporal fluctuations depends on the between‐generation autocorrelation in the environment in exactly the same way that genetic adaptation to spatial fluctuations depends on the probability of philopatry. This is because both measure the correlation in parent‐offspring environments and therefore the effectiveness of a genetic response to selection. If the capacity to genetically respond to selection is stronger in one dimension (e.g., space), then plasticity mainly evolves in response to fluctuations in the other dimension (e.g., time). If the relationships between the environments of development and selection are the same in time and space, the evolved plastic response to temporal fluctuations is useful in a spatial context and genetic differentiation in space is reduced. However, if the relationships between the environments of development and selection are different, the optimal level of plasticity is different in the two dimensions. In this case, the plastic response that evolves to cope with temporal fluctuations may actually be maladaptive in space, resulting in the evolution of hyperplasticity or negative plasticity. These effects can be mitigated by spatial genetic differentiation that acts in opposition to plasticity resulting in counter‐gradient variation. These results highlight the difficulty of making space‐for‐time substitutions in empirical work but identify the key parameters that need to be measured in order to test whether space‐for‐time substitutions are likely to be valid.

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

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

The conceptual structure of evolutionary biology: A framework from phenotypic plasticity

Abstract In this review, I approach the role of phenotypic plasticity as a key aspect of the conceptual framework of evolutionary biology. The concept of phenotypic plasticity is related to other relevant concepts of contemporary research in evolutionary biology, such as assimilation, genetic accommodation and canalization, evolutionary robustness, evolvability, evolutionary capacitance and niche construction. Although not always adaptive, phenotypic plasticity can promote the integration of these concepts to represent some of the dynamics of evolution, which can be visualized through the use of a conceptual map. Although the use of conceptual maps is common in areas of knowledge such as psychology and education, their application in evolutionary biology can lead to a better understanding of the processes and conceptual interactions of the complex dynamics of evolution. The conceptual map I present here includes environmental variability and variation, phenotypic plasticity and natural selection as key concepts in evolutionary biology. The evolution of phenotypic plasticity is important to ecology at all levels of organization, from morphological, physiological and behavioral adaptations that influence the distribution and abundance of populations to the structuring of assemblages and communities and the flow of energy through trophic levels. Consequently, phenotypic plasticity is important for maintaining ecological processes and interactions that influence the complexity of biological diversity. In addition, because it is a typical occurrence and manifests itself through environmental variation in conditions and resources, plasticity must be taken into account in the development of management and conservation strategies at local and global levels.

Genetic accommodation and the role of ancestral plasticity in the evolution of insect eusociality.

For over a century, biologists have proposed a role for phenotypic plasticity in evolution, providing an avenue for adaptation in addition to 'mutation-first' models of evolutionary change. According to the various versions of this idea, the ability of organisms to respond adaptively to their environment through phenotypic plasticity may lead to novel phenotypes that can be screened by natural selection. If these initially environmentally induced phenotypes increase fitness, then genetic accommodation can lead to allele frequency change, influencing the expression of those phenotypes. Despite the long history of 'plasticity-first' models, the importance of genetic accommodation in shaping evolutionary change has remained controversial - it is neither fully embraced nor completely discarded by most evolutionary biologists. We suggest that the lack of acceptance of genetic accommodation in some cases is related to a lack of information on its molecular mechanisms. However, recent reports of epigenetic transgenerational inheritance now provide a plausible mechanism through which genetic accommodation may act, and we review this research here. We also discuss current evidence supporting a role for genetic accommodation in the evolution of eusociality in social insects, which have long been models for studying the influence of the environment on phenotypic variation, and may be particularly good models for testing hypotheses related to genetic accommodation. Finally, we introduce 'eusocial engineering', a method by which novel social phenotypes are first induced by environmental modification and then studied mechanistically to understand how environmentally induced plasticity may lead to heritable changes in social behavior. We believe the time is right to incorporate genetic accommodation into models of the evolution of complex traits, armed with new molecular tools and a better understanding of non-genetic heritable elements.

Constraints on the evolution of phenotypic plasticity in the clonal plant Hydrocotyle vulgaris.

The evolution of phenotypic plasticity of plant traits may be constrained by costs and limits. However, the precise constraints are still unclear for many traits under different ecological contexts. In a glasshouse experiment, we grew ramets of 12 genotypes of a clonal plant Hydrocotyle vulgaris under the control (full light and no flood), shade and flood conditions and tested the potential costs and limits of plasticity in 13 morphological and physiological traits in response to light availability and flood variation. In particular, we used multiple regression and correlation analyses to evaluate potential plasticity costs, developmental instability costs and developmental range limits of each trait. We detected significant costs of plasticity in specific petiole length and specific leaf area in response to shade under the full light condition and developmental range limits in specific internode length and intercellular CO2 concentration in response to light availability variation. However, we did not observe significant costs or limits of plasticity in any of the 13 traits in response to flood variation. Our results suggest that the evolution of phenotypic plasticity in plant traits can be constrained by costs and limits, but such constraints may be infrequent and differ under different environmental contexts.

Unravelling the genetic differentiation among varieties of the Neotropical savanna tree Hancornia speciosa Gomes.

Spatial distribution of species genetic diversity is often driven by geographical distance (isolation by distance) or environmental conditions (isolation by environment), especially under climate change scenarios such as Quaternary glaciations. Here, we used coalescent analyses coupled with ecological niche modelling (ENM), spatially explicit quantile regression analyses and the multiple matrix regression with randomization (MMRR) approach to unravel the patterns of genetic differentiation in the widely distributed Neotropical savanna tree, Hancornia speciosa (Apocynaceae). Due to its high morphological differentiation, the species was originally classified into six botanical varieties by Monachino, and has recently been recognized as only two varieties by Flora do Brasil 2020. Thus, H. speciosa is a good biological model for learning about evolution of phenotypic plasticity under genetic and ecological effects, and predicting their responses to changing environmental conditions. We sampled 28 populations (777 individuals) of Monachino's four varieties of H. speciosa and used seven microsatellite loci to genotype them. Bayesian clustering showed five distinct genetic groups (K = 5) with high admixture among Monachino's varieties, mainly among populations in the central area of the species geographical range. Genetic differentiation among Monachino's varieties was lower than the genetic differentiation among populations within varieties, with higher within-population inbreeding. A high historical connectivity among populations of the central Cerrado shown by coalescent analyses may explain the high admixture among varieties. In addition, areas of higher climatic suitability also presented higher genetic diversity in such a way that the wide historical refugium across central Brazil might have promoted the long-term connectivity among populations. Yet, FST was significantly related to geographic distances, but not to environmental distances, and coalescent analyses and ENM predicted a demographical scenario of quasi-stability through time. Our findings show that demographical history and isolation by distance, but not isolation by environment, drove genetic differentiation of populations. Finally, the genetic clusters do not support the two recently recognized botanical varieties of H. speciosa, but partially support Monachino's classification at least for the four sampled varieties, similar to morphological variation.

The evolution of phenotypic plasticity under global change

Marine ecosystems are currently in a state of flux, with ocean warming and acidification occurring at unprecedented rates. Phenotypic plasticity underpins acclimatory responses by shifting the mean phenotype in a population, which may buffer the negative effects of global change. However, little is known about how phenotypic plasticity evolves across multiple generations. We tested this by reciprocally-transplanting the polychaete Ophryotrocha labronica between control and global change scenarios (ocean warming and acidification in isolation and combined) over five generations. By comparing the reaction norms of four life-history traits across generations, we show that juvenile developmental rate in the combined scenario was the only trait that changed its plastic response across generations when transplanted back to control conditions, and that adaptive plasticity was conserved in most traits, despite significant levels of selection and strong declines in individual fitness in the multi-generational exposure. We suggest the change in level of plasticity in the combined scenario is caused by differential allocation of energy between the mean and the plasticity of the trait along the multigenerational exposure. The ability to maintain within-generational levels of plasticity under global change scenarios has important eco-evolutionary and conservation implications, which are examined under the framework of assisted evolution programs.

Age- and environment-dependent changes in chemical defences of larval and post-metamorphic toads

BackgroundChemical defences are widespread in animals, but how their production is adjusted to ecological conditions is poorly known. Optimal defence theory predicts that inducible defences are favoured over constitutive defences when toxin production is costly and the need for it varies across environments. However, if some environmental changes occur predictably (e.g. coupled to transitions during ontogeny), whereas others are unpredictable (e.g. predation, food availability), changes in defences may have constitutive as well as plastic elements. To investigate this phenomenon, we raised common toad (Bufo bufo) tadpoles with ad libitum or limited food and in the presence or absence of chemical cues on predation risk, and measured their toxin content on 5 occasions during early ontogeny.ResultsThe number of compounds showed limited variation with age in tadpoles and was unaffected by food limitation and predator cues. The total amount of bufadienolides first increased and later decreased during development, and it was elevated in young and mid-aged tadpoles with limited food availability compared to their ad libitum fed conspecifics, whereas it did not change in response to cues on predation risk. We provide the first evidence for the active synthesis of defensive toxin compounds this early during ontogeny in amphibians. Furthermore, the observation of increased quantities of bufadienolides in food-restricted tadpoles is the first experimental demonstration of resource-dependent induction of elevated de novo toxin production, suggesting a role for bufadienolides in allelopathy.ConclusionsOur study shows that the evolution of phenotypic plasticity in chemical defences may depend on the ecological context (i.e. predation vs. competition). Our results furthermore suggest that the age-dependent changes in the diversity of toxin compounds in developing toads may be fixed (i.e., constitutive), timed for the developmental stages in which they are most reliant on their chemical arsenal, whereas inducible plasticity may prevail in the amount of synthesized compounds.