Matthew R. Walsh

Interactions between the direct and indirect effects of predators determine life history evolution in a killifish

The ecological impacts of the indirect effects of predators are well established, but the evolutionary consequences are unknown. Predators often decrease prey density, which indirectly increases the resources available to surviving prey. This ecological effect could provide a link to evolution because it is generally assumed that resource availability influences life history evolution. Yet, predictions from theory that consider food availability are inconsistent, and evidence for an important role of resources in shaping life history evolution is absent. We compared life history traits in a Trinidadian killifish, Rivulus hartii, from fish communities that differ in predation intensity; predators are associated with lower population density and faster growth rates. To determine whether the indirect effects of predators influence evolutionary change, we reared second-generation-born fish under two food levels that approximated natural differences in resources between communities. Rivulus from sites with predators are younger and smaller at maturity. They have increased reproductive investment and produce many small eggs and smaller hatchlings. Such divergence is predicted as a direct effect of predation. We also found significant interactions between predator community and food level for age and size at maturity, fecundity, and egg size. These interactions, whereby the differences between communities were more pronounced at high-food levels, argue that evolution in Rivulus has been influenced by the indirect effects of predators mediated through resource availability. Rivulus from sites with predators better exploit the higher resources in those habitats. Therefore, both direct and indirect effects of predators have evolutionary consequences.

PHENOTYPIC DIVERSIFICATION ACROSS AN ENVIRONMENTAL GRADIENT: A ROLE FOR PREDATORS AND RESOURCE AVAILABILITY ON THE EVOLUTION OF LIFE HISTORIES

Changes in age/size-specific mortality, due to such factors as predation, have potent evolutionary consequences. However, interactions with predators commonly impact prey growth rates and food availability and such indirect effects may also influence evolutionary change. We evaluated life-history differences in Trinidadian killifish, Rivulus hartii, across a gradient in predation. Rivulus are located in (1) “high predation” sites with large piscivores, (2) “Rivulus/guppy” sites with guppies, and (3) “Rivulus-only” sites with just Rivulus. Rivulus suffer higher mortality with large predators, and guppies may prey upon small/young Rivulus in Rivulus/guppy environments. In turn, population densities decline while growth rates increase in both localities compared to Rivulus-only sites. To explore how the direct and indirect effects of predators and guppies influence trait diversification in Rivulus, we examined life-history phenotypes across five rivers. High predation phenotypes exhibited a smaller size at reproduction, a greater number of eggs that were smaller, and increased reproductive allotment. Such changes are consistent with a direct response to predation. Rivulus from Rivulus/guppy sites were intermediate; they exhibited a smaller size at reproduction, increased fecundity, smaller eggs, and larger reproductive allotment than Rivulus-only fish. These changes are consistent with models that incorporate the impacts of growth and resources.

A cascade of evolutionary change alters consumer-resource dynamics and ecosystem function

It is becoming increasingly clear that intraspecific evolutionary divergence influences the properties of populations, communities and ecosystems. The different ecological impacts of phenotypes and genotypes may alter selection on many species and promote a cascade of ecological and evolutionary change throughout the food web. Theory predicts that evolutionary interactions across trophic levels may contribute to hypothesized feedbacks between ecology and evolution. However, the importance of ‘cascading evolutionary change’ in a natural setting is unknown. In lakes in Connecticut, USA, variation in migratory behaviour and feeding morphology of a fish predator, the alewife (Alosa pseudoharengus), drives life-history evolution in a species of zooplankton prey (Daphnia ambigua). Here we evaluated the reciprocal impacts of Daphnia evolution on ecological processes in laboratory mesocosms. We show that life-history evolution in Daphnia facilitates divergence in rates of population growth, which in turn significantly alters consumer-resource dynamics and ecosystem function. These experimental results parallel trends observed in lakes. Such results argue that a cascade of evolutionary change, which has occurred over contemporary timescales, alters community and ecosystem processes.

The evolutionary consequences of indirect effects

Indirect effects occur when the effect of one species on another is mediated by a third species or through other links in the ecosystem. Indirect interactions are widespread in nature with effects on the properties of populations, communities, and ecosystems. Despite their far-reaching ecological relevance, the evolutionary importance of indirect effects has only recently garnered attention. Here I highlight recent research illustrating significant evolutionary consequences of indirect effects. These studies span field and laboratory studies as well as terrestrial and aquatic ecosystems. I argue that these results provide a blueprint that future research can utilize to evaluate the link between the ecological community and evolution and, thereby, improve the predictive power of evolution in a natural setting.

Interpopulation variation in a fish predator drives evolutionary divergence in prey in lakes

Ecological factors are known to cause evolutionary diversification. Recent work has shown that evolution in strongly interacting predator species has reciprocal impacts on ecosystems. These divergent impacts of predators may alter the selective landscape and cause the evolution of prey. Yet, this link between intraspecific variation and evolution is unexplored. We compared the life history of a species of zooplankton (Daphnia ambigua) from lakes in New England in which the dominant planktivorous predator, the alewife (Alosa pseudoharengus), differs in feeding traits and migratory behaviour. Anadromous alewife (seasonal migrants) exhibit larger gapes, gill-raker spacing and target larger prey than landlocked alewife (year-round freshwater resident). In ‘anadromous’ lakes, Daphnia are abundant in the spring but extirpated by alewife predation in summer. Daphnia are rare year-round in ‘landlocked’ lakes. We show that Daphnia from lakes with anadromous alewife grew faster, matured earlier but at the same size and produced more offspring than Daphnia from lakes with landlocked or no alewife across multiple temperature and resource treatments. Our results are inconsistent with a response to size-selective predation but are better explained as an adaptation to colder temperatures and shorter periods of development (countergradient variation) mediated by seasonal alewife predation.

Influence of the indirect effects of guppies on life-history evolution in Rivulus hartii.

Early theories of life‐history evolution predict that increased predation on young/small individuals selects for delayed maturation and decreased reproductive effort, but such theory only considers changes in mortality. Predators reduce prey abundance and increase food to survivors. Theory that incorporates such indirect effects yields different predictions. Trinidadian killifish, Rivulus hartii, inhabit communities with and without guppies. Guppies prey on young Rivulus and Rivulus densities decline and growth rates increase when guppies are present. Prior work showed that Rivulus phenotypes from communities with guppies matured earlier and had higher fecundity, consistent with theories that incorporate indirect effects. Here we examined the genetic basis of these differences by rearing 2nd generation, laboratory‐born Rivulus from sites with and without guppies under two food levels that match natural differences in growth. Many locality × food interactions were significant, often reversing the relationship between communities. Such interactions imply that there are fitness trade‐offs associated with adaptation to high or low resource environments. On high food, Rivulus from localities with guppies matured earlier, produced many small eggs, and exhibited increased reproductive investment; these differences reversed on low food. Our results suggest that indirect effects mold Rivulus evolution and thereby highlight connections between community processes and evolutionary change.

Environmental and organismal predictors of intraspecific variation in the stoichiometry of a neotropical freshwater fish.

The elemental composition of animals, or their organismal stoichiometry, is thought to constrain their contribution to nutrient recycling, their interactions with other animals, and their demographic rates. Factors that affect organismal stoichiometry are generally poorly understood, but likely reflect elemental investments in morphological features and life history traits, acting in concert with the environmental availability of elements. We assessed the relative contribution of organismal traits and environmental variability to the stoichiometry of an insectivorous Neotropical stream fish, Rivulus hartii. We characterized the influence of body size, life history phenotype, stage of maturity, and environmental variability on organismal stoichiometry in 6 streams that differ in a broad suite of environmental variables. The elemental composition of R. hartii was variable, and overlapped with the wide range of elemental composition documented across freshwater fish taxa. Average %P composition was ∼3.2%(±0.6), average %N∼10.7%(±0.9), and average %C∼41.7%(±3.1). Streams were the strongest predictor of organismal stoichiometry, and explained up to 18% of the overall variance. This effect appeared to be largely explained by variability in quality of basal resources such as epilithon N∶P and benthic organic matter C∶N, along with variability in invertebrate standing stocks, an important food source for R. hartii. Organismal traits were weak predictors of organismal stoichiometry in this species, explaining when combined up to 7% of the overall variance in stoichiometry. Body size was significantly and positively correlated with %P, and negatively with N∶P, and C∶P, and life history phenotype was significantly correlated with %C, %P, C∶P and C∶N. Our study suggests that spatial variability in elemental availability is more strongly correlated with organismal stoichiometry than organismal traits, and suggests that the stoichiometry of carnivores may not be completely buffered from environmental variability. We discuss the relevance of these findings to ecological stoichiometry theory.

Predator-induced phenotypic plasticity within- and across-generations: a challenge for theory?

Much work has shown that the environment can induce non-genetic changes in phenotype that span multiple generations. Theory predicts that predictable environmental variation selects for both increased within- and across-generation responses. Yet, to the best of our knowledge, there are no empirical tests of this prediction. We explored the relationship between within- versus across-generation plasticity by evaluating the influence of predator cues on the life-history traits of Daphnia ambigua. We measured the duration of predator-induced transgenerational effects, determined when transgenerational responses are induced, and quantified the cues that activate transgenerational plasticity. We show that predator exposure during embryonic development causes earlier maturation and increased reproductive output. Such effects are detectable two generations removed from predator exposure and are similar in magnitude in response to exposure to cues emitted by injured conspecifics. Moreover, all experimental contexts and traits yielded a negative correlation between within- versus across-generation responses. That is, responses to predator cues within- and across-generations were opposite in sign and magnitude. Although many models address transgenerational plasticity, none of them explain this apparent negative relationship between within- and across-generation plasticities. Our results highlight the need to refine the theory of transgenerational plasticity.

EXPERIMENTALLY INDUCED LIFE‐HISTORY EVOLUTION IN A KILLIFISH IN RESPONSE TO THE INTRODUCTION OF GUPPIES

Life‐history theory predicts that increased predation on juvenile age/size‐classes favors delayed maturation and decreased reproductive investment. Although this theory has received correlative support, experimental tests in nature are rare. In 1976 and 1981, guppies (Poecilia reticulata) were transplanted into localities that previously only contained a killifish, Rivulus hartii. This situation presents an opportunity to experimentally test this life‐history prediction because guppies prey upon young Rivulus. We evaluated the response to selection in Rivulus by measuring phenotypic and genotypic divergence between introduction and upstream “control” localities that lack guppies. Contrary to expectations, Rivulus from the introduction sites evolved earlier maturation and increased reproductive investment within 25 years. Such evolutionary changes parallel previous investigations on natural communities of Rivulus, but do not comply with predictions of age/size‐specific theory. Guppies also caused reduced densities and increased growth rates of Rivulus, which are hypothesized indirect effects of predation. Additional life‐history theories show that changes in density and growth can interact with predator‐induced mortality to alter the predicted trajectory of evolution. We discuss how these latter frameworks improve the fit between theory and evolution in Rivulus.

EpiRADseq: scalable analysis of genomewide patterns of methylation using next‐generation sequencing

Summary Research addressing the role of epigenetics in a diversity of experimental and natural systems is rapidly accumulating. Diverse methods have been developed to study epigenetic states, including bisulphite sequencing and AFLP-based approaches. However, existing methods are sometimes difficult to apply to non-traditional model organisms that lack genomic resources (bisulphite sequencing), and can fail to be economical and readily scalable to diverse research questions because of reliance on traditional Sanger sequencing (AFLP approaches). Here we develop a reduced-representation library-based approach that is scalable and economical to quantitatively compare patterns of genomewide methylation. This approach shares substantial similarity to the now widely used double digest restriction-site associated DNA sequencing-based method (ddRADseq), except that it utilizes a methylation-sensitive restriction enzyme. This method therefore identifies changes in the genomic methylation state of cytosine (to 5-methylcytosine; 5mC) by sampling loci (via next-generation sequencing) that are not methylated within a sample. We test this method to identify shifts in the epigenome of clonal water fleas (Daphnia ambigua) in response to exposure to fish predator cues, which are known to induce transgenerational changes in life-history traits. We found evidence for differential transgenerational responses (inferred via significant shifts in the methylation state of sampled loci) to predator cues among our treatment groups and remarkably consistent responses within treatment groups. Our results demonstrate that this method is capable of producing highly repeatable results even without the use of a reference genome. Applications of this general method are broad and diverse and include the analysis of epigenetic shifts in both experimental and natural study systems.