Journal of Evolutionary Biology

Sperm competition can occur when sperm from two or more males compete for fertilization of a batch of eggs. While it has been extensively studied in internal fertilizers, it remains poorly under-stood in external fertilizers. To investigate how sperm competition affects fertilization under sperm limitation, we conducted controlled experiments on two sea urchin species: Paracentrotus lividus and Dendraster excentricus. Eggs from a single female were exposed to sperm from three individ-ual males separately (non-competitive treatment) or to a pooled sample from three males (competi-tive treatment). We measured fertilization success as proportion of eggs fertilized after two hours and assessed sperm motility as percentage of motile sperm and curvilinear velocity. For P. lividus , we estimated the sperm concentrations where fertilization success was 50% of maximum observed fertilization success (F50), and the fertilization efficiency of the sperm (Fe), i.e. the likelihood that a sperm - egg encounter results in a fertilizing contact, using the fertilization kinetics model of Styan (1998). For D. excentricus, fertilization success was measured at a sperm concentration expected to produce 50% fertilization success. Sperm competition reduced fertilization success and increased F50 compared to non-competitive treatments, suggesting antagonistic effects that may compromise fertilization under sperm-limited conditions in external fertilizers.

Whether sexual selection facilitates or hinders adaptation to novel environments is a question of fundamental importance due to current rapid environmental change. Using phylogenetic comparative analysis in 42 Drosophila species, we found that sperm length - a sexually selected ornament - positively associates with the ability of males to survive high temperatures. However, the ability to remain fertile does not increase with sperm length to the same extent, meaning that longer sperm species experience a larger gap between temperatures that kill and temperatures that sterilize males. The findings are consistent with the idea that strong sexual selection on condition-dependent exaggerated traits may facilitate somatic resilience to environmental stressors, but that susceptibility to heat-induced infertility may be a cost of extreme sperm ornamentation. Overall, the data suggests multifaceted links between postcopulatory sexual selection and climate change.

Parasites decrease host fitness, more so during physiologically challenging life stages such as migration. However, parasites are unevenly distributed among individuals, and hosts with different physiological condition may face different costs of migration with infections. We tested whether the relationship between infection by haemosporidian parasites and oxidative balance during autumn migration in blackcaps depends on individual age and sex, two major sources of variation in prevalence and physiological condition. We captured blackcaps at stopover in central Spain and measured infection status and physiological parameters. Blackcaps physiologically more prepared to depart from stopover had lower antioxidant capacity of plasma, probably because of physiological adjustments prior to the upcoming flight stage. Plasmodium parasites occurred only in coinfections, which were more frequent among young blackcaps. Regardless of individual sex and age, coinfected birds had lower antioxidant capacity of plasma, and blackcaps coinfected with Plasmodium had more oxidative damage to lipids in their red blood cells. Our results support the idea that haemosporidian infections impair the oxidative balance of blackcaps during autumn migration, which could impair individual performance and ultimately fitness, with increased population impacts during the first year of life when a larger proportion of individuals migrate coinfected with harmful Plasmodium parasites.

Vertebrates cope with rapid temperature fluctuations by plastically adjusting their behaviour and physiology. This phenotypic plasticity is partly mediated by plastic changes in glucocorticoid levels; however, whether individual differences in endocrine plasticity are associated with fitness remains unknown. Here, we examined the relationship between individual variation in glucocorticoid plasticity in response to air temperature changes and reproductive success in a wild great tit population (Parus major). Over nine years, we collected repeated measurements of baseline and stress-induced corticosterone during nestling rearing. We found consistent reaction norm patterns in hormone response to temperature variation -- as shown in a previous study conducted over a shorter period. This provides robust evidence of individual differences in average hormone levels (intercepts) and the strength of the plastic response (slopes), indicating potential for evolution. However, neither intercepts nor slopes were associated with reproductive success (fledgling number or brood mass), suggesting that individual variation in endocrine plasticity was not under natural selection, at least within the environmental conditions experienced. This study provides a first empirical test of the evolutionary implications of endocrine plasticity in the wild and highlights the need to assess its fitness relevance under more extreme conditions, such as those expected under future climate scenarios.

Animal behaviour is both innately constrained and shaped by learning. This mosaic organization has evolved in response to species-specific ecological demands and may differ between sensory modalities. Flower-visiting animals are a particularly useful system for investigating the relationship between sensory ecology and learning because they rely on multiple floral cues, particularly odour and colour, to locate food sources. However, it remains largely unexplored whether specialization on floral resources entails divergence in learning abilities across sensory modalities. Drosophila elegans is a flower-feeding species that depends heavily on floral resources throughout its life; adults spend much of their time on flowers and larvae develop on fallen flower leaves. Here, we compared odour-reward and colour-reward associative learning between the flower-feeding D. elegans and the fruit-feeding D. melanogaster. We found that, under conditions of equilibrated motivation, odour- and colour-preference, and using the same sugar reward, D. elegans exhibited poorer odour-reward learning performance but better colour-reward learning performance than D. melanogaster. These results suggest that the modality-specific eligibility of sensory information to enter into associations, known as the 'Garcia-effect' in experimental psychology, can evolve oppositely between species. This highlights the relationship between ecological specialization and mnemonic processing, and shows that biological 'intelligence' is not general.

The toxicity of chemical defences is well studied, but the potential contribution of compound structure to predator deterrence remains largely unexplored. Whether predation acts more strongly on toxicity or unpalatability remains largely untested, partly because few systems allow toxin structure to vary independently of quantity. Heliconius sara larvae provide such a system: those reared on Passiflora auriculata sequester cyclopentenyl cyanogenic glucosides (CGs), while those reared on P. biflora biosynthesise comparable quantities of aliphatic CGs. Using two invertebrate predators, Camponotus floridanus ants and Hierodula membranacea mantids, we tested whether this structural difference affects palatability independent of toxicity. Mantids rejected larvae with cyclopentenyl CGs more often than larvae with aliphatic CGs, despite no detectable difference in total CG content. This pattern was mirrored in extract-based assays with ants, independently of cyanide release: extracts with cyclopentenyl CGs remained deterrent, while extracts with aliphatic CGs did not differ in deterrence from water. Live larvae, by contrast, elicited similar responses from ants regardless of CG structure. These results show that variation in toxin structure can strongly affect palatability, with some compounds conferring greater protection than others. This demonstrates the importance of chemical structural diversity in the evolution of chemical defences.

Species interactions shape ecological dynamics, but adaptation to one interaction can trade off with others. Experiments that measure such evolved fitness changes across multiple interactions remain rare. We used experimental evolution to test whether recent selection under one ecological interaction changes fitness under other interactions in a fly-parasitoid system. We evolved a genetically diverse Drosophila melanogaster population for 15-17 generations under three regimes: benign conditions (control), high intraspecific competition, and infection by the parasitoid wasp Leptopilina boulardi. We then measured fly fitness (egg-to-adult survival and subsequent reproduction) under benign conditions, high intraspecific competition, and infection by three other parasitoid species and an additional conspecific L. boulardi strain. Selection history strongly influenced fitness across interactions. Populations maintained under high larval density showed reduced survival under infection by L. boulardi and L. victoriae, indicating a trade-off between past selection for competition and defence against some parasitoids. Populations evolved under parasitoid infection by L. boulardi showed increased resistance to a conspecific strain but reduced fitness when infected by another species, L. heterotoma. Our results show that rapid evolution in response to a single interaction can reshape multiple ecological interactions through fitness trade-offs, highlighting how recent evolutionary history may influence community dynamics and invasion susceptibility.

Social aggregation is widespread in vertebrate larvae, yet how genetic relatedness shapes group cohesion remains poorly understood. In particular, the association between genetic similarity and social aggregation may depend on the scale at which genetic relatedness is evaluated. Here, we quantitatively examined aggregation behaviour in tadpoles from three lineages of Japanese toads, Bufo japonicus, B. formosus, and B. gargarizans miyakonis . We experimentally combined tadpole cohorts differing in relatedness and lineage and quantified aggregation using spatial dispersion analyses. We then examined relationships between aggregation behaviour and genetic variation using mitochondrial DNA (mtDNA), major histocompatibility complex (MHC) class II variation, and microsatellite diversity. Overall degree of aggregation did not differ significantly between sibling and non-sibling cohorts, and Morisitas index provided no evidence that tadpoles formed separate clumps according to relatedness. However, the correlation between genetic variation and aggregation differed across scales of genetic relatedness: among non-sibling cohorts, greater mtDNA pairwise distance was associated with weaker aggregation, whereas among sibling cohorts, greater MHC amino-acid difference was associated with weaker aggregation. These results indicate that the degree of aggregation is related to genetic similarity across different scales of relatedness, rather than being explained by a simple sibling/non-sibling distinction. Our findings suggest that tadpole group cohesion is related to genetic similarity in a scale-dependent manner, highlighting the importance of considering the scale of genetic relatedness when investigating social discrimination in vertebrates, while the sensory and molecular mechanisms underlying these relationships remain to be determined.

The evolution of genome size and eukaryotic genome organisation has puzzled evolutionary biologists for decades. Are differences in DNA content and protein-coding capacity of eukaryotic genomes a result of natural selection, or neutral processes? If they were shaped by natural selection, which features might be connected with these differences: cell volume, ecological strategy, availability of nutrients, etc.? Kinetoplastids are protists with compact nuclear genomes that almost entirely lack introns, but whose mitochondrial DNA content (kDNA) can rival that of their nuclei. We hypothesised that the reduction in non-protein coding nuclear DNA content during the evolution of kinetoplastids is connected with an increase in kDNA content: species with larger fractions of total cellular DNA that is allocated to kDNA will have concomitantly smaller fractions of non-coding DNA in the nuclear genome. We developed a framework for estimation of DNA content in both the mitochondria and nuclei from quantitative fluorescence microscopy data, and inferred the fraction of non-coding DNA in nuclear genomes based on the combination of transcriptomic and fluorescence imaging data. Across 10 genetically distant isolates from diverse free-living kinetoplastid lineages, there is indeed a trend towards a lower proportion of non-protein-coding nuclear DNA when kDNA proportion is higher relative to the total DNA in the cell. This observation is consistent with the hypothesis that in free-living kinetoplastids, mitochondrial and nuclear genomes might co-evolve in terms of their DNA content. This finding raises the possibility that natural selection acts on genome size evolution in kinetoplastids, and contributes evidence and methodology to address this question more broadly in eukaryotes.

Male and female animals often differ remarkably in their reproductive behaviours, typified by competing males and caring females. One influential explanation traces sex-role divergence to anisogamy: sex differences in gamete size. This explanation remains controversial, because existing models have not fully resolved how anisogamy leads to secondary sex-role divergence, and exceptions to typical patterns are empirically well-documented. Here, we develop mathematical models linking anisogamy to secondary sex-role evolution, showing that anisogamy drives sex-role divergence along contrasting evolutionary trajectories depending on ecological context. When the sexes differ only in gamete size, male-biased mating competition and offspring care evolve under scramble competition, whereas uncertain paternity shifts the population towards typical patterns. By contrast, anisogamy generally drives typical sex roles under contest competition, although sex-role reversal can emerge as an alternative stable state when care must be apportioned among offspring. Anisogamy can thus explain the striking diversity of sex differences across taxa.

Artificial light at night (ALAN) disrupts the natural light-dark cycle, which is the most important cue for synchronizing circadian clocks. If populations most exposed to ALAN adapt to get less disturbed by it, we expect forest and city populations to differ genetically. We tested this prediction in a common garden experiment using a songbird, the great tit (Parus major), originating from forest and city populations. We simulated ALAN exposure during the breeding season by keeping the birds in pairs under ALAN or dark nights for three months and then repeated the experiment in the following year with the same pairs under the opposite treatment. The advancement of daily onset under ALAN disappeared over time for city birds, but not for their forest counterparts. For these same birds, we measured their onset of daily activity under constant dim light and shift in activity in response to light pulses and showed that the city/forest difference in behavioural response was not reflected in any clock property measured. This indicates that city and forest birds differ genetically in the downstream behavioural response to ALAN, and not in the circadian clock. Thus, urbanization is unlikely to select for divergence in circadian clock properties in songbirds.

Regular environmental cycles such as day and night or the lunar cycle are ubiquitous and drive many biological processes. Many organisms have evolved endogenous time-keeping mechanisms to anticipate these cycles to accurately time their behavior or synchronize physiological processes. The evolution of such synchronization to environmental cycles and their variability is not very well understood. As an example, we use the marine insect Clunio (Diptera, Chironomidae), which synchronize adult emergence and reproduction with the lunar cycle, and show different emergence patterns across populations. We use a model constructed with replicator-mutator dynamics to provide a novel outlook at the evolution of these emergence patterns. In particular, we investigate the effect of two traits of individual phenotypic variability of emergence, the spread and shape, on population-wide emergence patterns. We find that the semilunar and lunar emergence patterns are alternatively stable and occur when the phenotypic distribution has a narrow and is roughly flat. Arrhythmic emergence occurs when the phenotypic distribution is wide and nearly flat. The semi-lunar pattern can only be observed when the parameter of rhythmicity is changed. With our model, we introduce the concepts of frequency-dependent selection and individual phenotypic variability into the theory of chronobiology, which gives us insights into the evolution of rhythmicity in these organisms.

Self-fertilization results in reduction of heterozygosity over time, accelerated by small population sizes, restricted ranges, and fragmented habitats. Yet some species maintain heterozygosity despite these factors. One is Corallorhiza bentleyi, a threatened, mycoheterotrophic, self-pollinating orchid restricted to Virginia and West Virginia, USA. Reduced representation sequencing was employed across five of the ~12 known localities of C. bentleyi, including representatives of sister species C. involuta from Mexico, to quantify patterns of population structure and genomic diversity. Surveys of C. bentleyi revealed a predominance of the cleistogamous (closed-flowered) growth form. Differentiation was observed between the two species but with weak population structure in C. bentleyi. Surprisingly, excess heterozygosity and negative inbreeding were recovered in both species, counter to expectations. Analyses provided evidence against bioinformatic artifacts (collapsed paralogs), small-sample bias (Robertson effect), and accumulation of mutations along chromosomes due to low recombination associated with selfing (Meselson effect) as explanations for excess heterozygosity. Demographic analyses favored a model of post-glacial divergence among C. bentleyi and C. involuta lacking bottlenecks. Finally, simulations under alternative selection scenarios favored models of overdominance vs. those of negative frequency dependent selection. It was concluded that overdominance, namely selection against selfed homozygotes, is the most likely explanation for elevated heterozygosity in C. bentleyi. Conservation concerns are discussed in light of these findings and the threats of extinction for one of the most vulnerable orchid species in North America.

A major transition in the evolution of eukaryotic cells was phagocytosis: the ability of one cell to engulf another. In some cases, the engulfed cell persists, forming an endosymbiosis which is often mutually beneficial. However, it is unclear whether the interaction between the ancestors was also cooperative. There are clear benefits to developing the ability to eat a competitor, but can it ever be beneficial to eat a cooperator? We use a dynamical systems framework to identify the ecological conditions under which phagocytosis can evolve from a system of cooperating microbes.Beginning with a system of two cooperating species, we use invasion analysis to identify the parameter regimes under which a phagocytosing mutant can invade and the subsequent population dynamics. We identify a critical threshold that determines whether an invading predator will coexist with the cooperative community or replace members. Repeatedly invading with different predators, we find that the system saturates when there are three coexisting species, though this community does not have a single optimal composition. However, if we incorporate additional costs, we find there is a single evolutionarily stable community of two cooperators and a pure predator. Ultimately, we show that it is possible for phagocytosis to evolve from a cooperative community and, moreover, coexist with the original cooperating species.

Evolution has given rise to the diversity of life on Earth, but it can also cause species extinctions. While major extinction events are usually associated with external drivers, in this study, we show mechanistically how evolution can trigger abrupt extinction avalanches in ecological networks without any environmental change. In particular, we investigate an eco-evolutionary simulation model of a tripartite mutualistic-antagonistic network, such as a plant-pollinator-herbivore network, with additional intraguild competition. The prevalence of self-organised extinction avalanches depends on a subtle balance of the different interaction types, where antagonism and similarity-based competition are needed for initial diversification while mutualism and trait-independent competition foster evolutionarily driven extinction events. Extinction avalanches are triggered by a mutualist evolving to interact with plants that are not subject to sufficient antagonistic control. Subsequently, the mutualist, its plant partners and their main antagonist potentially outcompete the rest of the community. Our findings point out that evolutionary murder may claim many victims simultaneously and highlight the need to take different interaction types into account to gain a comprehensive understanding of the links between evolution and biodiversity in ecological communities.

Flowering plants vary in sex allocation (resource division between male and female function) and in selfing rate. These traits should evolve together: selfing changes fitness returns from pollen and ovules, whereas sex allocation changes the pollen available for selfing and export. Yet most models allow only one trait to evolve, leaving their evolutionary feedback unresolved. Here we model the joint evolution of sex allocation, pollen export and maternal control of competition between self- and cross-pollen. We show that selfing and sex allocation strongly influence one another, as selection favours associations between sex allocation and mating-system traits. When maternal control is fixed to recover classical mass-action, evolution can lead to the stable coexistence of selfing hermaphrodites and males specialised in pollen export. This androdioecy persists because hermaphrodites cannot prevent fertilisation by males. When maternal control of pollen competition also evolves, we find that where a mixed-mating equilibrium exists, selection again favours the emergence of androdioecy. As hermaphrodites evolve to favour their own pollen, however, they progressively exclude pollen from males, driving them extinct. The population may then return to mixed mating and repeat the process, or evolve complete selfing. We discuss these results with reference to known variation in the mating and sexual systems of plants.

Assortative mating for a trait under selection can increase the rate of adaptive evolution and limit introgression of detrimental alleles. We therefore hypothesized that assortative mating facilitated the rapid evolution of flowering phenology observed in the self-incompatible perennial Lythrum salicaria (purple loosestrife) as it invaded eastern North America. We characterized flowering schedules of 600 individuals representing 20 populations sampled along a latitudinal gradient of 10 degrees and grown in a common garden at mid-latitude. Consistent with our hypothesis, we found that flowering schedules were less synchronous in the mixed population of the common garden (s = 0.50) relative to the 20 locally adapted populations comprising it (s = 0.56 to 0.86), and flowering synchrony decreased towards higher latitudes of origin. Despite high synchrony, assortment potential ({rho}) by growth and phenology varied by population ({rho} = 0.03 to 0.61), but the predicted negative correlation between s and {rho} was masked by among-individual fitness variation in the common garden. We show how fitness variation and asynchrony associated with locally adapted phenology can reduce the fitness impacts of maladaptive gene flow and accelerate the rate of adaptive evolution during invasion.

Extreme climatic events (ECEs) could potentially induce episodes of rapid adaptive evolution, altering the evolutionary trajectories and viability of wild populations experiencing increasingly stochastic environments. Adaptive potential stems substantially from additive genetic variance (V) in survival. However, the degree to which ECEs impact this variance, and the resulting evolutionary implications, have not been fully conceptualised or quantified in nature. These aims are highly challenging because underlying genetic and environmental variances have fundamentally non-linear effects on the dichotomous phenotypic outcome of survival versus death, while substantial data are required for estimation. We present a conceptual and analytical framework that formalises biological hypotheses for how V depends on environmental severity, how such dependences are reshaped by non-linear genotype-environment-phenotype relationships and demographic structure, and the implications for evolutionary responses. We then use 25 years of survival and pedigree data from European shags (Gulosus aristotelis) to decompose variance in adult survival across diverse environmental conditions including ECEs. Using a capture-recapture animal model, we demonstrate non-negligible V underlying adult survival across all years combined, implying notable adaptive potential. Yet, partitioning by environmental severity revealed substantial V in good years with high survival but negligible V in bad years with ECE-induced low survival, implying a genotype-by-environment interaction that constrains adaptive potential under severe conditions. However, scale transformations showed how the non-linear genotype-environment-phenotype relationship can distort or even reverse such patterns, by reducing the extent to which V is phenotypically expressed in good conditions with high mean survival and amplifying it in more severe conditions with lower mean survival. Evolutionary responses to severe conditions could consequently be non-negligible even given small underlying V. We thereby demonstrate how ECEs can reshape adaptive potential, highlighting the need to integrate impacts of environmental severity on genetic variation in fitness components into predictions of population resilience under climate change.

Dominance is an important and evolvable property of the genotype-phenotype map. A longstanding debate in evolutionary genetics concerns whether dominance is determined primarily by intrinsic properties of alleles at the focal locus or by modifiers elsewhere in the genome. Theoretical models show that second-site modifiers of dominance can evolve under some conditions. However, direct empirical tests of these models are rare because they require holding the focal heterozygous genotype constant while changing the rest of the genome. In the Drosophila montium species subgroup, female pigmentation is a Mendelian trait controlled by a single gene, POU domain motif 3 (pdm3), but the dominance relationships between pdm3 alleles differ among species. Using reciprocal introgressions among three closely related species, we compared females carrying the same Light/Dark pdm3 allele combinations in different homozygous or heterozygous genomic backgrounds and across different temperatures. In both species-pair comparisons, changing genomic background significantly altered the phenotype of identical pdm3 heterozygotes, demonstrating the existence of second-site modifiers of dominance. The effects of dominance modifiers depend on the particular pair of pdm3 alleles and on rearing temperature. These results show that dominance in this system is a genotype-by-genotype-by-environment trait, rather than a fixed attribute of individual alleles. The finding that second-site modifiers can alter dominance by reshaping reaction norms provides a developmental mechanism that links the evolution of dominance to phenotypic plasticity.

Populations that have detectable additive genetic variance for fitness (VA(W)) have the capacity for increase in their mean fitness through natural selection. The extent to which the magnitude of VA(W) in wild populations is predictive of the rate of adaptation is not well known, because few empirical field-based studies have been designed to address this question. Here, we report a two-year field experiment with pedigreed seeds embedded in four natural populations of the annual wildflower Baby Blue Eyes (Nemophila menziesii H. & A.), a species native to western North America. We first estimated VA(W) in a parental generation and then evaluated whether that estimate accurately predicted the adaptive response observed in the subsequent offspring generation grown under natural field conditions. We achieved this by directly comparing the mean fitnesses of the parental generation and their progeny grown under the same environmental conditions. We detected VA(W) and evidence of adaptation in all populations despite inter-annual differences in environmental conditions that directly affected fitness. Genotype-by-year interactions in fitness indicate that the fitness ranking of families differed between years, implying that the selective environment shifted over time. Additionally, although the environment in the second year reduced population mean fitness in three sites, the offspring generation maintained mean fitness at or above replacement, suggesting that adaptive evolution may have buffered the effects of environmental change. Overall, our results suggest that the four populations harbored sufficient VA(W) for adaptation through natural selection to the environmental conditions observed during the two years of this study.