BMC Ecology and Evolution

Allopatric isolation under contrasting environments can drive rapid phenotypic divergence, even over contemporary timescales. Rapid changes in morphology or physiology can allow organisms to adapt to biotic and abiotic characteristics of their habitats. While studying metabolism, growth and resources needs may allow to understand adaptation to several selective pressures, these traits are rarely jointly considered. We investigated morphological, growth, and metabolic divergence in two allopatric populations of Arctic charr (Salvelinus alpinus) sharing a common evolutionary origin but inhabiting contrasting environments. We combined field observations, common garden and quantitative genetic approaches to disentangle contributions of genetic divergence and plasticity to phenotypic variability. Wild adults differed in body shape and growth trajectories, potentially reflecting plasticity related to resource availability and temperature variations. Under common garden conditions, juveniles displayed inter-population differences in routine metabolic rate, its allometric scaling with body mass. These patterns suggest divergent selection on physiological traits. Despite low neutral genetic differentiation, phenotypic divergence unfolded in fewer than 100 years, suggesting that plasticity and selection can promote rapid multi-trait changes. These findings highlight that considering changes in physiological, growth and morphological traits can reveal the adaptive potential of small, isolated populations facing rapid environmental change.

Although it is well established that certain stages of development are molecularly more conserved than others, the reasons for this phenomenon remain largely unknown. We study molecular conservation in the development of an organ, the molar, by comparing the temporal profiles of expression in mice and hamsters. We find that the cause of conservation of expression and of coding sequences changes over molar development. Gene expression levels display a classical increase of divergence as development progresses. In terms of genes expressed, the composition of early and late stages is better conserved and enriched in pleiotropic genes, yet each stage mobilizes different sets of pleiotropic genes, cell division for bud growth and secretion for tooth mineralization. Moreover similar patterns of higher divergence of gene sets and of coding sequences at mid development, are caused by different biological phenomena, in that case heterochronies and blood colonisation respectively. In conclusion, the patterns of molecular conservation in developing molars are shaped by a combination of processes intrinsic to the teeth, and by negative and positive selection on functions which are mostly extrinsic to the teeth. This is likely translatable to explain molecular conservation patterns in many other biological systems. AUTHOR SUMMARYFor species to evolve different adaptations to different life styles, their anatomy has to evolve correspondingly. This in turn implies evolution of the embryonic development of anatomical structures. Notably, tooth shape can evolve rapidly as an adaptation to different diets. Mice and hamsters are closely related rodents who yet differ in the shape of their molars, and thus in their development. In this study, we investigated why the genes active in molar development are more or less similar between the two species from early tooth bud to fully formed embryo molar. We found that early and late molar development were slow evolving, while mid-development was evolving faster. But surprisingly, this was in part due not to tooth evolution, but to the involvement of genes which are active in other processes in the body. For example an influx of immune cells also brings fast evolving immune genes. This helps us understand better the complexity of causes of apparently simple evolutionary patterns.

Accurate biological sex estimation is a key objective in archaeological and bioanthropological research but remains challenging when skeletal remains are fragmented, juvenile, or poorly preserved. Paleoproteomics approaches based on the detection of sex-specific amelogenin peptides (AMELX/AMELY) have emerged as a powerful alternative to osteological and genetic methods. However, current workflows often lack standardized criteria for peptide-level confidence assessment, potentially affecting the reproducibility and reliability of sex assignments. In this study, I evaluated the impact of peptide-level confidence filtering on paleoproteomics-based sex estimation through the reanalysis of 164 Homo sapiens individuals from 10 published datasets and 26 Bos taurus individuals from 3 datasets, spanning contexts from the Pleistocene to the present. To address methodological inconsistencies, I developed SexPeptID, an R/Shiny-based framework that integrates Posterior Error Probability (PEP) filtering, standardized peptide selection, and explicit uncertainty assessment. Application of SexPeptID revealed that peptide-level filtering substantially affects sex assignment outcomes: 17 previously classified males (10.4%) were reclassified as non-conclusive, while 5 individuals (3.1%) were identified as potentially female. Despite this sensitivity, AMELX/AMELY-based sex estimation remained robust overall, with stable signal ratios observed across archaeological periods. Variability in peptide intensities was primarily associated with dataset-specific factors rather than temporal differences, highlighting the influence of analytical workflows and preservation conditions. By incorporating confidence-based filtering and a non-conclusive classification category, SexPeptID improves the transparency, reproducibility, and reliability of palaeoproteomics sex estimation, providing a standardized framework for future archaeological and bioanthropological studies. HighlightsO_LISexPeptID provides a reproducible framework for amelogenin-based sex estimation. C_LIO_LIPeptide-level confidence filtering significantly affects paleoproteomics sex estimates. C_LIO_LI13.4% of published male assignments were revised after confidence filtering. C_LIO_LIAMELX/AMELY ratios show temporal stability from modern to Pleistocene samples. C_LIO_LIStandardized uncertainty assessment strengthens palaeoproteomics inference. C_LI

Mitochondrial loci were for long considered as markers of choice to reconstruct phylogenies. The development of high-throughput sequencing over the past two decades fostered the sequencing of mitogenomes, allowing further macroevolutionary questions to be tested. Several biological traits of birds (e.g. body mass, migration distances) have been related to mitochondrial substitution rates. Environmental parameters in ectothermics vertebrates, and potentially in endotherms, have been further suggested to impact substitution rates for specific taxa or loci. Yet, the relative importance of biological traits versus bioclimatic variables is unknown because the former were not systematically controlled for in studies that underlined the effect of the bioclimatic variables. To assess the importance of bioclimatic variables on selection regimes, we analysed the thirteen mitochondrial protein-coding genes for 176 Piciformes (toucans, honeyguides, woodpeckers), a clade with homogeneous life-history traits that can be found in diverse bioclimatic environments. Our analyses highlighted a negative relationship between temperature annual range and the non synonymous to synonymous substitutions ratio. The higher purifying selection in temperate environments may be a result of the strong constraints on maintaining an optimal metabolism in broader climatic variations. Our results further highlight that care should be taken when applying general mitochondrial clocks to estimate divergence times among avian lineages distributed in different climatic conditions.

AbstractsThe Japanese giant salamander (Andrias japonicus), an apex predator and a Special Natural Monument in Japan, is threatened by hybridization with introduced Chinese giant salamanders (Andrias davidianus). This hybridization has caused genetic introgression and expansion of hybrid populations, posing a serious conservation risk. Because morphological identification of hybrids is occasionally unreliable and current genetic methods rely on limited markers, a genome-wide approach is required. However, the extremely large genome ([~]50 Gb) of giant salamanders has hindered whole-genome analyses. In this study, we conducted transcriptome-based analyses of Japanese giant salamanders, Chinese giant salamanders, and their hybrids, generating RNA-seq data from 34 individuals. A total of over 419,000 SNP candidates were identified, from which 4,457 high-confidence SNPs in highly expressed genes were selected for analysis. Population structure analyses for Nabari colony revealed that hybrid individuals form two major groups, corresponding to different degrees of genetic contribution from Japanese and Chinese lineages. Most hybrids were inferred to be F2 or backcross individuals, while F1 hybrids were rare. Mitochondrial analysis indicated that all hybrids possessed Japanese-type mitochondrial genome, suggesting male-mediated introgression from Chinese salamanders. Differential expression analysis revealed enhanced stress-response pathways in hybrids and stronger antiviral responses in Japanese individuals. Using the axolotl genome as a reference, we constructed a virtual chromosomal map, identifying large haplotype blocks and supporting recent hybridization with limited recombination. This study provides a genome-wide framework for understanding hybridization dynamics and supports future conservation and evolutionary studies.

The origin of novel phenotypes can influence access to new ecological resources, which may have positive, neutral, or negative effects on subsequent phenotypic diversification. In this study, we tested the macroevolutionary consequences of a pair of putative functional innovations occurring in deep-sea fishes of the order Stomiiformes. Integrating phylogenetic comparative methods, micro-CT scans, and external body measurements, we recover a mosaic of diversification trends associated with these innovations. We found some evidence for elevated evolutionary rates in tooth morphology associated with the predatory dragonfishes, which possess a gap between their vertebral column and skull that exposes the notochord and enables neck-like flexibility. However, a second novelty building upon the first, a functional neck joint enabling extreme cranial kinesis, was linked to faster rates of skull evolution. Our results suggest that innovations that help shift ecological roles and overcome functional constraints related to those roles, like gape-limitation in prey depauperate habitats, may play an important role in promoting phenotypic diversification. This work builds on a growing body of evidence highlighting how the deep sea promotes phenotypic diversity, generating the extreme forms that are celebrated by scientists and the public alike.

Given the ever-increasing availability of highly detailed phenotypes, modelling trait evolution in a multivariate framework is becoming a challenging task. Current phylogenetic comparative methods often struggle with high-dimensional datasets because they suffer computational limitations and interpretability. Here, we propose a maximum likelihood-based approach called Probabilistic and Phylogenetic Principal Components Analysis (P3CA) to circumvent current limitations. This approach is based on a continuous latent variable model, whereby observed traits are explained by a smaller number of unobserved variables that evolve according to a given evolutionary model. We implement the approach under Pagels lambda model using an Expectation-Maximisation algorithm that makes it computationally efficient and allows missing values. Using simulations, we demonstrate that evolutionary parameters are accurately estimated, regardless of phylogenetic signal, the number of traits or the proportion of missing values. The reconstruction of the reduced space is more accurate than the one obtained using other dimensionality reduction approaches, such as phylogenetic and conventional PCA. Likewise, the estimated values for missing data are more accurate than those obtained using current phylogenetic data imputation approaches. We illustrate the approach on a 3D geometric morphometric dataset describing Crocodyliformes skull shapes and containing around 4% of missing data. Our P3CA method unlocks the possibility to analyse and more easily interpret the large-scale multivariate datasets generated in recent decades within a phylogenetic comparative framework.

The introduction of alien species into new habitats stands as a pressing economic and ecological challenge but it is also essential for unveiling evolutionary processes. The introduction of the Argentine ant (Linepithema humile) led to the spread of a single supercolony through different continents and thousands of kilometres like in Europe, from Northwest Spain to Greece. It was assumed that the high invasiveness of the species mainly relied on the lack of agonism among colonies, an effect derived from its introduction. However, recent studies suggest that local adaptations and evolutionary divergence could involve the disruption of the Argentine ant "empire" into a mosaic of competitive colonies. We investigated how isolation affects population divergence by comparing mainland and island populations in two distant regions colonized in Spain and in Greece with morphology, agonism, cuticular hydrocarbons, and genetic diversity of ant workers. Our results showed that all colonies sampled belonged to the most spread supercolony in Europe (main supercolony) except one sampled in Crete (Heraklion; Greece), which resulted to be a supercolony not registered in Europe. The Heraklian supercolony showed a different chemical and genetic profile and hostile agonism towards the other Greek colonies. Differences between islands and mainland colonies belonging to the main supercolony were higher in Galiza than in Greece. Surprisingly, the chemical profile of the Cretan colony belonging to the main supercolony showed more similarity with the Galizan colonies than with the Greek mainland, suggesting that L. humile may have been introduced into Greece through this island instead of the mainland. Our study suggests that local adaptations in Argentine ant colonies can trigger competition between colonies. Our data strongly support the existence of a candidate supercolony which highlights either ongoing introductions of L. humile in Europe or gaps in our understanding of its metapopulation dynamics.

The diversity of body shapes is one of the most prominent features of phenotypic variation in mammals. Yet, mammalian body shapes are poorly quantified and the underlying components contributing to its diversity as well as its relationship to other components of the skeleton are rarely tested. Here, we use lagomorphs (hares, rabbits and pikas) as a model system to (1) investigate which components of the skeleton contributed the most to body shape diversity, (2) examine the relationships between body shape and relative limb lengths, and (3) test how body size, ecotype, burrowing behavior, and locomotor mode influenced variation in lagomorph body shape and appendicular morphology. We quantified the body shape and functional proxies of the appendicular skeleton in 40 lagomorph species from osteological specimens held at museum collections. Using phylogenetic comparative methods, we found the relative length of the ribs and elongation or shortening of the thoracic and lumbar regions contributed the most to body shape evolution across lagomorphs. Second, we found that only leporids (hares and rabbits) exhibited a significant relationship between limb length and body shape, where more elongate species exhibit relatively shorter forelimbs and hindlimbs. Lastly, we found that models incorporating body size were the best predictors of lagomorph body shape and the majority of the appendicular traits, whereas models incorporating burrowing behavior and locomotor mode were largely poor fits. Broadly, these results indicate that larger lagomorphs tend to exhibit more robust body shapes with longer, more gracile forelimbs, whereas smaller lagomorphs tend to exhibit more elongate body shapes with shorter, more robust forelimbs. Overall, this work contributes to the growing understanding of mammalian body shape evolution and demonstrates the importance of not omitting body size in ecomorphological analyses.

This investigation aimed at compiling all phylogenetic lineages within and around genus Cyanoboletus. The evolutionary inference obtained from the nuclear ribosomal genes internal transcribed spacer region (ITS) suggests that part of the species currently classified in Cyanoboletus belong in lineages separate from the genus, thus suggesting a narrower boundary that includes only the species that develop a strong staining reaction to touch and to air exposure of the context. The separate lineages are the monotypic Cupreoboletus genus and a few species that do not develop such reaction, which are part of a clade together with genera Cacaoporus and Acyanoboletus, thus broadening the concept of Cacaoporus to encompass all of them. The emerging 3C perspective of Cupreoboletus, Cacaoporus and Cyanoboletus offers a remarkably consistent morphological diagnosis, overcoming the problems of a too broad concept for Cyanoboletus. This work reveals that Boletus neotropicus, B. novae-zelandiae and B. sensibilis belong respectively in Cyanoboletus, Cacaoporus and Lanmaoa, and by studying multigene alignment concatenates it identifies lineages that probably represent undescribed species: at least four in Cacaoporus and at least five in Cyanoboletus. Diagnostic tables and dichotomic keys are presented by geographic region. The present work also includes a study of the phylogenetic position of Neoboletus flavosanguineus, a species once classified in Cyanoboletus. The complexity of assigning species epithets in some lineages is addressed, namely for the boundaries between Cacaoporus instabilis and Ca. fagaceophilus as well as the diversity under the names Cyanoboletus sinopulverulentus and Cy. pulverulentus. The overall picture of evolutionary lineages sets a framework for the choice of reference data that can provide, in future phylogenetic studies that involve the 3C, a balanced and efficient coverage. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=197 SRC="FIGDIR/small/724631v1_ufig1.gif" ALT="Figure 1"> View larger version (23K): org.highwire.dtl.DTLVardef@7f618corg.highwire.dtl.DTLVardef@dd6a14org.highwire.dtl.DTLVardef@5f7399org.highwire.dtl.DTLVardef@9e7443_HPS_FORMAT_FIGEXP M_FIG C_FIG

In this study, I implemented an agent-based model aimed at exploring the competition between plastic and non-plastic genotypes in a digital environment with periodic fluctuations in food resources, using the dimorphic nematode Pristionchus pacificus as a proxy for mixed-strategy systems showing a combination of stochastic and conditional phenotype production. Emerging behaviours generated in response to variation in three main variables, that is, i) intrinsic cost of plasticity, ii) timescale of environmental fluctuation, and iii) degree of plasticity, were monitored in terms of frequency and time to fixation of two alleles of the developmental switch gene eud-1, one enabling mouth-form dimorphism and predation, the other leading to the constitutive expression of a single, bacterivorous morph. Interestingly, while intermediate-to-long periods of environmental stability (in terms of generations) and a higher level of plasticity might favour the evolution of plastic strategies in a "cost-free" condition, the introduction and increase of inherent costs of plasticity could make pure bet-hedging more advantageous, induce a sequential collapse in the frequency of fixation of plastic strains and time of coexistence between strains, and make the invasion by non-plastic mutants more likely until a plateau is reached. In addition, asymmetries in fitness between the two morphs might be an almost necessary condition to enable the invasion of a non-plastic population by plastic genotypes. Collectively, while confirming some previous theoretical findings, these outcomes could uncover the sensitivity of a mixed-strategy system to even small changes in key variables, suggesting the existence of phase transitions and critical evolutionary constraints.

BackgroundSynonymous mutations, once considered neutral, can affect translation efficiency through mRNA folding and splicing, generating codon usage bias. This bias is often linked to genomic GC content, which also influences gene regulation. In the parasitoid wasp Lysiphlebus fabarum, GC content was previously shown to shift between developmental stages, with larvae showing higher GC than adults. Whether this phenomenon is widespread among insects remains unknown. ResultsTranscriptomic data from six insect species spanning Diptera, Hymenoptera, and Lepidoptera was used to compare GC content between expressed genes in larvae and adults. In five species, larval transcripts exhibited higher GC content than adult transcripts. Differential expression analysis revealed that stage-biased genes displayed consistent GC shifts, and orthologous gene families with representatives across species showed particularly GC-rich larval-biased genes in Hymenoptera and Diptera. At the genome scale, modeling in 317 insect species demonstrated an association between parasitic lifestyle and reduced mean GC content in Hymenoptera and Diptera, providing a possible ecological explanation for AT-rich genomes. ConclusionsOur results show that GC content is dynamic across developmental stages, independent of overall genome composition. Stage-specific GC enrichment may reflect adaptive codon usage optimizing translation during energetically demanding life-history stages such as larval development. Furthermore, the association between parasitism and reduced genomic GC highlights how ecological lifestyle might with genome content and evolution. Lastly, this work identifies candidate genes underlying stage-specific GC bias and provides new insights into the interplay between molecular evolution, development, and parasitic adaptation in insects.

Sharks exhibit extraordinary morphological diversity across a wide range of ecological niches, yet large-scale, high-resolution digital datasets of their internal anatomy remain limited. Here we present an open-access 3D shark anatomical repository derived from published X-ray computed tomography (CT) data, featuring manually segmented and systematically annotated models of the chondrocranium, visceral arches, axial skeleton, musculature, and viscera in standard STL format. The dataset comprises 117 individuals, representing 72 species across 25 families and all nine extant shark orders, with 115 full-body reconstructions and two head-only models. This open-access dataset offers a comprehensive resource for comparative anatomy, biomechanical simulations, evolutionary developmental biology and biomimetics research of extant sharks.

Life history strategies such as rapid growth and high population turnover rates observed in vertebrates have been thought to have emerged relatively late in evolution. However, very little direct evidence exists at the species level for early vertebrates. In this study, a large fossil collection of over 450 specimens of Protaspis spp, heterostracan agnathans from the Cottonwood Canyon Formation at Beartooth Butte, Wyoming, from the Early Devonian, was analyzed. Morphological observations, analysis of bone plate completeness, and length-frequency analyses using ELEFAN--commonly used in recent fish studies--were applied to reconstruct growth rates, cohort structure, and ontogenetic processes. Protaspis specimens exhibited a continuous growth series from juvenile to adult stages, and a clear cohort structure was identified from the length-frequency distributions. The ELEFAN analysis suggested a life-history characterized by rapid growth and a short life span, and these features remained consistent in subset analyses restricted by species or locality, confirming the robustness of the estimates. Furthermore, the integration of the dermal bone plates progressed during the late stages of ontogeny, revealing that the rapid growth during the juvenile stage preceded the completion of this defensive structure. Comparisons of their growth parameters with those of extant fishes show that Protaspis does not align with slow-growing, long-lived "living fossil" taxa, but instead clusters with small-bodied, fast-growing species. These findings suggest that life-history strategies involving rapid growth and high population turnover were already established in early jawless vertebrates, much earlier than previously assumed.

Cuticular hydrocarbons (CHCs) are essential for insect waterproofing, yet how they change seasonally in social insects remains poorly understood. Due to its distinct seasonal worker phenotypes (summer and winter bees) and diverse subspecies, the western honey bee (Apis mellifera) is an ideal model to study seasonal CHC plasticity across populations with distinct local adaptations. We performed a common garden experiment to investigate the seasonal plasticity in CHC profiles across five European subspecies (A. m. carnica, A. m. iberiensis, A. m. ligustica, A. m. macedonica, A. m. ruttneri). We compared the CHC composition of workers performing tasks inside ("in-hive") or outside ("out-hive") the colony during summer and winter. Notably, out-hive workers consistently exhibited more waterproofing CHC profiles compared to in-hive workers, regardless of season or subspecies. The persistence of this stereotypical task-related differentiation in long-lived winter bees, which largely lack an age-based division of labor, indicates a robust, age-independent regulatory mechanism linked to the environment faced by the workers rather than a simple response to seasonal desiccation pressure. Moreover, we demonstrate CHC seasonal plasticity for the first time in honey bees. However, these seasonal shifts in hydrocarbon classes and chain length were not uniform; they varied across subspecies and critically depended on the task the workers performed.

Hearing in Lepidoptera mediates a range of ecologically important behaviours, including mate communication, predator avoidance, and acoustic signalling. In moths, the evolution of predator-prey interactions with bats has further shaped hearing through a sensory arms race, with repeated co-option of auditory organs to detect and evade echolocating predators. Despite significant prior characterization of the neurophysiology and behaviour of hearing in moths, the genetic basis of hearing is poorly understood in most insects. In this study, we identify a core set of putative auditory genes in Lepidoptera using a combination of homology-based searches from Drosophila and evolutionary rate analyses. We find 56 genes present across all species and investigate whether gene copy number varies among non-hearing and hearing lineages and among 3 different ear types. We discovered seven genes associated with ear type and one with ear presence, but did not find significant losses in gene copy number in non-hearing species. We identified three genes (btv, Dnai2, and nompB) with strong evidence of selection in hearing clades and five genes with weaker evidence of selection. We discuss the potential roles of btv, nompB, and Dnai2 in ciliary transport and the aging of hair cells, as well as the possibility of actively amplified hearing. Our study serves as a primer and resource for further gene mining and functional testing of auditory genes in moths and other insects.

The presence of multiple discrete color patterns within a species has captivated evolutionary biologists for more than a century, especially when such polymorphism is confined to one sex. The brown anole Anolis sagrei exhibits a female-limited polymorphism in dorsal patterning, which is controlled by allelic variation at the autosomal gene CCDC170. Here, we present and test a threshold model that can explain why the polymorphism is female-limited. We hypothesize that allelic variation at the CCDC170 locus affects only female color pattern because this gene is co-expressed with its neighboring gene ESR1, highly expressed in female, but not male, embryos. By manipulating embryonic estradiol levels, we show that genetic males can be induced to express the polymorphism according to allelic variation at the CCDC170 locus, which is naturally masked by low expression levels of this gene. Inversely, treating genetic females with fadrozole, which depletes estradiol, leads to monomorphic patterns irrespective of genotype, as for natural males. Using RT-qPCR, we demonstrate that these effects are accompanied by a direct influence of estradiol and fadrozole on gene expression levels of CCDC170 and ESR1, thereby validating the threshold model. Our results suggest that the CCDC170-ESR1-locus is part of a mechanistic link between the morph-determining and the sex differentiation systems and provide a causal explanation for the developmental origin of a sex-limited color polymorphism.

We investigated the influence of small-scale topographic and associated microclimatic drivers on the elevational patterns of functional alpha and beta diversity in ant communities of eastern Himalayas. Ants were sampled along three elevational transects spanning [~]1000-2000 m on two nearby mountain slopes, but differing in aspect, inclination and summit height. Ant genera were quantified at the quadrat level, yielding an abundance of 3090 genus-quadrats (from 22577 individuals in 1215 quadrats), almost evenly split between the two mountain slopes. Each ant was assigned to guilds along functional axes of thermal niche, trophic position and competitive abilities. Comparative analyses of functional diversity profiles along the two mountain slopes revealed no differences in the composite multi-guild alpha and beta diversity but significant differences amongst individual guild profiles. Seven of the eleven functional guilds showed strong elevational trends along the steeper north-facing slope, compared to only two of eleven on the shallower south-facing slope. These results indicate that similar functional space can be maintained despite taxonomic turnover within guilds or shifts in their relative abundances highlighting the influence of fine-scale topographic variation on community assembly. Alternatively, these findings demonstrate that the pattern of multivariate functional diversity across elevation does not necessarily provide an understanding of changing composition of the communities therein.

Genetically identical cells grown in the same environment show variation in gene expression known as expression noise. Expression noise can be heritable and impact fitness, making it subject to natural selection. Increasing expression noise for the Saccharomyces cerevisiae TDH3 gene was shown to be beneficial in glucose-based media when mean TDH3 expression was far from the fitness optimum but deleterious when it was close to this optimum. Here, we show that growth on different carbon sources alters the effects of new mutations on TDH3 expression noise and examine the fitness effects of changing expression noise. In galactose-based media, we observed the same relationship between expression noise and fitness seen in glucose-based media, but in glycerol- and ethanol-based media, we observed the opposite relationship or no significant relationship, respectively. Using simulations of single-cell organisms, we found that these differences were most likely explained by environment-specific relationships between gene expression and fitness. We also found that, far from the optimum, the fitness effects of noise were greatest when expression was highly heritable between mother and daughter cells. The empirical observations and simulations reported in this study show how environments influence both the production of expression noise and its impacts on fitness.

O_LIIslands are priority areas for the study and conservation of biodiversity, as they frequently harbour distinct lineages. Yet they are particularly threatened by the global change. In the frame of an accelerated biodiversity crisis, many insular species risk disappearing before they are even discovered. C_LIO_LIFireflies, despite their ecological and cultural significance, remain poorly investigated, especially in temperate regions, where they have shown marked decline over the last decades. C_LIO_LIWe investigated firefly diversity across a Mediterranean biodiversity hotspot by genotyping 114 fireflies of the genus Luciola from 29 locations across the Tuscan Archipelago and the adjacent Italian peninsula and applied phylogenetic and species delimitation methods to characterise genetic differentiation and its geographic structure. C_LIO_LIWe found an unexpectedly high level of genetic differentiation in this area. Phylogenetic inference recovered six deeply divergent and geographically structured mitochondrial lineages: three restricted to Elba Island and three to the Italian Peninsula. Genetic divergence among these lineages ranged from 1.97% to 4.58%, values comparable to or exceeding those typically observed among distinct species. Accordingly, species delimitation methods consistently supported their status as distinct species. C_LIO_LIThe coexistence of three divergent lineages on Elba Island suggests a biogeographic scenario characterised by ancient island colonisations and possible in situ diversification. These findings reveal a previously unrecognised depth of evolutionary diversity in Italian fireflies and identify the Tuscan Archipelago as a priority area for future research on firefly evolution and conservation, emphasizing that fireflies are a major gap in our knowledge of insect biodiversity in Europe. C_LI