Organisms often adapt to new conditions by means of beneficial mutations that become fixed in the population. Often, full adaptation requires several different mutations in the same cell, each of which may affect a different aspect of the behavior. Can one predict order in which these mutations become fixed? To address this, we experimentally studied evolution of Escherichia coli in a growth medium in which the effects of different adaptations can be easily classified as affecting growth rate or the lag-phase duration. We find that adaptations are fixed in a defined and reproducible order: first reduction of lag phase, and then an increase of the exponential growth rate. A population genetics theory explains this order, and suggests growth conditions in which the order of adaptations is reversed. We experimentally find this order reversal under the predicted conditions. This study supports a view in which the evolutionary path to adaptation in a new environment can be captured by theory and experiment.

We revisited a classic study of morphological variation in the oldfield mouse (Peromyscus polionotus) to estimate the strength of selection acting on pigmentation patterns and to identify the underlying genes. We measured 215 specimens collected by Francis Sumner in the 1920s from eight populations across a 155-km, environmentally variable transect from the white sands of Florida's Gulf coast to the dark, loamy soil of southeastern Alabama. Like Sumner, we found significant variation among populations: mice inhabiting coastal sand dunes had larger feet, longer tails, and lighter pigmentation than inland populations. Most striking, all seven pigmentation traits examined showed a sharp decrease in reflectance about 55 km from the coast, with most of the phenotypic change occurring over less than 10 km. The largest change in soil reflectance occurred just south of this break in pigmentation. Geographic analysis of microsatellite markers shows little interpopulation differentiation, so the abrupt change in pigmentation is not associated with recent secondary contact or reduced gene flow between adjacent populations. Using these genetic data, we estimated that the strength of selection needed to maintain the observed distribution of pigment traits ranged from 0.0004 to 21%, depending on the trait and model used. We also examined changes in allele frequency of SNPs in two pigmentation genes, Mc1r and Agouti, and show that mutations in the cis-regulatory region of Agouti may contribute to this cline in pigmentation. The concordance between environmental variation and pigmentation in the face of high levels of interpopulation gene flow strongly implies that natural selection is maintaining a steep cline in pigmentation and the genes underlying it.

We investigate how the intensity of competition for resources affects the strength of disruptive selection on a resource acquisition trait. This is done by analyzing several consumer–resource models in which consumers use a linear array of resources. We show that disruptive selection can be diminished under both strong and weak competition, making disruptive selection a unimodal function of the strength of competition. Weak selection under strong competition arises when competition causes the extinction (for self-reproducing resources) or depletion (for abiotic resources) of the most rapidly caught resources. Weak selection under weak competition is a consequence of minimal effects of consumers on resources. The precise relationship between intensity of competition and strength of disruptive selection is sensitive to the shape of the consumer's resource utilization curve and the nature of resource growth. The most strongly unimodal competition–selection relationships result from utilization curves with long tails. Our results show that a simple comparison of the width of the resource abundance distribution and the consumer's utilization function is not sufficient to determine whether selection is disruptive. The results may explain some contradictory experimental findings regarding the effect of consumer mortality on the strength of disruptive selection.

Morphological convergence provides strong evidence that evolution is adaptive. However, putatively convergent morphology is often examined in two dimensions with no explicit model of function. In this study, we investigated structural and mechanical similarities of the lower pharyngeal jaw (LPJ) in cichlid fish that have evolved the ability to crush hard-shelled molluscs. Using a novel phylogeny, we demonstrated molluscivory has been gained and/or been lost numerous times in Heroine cichlids. Within this comparative framework, we produced three-dimensional computed tomography (CT) scans for LPJs of both morphotypes in the trophically polymorphic Herichthys minckleyi and six evolutionarily independent pairs of closely related species. Like H. minckleyi, these species exhibit divergence between a molluscivore and a nonmolluscivore. Using the CT scans, we generated finite element models of papilliform H. minckleyi LPJs to determine where stress would concentrate in a jaw not modified to crush molluscs. Then, we examined whether stress in the papilliform LPJ predicted structural modifications in molariform H. minckleyi and other molluscivorous species. Despite potential constraints, stresses imposed during prey processing explain 40% of LPJ variation in mollusc crushing species. The structural and mechanical analyses also suggest divergence found in polymorphic species could provide the substrate for trophic differences found in reproductively isolated cichlids.

The burst-death model has been developed to describe the life history of organisms with variable generation times and a burst of a fixed number of offspring. The model also includes an optional constant clearance rate, such as washout from a chemostat, and the possibility of sustained periods of population growth followed by severe bottlenecks, as in serial passaging. In this model, a beneficial mutation can either increase the burst rate or the burst size, or reduce the clearance rate, thus increasing survival. In this article we examine the effects of these three possible mechanisms on both the Malthusian fitness and the fixation probability of the lineage. We find that equivalent relative increases in the burst rate or burst size confer equivalent increases in the Malthusian fitness of a lineage, whereas increasing survival typically has a more moderate effect on Malthusian fitness. In contrast, for beneficial mutations that confer the same increase in fitness, mutations that increase survival are the most likely to fix, followed by mutations that increase the burst rate. Mutations that increase the burst size are the least likely to fix. These results imply that mutant lineages with the highest Malthusian fitness are not, in many cases, the most likely to escape extinction.

Larvae of the pipevine swallowtail (Battus philenor) sequester toxic alkaloids called aristolochic acids from their Aristolochia host plants, rendering both larvae and adults chemically defended against most predators. Using a chemically controlled artificial diet, we observed substantial among-family variation in sequestration ability and larval developmental rate in a population occurring in central Texas. Early instar larvae from families that sequester greater amounts of aristolochic acid showed increased survivorship in a field experiment in which cohorts from each family were exposed to natural predators, whereas among-family variation in growth rate did not predict survivorship. Conversely, the aristolochic acid content of adult butterflies was negatively correlated with adult fat content, a fitness correlate. Sequestration ability positively affects the probability of larval survivorship, but at the cost of adult fat content. The costs and benefits of aristolochic acid sequestration vary during the course of the butterfly's development, and these antagonistic selection pressures may explain why variation in sequestration ability persists in wild populations.

Analyses of molecular phylogenies of three unrelated tropical marine gastropod genera, Turbo, Echinolittorina, and Conus, reveal an increase in the rate of cladogenesis of some Indo-West Pacific (IWP) clades beginning in the Late Oligocene or Early Miocene between 23.7 and 21.0 million years ago. In all three genera, clades with an increased rate of diversification reach a maximum of diversity, in terms of species richness, in the central IWP. Congruence in both the geographical location and the narrow interval of timing suggests a common cause. The collision of the Australia and New Guinea plate with the southeast extremity of the Eurasian plate approximately 25 Mya resulted in geological changes to the central IWP, including an increase in shallow-water areas and length of coastline, and the creation of a mosaic of distinct habitats. This was followed by a period of rapid diversification of zooxanthellate corals between 20 and 25 Mya. The findings reported here provide the first molecular evidence from multiple groups that part of the present-day diversity of shallow-water gastropods in the IWP arose from a rapid pulse of speciation when new habitats became available in the Late Oligocene to Early Miocene. After the new habitats were filled, the rate of speciation likely decreased and this combined with high levels of extinction (in some groups), resulted in a slow down in the rate of diversification in the genera examined.

The establishment of new species by hybridization is difficult because it requires the development of reproductive isolation (RI) in sympatry to escape the homogenizing effects of gene flow from the parental species. Here we investigated the role of two pre- and two postzygotic mechanisms of RI in a system comprising two interdependent Pogonomyrmex harvester ant lineages (the H1 and H2 lineages) of hybrid origin and one of their parental species (P. rugosus). Similar to most other ants, P. rugosus is characterized by an environmental system of caste determination with female brood developing either into queens or workers depending on nongenetic factors. By contrast, there is a strong genetic component to caste determination in the H1 and H2 lineages because the developmental fate of female brood depends on the genetic origin of the parents, with interlineage eggs developing into workers and intralineage eggs developing into queens. The study of a mixed mating aggregation revealed strong differences in mating flight timing between P. rugosus and the two lineages as a first mechanism of RI. A second important prezygotic mechanism was assortative mating. Laboratory experiments also provided support for one of the two investigated mechanisms of postzygotic isolation. The majority of offspring produced from the few matings between P. rugosus and the lineages aborted at the egg stage. This hybrid inviability was under maternal influence, with hybrids produced by P. rugosus queens being always inviable whereas a small proportion of H2 lineage queens produced large numbers of adult hybrid offspring. Finally, we found no evidence that genetic caste determination acted as a second postzygotic mechanism reducing gene flow between P. rugosus and the H lineages. The few viable P. rugosus-H hybrids were not preferentially shunted into functionally sterile workers but developed into both workers and queens. Overall, these results reveal that the nearly complete (99.5%) RI between P. rugosus and the two hybrid lineages stems from the combination of two typical prezygotic mechanisms (mating time divergence and assortative mating) and one postzygotic mechanism (hybrid inviability).

Selfish genetic elements (SGEs) are ubiquitous in animals and often associated with low male fertility due to reduced sperm number in male carriers. In the fruit fly Drosophila pseudoobscura, the meiotic driving X chromosome “sex ratio” kills Y-bearing sperm in carrier males (SR males), resulting in female only broods. We competed SR males against the ejaculates of noncarrying standard males (ST males), and quantified the number of sperm transferred by SR and ST males to females. We show that SR males are very poor sperm competitors, which is partly related to transfer of fewer sperm during mating. However, sperm numbers alone cannot explain the observed paternity reduction, indicating SR males’ sperm may be of reduced quality, possibly due to damage during the killing of the noncarrying Y-sperm. The reduction in sperm competitive ability due to SR is large enough to potentially stabilize the spread of sex ratio drive through populations. The poor sperm competitive ability of SR males coupled with their low fitness as mates could favor increased remating by females to reduce paternity by SR males. Given the generally poor performance of SGE-carrying males in sperm competition, this may generate strong selective pressure favoring polyandry in many species.

Studies of postcopulatory sexual selection typically estimate a male's fertilization success from his paternity success (P₂) calculated at hatching or birth. However, P₂ may be affected by differential embryo viability, thereby confounding estimations of true fertilization success (F₂). This study examines the effects of variation in the ability of males to influence embryo viability upon the inequality between P₂ and F₂. It also investigates the consequences of this inequality for testing the hypothesis that polyandrous females accrue viability benefits for their offspring through facilitation of sperm competition (good-sperm model). Simulations of competitive mating trials show that although relative measures of male reproductive success tend to underestimate the strength of underlying good-sperm processes, good-sperm processes can be seriously overestimated using P₂ values if males influence the viability of the embryos they sire. This study cautions the interpretation of P₂ values as a proxy for fertilization success or sperm competitiveness in studies of postcopulatory sexual selection, and highlights that the good-sperm hypothesis needs empirical support from studies able to identify and separate unequivocally the males' ability to win fertilizations from their ability to influence the development of embryos.

Understanding the evolution and maintenance of female mate choice requires information on both the benefits (the sum of direct and indirect benefits) and costs of selective mating. In this study, I assessed the fitness consequences of female mate choice in a freshwater crustacean. In Hyalella amphipods, males attempt to form precopulatory pairs with females. Large males, bearing large posterior gnathopods, tend to be over-represented in precopulatory pairs. I show that females receive both direct (reduced risk of predation while paired) and indirect (sexy sons) benefits from mating with these males. Furthermore, the behavioral mechanisms used to filter male phenotypes carry no detectable energetic cost for females. Thus, females that choose males with successful phenotypes are expected to have higher Darwinian fitness than females that mate at random. This study shows that direct and indirect selection act together to favor large male size, which explains the sexual size dimorphism and size-based mating biases observed in this species.

In sexually polymorphic species, the morphs are maintained by frequency-dependent selection through disassortative mating. In heterodichogamous populations in which disassortative mating occurs between the protandrous and protogynous morphs, a decrease in female fitness in one morph is hypothesized to drive sexual specialization in the other morph, resulting in dimorphic populations. We test these ideas in a population of the heterodichogamous species, Acer opalus. We assessed both prospective gender of individuals in terms of their allocations and actual parentage using microsatellites; we found that most matings in A. opalus occur disassortatively. We demonstrate that the protogynous morph is maintained by frequency-dependent selection, but that maintenance of males versus protandrous individuals depends on their relative siring success, which changes yearly. Seeds produced later in the reproductive season were smaller than those produced earlier; this should compromise reproduction through ovules in protandrous individuals, rendering them male biased in gender. Time-dependent gender and paternity analyses indicate that the sexual morphs are specialized in their earlier sexual functions, mediated by the seasonal decrease in seed size. Our results confirm that mating patterns are context-dependent and change seasonally, suggesting that sexual specialization can be driven by seasonal effects on fitness gained through one of the two sexual functions.

Coevolution between parasites and their hosts typically leads to increasing specialization on host species by the parasite. Where multiple hosts are parasitized, specialization on each host can result in genetic divergence within the parasite population to create host races, and, ultimately, new species. We investigate how host-specific traits arise in Horsfield's bronze-cuckoo Chalcites basalis nestlings. Newly hatched cuckoos evict host young from the nest, yet in the absence of a model they accurately mimic the different begging calls of a primary host (superb fairy-wren, Malurus cyaneus) and a secondary host (buff-rumped thornbill, Acanthiza reguloides). Using cross-fostering experiments, we show that begging calls are modified after parasitism, through experience. Further, we demonstrate the mechanism by which mimetic calls are acquired. All cuckoo nestlings initially produced the call of their primary host. When cross-fostered as eggs to a secondary host, calls increased in variability and were rapidly modified to resemble those of the secondary host through shaping by host parents. We suggest that plasticity in the development of host-specific traits after parasitism is likely to reduce selection for host race formation.

Multiple infections of a host by different strains of the same microparasite are common in nature. Although numerous models have been developed in an attempt to predict the evolutionary effects of intrahost competition, tests of the assumptions of these models are rare and the outcome is diverse. In the present study we examined the outcome of mixed-isolate infections in individual hosts, using a single clone of the waterflea Daphnia magna and three isolates of its semelparous endoparasite Pasteuria ramosa. We exposed individual Daphnia to single- and mixed-isolate infection treatments, both simultaneously and sequentially. Virulence was assessed by monitoring host mortality and fecundity, and parasite spore production was used as a measure of parasite fitness. Consistent with most assumptions, in multiply infected hosts we found that the virulence of mixed infections resembled that of the more virulent competitor, both in simultaneous multiple infections and in sequential multiple infections in which the virulent isolate was first to infect. The more virulent competitor also produced the vast majority of transmission stages. Only when the less virulent isolate was first to infect, the intrahost contest resembled scramble competition, whereby both isolates suffered by producing fewer transmission stages. Surprisingly, mixed-isolate infections resulted in lower fecundity-costs for the hosts, suggesting that parasite competition comes with an advantage for the host relative to single infections. Finally, spore production correlated positively with time-to-host-death. Thus, early-killing of more competitive isolates produces less transmission stages than less virulent, inferior isolates. Our results are consistent with the idea that less virulent parasite lines may be replaced by more virulent strains under conditions with high rates of multiple infections.

Pollination systems frequently reflect adaptations to particular groups of pollinators. Such systems are indicative of evolutionary specialization and have been important in angiosperm diversification. We studied the evolution of pollination systems in the large genus Ruellia. Phylogenetic analyses, morphological ordinations, ancestral state reconstructions, and a character mapping simulation were conducted to reveal key patterns in the direction and lability of floral characters associated with pollination. We found significant floral morphological differences among species that were generally associated with different groups of floral visitors. Floral evolution has been highly labile and also directional. Some specialized systems such as hawkmoth or bat pollination are likely evolutionary dead-ends. In contrast, specialized pollination by hummingbirds is clearly not a dead-end. We found evidence for multiple reverse transitions from presumed ancestral hummingbird pollination to more derived bee or insect pollination. These repeated origins of insect pollination from hummingbird-pollinated ancestors have not evolved without historical baggage. Flowers of insect-pollinated species derived from hummingbird-pollinated ancestors are morphologically more similar to hummingbird flowers than they are to other more distantly related insect-pollinated flowers. Finally, some pollinator switches were concomitant with changes in floral morphology that are associated with those pollinators. These observations are consistent with the hypothesis that some transitions have been adaptive in the evolution of Ruellia.

The heritability and genetic basis of nectar traits have been rarely studied in the field, where plants are exposed to environmental factors that could mask underlying genetic effects. Heritabilities and variance components were estimated for nectar and morphological traits of Nicotiana alata, using a partial diallel design. The main experiment was conducted in a Missouri experimental garden using a randomized block design with three plant density treatments, whereas a smaller experiment was conducted near native Brazil habitat to compare the environmental variance in traits between Missouri and Brazil. Significant heritability was detected for nectar volume and energy content, and for corolla tube length. Phenotypic correlations were significant between all traits investigated, whereas significant genetic correlations were only found between nectar volume and energy and between corolla limb width and mouth diameter. There were no significant family-by-density interactions detected in the Missouri field environment. All traits differed significantly between Missouri and Brazil environments, but significant genetic by environment (G × E) interactions between Missouri and Brazil were detected for only one trait. This study shows that nectar traits can be heritable despite considerable environmental variation.

Measurements of natural selection in hermaphrodite populations require the analysis of performance through both female and male sex functions. Here, we investigate selection on three floral traits: flower number, flower length, and corona width through both sex functions in natural populations of the tristylous daffodil Narcissus triandrus. Selection through female function was examined in six populations, and in two of these we also estimated male selection gradients using multilocus microsatellite genotyping of parents and offspring. We detected significant directional selection for flower number through female function, and significant stabilizing selection for corona width and flower length through male function. Variation in male reproductive success was strongly influenced by the distance between mates and was significantly higher than variation in female reproductive success in one population, a result consistent with Bateman's principle. However, variation through both sex functions was similar in the other population and there was a significant negative correlation between female and male fitness indicating sex-specific trade-offs in reproductive success. Selection on floral design in N. triandrus was stronger through male than female function probably because floral morphology plays an important role in promoting effective cross-pollen transfer in populations of this heterostylous species.

The amount of time asleep varies greatly in mammals, from 3 h in the donkey to 20 h in the armadillo. Previous comparative studies have suggested several functional explanations for interspecific variation in both the total time spent asleep and in rapid-eye movement (REM) or “quiet” (non-REM) sleep. In support of specific functional benefits of sleep, these studies reported correlations between time in specific sleep states (NREM or REM) and brain size, metabolic rate, and developmental variables. Here we show that estimates of sleep duration are significantly influenced by the laboratory conditions under which data are collected and that, when analyses are limited to data collected under more standardized procedures, traditional functional explanations for interspecific variation in sleep durations are no longer supported. Specifically, we find that basal metabolic rate correlates negatively rather than positively with sleep quotas, and that neither adult nor neonatal brain mass correlates positively with REM or NREM sleep times. These results contradict hypotheses that invoke energy conservation, cognition, and development as drivers of sleep variation. Instead, the negative correlations of both sleep states with basal metabolic rate and diet are consistent with trade-offs between sleep and foraging time. In terms of predation risk, both REM and NREM sleep quotas are reduced when animals sleep in more exposed sites, whereas species that sleep socially sleep less. Together with the fact that REM and NREM sleep quotas correlate strongly with each other, these results suggest that variation in sleep primarily reflects ecological constraints acting on total sleep time, rather than the independent responses of each sleep state to specific selection pressures. We propose that, within this ecological framework, interspecific variation in sleep duration might be compensated by variation in the physiological intensity of sleep.

The study of chalcid wasps that live within syconia of fig trees (Moraceae, Ficus), provides a unique opportunity to investigate the evolution of specialized communities of insects. By conducting cospeciation analyses between figs of section Galoglychia and some of their associated fig wasps, we show that, although host switches and duplication have evidently played a role in the construction of the current associations, the global picture is one of significant cospeciation throughout the evolution of these communities. Contrary to common belief, nonpollinating wasps are at least as constrained as pollinators by their host association in their diversification in this section. By adapting a randomization test in a supertree context, we further confirm that wasp phylogenies are significantly congruent with each other, and build a “wasp community” supertree that retrieves Galoglychia taxonomic subdivisions. Altogether, these results probably reflect wasp host specialization but also, to some extent, they might indicate that niche saturation within the fig prevents recurrent intrahost speciation and host switching. Finally, a comparison of ITS2 sequence divergence of cospeciating pairs of wasps suggests that the diversification of some pollinating and nonpollinating wasps of Galoglychia figs has been synchronous but that pollinating wasps exhibit a higher rate of molecular evolution.

Batesian mimics are predicted to lose their fitness advantage not only in the absence of an unpalatable model, but also when the mimic becomes relatively abundant. The phenotypic hybrid zone between mimetic and nonmimetic admiral butterflies, comprising the polytypic Limenitis arthemis species complex, offers an ideal opportunity to test these predictions because the position of the hybrid zone is hypothesized to be controlled by the geographic range of Battus philenor, the chemically defended model. We used 29 years of observational field data from a continental-scale butterfly monitoring program, the 4th of July Butterfly Counts, to show that (1) the advantage of mimicry does not extend beyond the range of the model, (2) in contrast to expectations, the mimicry complex is maintained even where the model is rare and (3) the sharp phenotypic transition between mimetic and nonmimetic admiral populations occurs over a very narrow spatial scale corresponding to the limit of the model's range. These results suggest that, even at very low densities, there is selection for Batesian mimicry and it maintains the geographic position of this hybrid zone. Our findings highlight the value of large-scale, long-term citizen science monitoring programs for answering basic ecological and evolutionary questions.

Sexual selection requires social interactions, particularly between the sexes. When trait expression is influenced by social interactions, such traits are called interacting phenotypes and only recently have the evolutionary consequences of interacting phenotypes been considered. Here we investigated how variation in relative fitness, or the opportunity for sexual selection, affected the evolutionary trajectories of interacting phenotypes. We used experimentally evolved populations of the naturally promiscuous Drosophila pseudoobscura, in which the numbers of potential interactions between the sexes, and therefore relative fitness, were manipulated by altering natural levels of female promiscuity. We considered two different mating interactions between the sexes: mating speed and copulation duration. We investigated the evolutionary trajectories of means and (co)variances (P) and also the influence of genetic drift on the evolutionary response of these interactions. Our sexual selection treatments did not affect the means of either mating speed or copulation duration, but they did affect P. We found that the means of both traits differed among replicates within each selection treatment whereas the Ps did not. Changes as a consequence of genetic drift were excluded. Our results show that although variable potential strengths of sexual interactions influence the evolution of interacting phenotypes, the influence may be nonlinear.