Showing papers in "Biological Journal of The Linnean Society in 2019"

High-density three-dimensional morphometric analyses support conserved static (intraspecific) modularity in caecilian (Amphibia: Gymnophiona) crania

Abstract: Ashleigh F. Marshall, Carla Bardua, David J. Gower, Mark Wilkinson, Emma Sherratt and Anjali Goswami

Raising your voice: evolution of narrow-band high-frequency signals in toothed whales (Odontoceti)

Abstract: 1Department of Bioscience, Aarhus University, 4000 Roskilde, Denmark 2Evolutionary Genomics, Natural History Museum of Denmark, University of Copenhagen, 2100 Copenhagen O, Denmark 3Museum of Southern Jutland, Section of Natural History, 6510 Gram, Denmark 4W. M. Keck Science Department, Claremont McKenna, Pitzer, and Scripps Colleges, Claremont, CA, 91711, USA 5The Dinosaur Institute, Natural History Museum of Los Angeles County, Los Angeles, CA 90007, USA 6Department of Earth and Environmental Sciences, Vanderbilt University, Nashville, TN 37240, USA 7School of Geographical Sciences, University of Bristol, Bristol BS8 1TH, UK 8Met Office Hadley Centre, Exeter EX1 3PB, UK 9Department of Biology, University of Southern Denmark, 5230 Odense, Denmark

Nematodes as evolutionary commuters between marine, freshwater and terrestrial habitats

Abstract: Nematodes are the only major metazoan group which is persistently abundant and diverse across marine, freshwater and terrestrial ecosystems. This could be the result of a few major habitat transitions followed by extensive diversification, or numerous habitat transitions followed by moderate diversification. To pinpoint habitat transitions, we superposed nematode habitat associations on an available phylum-wide phylogenetic tree based on small subunit ribosomal DNA sequences (≈2730 SSU rDNA sequences covering ≈1750 nematode taxa). Our analysis revealed at least 30 major habitat transitions within the phylum Nematoda. These transitions as well as their directionality were unevenly spread over the 12 major clades. Most transitions reside in Clades 1–6, and these transitions are bidirectional. Members of Clades 8–12 showed five full transitions, and these took place exclusively from terrestrial to marine systems. We relate our results to the distinct secretory–excretory systems in Clades 1–6 and Clades 8–12, as well as to differences in water permeability of the nematode cuticle. Hence, the phylum Nematoda is characterized by a relatively large number of habitat transitions followed by moderate diversification. The identification of multiple habitat transitions at a low taxonomic level will facilitate future investigations into the mechanisms underlying this unusual ecological flexibility.

The repeated evolution of dental apicobasal ridges in aquatic-feeding mammals and reptiles

Abstract: 1Australian Museum Research Institute, Sydney, NSW 2010, Australia 2PANGEA Research Centre, School of Biological, Earth and Environmental Sciences, University of New South Wales, Sydney, NSW 2052, Australia 3School of Biological Sciences, Monash University, Melbourne, VIC 3800, Australia 4Geosciences, Museums Victoria, Melbourne, VIC 3001, Australia 5School of Engineering, University of Newcastle, Newcastle, NSW 2308, Australia 6School of Environmental and Life Sciences, University of Newcastle, Newcastle, NSW 2308, Australia 7Department of Anatomy and Developmental Biology, Monash University, Melbourne, VIC 3800, Australia 8Australian Centre for Neutron Scattering, Australian Nuclear Science and Technology Organisation, Sydney, NSW 2234, Australia 9Paleobiology, National Museum of Natural History, Smithsonian Institution, Washington, DC 20560, USA 10Mammalogy and Paleontology, Burke Museum of Natural History and Culture, University of Washington, Seattle, WA 98195, USA

Macroevolutionary trends of brain mass in Primates

Abstract: A distinctive trait in primate evolution is the expansion in brain mass. The potential drivers of this trend and how and whether encephalization influenced diversification dynamics in this group are hotly debated. We assembled a phylogeny accounting for 317 primate species, including both extant and extinct taxa, to identify macroevolutionary trends in brain mass evolution. Our findings show that Primates as a whole follow a macroevolutionary trend for an increase in body mass, relative brain mass and speciation rate over time. Although the trend for increased encephalization (brain mass) applies to all Primates, hominins stand out for their distinctly higher rates. Within hominins, this unique trend applies linearly over time and starts with Australopithecus africanus. The increases in both speciation rate and encephalization begin in the Oligocene, suggesting the two variables are causally associated. The substitution of early, stem Primates belonging to plesiadapiforms with crown Primates seems to be responsible for these macroevolutionary trends. However, our findings also suggest that cognitive capacities favoured speciation in hominins.

Cloudy with a chance of speciation: integrative taxonomy reveals extraordinary divergence within a Mesoamerican cloud forest bird

Abstract: This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs licence (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial reproduction and distribution of the work, in any medium, provided the original work is not altered or transformed in any way, and that the work is properly cited. For commercial re-use, please contact journals.permissions@oup.com Cloudy with a chance of speciation: integrative taxonomy reveals extraordinary divergence within a Mesoamerican cloud forest bird

The Extended Evolutionary Synthesis: what is the debate about, and what might success for the extenders look like?

Abstract: Debate over the Extended Evolutionary Synthesis (EES) ranges over three quite different domains of enquiry. Protagonists are committed to substantive positions regarding (1) empirical questions concerning (for example) the properties and prevalence of systems of epigenetic inheritance; (2) historical characterizations of the modern synthesis; and (3) conceptual/philosophical matters concerning (among other things) the nature of evolutionary processes, and the relationship between selection and adaptation. With these different aspects of the debate in view, it is possible to demonstrate the range of cross-cutting positions on offer when well-informed evolutionists consider their stance on the EES. This overview of the multiple dimensions of debate also enables clarification of two philosophical elements of the EES debate, regarding the status of niche-construction and the role of selection in explaining adaptation. Finally, it points the way to a possible resolution of the EES debate, via a pragmatic approach to evolutionary enquiry.

Lizard nest environments differ between suburban and forest habitats

Abstract: Nesting success is critical for oviparous species to maintain viable populations. Many species often do not provide parental care (e.g. oviparous reptiles), so embryos are left to develop in the prevailing conditions of the nest. For species that occupy diverse habitats, embryos must be able to complete development across a broad range of environmental conditions. Although much research has investigated how environmental conditions influence embryo development, we know little about how nest conditions differ between diverse habitats. Anolis lizards are commonly found in various habitats including those heavily modified by humans (e.g. cities). We describe nest sites of anoles in two different habitat types: a suburban area and a nearby forest. The suburban area had less total nesting habitat but a greater variety of microenvironment conditions for females to use for nesting, compared to the forest. Suburban nests were warmer and drier with greater thermal variance compared to forest nests. Finally, we use data from the literature to predict how nest conditions may influence development. Our study provides the first quantitative assessment of anole nest sites in human-modified environments and shows how suburban habitats may generate variation in developmental rate.

Spatiotemporal variation in thermal niches suggests lability rather than conservatism of thermal physiology along an environmental gradient

Abstract: Temperature variation throughout a species range can be extensive and exert divergent spatiotemporal patterns of selection. The estimation of phenotypic differences of populations along environmental gradients provides information regarding population-level responses to changing environments and evolutionary lability in climate-relevant traits. However, few studies have found physiological differentiation across environmental gradients attributable to behavioural thermoregulation buffering physiological evolution. Here, we compared thermal sensitivity of physiological performance among three populations of the ornate tree lizard (Urosaurus ornatus) along a 1100 m elevational gradient in southeastern Arizona across years in order to determine whether spatial differences in thermal environments are capable of driving local physiological differentiation. Lizards exhibited significant population-level differences in thermal physiology. The thermal traits of lizards at low elevations included warmer body temperatures and higher preferred and critical thermal temperatures. In contrast, lizards at higher elevations had cooler body temperatures and lower preferred and critical thermal temperatures. Populations also exhibited differences in the optimal temperature for performance and thermal performance breadth. The direction of population variation was consistent across years. Environmental gradients can provide model systems for studying the evolution of thermal physiology, and our study is one of the first to suggest that population differentiation in thermal physiology could be more prominent than previously thought.

A simple skeletal measurement effectively predicts climbing behaviour in a diverse clade of small mammals

Abstract: Arboreal locomotion allows access to above-ground resources and might have fostered the diversification of mammals. Nevertheless, simple morphological measurements that consistently correlate with arboreality remain indefinable. As such, the climbing habits of many species of mammals, living and extinct, remain speculative. We collected quantitative data on the climbing tendencies of 20 species of murine rodents, an ecologically and morphologically diverse clade. We leveraged Bayesian phylogenetic mixed models (BPMMs), incorporating intraspecific variation and phylogenetic uncertainty, to determine which, if any, traits (17 skeletal indices) predict climbing frequency. We used ordinal BPMMs to test the ability of the indices to place 48 murine species that lack quantitative climbing data into three qualitative locomotor categories (terrestrial, general and arboreal). Only two indices (both measures of relative digit length) accurately predict locomotor styles, with manus digit length showing the best fit. Manus digit length has low phylogenetic signal, is largely explained by locomotor ecology and might effectively predict locomotion across a multitude of small mammals, including extinct species. Surprisingly, relative tail length, a common proxy for locomotion, was a poor predictor of climbing. In general, detailed, quantitative natural history data, such as those presented here, are needed to enhance our understanding of the evolutionary and ecological success of clades.

Allopatric divergence and secondary contact with gene flow: a recurring theme in rattlesnake speciation

Abstract: The study of recently diverged lineages whose geographical ranges come into contact can provide insight into the early stages of speciation and the potential roles of reproductive isolation in generating and maintaining species. Such insight can also be important for understanding the strategies and challenges for delimiting species within recently diverged species complexes. Here, we use mitochondrial and nuclear genetic data to study population structure, gene flow and demographic history across a geographically widespread rattlesnake clade, the western rattlesnake species complex (Crotalus cerberus, Crotalus viridis, Crotalus oreganus and relatives), which contains multiple lineages with ranges that overlap geographically or contact one another. We find evidence that the evolutionary history of this group does not conform to a bifurcating tree model and that pervasive gene flow has broadly influenced patterns of present-day genetic diversity. Our results suggest that lineage diversity has been shaped largely by drift and divergent selection in isolation, followed by secondary contact, in which reproductive isolating mechanisms appear weak and insufficient to prevent introgression, even between anciently diverged lineages. The complexity of divergence and secondary contact with gene flow among lineages also provides new context for why delimiting species within this complex has been difficult and contentious historically.

Escape dynamics of free-ranging desert kangaroo rats (Rodentia: Heteromyidae) evading rattlesnake strikes

Abstract: 1Department of Biology, San Diego State University, 5500 Campanile Drive, San Diego, CA 92182, USA 2Department of Evolution, Ecology, and Organismal Biology, University of California, Riverside, 900 University Avenue, Riverside, CA 92521, USA 3Graduate Group in Ecology, University of California, Davis, 1 Shields Avenue, Davis, CA 95616, USA 4Chiricahua Desert Museum, 4 Rattlesnake Canyon Road, Rodeo, NM 88056, USA

Extreme genetic structure and dynamic range evolution in a montane passerine bird: implications for tropical diversification

Abstract: Aim: Employ phylogeographic analyses of a widespread species complex to examine the role of historical and evolutionary processes in the origin and maintenance of high species diversity in the Neotropical montane region. Location: Neotropical highlands. Taxon: Henicorhina wood-wrens (Aves, Troglodytidae). Methods: We collected mtDNA sequence data for 288 individuals thoroughly covering the range of the Henicorhina leucophrys complex from Mexico to Bolivia. Sequences were employed to characterize population structure, infer phylogenetic relationships among populations and their divergence times, examine lineage accumulation through time, and identify presumptive species using coalescent methods. We also explored the origin of elevational and latitudinal replacements involved in spatial changes in species assemblages in the Andes. Results: We found remarkable genetic structure within the complex, which consists of numerous lineages reaching >12% sequence divergence; most divergent populations occur in areas separated by topographic barriers but several of them, typically not sister to each other, co-occur with elevational segregation on mountain slopes or replace each other with latitude along the Andes. Some close relatives occur in areas separated by thousands of kilometers, with more distant relatives occupying intervening areas. The complex likely originated in the Mexican highlands and expanded extensively in South America while diverging rapidly at a constant rate into many different lineages which have persisted for millions of years. Coalescent analyses consistently revealed that the complex may comprise more than 30 species; while we do not suggest these presumptive species should be recognized by taxonomists in the absence of additional data, H. leucophrys is a distant outlier among New World birds in terms of high lineage diversity within a single recognized species. Main Conclusions: Our study captured wood-wren lineages in the act of building up diversity via divergence and persistence in allopatry, achievement of secondary sympatry, and coexistence at the landscape scale mediated by ecological and evolutionary divergence. Although dispersal by wood-wrens is restricted at present and this likely accounts for strong population structure across topographic barriers, their ranges have been dynamic, managing to disperse over much of the montane Neotropics. Phases of expansion and contraction of ranges and localized extinctions of populations likely account for phylogeographic patterns which are precursors to the origin of new species and the accumulation of diversity in tropical mountains.

Manual skills for food processing by mountain gorillas (Gorilla beringei beringei) in Bwindi Impenetrable National Park, Uganda

Abstract: Although gorillas rarely use tools in the wild, their manipulative skills during plant processing may be similar to those of other tool-using great apes. Virunga mountain gorillas are known for the complexity in their methods of thistle and nettle plant preparation in the wild. However, there has been no comparable data on food processing in the population of mountain gorillas from the Bwindi Impenetrable National Park, Uganda. We investigated the manual actions and hand grips used when accessing edible parts of two hard-to-process plants defended by stinging hairs, epidermis or periderm (i.e., peel of Urera hypselodendron and pith of Mimulopsis arborescens) and one undefended plant (i.e., leaves of Momordica foetida) in 11 Bwindi wild mountain gorillas (Gorilla beringei beringei) using video records ad libitum. Similar to thistle feeding by Virunga gorillas, Bwindi gorillas used the greatest number of manual actions for the most hard-to-process plant (U. hypselodendron), the actions were ordered in several key stages and organised hierarchically. The demands of processing plant material elicited 19 different grips and variable thumb postures, of which three grips were new and 16 grips have either been previously reported or show clear similarities to grips used by other wild and captive African apes and humans. Moreover, our study only partly supports a functional link between diet and hand morphology in mountain gorillas and suggests that the gorilla hand is best adapted to forceful grasping that is required for both manipulation and arboreal locomotion.

Evolution of the eyes of vipers with and without infrared-sensing pit organs

Abstract: We examined lens and brille transmittance, photoreceptors, visual pigments, and visual opsin gene sequences of viperid snakes with and without infrared-sensing pit organs. Ocular media transmittance is high in both groups. Contrary to previous reports, small as well as large single cones occur in pit vipers. Non-pit vipers differ from pit vipers in having a twotiered retina, but few taxa have been examined for this poorly understood feature. All vipers sampled express rh1, sws1 and lws visual opsin genes. Opsin spectral tuning varies but not in accordance with the presence/absence of pit organs, and not always as predicted from gene sequences. The visual opsin genes were generally under purifying selection, with positive selection at spectral tuning amino acids in RH1 and SWS1 opsins, and at retinal pocket stabilization sites in RH1 or LWS (and without substantial differences between pit and nonpit vipers). Lack of evidence for sensory trade-off between viperid eyes (in the aspects examined) and pit organs might be explained by the high degree of neural integration of vision and infrared detection; the latter representing an elaboration of an existing sense with addition of a novel sense organ, rather than involving the evolution of a wholly novel sensory system.

The early wasp plucks the flower : Disparate extant diversity of sawfly superfamilies (Hymenoptera: 'Symphyta') may reflect asynchronous switching to angiosperm hosts

Abstract: The insect order Hymenoptera originated during the Permian nearly 300 Mya. Ancestrally herbivorous hymenopteran lineages today make up the paraphyletic suborder 'Symphyta', which encompasses c. 8200 species with very diverse host-plant associations. We use phylogeny-based statistical analyses to explore the drivers of diversity dynamics within the 'Symphyta', with a particular focus on the hypothesis that diversification of herbivorous insects has been driven by the explosive radiation of angiosperms during and after the Cretaceous. Our ancestral-state estimates reveal that the first symphytans fed on gymnosperms, and that shifts onto angiosperms and pteridophytes - and back - have occurred at different time intervals in different groups. Trait-dependent analyses indicate that average net diversification rates do not differ between symphytan lineages feeding on angiosperms, gymnosperms or pteridophytes, but trait-independent models show that the highest diversification rates are found in a few angiosperm-feeding lineages that may have been favoured by the radiations of their host taxa during the Cenozoic. Intriguingly, lineages-through-time plots show signs of an early Cretaceous mass extinction, with a recovery starting first in angiosperm-associated clades. Hence, the oft-invoked assumption of herbivore diversification driven by the rise of flowering plants may overlook a Cretaceous global turnover in insect herbivore communities during the rapid displacement of gymnosperm- and pteridophyte-dominated floras by angiosperms. (Less)

Phenotypic and genetic divergence in reed frogs across a mosaic hybrid zone on São Tomé Island

Abstract: Although naturally heterogeneous environments can lead to mosaic hybrid zones, human-induced habitat fragmentation can also lead to environmental heterogeneity and hybridization. Here we quantify phenotypic and molecular divergence across a reed frog mosaic hybrid zone on São Tomé Island as a first step towards understanding the consequences of hybridization across this heterogeneous landscape. The São Tomé giant reed frog (Hyperolius thomensis) is strongly tied to cool, wet, forest habitats whereas the distribution of Moller’s reed frog (H. molleri) spans cool, wet, forests to warm, dry, disturbed habitats. Correspondingly, hybridization is concentrated in the more forested, cool, wet sites relative to non-forested, warmer, drier habitats. Four of six sites with hybrid frogs are artificial water bodies near the forest edge, indicating that both breeding habitat and broader scale environmental variation are probably important for understanding interspecific interactions and the extent of hybridization in this system. Phenotypic variation (body size and ventral coloration) largely tracks genetic and environmental variation across the hybrid zone with larger and more pigmented frogs occurring in forested, cool, wet habitats. Understanding whether human-induced changes in habitat break down reproductive barriers will be essential for conservation management of the less abundant, forest-associated H. thomensis in the face of rampant hybridization.

Functional diversity in biters: the evolutionary morphology of the oral jaw system in pacus, piranhas and relatives (Teleostei: Serrasalmidae)

Abstract: Serrasalmid fishes form a highly specialized group of biters that show a large trophic diversity, ranging from pacus able to crush seeds to piranhas capable of cutting flesh. Their oral jaw system has been hypothesized to be forceful, but variation in bite performance and morphology with respect to diet has not previously been investigated. We tested whether herbivorous species have higher bite forces, larger jaw muscles and more robust jaws than carnivorous species. We measured in vivo and theoretical bite forces in 27 serrasalmid species. We compared the size of the adductor mandibulae muscle, the jaw mechanical advantages, the type of jaw occlusion, and the size and shape of the lower jaw. We also examined the association between bite performance and functional morphological traits of the oral jaw system. Contrary to our predictions, carnivorous piranhas deliver stronger bites than their herbivorous counterparts. The size of the adductor mandibulae muscle varies with bite force and muscles are larger in carnivorous species. Our study highlights an underestimated level of functional morphological diversity in a fish group of exclusive biters. We provide evidence that the trophic specialization towards carnivory in piranhas results from changes in the configuration of the adductor mandibulae muscle and the lower jaw shape, which have major effects on bite performance and bite strategy.

Off-target capture data, endosymbiont genes and morphology reveal a relict lineage that is sister to all other singing cicadas

Abstract: Phylogenetic asymmetry is common throughout the tree of life and results from contrasting patterns of speciation and extinction in the paired descendant lineages of ancestral nodes. On the depauperate side of a node, we find extant ‘relict’ taxa that sit atop long, unbranched lineages. Here, we show that a tiny, pale green, inconspicuous and poorly known cicada in the genus Derotettix, endemic to degraded salt-plain habitats in arid regions of central Argentina, is a relict lineage that is sister to all other modern cicadas. Nuclear and mitochondrial phylogenies of cicadas inferred from probe-based genomic hybrid capture data of both target and non-target loci and a morphological cladogram support this hypothesis. We strengthen this conclusion with genomic data from one of the cicada nutritional bacterial endosymbionts, Sulcia, an ancient and obligate endosymbiont of the larger plant-sucking bugs (Auchenorrhyncha) and an important source of maternally inherited phylogenetic data. We establish Derotettiginae subfam. nov. as a new, monogeneric, fifth cicada subfamily, and compile existing and new data on the distribution, ecology and diet of Derotettix. Our consideration of the palaeoenvironmental literature and host-plant phylogenetics allows us to predict what might have led to the relict status of Derotettix over 100 Myr of habitat change in South America.

Marsupial versus placental: assessing the evolutionary changes in the scapula of didelphids and sigmodontines

Abstract: It is not a new concept that marsupials and placentals are distant and distinct clades among mammals. In South America, these animals coexist, occupy similar niches and, in some cases, are similar in appearance. This is especially true with respect to the locomotor categories of smaller rodents belonging to the family Cricetidae or, more specifically, the subfamily Sigmodontinae, compared with the marsupials of the Didelphidae family. In this study, we have investigated both the similarities and the differences between the two clades by examining locomotion-dependent adaptation, a crucial survival mechanism that has affected the morphology of both clades. We applied geometric morphometrics to quantify the shape of the scapula, which is a very adaptable structure. We found similar morphological adaptations between the clades, especially with respect to adaptation to life in trees. Moreover, Didelphidae are influenced by phylogenetic history to a greater extent than Sigmodontinae with regard to variation of scapula shape and allometry. These differences can be explained by the greater degree of body size variation that exists within the Didelphidae. Didelphidae have an ancient evolutionary history in South America compared with the Sigmodontinae, which have undergone a very successful and rapid diversification more recently.

Echiniscus virginicus complex: the first case of pseudocryptic allopatry and pantropical distribution in tardigrades

Abstract: Mainly because of the problems with species delineation, the biogeography of microscopic organisms is notoriously difficult to elucidate. In this contribution, variable nuclear and mitochondrial DNA markers were sequenced from individual specimens representing the Echiniscus virginicus complex that are morphologically indistinguishable under light microscopy (five populations from the temperate Eastern Nearctic and 13 populations from the subtropical and tropical zone). A range of methods was used to dissect components of variability within the complex (Bayesian inference, haplotype networks, Poisson tree processes, automatic barcode gap discovery delineations, principal components analysis and ANOVA). We found deep divergence between the temperate Eastern Nearctic E. virginicus and pantropical Echiniscus lineatus in all three genetic markers. In contrast, intraspecific genetic variation was very low, regardless of the geographical distance between the populations. Moreover, for the first time, statistical predictions of tardigrade geographical distributions were modelled. The factor determining the allopatric geographical ranges of deceptively similar species analysed in this study is most likely to be the type of climate. Our study shows that widespread tardigrade species exist, and both geographical distribution modelling and the genetic structure of populations of the pantropical E. lineatus suggest wind-mediated (aeolian) passive long-distance dispersal.

Life history and age-dependent mortality processes in tropical reptiles

Abstract: Actuarial senescence appears to be a common process, and senescence patterns are highly variable across the tree of life. To date, studies on animal senescence have largely focused on model species, such as as fruit flies, humans and a few other endotherms. In contrast, our knowledge about ageing remains fragmentary in ectotherm vertebrates, such as reptiles. Here, we examined life history and age-dependent mortality patterns in three tropical tortoises (Kinixys erosa, Kinixys homeana and Kinixys nogueyi) and snakes (Bitis gabonica, Bitis nasicornis and Causus maculatus). Our study revealed that tortoises of the genus Kinixys had a higher survival and a lower recruitment than snakes of the genera Bitis and Causus, indicating a slower life history. Furthermore, we confirmed that survival decreased more slowly with age in tortoises than in snakes. In addition, we highlighted contrasting patterns of age-dependent mortality among the three genera. In Kinixys, the relationship between mortality rate and age was positive and linear, suggesting gradual senescence over tortoise lifetime. In contrast, the relationship between mortality rate and age was negative and sharp in Bitis and Causus, possibly owing to a ‘negative senescence’. Our study is one of the few to have examined and compared the demography and age-dependent mortality patterns of tropical reptiles. Among other things, our results suggest that although negative senescence has never been reported in endotherm vertebrates, it could be a common phenomenon in ectotherms.

Reaching the edge of the speciation continuum: hybridization between three sympatric species of Hyla tree frogs

Abstract: Alloparapatric species meeting in secondary contact zones are evolutionary witnesses to how reproductive isolation progresses over time and space. Western Palearctic tree frogs (Hyla) are phenotypically similar and all the species pairs tested can hybridize and eventually admix at range margins. All except one. The early-diverged Hyla meridionalis exhibits sharp phenotypic differences: a “long” breeding call and the absence of a lateral stripe. In southwestern Europe, this species co-occurs with the “short-call” striped tree frogs H. arborea and H. molleri, two expanding lineages that admix at their parapatric margins. We estimated local gene flow between these three taxa at several syntopic breeding sites in western France. We congruently matched genotypes to phenotypes: the “short-call” striped individuals were a nuclear mixture of H. arborea and H. molleri; the “long-call” stripeless individuals all featured pure H. meridionalis nuclear ancestry and mtDNA, confirming complete genetic isolation from H. arborea/molleri. Yet, we documented an F1 hybrid between a female H. arborea/molleri and a male H. meridionalis: an incompletely-striped male with an intermediate breeding call. These findings suggest H. meridionalis is still able to reproduce with parapatric congeneric species despite 20My of divergence and strong phenotypic differentiation, but that intrinsic incompatibilities (sterility) prevent genetic introgression.