Allometry, the relationship between body size and the size of other body parts, explains a significant portion of morphological variation across biological levels, at the individual level, within and between species. We used external morphology measurements of 6 Triturus (sub)species, focussing on the T. marmoratus species group, to explore allometric parameters within and between taxa. We tested for allometry of sexual size dimorphism in body, head, and limb dimensions and examined whether intraspecific allometry directed evolutionary allometry, as described by Rensch's rule. Our findings indicated that female-biased trunk and head dimensions exhibited positive allometry, whereas male-biased limb dimensions showed isometric relationships or weak correlations with body size. Morphological divergences between sexes occurred along common allometric slopes, most often through changes in the intercepts. Among taxon, comparisons revealed that (sub)species diverged in the direction of the allometric slopes. In line with Rensch's rule, sexual size dimorphism in female-biased traits significantly decreased as overall body size increased. However, the observed intraspecific allometric parameters deviated from theoretical expectations because the steepest allometric slopes for female-biased traits were recorded in the larger species. Our results contribute to understanding the dynamics of allometric relationships and sexual dimorphism in amphibians and provide a robust baseline for future comparative analyses.

Plant hosts can gain significant growth benefits from symbiosis with microbes, but these benefits could be threatened by divergent fitness interests among partners. Here, we measured fitness outcomes in symbiosis, by varying the genotypes of both microbes and hosts, to examine scenarios that might favour uncooperative symbionts. We studied associations between Acmispon strigosus, an annual legume native to California, and its nitrogen-fixing symbionts in the genus Bradyrhizobium. Bradyrhizobium symbionts form root nodules on compatible hosts, with strains varying from effective, fixing substantial nitrogen for the host, to ineffective strains that do not fix nitrogen and provide no benefit to host growth. We co-inoculated four A. strigosus plant lines with nine combinations of effective and ineffective Bradyrhizobium strains and measured the relative fitness of ineffective strains within individual nodules, as hosts must select against uncooperative symbionts to maintain benefits. In mixed infections, ineffective strains always had lower relative fitness in nodules compared to beneficial strains, consistent with efficient punishment of non-fixing rhizobia. However, ineffective strains exhibited genotypic variation in their fitness in nodules within individual nodules co-infected with a beneficial strain, suggesting a role for symbiont competitiveness in shaping this joint phenotype. Variation in symbiont fitness during co-inoculations did not measurably affect plant performance, suggesting that predicted conflict over the joint phenotype of rhizobia fitness has negligible effect on the host.

Mitochondrial paraphyly between arthropod species is not uncommon and has been speculated to largely be the result of incomplete lineage sorting (ILS) of ancestral variation within the common ancestor of both species, with hybridization playing only a minor role. However, in the absence of comparable nuclear genetic data, the relative roles of ILS and hybridization in explaining mitochondrial DNA (mtDNA) paraphyly remain unclear. Hybridization itself is a multifaceted gateway to mtDNA paraphyly, which may lead to paraphyly across both the nuclear and mitochondrial genomes, or paraphyly that is largely restricted to the mitochondrial genome. These different outcomes will depend upon the frequency of hybridization, its demographic context, and the extent to which mtDNA is subject to direct selection, indirect selection, or neutral processes. Here, we describe extensive mtDNA paraphyly between two species of iron-clad beetle (Zopheridae) and evaluate competing explanations for its origin. We first test between hypotheses of ILS and hybridization, revealing strong nuclear genetic differentiation between species, but with the complete replacement of Tarphius simplex mtDNA through the introgression of at least 5 mtDNA haplotypes from T. canariensis. We then contrast explanations of direct selection, indirect selection, or genetic drift for observed patterns of mtDNA introgression. Our results highlight how introgression can lead to complex patterns of mtDNA paraphyly across arthropod species, while simultaneously revealing the challenges for understanding the selective or neutral drivers that underpin such patterns.

This paper evaluates recent work purporting to show that the "agency" of organisms is an important phenomenon for evolutionary biology to study. Biological agency is understood as the capacity for goal-directed, self-determining activity-a capacity that is present in all organisms irrespective of their complexity and whether or not they have a nervous system. Proponents of the "agency perspective" on biological systems have claimed that agency is not explainable by physiological or developmental mechanisms, or by adaptation via natural selection. We show that this idea is theoretically unsound and unsupported by current biology. There is no empirical evidence that the agency perspective has the potential to advance experimental research in the life sciences. Instead, the phenomena that the agency perspective purports to make sense of are better explained using the well-established idea that complex multiscale feedback mechanisms evolve through natural selection.

Host-pathogen infections and possible effects on co-evolutionary patterns depend on the genotypes of both host and pathogen. Obligate fungal pathogens of plants are often characterized by host-pathogen genotype-by-genotype (GxG) interactions, but whether these patterns exist in obligate insect fungal pathogens is unclear. We take advantage of the obligate insect pathogenic fungus Entomophthora muscae, where individual isolates are specific to different dipteran host species in nature but can cross-infect multiple fly species in the laboratory. We collected three new isolates of E. muscae from Drosophila species. Phylogenetic analysis showed that Drosophila-isolated E. muscae represents a distinct geographically widespread Drosophila lineage compared to the house fly (Musca domestica) or Delia species-isolated E. muscae. We used the three new E. muscae isolates from Drosophila spp. together with a genetically distinct E. muscae isolate from house flies and assessed their virulence in a cross-infection experiment using one house fly, three Drosophila suzukii, and two D. melanogaster genotypes as hosts. All fungal isolates successfully infected hosts, induced behavioural manipulation, sporulated in all fly hosts, and differed in virulence between host genotypes, revealing GxG interactions. While house flies were most susceptible to fungal infection with 99% mortality, we found a lower virulence of 49% and 25% mortality in D. melanogaster and D. suzukii genotypes, respectively. Furthermore, all isolates harboured a specific mycovirus (family Iflaviridae), but co-phylogenetic branching patterns did not support fungus-virus co-speciation. We show that the genetic makeup of both fungal pathogen and fly host influence E. muscae infectivity, confirming GxG interactions in obligate fly fungal pathogens.

Rensch's rule describes a pattern of interspecific allometry in which sexual size dimorphism (SSD) increases with size among closely related species (i.e., among a group of related species, the largest ones tend to show more male-biased SSD). Sexual selection is often invoked to explain Rensch's rule, as larger male body size is assumed to be favoured by sexual selection for increased fighting performance in contests for mating opportunities. Often, however, the correlation between size and performance is not well described. We studied a sexually selected performance trait, bite force in Anolis lizards, to determine whether patterns of SSD are linked to size-associated patterns of performance dimorphism at the macroevolutionary level, as expected under the sexual selection hypothesis for Rensch's rule. Additionally, we tested whether allometric patterns of performance dimorphism differ between mainland and island species, as the latter have likely evolved under a stronger sexual selection regime. We found that SSD overwhelmingly explains the relationship between performance dimorphism and size in anoles, as expected under a sexual selection model for Rensch's rule. However, residual performance dimorphism was higher in island than in mainland species, suggesting that these groups differ in performance dimorphism for reasons unrelated to size. Head size dimorphism was associated with residual performance dimorphism, but did not fully explain the difference in performance dimorphism between island and mainland species. Together, these findings highlight the need to interpret Rensch's rule patterns of body size evolution cautiously, as allometric patterns of performance dimorphism and size dimorphism might not be equivalent.

Polymorphic short insertions and deletions (INDELs ≤ 50 bp) are abundant, although less common than single nucleotide polymorphisms (SNPs). Evidence from model organisms shows INDELs to be more strongly influenced by purifying selection than SNPs. Partly for this reason, INDELs are rarely used as markers for demographic processes or to detect divergent selection. Here, we compared INDELs and SNPs in the intertidal snail Littorina saxatilis, focusing on hybrid zones between ecotypes, in order to test the utility of INDELs in the detection of divergent selection. We computed INDEL and SNP site frequency spectra (SFS) using capture sequencing data. We assessed the impact of divergent selection by analysing allele frequency clines across habitat boundaries. We also examined the influence of GC-biased gene conversion because it may be confounded with signatures of selection. We show evidence that short INDELs are affected more by purifying selection than SNPs, but part of the observed SFS difference can be attributed to GC-biased gene conversion. We did not find a difference in the impact of divergent selection between short INDELs and SNPs. Short INDELs and SNPs were similarly distributed across the genome and so are likely to respond to indirect selection in the same way. A few regions likely affected by divergent selection were revealed by INDELs and not by SNPs. Short INDELs can be useful (additional) genetic markers helping to identify genomic regions important for adaptation and population divergence.

In ovipositing animals, egg placement decisions can be key determinants of offspring survival. One oviposition strategy reported across taxa is laying eggs in clusters. In some species, mothers provision eggs with diffusible defence compounds, such as antimicrobials, raising the possibility of public good benefits arising from egg clustering. Here we report that Drosophila melanogaster females frequently lay eggs in mixed maternity clusters. We tested two hypotheses for potential drivers of this oviposition behaviour: (i) the microbial environment affects fecundity and egg placement in groups of females; (ii) eggs exhibit antimicrobial activity. The results partially supported the first hypothesis. Females reduced egg laying, but did not alter egg clustering, on non-sterile substrates that had been naturally colonised with microbes from the environment. However, oviposition remained unaffected when the substrate community consisted of commensal (fly-associated) microbes. The second hypothesis was not supported. There was no evidence of antimicrobial activity, either in whole eggs or in soluble egg surface material. In conclusion, while we found no behavioural or physiological evidence that egg clustering decisions are shaped by the opportunity to share antimicrobials, females are sensitive to their microbial environment and can adjust egg laying rates accordingly.

Walnut is a significant woody oil tree species that has been increasingly affected by anthracnose in recent years. Effective anthracnose control is crucial for walnut yield and quality, which requires a comprehensive understanding of the response mechanisms to Colletotrichum gloeosporioides. The PR10/Bet v1-like proteins are involved in defense against various disease, therefore, in this study, 7 JrBet v1s were identified from the walnut transcriptome (named JrBet v1-1~1-7), whose open reading frame (ORF) was 414~483 bp in length with isoelectric point ranging from 4.96 to 6.11. These JrBet v1s were classified into three groups, with the MLP/RRP and Dicot PR-10 subfamilies each comprising three members (the largest ones), indicating that the proteins within these two subfamilies may have evolved from a shared ancestral gene within the broader PR10/Bet v1 protein family. The cis-elements in the promoters of JrBet v1s were involved in response to hormones, coercive, and plant growth metabolism. Most JrBet v1s could be significantly upregulated by walnut anthracnose, JrBet v1-1, JrBet v1-2, JrBet v1-4, and JrBet v1-6 peaked at 12 days of anthracnose stress, showing a 2.85- to 63.12-fold increase compared to the control, while JrBet v1-3, JrBet v1-5 and JrBet v1-7 peaked at 9 days, with a 3.23- to 27.67-fold increase. Furthermore, the significant corelation of the upregulation under anthracnose stress of JrBet v1s and JrChit02-1 as well as JrChit02-2, the genes encoding chitinase, suggesting that during the long process of microevolution in walnut-C. gloeosporioides interactions, walnut has developed a Bet v1-chitinase defense mechanism to counteract pathogen invasion.

Modularity and developmental (in)stability have the potential to influence phenotype production and, consequently, the evolutionary trajectories of species. Depending on the environmental factors involved and the buffering capacity of an organism, different developmental outcomes are expected. Cactophilic Drosophila species provide an established eco-evolutionary model with well-studied ecological conditions, making them ideal for studying these phenomena. Here, we investigated how variations in larval diet and exposure to alkaloids on primary and secondary host plants affect the degree of integration/modularity and fluctuating asymmetry (a proxy for developmental instability) of wing shape in two sibling species with different degrees of specialization: D. buzzatii (generalist) and D. koepferae (specialist). Additionally, we compared the anterior-posterior modular configuration with a recently proposed proximal-distal modular configuration. Our results revealed greater independence among proximal-distal modules compared to anterior-posterior modules. Moreover, we observed sex-specific responses, with males exhibiting greater susceptibility to stressful environments than females. Each species showed a particular trait pattern across treatments: D. buzzatii showed increased integration and fluctuating asymmetry when reared in a nutrient-poor, alkaloid-rich secondary host, while D. koepferae displayed similar responses in novel environments characterized by double doses of alkaloids on the secondary host plant. These findings align with the generalist-specialist paradigm, suggesting that specialists may be challenged by novel environments, whereas generalists may be more affected by stressful conditions. Our study highlights the importance of considering each part of the proximal-distal wing axis independently, and the need to consider ecological-evolutionary history when investigating the relationship between complex phenotypic traits and environmental stress.

Differences in habitat use impose ecological constraints which in turn lead to functional and morphological differences through adaptation. In fact, a convergent evolutionary pattern is evident when species exhibit similar responses to similar environments. In this study we examine how habitat use influences the evolution of body shape in lizards from the family Lacertidae. We divided our species set into two categories: ground-dwellers and climbers, which encompasses the verticality and horizontality aspects of the habitat. We performed phylogenetic comparative analyses employing 186 species and seven linear morphological traits. Our results show contrasting patterns between head and limb shape, which are considered distinct functional blocks. We observed differences in forelimb proportions, but not in hindlimb length, contrary to what was documented in other lizard groups, demonstrating a novel axis in the limb-locomotion-habitat relationship in this family. In addition, a clear effect of habitat use on head shape was detected. We observed that climbing species present on average flatter heads than ground-dwelling species, as well as different evolutionary trajectories. These findings suggest the complex interplay between habitat use and morphological evolution in lizards, highlighting how distinct selective pressures drive divergent adaptations in different functional traits.

Sex allocation theory predicts the adaptive allocation of resources to male versus female reproduction in simultaneous hermaphrodites in response to individual characteristics or environmental factors. Because parasites uptake resources from their hosts, their presence could affect the sex allocation of the hosts. We investigated the effects of infestation status and infestation intensity by the rhizocephalan barnacle Boschmaella japonica on reproduction, including sex allocation, of the host intertidal barnacle Chthamalus challengeri. Feeding activity was also examined as a factor related to resource intake. Both male and female reproductive investment decreased with increasing parasite infestation, and the sex allocation of large infested hosts was more male-biased than that of large uninfested hosts. Moreover, in contrast to the model prediction that male investment does not change under resource limitation, male investment decreased in infested hosts whose resources were taken by parasites. This reduction in male investment could be explained by changes in mating group size, since infested hosts have shorter penises and consequently are able to access fewer mating partners.

In eutherians, one of the X chromosomes in each cell of the early female embryo is rendered transcriptionally silent through X chromosome inactivation. The choice of which X chromosome to inactivate takes place independently in each cell and is stably inherited through development, leading to a roughly 50:50 ratio of cells in the adult body expressing one or the other X chromosome. However, X chromosome inactivation can be skewed, with certain X chromosomes showing a heritable tendency to avoid inactivation. Using population genetic models, we test whether genetic variation for this trait can be maintained by linked sexually antagonistic selection. In favor of this hypothesis, we find that a neutral modifier that affects the chances of its chromosome's inactivation-e.g., a variant of the X controlling element (Xce)-can spread when linked to a sexually antagonistic gene. We explore the logic of this modifier's spread, which we find to be similar in many respects to that of a modifier of dominance. We also test for the presence of a "drift barrier"-i.e., a population size below which the indirect selective force favoring the modifier becomes too weak to overcome drift. On balance, we find that sexual antagonism may encourage the spread of skewed X chromosome inactivation, but only under favorable conditions.

Characterizing molecular underpinnings of plastic traits and balanced polymorphisms represent two important goals of evolutionary biology. Fire ant gynes (pre-reproductive queens) provide an ideal system to study potential links between these phenomena because they exhibit both supergene-mediated polymorphism and nutritional plasticity in weight and colony-founding behavior. Gynes with the inversion supergene haplotype are lightweight and depend on existing workers to initiate reproduction. Gynes with only the ancestral, non-inverted gene arrangement accumulate more nutrient reserves as adults and, in a distinct colony-founding behavior, initiate reproduction without help from workers. However, when such gynes overwinter in the natal nest they develop an environmentally induced lightweight phenotype and colony-founding behavior, similar to gynes with the inversion haplotype that have not overwintered. To evaluate the extent of shared mechanisms between plasticity and balanced polymorphism in fire ant gyne traits, we assessed whether genes with expression variation linked to overwintering plasticity may be affected by evolutionary divergence between supergene haplotypes. To do so, we first compared transcriptional profiles of brains and ovaries from overwintered and non-overwintered gynes to identify plasticity-associated genes. These genes were enriched for metabolic and behavioral functions. Next, we compared plasticity-associated genes to those differentially expressed by supergene genotype, revealing a significant overlap of the two sets in ovarian tissues. We also identified sequence substitutions between supergene variants of multiple plasticity-associated genes, consistent with a scenario in which an ancestrally plastic phenotype responsive to an environmental condition became increasingly genetically regulated.

The time needed for the evolution of mating cues that distinguish species, such as species-specific songs or plumage coloration in birds, has received little attention. Aiming to gain some understanding of the timing of the evolutionary process we here present models of how mating cues evolve in populations split into subpopulations between which there may (parapatry) or may not (allopatry) be migration. Mating cues can be either neutral or directly selected. In models in which evolution commences with a substitution at a neutral mating-cue locus, under allopatry there is no selection on the mating cue, but under parapatry selection may be induced on the mating cue by the selective conditions in the subpopulations and the migration rates between them. We use simulation to calculate how selection pressures on mating cues then depend on selective conditions in subpopulations and migration rates between them. In the second part of the paper we demonstrate quantitatively how the resulting selection pressures on new mating cues together with mutation rate affect speciation time. Our results suggest that species-specific songs or plumage colorations that are selectively neutral evolve faster under parapatry than under allopatry, and this may explain the short speciation times that are sometimes reported. Although our modelling assumptions are restrictive so that caution is needed in comparing the results to empirical data, we hope that our main results, showing quantitatively how parapatry can reduce speciation times, will encourage further work relaxing model assumptions or studying different models of mate choice.

Recombination diversifies the genomes of offspring, influences the evolutionary dynamics of populations, and ensures that chromosomes segregate properly during meiosis. Individuals recombine at different rates but observed levels of variation in recombination rate remain mostly unexplained. Genetic dissection of differences in recombination rate within and between species provides a powerful framework for understanding how this trait evolves. In this Perspective, I amalgamate published findings from genetic studies of variation in the genome-wide number of crossovers within and between species, and I use exploratory analyses to identify preliminary patterns. The narrow-sense heritability of crossover count is consistently low, indicating limited resemblance among relatives and predicting a weak response to short-term selection. Variants associated with crossover number within populations span the range of minor allele frequency. The size of the additive effect of recombination-associated variants, along with a negative correlation between this effect and minor allele frequency, raises the prospect that mutations inducing phenotypic shifts larger than a few crossovers are deleterious, though the contributions of methodological biases to these patterns deserve investigation. Quantitative trait loci that contribute to differences between populations or species alter crossover number in both directions, a pattern inconsistent with selection toward a constant optimum for this trait. Building on this characterization of genetic variation in crossover number within and between species, I describe fruitful avenues for future research. Better integrating recombination rate into quantitative genetics will reveal the balance of evolutionary forces responsible for genetic variation in this trait that shapes inheritance.

Wing reduction is a common feature of upland insect communities. This phenomenon is thought to be primarily driven by selection against flight, which is typically unfavorable in upland environments due to high winds and cold temperatures. In some insect taxa, wing-reduction has been directly linked to increased fecundity. However, few studies have directly tested for shifts in fecundity linked to flight musculature. Here we test for dispersal-fecundity trade-offs in the widespread subalpine stonefly Zelandoperla fenestrata. Our analysis of 450 stoneflies across 81 localities reveals significant dispersal-fecundity tradeoffs. Specifically, we identify a positive association between the size of their flight muscles and the length of their wings, and a negative association between wing length and ovarian mass. Furthermore, we found a significant negative relationship between flight musculature and ovary mass. These results represent a rare example of a dispersal-fecundity tradeoff in the wild, and illustrate that such tradeoffs can potentially involve corresponding reductions in both flight musculature and wing development. Our findings suggest that widespread taxa subject to variable environmental conditions may benefit from flexible allocation of energetic resources.

Individuals living in heterogeneous environments often choose microenvironments that provide benefits to their fitness. Theory predicts that such niche choice can promote rapid adaptation to novel environments and help maintain genetic diversity. An open question of large applied importance is how niche choice and niche choice evolution affect the evolution of insecticide resistance in phytophagous insects. We, therefore, developed an individual-based model based on phytophagous insects to examine the evolution of insecticide resistance and niche choice via oviposition preferences. To find biologically realistic parameter ranges, we performed an empirical literature survey on insecticide resistance in major agricultural pests and also conducted a density-dependent survival experiment using potato beetles. We find that, in comparison to a scenario where individuals randomly oviposit eggs on toxic or non-toxic plants, the evolution of niche choice generally leads to slower evolution of resistance and facilitates the coexistence of different phenotypes. Our simulations also reveal that recombination rate and dominance effects can influence the evolution of both niche choice and resistance. Thus, this study provides new insights into the effects of niche choice on resistance evolution and highlights the need for more studies on the genetic basis of resistance and choice.

Several studies have emphasized that phenotypic plasticity should be a key mechanism to cope with current rapid environmental changes by allowing individuals to quickly express new adaptive phenotypes. Yet, few studies have investigated the evolutionary potential of plasticity for multiple traits simultaneously and using several different environmental variables. Here, we assess the extent of variation in, and the selection acting on phenotypic plasticity of key ecological traits, laying date and clutch size, using five environmental variables, in a Tree swallow (Tachycineta bicolor) population monitored since 2004. While we found some variation among females in their mean laying date and plasticity, we found evidence of selection acting only on mean laying date. We found no variation among females in mean clutch size or plasticity, such that we could not assess selection acting on either. Our results suggest that the evolutionary potential of plasticity in the population under study is limited, especially for clutch size. More studies investigating plasticity in wild populations and incorporating multiple traits and environmental variables are needed to understand future responses of animal populations to environmental changes.