The definition of biological sex has become a renewed focus of societal debate, fuelled by the conflation of biological principles with discussions of human gender diversity. Here, we argue that conceptual clarity critically depends on separating these domains. Drawing on evolutionary theory and empirical evidence, we maintain that biological sex is best defined as a binary classification of male and female reproductive strategies rooted in anisogamy, characterised by the production of two discrete gamete types of different sizes. We stress that gamete size constitutes the ultimate criterion for biological sex and that this definition applies consistently across sexual systems, from separate-sexed species to hermaphrodites, irrespective of variation in karyotype, hormonal profile, somatic phenotype, or behaviour. Further, we emphasise that evolutionary insights offer a coherent explanation for recurring, though not universal, associations between biological sex and patterns of sex-specific selection, sexual dimorphism and parental care. We conclude that the definition of biological sex as a binary classification based on gamete size is a powerful scientific framework compatible with the diversity of sexual phenotypes found in anisogamous organisms and distinct from the concept of human gender.

Sexual selection is a cornerstone of evolutionary biology in animals but remains underexplored in plants, where empirical work is often disconnected from sexual selection theory. A major barrier to crosstaxonomic synthesis is the inconsistent definition of core concepts, especially mating success. We advocate for a unified, taxon-neutral three-phase framework of sexual selection (premating, postmating, and postfertilization), grounded in traditional practice and applicable to both animals and plants. Parallels and differences across taxa are highlighted, capturing universal processes alongside biological diversity. Traits common in plants and historically seen as incompatible with sexual selection—such as modularity, pollen dispersal by third-party vectors, and lack of physical contact between mates—do not preclude its action and instead offer new biological processes to study.

Inbreeding depression (ID)-the reduction in fitness with increasing parental relatedness-is classically attributed to the expression of recessive deleterious mutations in homozygous individuals. Yet, the assumption that ID can only arise from changes in genetic heterozygosity has rarely, if ever, been directly tested. To test this, we produced highly inbred lines (F = 0.99999997) of the freshwater snail Physa acuta and generated offspring that differed in parental relatedness (self-fertilization, sib or cousin matings) while their parents, produced by crossing two inbred lines, all shared an identical genome. Several fitness traits showed significant declines with increasing parental relatedness. These traits included juvenile survival, body size, and self-fertility, and the magnitude of their decline was equivalent to a substantial fraction of the ID observed in natural, genetically polymorphic populations of P. acuta. Individual-based simulations demonstrated that spontaneous mutation rates compatible with natural levels of ID are far too low to account for the magnitude of ID observed here. These findings suggest that non-genetic mechanisms, most plausibly involving heritable epigenetic changes, can generate ID even in genetically uniform populations. This challenges the long-standing view that ID arises exclusively from genetic homozygosity and highlights the need to investigate epigenetic contributions to ID.

Environmental heterogeneity across freshwater systems often promotes phenotypic variation among populations. Yet, the respective contributions of environmentally-induced and heritable variation to population differences are rarely known. We investigated the geographic distribution and morphological differentiation, and heritability of shell traits among populations of the freshwater lymnaeid snail Pectinidens diaphanus in Patagonia. Extensive field surveys across 193 freshwater sites revealed that P. diaphanus is the sole lymnaeid inhabiting southern Patagonia and occupies a broader range of lentic and lotic habitats than previously documented. While reproductive anatomical structures were conserved across populations, shell shape differed markedly among populations from contrasting habitat types, with population identity explaining nearly 50% of total shape variation. Snails from hydrologically unstable habitats (ponds and streams) exhibited more elongated shells and relatively smaller apertures than lagoon snails, a pattern consistent with functional responses to hydroperiod variability and desiccation risk. To further investigate whether this differentiation was heritable, we conducted a common-garden experiment across two generations. Shell shape differences between permanent- (lagoon) and temporary- (pond) habitat-derived populations persisted into the G2 generation reared under standardized laboratory conditions, indicating that the observed variation is not solely a response to local environmental conditions but includes a heritable component. Together, our findings demonstrate that P. diaphanus constitutes the sole lymnaeid across southern Patagonia, occupying a broader range than previously documented, and that populations show heritable shell differentiation potentially associated with contrasting freshwater habitats. By integrating large-scale biogeographic surveys with morphometric and experimental approaches, this study provides new insight into how habitat variation may contribute to ecological and evolutionary differentiation in freshwater gastropods.

Cryptic female choice - female-mediated bias in fertilization after mating - is well established in animals and can also occur in plants when multiple pollens compete on the same pistil. However, whether interactions between pollen and pistil tissues after pollen deposition contribute to this process remains unknown. Here, we experimentally test whether such interactions mediate cryptic female choice in the angiosperm Brassica rapa. We quantified fertilization success of pollen donors competing on the same pistil using paternity analyses, and in parallel, made semi-in vivo assays to measure pollen tubes trajectories emerging from the excised styles and growing toward unfertilized ovules for each donor-recipient pair. We show that pollen tube growth towards ovules predicts higher fertilization success under pollen competition. Thus, we document a previously unobserved mechanism of cryptic female choice based on physical interactions between male and female components of reproduction. In addition, different recipient plants favour different pollen donors, consistent with non-directional female choice. Plants with longer styles bias paternities more strongly towards the most successful pollen donor. Overall, our study demonstrates that interactions between pollen tubes and pistil tissues after pollen germination enable plants to bias paternity toward particular donors.

Cytoplasmic male sterility (CMS) originates from a mito-nuclear conflict where mitochondrial genes induce male sterility and nuclear genes restore male fertility in hermaphrodites. The first observation of CMS in animals was reported recently in the freshwater snail Physa acuta where it is associated with two extremes divergent mitotypes D and K. The D individuals are male-steriles while male fertility is restored by nuclear genes in K and are found mixed with the most common male-fertile N mitotype in natural populations (i.e. gynodioecy). We compared male and female fitness, growth rate and metabolism between the three mitotypes at two temperatures as this factor influences CMS in gynodioecious plants via alteration of mitochondrial functioning. Temperature did not affect male sterility which depended only on the mitotype and the presence of restorers. Our results provided evidence that CMS is beneficial to female fitness in the absence of restorers while it is costly in their presence, and furthermore driven by body mass, fulfilling a key theoretical condition for the long-term maintenance of gynodioecy. Fitness benefits and costs mediated by differences in body mass are enhanced at cold temperature, suggesting that the system dynamics may vary according to thermal conditions in nature.

Inbreeding depression (ID), the decline in fitness upon inbreeding, is thought to result from a decrease in genetic heterozygosity enhancing phenotypic effects of recessive deleterious mutations. However, emerging evidence suggests that mutations may not explain ID completely. In this study, we test whether ID can emerge even in contexts where genetic heterozygosity does not vary. To that end, highly inbred lines (F=0.99999997) of the freshwater snail Physa acuta were used to produce individuals with varying levels of parental relatedness (self-fertilization, sibling crosses, and cousin crosses), though with identical genomic heterozygosity. Several fitness traits declined significantly with increasing parental relatedness, a pattern characteristic of ID, and quantitatively representing a non-negligible fraction of the ID usually observed in natural, genetically diverse populations of Physa acuta. Individual-based simulations showed that mutation rates compatible with values of ID found in natural populations are way too low to generate as much ID as observed in our experiment. These findings are consistent with the hypothesis that epigenetic changes, in addition to mutations, could contribute to a rapid regeneration of ID and explain the persistence of detectable ID in sets of genetically identical individuals. ### Competing Interest Statement The authors have declared no competing interest.

Male fertility in plants is often controlled by the interaction between mitochondrial and nuclear genes. Some mitotypes confer cytoplasmic male sterility (CMS), making the individual male-sterile, unless the nuclear background contains alleles called restorers, which suppress the effects of CMS and restore the hermaphroditic phenotype. Restorers in cultivated crops are often alleles with strong and dominant effect, but in wild plants, data often suggest more complex systems. Here, we characterized the inheritance and specificity of restoration in a new CMS model, the freshwater snail Physa acuta. We explored two different populations: (i) a naive population, i.e., without contact with CMS in the past 80 generations, and (ii) a non-naive population, where CMS is present and largely restored. Although we found male fertility of individuals with CMS mitogenomes to be heritable in both contexts, this genetic determinism was of a different nature depending on population history. In naive populations not coevolved with CMS, the background variation may include alleles that happen to act as weak quantitative modifiers of the penetrance of CMS, while in populations coevolved with CMS, selection may have favoured, when such variants were available, the emergence of strong alleles with a dominant effect.

AbstractGenomic conflicts arise when different genes in a genome are selected for opposite phenotypic effects. One well-known conflict occurs in plants, between mitochondrial genes causing cytoplasmic male sterility (CMS) and their nuclear suppressors, called restorers of male fertility. The evolution of CMS-restorer polymorphisms has been modeled many times, but empirical validations remain indirect. Here we use a new biological model, a freshwater snail, to directly observe evolutionary trajectories. In this species, CMS-associated mitogenomes coexist with male-fertile ones in populations. Models predict such a coexistence when nuclear restorers make CMS mitogenomes less fit than male-fertile ones, thus preventing the fixation of CMS. During 11 generations of experimental evolution, we observed rapid decreases in the frequency of CMS mitogenomes in a restorer-rich nuclear background, with an estimated ∼20% fitness disadvantage, consistent with theoretical conditions for the maintenance of cytonuclear polymorphism. In parallel, in an ancillary experiment, eggs laid by isolated snails carrying CMS showed a reduced hatching rate. Although significant, this reduction did not reach 20%, suggesting that fitness differentials in populations are enhanced by competition or rely on unmeasured traits. Our study illustrates the speed at which evolution can proceed in the context of cytonuclear conflicts over sex allocation.

Biological invasions by phylogenetically and ecologically similar competitors pose an evolutionary challenge to native species. Cases of character displacement following invasions suggest that they can respond to this challenge by shifting their traits. However, the actual impact of such shifts on competition are seldom studied. Here, we study competition between two freshwater snails from Guadeloupe (French Antilles), the native Aplexa marmorata and the introduced Physa acuta. The former has responded to invasion by rapid life-history evolution towards earlier maturity, higher fecundity and higher juvenile survival, traits that might favor rapid population growth in a noncompetitive context, but not necessarily in a competitive one. We here observe negative impacts of competition by both species on each other, though P. acuta is dominant and over generations largely displaces A. marmorata from co-cultures. In addition, our experiments suggest that A. marmorata populations having experienced competition by P. acuta for sufficient time in nature, have evolved to become even less tolerant to it. Though apparently paradoxical, this result supports the hypothesis that rapid life-history evolution has allowed A. marmorata not to resist competition, but to avoid it by increasing its specialization into the colonizer lifestyle previously documented by long-term field surveys. This example illustrates how evolution, in accordance with metacommunity coexistence theory, sometimes takes other ways than specialization into distinct types of resources or habitats to ensure coexistence between related species inhabiting the same landscape.

There exists a controversy in the literature concerning the values of coercive and bias fields in antidots magnetic structures formed by a hexagonal network of nanoholes. The coercive fields (HC) and the exchange bias fields (∣HEXC∣) for antidots (deposited on ultrathin anodic aluminum oxide, namely, AAO) are either increased or diminished by comparison with the same magnetic nanostructures grown on continuous substrates (namely, CML). We propose to elucidate these debates by showing the importance of the easy axis of the magnetization, the direction of the applied magnetic field, the thicknesses of the layers, and the 3D-topology of nanoholes, as well as the magnetic and thermal history of the magnetic measurements. Here, biased Ta(5 nm)/Pt(5 nm)/Co(0.6 nm)/Fe50Mn50(X)/Ta(5 nm) antidots are investigated by extraordinary Hall effect measurements at 5 K, where X varies in the (0–5.5) nm range. The substrate consists in a hexagonal array of holes, described by the pair of (p,d) values, respectively, the period as the distance from center to center of two consecutive holes and the hole diameter. The dimensions of antidots are (p≈100 and d≈40 nm) for X=(2–5.5) nm, (p≈150 and d≈60 nm) for X=3.5 nm, and (p≈100 and d≈60 nm) for X=0. A continuous stack using Si/SiO2(100 nm) is used for comparison. HC and ∣HEXC∣ gradually increase when X is enhanced for both substrates, with nevertheless a weak decrease at high X for the continuous system. Perpendicular magnetic anisotropy is only observed for both unbiased samples, the X=2 nm continuous sample, and both X=5 nm samples that have undergone field cooling treatment from 500 to 5 K under −2 T. Usually, HC(AAO)>HC(CML), ∣HEXC(AAO)∣>∣HEXC(CML)∣, and ∣HA(AAO)∣<∣HA(CML)∣ (HA designating the anisotropy field). However, for certain conditions, as, for instance, for FC-procedures starting from high temperatures and/or strong magnetic field, other situations might be observed. A discussion pertaining to the amplitudes of HC, ∣HEXC∣ and the anisotropy field (∣HA∣) of continuous and discontinuous samples is given for our experimental results as well as for published data in the literature, in the light of structural characteristics (wedge-to-wedge distance, porosity, or coverage ratio). Such biased perpendicular antidots might be particularly used in specific nanomaterials devoted to spintronics.

Gynodioecy, the coexistence of hermaphrodites with females, often reflects conflicts between cytoplasmic male sterility (CMS) genes and nuclear genes restoring male fertility. CMS is frequent in plants and has been recently discovered in one animal: the freshwater snail, Physa acuta. In this system, CMS was linked to a single divergent mitochondrial genome (D), devoid of apparent nuclear restoration. Our study uncovers a second, novel CMS-associated mitogenome (K) in Physa acuta, demonstrating an extraordinary acceleration of molecular evolution throughout the entire K mitochondrial genome, akin to the previously observed pattern in D. This suggests a pervasive occurrence of accelerated evolution in both CMS-associated lineages. Through a 17-generation introgression experiment, we further show that nuclear polymorphisms in K-mitogenome individuals contribute to the restoration of male function in natural populations. Our results underscore shared characteristics in gynodioecy between plants and animals, emphasizing the presence of multiple CMS mitotypes and cytonuclear conflicts. This reaffirms the pivotal role of mitochondria in influencing male function and in generating genomic conflicts that impact reproductive processes in animals.

The pace of biological invasions has increased in recent decades, leading to multiple invasions and the potential dominance of invasive species, destabilizing local ecological networks. This provides opportunities to study new ecological species interactions, including predation. Tropical freshwaters have been particularly concerned by aquatic invasions and we focused here on the Martinique island (Lesser Antilles). We examined the predator-prey relationships involving invasive Thiarid snails (Tarebia granifera and Melanoides tuberculata) and the native Neritina punctulata, both confronted with a newcomer predator, the redclaw crayfish (Cherax quadricarinatus). We conducted several mesocosm experiments to assess the impact of crayfish predation on snail survival and the passive and active antipredator responses of snails. A first experiment indicated snail survival rates between 50% and 80%, depending on crayfish size and sex. Notably, there was a negative correlation between snail survival and male crayfish size and the predation method (shell crushing vs. "body sucking") varied with crayfish size. The second experiment suggested no refuge size for snails, with both very small (<5 mm) and very large (>5 mm) unable to escape predation, regardless of crayfish size (from 77 to 138 mm) or sex. Finally, we investigated the escape behavior of Thiarids regarding three crayfish cues. Melanoides tuberculata tend to bury in the substrate and T. granifera to climb up aquarium walls, what was expected from their shell morphologies, and both responding to crayfish cues within minutes. Overall, C. quadricarinatus proves to be an efficient snail predator with limited escape options for snails, potentially contributing to the decline of certain snail populations in Martinique. This omnivorous predator might impact other native species across different groups, including shrimps and fish. Our study underscores the urgent need for monitoring efforts, solidifying the redclaw crayfish reputation as a dangerous invasive species for freshwater macrobenthic faunas worldwide.

Under certain experimental conditions, the deposition of C60 molecules onto an atomically flat copper surface gives rise to the formation of corrugated islands. This corrugation, which reflects a molecular displacement perpendicular to the surface plane, presents an astonishing pattern: it is well described by a frustrated Ising spin Hamiltonian whose thermodynamics is compatible with a spin liquid about to transit towards an ordered zigzag state. Here we study the statistical properties of such a molecular corrugation using tools generally employed in frustrated magnetism. More specifically, the real and reciprocal space analysis of pairwise molecule correlations allows us to demonstrate that the C60/Cu system, in which magnetism is totally absent, has all the characteristics of a triangular Ising antiferromagnet. Our results indicate that the organization of two-dimensional matter, at the molecular length scale, sometimes turns out to be particularly close to that encountered in highly frustrated magnets.

Lymnaeid snails are simultaneous hermaphrodites that have a worldwide distribution, inhabiting freshwater areas from almost all continents ranging from tropical to arctic regions and from sea level to very high altitudes. In this chapter, we review the reproductive anatomy, behavioral and physiological traits, and mating strategies associated with increased survival and invasiveness of lymnaeids across different ecosystems around the globe. We also discuss the biotic and abiotic factors that can affect mating systems in this family, and how they have expanded their geographical range by natural, as well as human-mediated ways, likely promoting the spread of infectious diseases. Finally, we discuss why we believe that lymnaeids are suitable model organisms for studying mechanisms and processes involved in the ecology and evolution of mating systems and biological invasions.

Conspicuous body colours and colour polymorphism have been hypothesized to increase rates of speciation. Conspicuous colours are evolutionary labile, and often involved in intraspecific sexual signalling and thus may provide a raw material from which reproductive isolation can easily evolve, while polymorphism could favour rapid evolution of new lineages through morphic speciation. Here, we investigated the influence of the presence/absence of conspicuous colourations, and of colour polymorphism on the speciation of Lacertids. We used several state-dependent diversification models, and showed that, regardless of the methods, conspicuous colourations and colour polymorphism were not related to species speciation. While the lack of correlation between conspicuous colourations and speciation rates is in line with most of the literature testing this hypothesis, the results for colour polymorphism contradict previous studies, and question the generality of the morphic speciation hypothesis.

How the complexity of food webs depends on environmental variables is a long-standing ecological question. It is unclear though how food-chain length should vary with adaptive evolution of the constitutive species. Here we model the evolution of species colonisation rates and its consequences on occupancies and food-chain length in metacommunities. When colonisation rates can evolve, longer food-chains can persist. Extinction, perturbation and habitat loss all affect evolutionarily stable colonisation rates, but the strength of the competition-colonisation trade-off has a major role: weaker trade-offs yield longer chains. Although such eco-evo dynamics partly alleviates the spatial constraint on food-chain length, it is no magic bullet: the highest, most vulnerable, trophic levels are also those that least benefit from evolution. We provide qualitative predictions regarding how trait evolution affects the response of communities to disturbance and habitat loss. This highlights the importance of eco-evolutionary dynamics at metacommunity level in determining food-chain length.

When environmental conditions are unpredictable, expressing alternative phenotypes spreads the risk of failure, a mixed strategy called bet-hedging. In the southern part of its range, the Parsley Frog Pelodytes punctatus breeds both in autumn and in spring. Our aim was to study the breeding phenology and reproductive success associated with the use of those two seasonal niches to understand how this breeding strategy can be maintained. Field surveys revealed that breeding phenology was typically bimodal with a higher breeding effort in autumn. More importantly, in spring, the survival rate of offspring was severely reduced by the presence of autumn tadpoles, indicating a clear priority effect. However, the autumn cohort often failed to survive over winter, in which case spring cohorts were often successful. Based on those results, we constructed a model in which females can allocate a variable portion of eggs to each season and added a priority effect. We conclude that the existence of the two breeding seasons may indeed constitute a bet-hedging strategy.