Premating Isolation Research Articles

Contrast and Genomic Characterisation of Ancient and Recent Interspecific Introgression Between Deeply Diverged Moustache Toads (Leptobrachium).

Recent genomic analyses have provided new insights into the process of interspecific introgression and its consequences on species evolution. Most recent studies, however, focused on hybridization between recently radiated species, with few examining the genomic outcomes of ancient hybridization across deeply diverged species. Using whole genome data of moustache toads (Leptobrachium), we identified signals of three hybridization events among nine species that diverged at the Eocene. An ancient introgression from L. leishanense to the ancestral branch (C1) of L. liui introduced adaptive variants. The highly introgressed regions include genes with important functions in odorant detection and immune responses. These genes are preserved in all three descendent populations of L. liui_C1, and these regions likely have been positively selected over a long filtering process. A recent introgression occurred from L. huashen to L. tengchongense, with the introgressed regions being mostly neutral. Furthermore, one F1 hybrid individual was detected between sympatric L. ailaonicum and L. promustache. The signals of introgression largely disappeared after removing the hybrid individual, indicating an occasional hybridization but minimal introgression. Further examination of highly divergent but low introgressed genomic regions revealed both pre-mating isolation and genetic incompatibility as potential mechanisms of resisting introgression and maintaining species boundaries. Additionally, no large X-effect was found in these introgression events. Hybridization between deeply diverged amphibian species may be common, but detectable introgressions are likely less so, with recent introgression being mostly neutral and the rare ancient one potentially adaptive. Our findings complement recent genomic work, and together they provide a better understanding of the genomic characteristics of interspecific introgression and its significance in species adaptation and evolution.

A consequential one-night stand: Episodic historical hybridization leads to mitochondrial takeover in sympatric desert ant-eating spiders

Disentangling the genomic intricacies underlying speciation and the causes of discordance between sources of evidence can offer remarkable insights into evolutionary dynamics. The ant-eating spider Zodarion nitidum, found across the Middle East and Egypt, displays yellowish and blackish morphs that co-occur sympatrically. These morphs additionally differ in behavioral and physiological features and show complete pre-mating reproductive isolation. In contrast, they possess similar sexual features and lack distinct differences in their mitochondrial DNA. We analyzed both Z. nitidum morphs and outgroups using genome-wide and additional mitochondrial DNA data. The genomic evidence indicated that Yellow and Black are reciprocally independent lineages without signs of recent admixture. Interestingly, the sister group of Yellow is not Black but Z. luctuosum, a morphologically distinct species. Genomic gene flow analyses pinpointed an asymmetric nuclear introgression event, with Yellow contributing nearly 5 % of its genome to Black roughly 320,000 years ago, intriguingly aligning with the independently estimated origin of the mitochondrial DNA of Black. We conclude that the blackish and yellowish morphs of Z. nitidum are long-diverged distinct species, and that the ancient and modest genomic introgression event registered resulted in a complete mitochondrial takeover of Black by Yellow. This investigation underscores the profound long-term effects that even modest hybridization events can have on the genome of organisms. It also exemplifies the utility of phylogenetic networks for estimating historical events and how integrating independent lines of evidence can increase the reliability of such estimations.

The Ortholog Receptor Or67d in Drosophila Bipectinata is able to Detect Two Different Pheromones.

Sex pheromones play a crucial role in species recognition and reproductive isolation. Despite being largely species-specific in drosophilids, the mechanisms underlying pheromone detection, production, and their influence on mating behavior remain poorly understood. Here, we compare the chemical profiles of Drosophila bipectinata and D. melanogaster, the mating behaviors in both species, as well as the tuning properties of Or67d receptors, which are expressed by neurons in antennal trichoid sensilla at1. Through single sensillum recordings, we demonstrate that the D. bipectinata Or67d-ortholog exhibits similar sensitivity to cis-vaccenyl acetate (cVA) as compared to D. melanogaster but in addition also responds uniquely to (Z)-11-eicosen-1-yl-acetate (Z11-20:Ac), a compound exclusively produced by D. bipectinata males. Through courtship behavior assays we found that, surprisingly, perfuming the flies with Z11-20:Ac did not reveal any aphrodisiacal or anti-aphrodisiacal effects in mating assays. The behavioral relevance of at1 neuron channels in D. bipectinata compared to D. melanogaster seems to be restricted to its formerly shown function as an aggregation pheromone. Moreover, the non-specific compound cVA affected copulation negatively in D. bipectinata and could potentially act as a premating isolation barrier. As both ligands of Or67d seem to govern different behaviors in D. bipectinata, additional neurons detecting at least one of those compounds might be involved. These results underscore the complexity of chemical signaling in species recognition and raise intriguing questions about the evolutionary implications of pheromone detection pathways in Drosophila species.

Wolbachia strain diversity in a complex group of sympatric cryptic parasitoid wasp species

BackgroundMaternally-inherited symbionts can induce pre-mating and/or post-mating reproductive isolation between sympatric host lineages, and speciation, by modifying host reproductive phenotypes. The large parasitoid wasp genus Cotesia (Braconidae) includes a diversity of cryptic species, each specialized in parasitizing one to few related Lepidoptera host species. Here, we characterized the infection status of an assemblage of 21 Cotesia species from 15 countries by several microbial symbionts, as a first step toward investigating whether symbionts may provide a barrier to gene flow between these parasitoid host lineages.ResultsThe symbiotic microbes Arsenophonus, Cardinium, Microsporidium and Spiroplasma were not detected in the Cotesia wasps. However, the endosymbiotic bacterium Wolbachia was present in at least eight Cotesia species, and hence we concentrated on it upon screening additional DNA extracts and SRAs from NCBI. Some of the closely related Cotesia species carry similar Wolbachia strains, but most Wolbachia strains showed patterns of horizontal transfer between phylogenetically distant host lineages.ConclusionsThe lack of co-phylogenetic signal between Wolbachia and Cotesia suggests that the symbiont and hosts have not coevolved to an extent that would drive species divergence between the Cotesia host lineages. However, as the most common facultative symbiont of Cotesia species, Wolbachia may still function as a key-player in the biology of the parasitoid wasps. Its precise role in the evolution of this complex clade of cryptic species remains to be experimentally investigated.

Asymmetric mate preference and reproductive interference mediate climate-induced changes in mate availability in a small mammal hybrid zone.

Range expansion and contraction are among the most common biotic responses to changing environmental conditions, yet much is to be learned about the mechanisms that underlie range-edge population dynamics, especially when those areas are points of secondary contact between closely related species. Here, we present field-measured parentage data that document the reproductive outcomes of changes in mate availability at a secondary contact zone between two species of woodrat in the genus Neotoma. Changes in mate availability resulted from drought-driven differential survival between the species and their hybrids. As the availability of conspecific mates declined, rates of hybridization increased, leading to the accumulation of admixed individuals in the zone of contact. Patterns of reproductive success in the wild appear to be the result of a combination of both pre-mating isolation and post-zygotic selection resulting from genomic incompatibilities between the parental lineages. Evidence of asymmetric mate preference between the parental lineages came from both skewed reproductive output in the field and laboratory preference trials. Moreover, partial genomic incompatibility was evident from the near-zero reproductive success of F1 males and because nearly all surviving hybrids had one pure parent. Nonetheless, high reproductive success of F1 females and backcrossing in both parental directions allow for introgression between the parental species. These findings reveal how climate change may alter evolutionary outcomes for species at the edge of their ranges through an interplay of behavioral, demographic, and genetic mechanisms.

Mating preferences act independently on different elements of visual signals in Heliconius butterflies

Abstract Mating cues are often comprised of several elements, which can act independently, or in concert to attract a suitable partner. Individual elements may also function in other contexts, such as anti-predator defense or camouflage. In Heliconius butterflies, wing patterns comprise several individual color pattern elements, which advertise the butterflies’ toxicity to predators. These wing patterns are also mating cues and males predominantly court females that possess the same wing pattern as their own. However, it is not known whether male preference is based on the full wing pattern or only individual pattern elements. We compared preferences of male H. erato lativitta between female models with the full wing pattern and those with some pattern elements removed. We found no differences in preference between the full wing pattern model and a model with pattern elements removed, indicating that the complete composition of all elements is not essential to the mating signal. Wing pattern preferences also contribute to pre-mating isolation between two other Heliconius taxa, H. erato cyrbia, and H. himera; therefore, we next compared preferences for the same models in these species. H. erato cyrbia and H. himera strongly differed in preferences for the models, potentially providing a mechanism for how pre-mating isolation acts between these species. These findings suggest that contrasting levels of selective constraint act on elements across the wing pattern.

Ecotype variation in the endemic tree Callicarpa subpubescens on small oceanic islands: genetic, phenotypic, and environmental insights.

Callicarpa subpubescens, endemic to the Ogasawara Islands, is suggested to have multiple ecotypes in the Hahajima Islands, specifically in the central part of the Ogasawara Islands. In this study, associations between genetic groups and spatial distribution, habitat, leaf morphology, size structure, and flowering time of each genetic group were investigated on Hahajima and the satellite Imoutojima Islands. Genetic groups were identified using EST-SSR markers, revealing four ecotypes named based on morphological features: Dwarf (D), Glabrescent (G), Tall (T), and Middle (M), with M being a result of the hybridization of G and T. Ecotype D, adapted to dry environments, is characterized by small tree size, dense thick leaves with abundant hairs, and is distributed in dry scrub. Ecotype G, adapted to understory of mesic forests, lacks leaf hairs. Ecotype T, adapted to the canopy of mesic forests, has hairy leaves and is tall in tree height. Ecotype M, adapted to the canopy of mesic scrub or edges of mesic forests, has hairy leaves but with a shorter tree height than ecotype T. Flowering peaks differed among all ecotype pairs except G and M, but the flowering times more or less overlapped among all ecotypes, suggesting that pre-mating isolation among ecotypes is not perfect. Post-mating isolation is considered absent, as there were no differences in the results, germination, and survival rates of one-year seedlings among inter- and intra-ecotype crossings. The existence of such ecotypes provides valuable insights into the ongoing speciation processes adapting to the oceanic island environments.

Differential pheromone profile as a contributor to premating isolation between two sympatric sibling fruit fly species.

Bactrocera tryoni (Froggatt) and Bactrocera neohumeralis (Hardy) are sibling fruit fly species that are sympatric over much of their ranges. Premating isolation of these close relatives is thought to be maintained in part by allochrony-mating activity in B. tryoni peaks at dusk, whereas in B. neohumeralis, it peaks earlier in the day. To ascertain whether differences in pheromone composition may also contribute to premating isolation between them, this study used solid-phase microextraction and gas chromatography-mass spectrometry to characterize the rectal gland volatiles of a recently collected and a more domesticated strain of each species. These glands are typical production sites and reservoirs of pheromones in bactrocerans. A total of 120 peaks were detected and 50 were identified. Differences were found in the composition of the rectal gland emissions between the sexes, species, and recently collected versus domesticated strains of each species. The compositional variation included several presence/absence and many quantitative differences. Species and strain differences in males included several relatively small alcohols, esters, and aliphatic amides. Species and strain differences in females also included some of the amides but additionally involved many fatty acid esters and 3 spiroacetals. While the strain differences indicate there is also heritable variation in rectal gland emissions within each species, the species differences imply that compositional differences in pheromones emitted from rectal glands could contribute to the premating isolation between B. tryoni and B. neohumeralis. The changes during domestication could also have significant implications for the efficacy of Sterile Insect Technique control programs.

Incomplete isolation in the nonbreeding areas of two genetically separated but sympatric short-tailed albatross populations

The short-tailed albatross Phoebastria albatrus, a globally Vulnerable species recovering from near-extinction, breeds mainly on 2 island groups in the north-western Pacific: Torishima and Senkaku Islands. Recently, it became clear that this is a species complex, composed of 2 populations (‘Torishima’ and ‘Senkaku’ types) that are distinct from both genetic and morphological perspectives, and which mate assortatively on Torishima. We tested the hypothesis of premating isolation as a possible mechanism for genetic differentiation and shifted breeding date in short-tailed albatrosses. Using light-based geolocation and molecular analysis, we examined whether adults from the 2 populations breeding on Torishima differed in their at-sea distribution during the nonbreeding period (June-September) and in their return dates. From 22 bird tracks collected from 12 birds over 4 yr, we identified 2 distinct nonbreeding areas: around the Aleutian Islands in the Bering Sea and near the Kuril Islands in the Okhotsk Sea. All 6 tracks of Torishima-type birds migrated directly to the Bering Sea in all years. In contrast, most of the Senkaku-type birds migrated along northeast Japan to the Okhotsk Sea (9 tracks), with the others moving to the Bering Sea (7 tracks). There was no clear difference in the dates of return to Torishima between the 2 bird types. Overall, no absolute pre-mating isolation in space or time was observed in short-tailed albatrosses, but we highlighted notable divergences in the nonbreeding distribution of both types. This result suggests that Torishima- and Senkaku-type populations should be treated as separate management units.

Complexity of the Male Perfume of Eulaema nigrita from Forest and Woody Physiognomies of the Brazilian Savanna: Is There a Relationship with Body Size and Wing Wear?

Male orchid bees collect volatile and semi-volatile compounds from the environment for storage and accumulation in specialized hind legs. Later, these compounds form a perfume blend used during courtship to lure conspecific females for mating. Male orchid bees perfume has been suggested to play an important role as a sexual signaling trait involved in pre-mating isolation of species, functioning as an indicator of male genetic quality. Eulaema nigrita Lepeletier (Apidae: Euglossini) is a common species in both forested and woody savanna (Cerrado stricto sensu) physiognomies of the Brazilian savanna biome. By identifying the chemical composition of male E. nigrita perfume, we tested for differences in the bouquet chemical profile in populations from remnants of seasonal semideciduous forest and woody savanna. In addition, we assessed the relation between perfume complexity and morphological traits associated with size and age of males. Our analysis showed a low effect of physiognomies on differences in the perfume chemical profile of sampled males. Nevertheless, we observed significant differences in the chemical profile of individuals from two seasonal semideciduous forest remnants, which suggests an environmental effect in individual bouquet. Wing wear measurements were positively related to perfume complexity, consistent with the premise that perfumes from older individuals are indicators of survival capacity in male orchid bees.

Characterization of the swarming behavior of Anopheles coluzzii and Anopheles gambiae (Diptera: Culicidae) populations in a hybrid zone of Senegal.

Anopheles gambiae and Anopheles coluzzii, often found in sympatry and synchronous, have undergone a premating reproductive isolation across their distribution range. However, in the Western coast of Africa, unexpected hybridization zones have been observed, and little is known about swarming behavior of these cryptic taxa. Here, we characterized the swarming behavior of An. coluzzii and An. gambiae to investigate its role in the high hybridization level in Senegal. The study was conducted in the south and central Senegal during the 2018 rainy season. Mating swarms of malaria vectors were surveyed at sunset and collected using an insect net. Meanwhile, indoor resting populations of malaria vectors were collected by pyrethrum spray catches. Upon collection, specimens were identified morphologically, and then members of the An. gambiae complex were identified at the species level by polymerase chain reaction (PCR). An. gambiae swarmed mainly over bare ground, whereas An. coluzzii were found swarming above various objects creating a dark-light contrast with the bare ground. The swarms height varied from 0.5 to 2.5 m. Swarming starting time was correlated with sunset whatever the months for both species, and generally lasted about 10 min. No mixed swarm of An. gambiae and An. coluzzii was found even in the high hybridization area. These results indicated a premating isolation between An. coluzzii and An. gambiae. However, the high hybridization rate in the sympatric area suggests that heterogamous mating is occurring, thus stressing the need for further extensive studies.

Description of Calling, Courtship and Mating Behaviour of Six Species of the Genus Zyginidia (Auchenorrhyncha: Cicadellidae).

The vibrational communication and mating behaviour of the graminicolous leafhoppers Zyginidia pullula, Z. ribauti, Z. scutellaris, Z. serpentina, Z. sohrab, and Z. biroi were investigated to explain why the first five species hybridize. Z. biroi was used as a control species. All species behaved in the same way and no significant statistical differences were detected with regard to male calls, while female calls and the male courtship song differed in Z. biroi, thus showing that a specific pre-mating isolation mechanism was used by the latter species and the first five ones lacked such a mechanism. In addition, Z. sohrab is missing in Italy, while the other species live allopatrically in Italy, with the only exceptions being Z. serpentina and Z. biroi, which live in Sicily and are often found in sympatry, and Z. scutellaris and Z. biroi, which live in Sardinia. All these species can be distinguished by means of male genital appendages; however, Z. biroi is longer and has a different body colour. The existence of natural hybrids of Z. pullula, Z. ribauti, and Z. scutellaris in the Italian peninsula and their hybridization in the laboratory with Z. serpentina and Z. sohrab require the investigation of possible post-mating reproductive barriers before re-considering their systematic validity.

Hybridisation between host races broadens the host range of offspring in Eotetranychus asiaticus (Acari: Tetranychidae).

Host adaptation in herbivorous arthropods is one of the first steps to sympatric speciation, and spider mites (Acari: Tetranychidae) are useful model organisms for studying this phenomenon. Many researchers have studied the process of host adaptation via artificial selection experiments. Recent analyses suggest that hybridisation has diversified host ranges, although empirical evidence is scarce. We explored the host exploitation of two strains of Eotetranychus asiaticus established from Ternstroemia gymnanthera (Pentaphylacaceae) and Japanese cinnamon, Cinnamomum yabunikkei (Lauraceae), and evaluated the effect of hybridisation on offspring host use. Transplant experiments showed that females oviposited and immature mites developed only on their native hosts, suggesting specialisation to the secondary metabolites of each host plant. However, F1 hybrids from reciprocal crosses developed on both host plants (survival rate: 92-100%) with normal female-biased sex ratios. Furthermore, all backcrosses to the parental strains yielded B1 offspring that were also viable on both host plants with normal sex ratios (69-87% and39-92%femalesonT. gymnanthera andC. yabunikkei, respectively). B1 survival rates in interstrain crosses were varied (11-63%) and lower than those in intrastrain crosses (88-93%). We could not detect any reproductive barriers in these experiments, and host preference may be the sole factor determining pre-mating isolation. The survival rates and sex ratios we observed suggest cytochromosome interactions. In conclusion, hybridisation, which results in heterozygotes and recombination, is an underexplored way to provide spider mites with a novel host plant.

Hybrid origin of a primate, the gray snub-nosed monkey.

Hybridization is widely recognized as promoting both species and phenotypic diversity. However, its role in mammalian evolution is rarely examined. We report historical hybridization among a group of snub-nosed monkeys (Rhinopithecus) that resulted in the origin of a hybrid species. The geographically isolated gray snub-nosed monkey Rhinopithecus brelichi shows a stable mixed genomic ancestry derived from the golden snub-nosed monkey (Rhinopithecus roxellana) and the ancestor of black-white (Rhinopithecus bieti) and black snub-nosed monkeys (Rhinopithecus strykeri). We further identified key genes derived from the parental lineages, respectively, that may have contributed to the mosaic coat coloration of R. brelichi, which likely promoted premating reproductive isolation of the hybrid from parental lineages. Our study highlights the underappreciated role of hybridization in generating species and phenotypic diversity in mammals.

Detecting introgressive hybridization to maintain genetic integrity in endangered large waterbird: a case study in milky stork

Hybridization between milky stork (Mycteria cinerea) and painted stork (M. leucocephala) occurs frequently in captivity. Dusit Zoo is a captive breeding facility where storks with phenotypically ambiguous patterns have recently been observed, and their status remaining inconclusive. Here, we used a combination of phenotypic characters and genetic markers (cytochrome b and 14 microsatellite markers) to distinguish and identify hybrids from the two parental species (n = 114). Haplotype analysis revealed asymmetric mtDNA introgression from M. cinerea to M. leucocephala, with twelve morphologically classified M. leucocephala individuals carrying heterospecific mtDNA. Comprehensive biparental genetic assessments identified 33% of all three genetic clusters as admixed individuals, of which most were either F2 hybrids, backcrosses with M. leucocephala, or hybrids of unknown generation, implying weak premating isolation with the absence of intrinsic postzygotic isolation between parentals. Morphological analysis demonstrated that the absence or indistinctness of a black bar across the breast is the most noticeable trait to identify these hybrids. The endangered M. cinerea was found to have genomic contamination from M. leucocephala and vice versa, with at least 41 hybrid individuals being identified. These findings provide critical information for detecting hybrids and identifying suitable breeding stocks with genetic purity for future reintroduction and conservation management.

The effectiveness of pseudomagic traits in promoting premating isolation.

Upon the secondary contact of populations, speciation with gene flow is greatly facilitated when the same pleiotropic loci are both subject to divergent ecological selection and induce non-random mating, leading to loci with this fortuitous combination of functions being referred to as 'magic trait' loci. We use a population genetics model to examine whether 'pseudomagic trait' complexes, composed of physically linked loci fulfilling these two functions, are as efficient in promoting premating isolation as magic traits. We specifically measure the evolution of choosiness, which controls the strength of assortative mating. We show that, surprisingly, pseudomagic trait complexes, and to a lesser extent also physically unlinked loci, can lead to the evolution of considerably stronger assortative mating preferences than do magic traits, provided polymorphism at the involved loci is maintained. This is because assortative mating preferences are generally favoured when there is a risk of producing maladapted recombinants, as occurs with non-magic trait complexes but not with magic traits (since pleiotropy precludes recombination). Contrary to current belief, magic traits may not be the most effective genetic architecture for promoting strong premating isolation. Therefore, distinguishing between magic traits and pseudomagic trait complexes is important when inferring their role in premating isolation. This calls for further fine-scale genomic research on speciation genes.

Evidence of historical pairing between two cryptic species of Short-tailed Albatross

When secondary contact occurs between allopatric sister species, several evolutionary consequences are expected, such as reinforcement of reproductive isolation, hybrid speciation, de-speciation, introgressive hybridization, or formation of a stable hybrid zone. The Short-tailed Albatross (<em>Phoebastria albatrus</em>) is a vulnerable seabird that breeds mainly in Torishima, the Izu Islands, and two islets in the Senkaku Islands in the western North Pacific. Recent studies revealed that Short-tailed Albatross comprises two cryptic species (Senkaku-type and Torishima-type) that breed sympatrically on Torishima. Ringed (hatched in Torishima) and unringed (probably hatched in the Senkaku Islands) birds mate in a mutually assortative manner at the Hatsunezaki colony (artificially established in 1995) on Torishima. However, observations of some ringed–unringed pairs suggest possible hybridization between the two cryptic species. To clarify the degree of hybridization, we analyzed microsatellite DNA and mitochondrial DNA control region 2 (CR2) sequences of chicks from Hatsunezaki and Tsubamezaki (original colony discovered in 1951) colonies and of unringed birds from Hatsunezaki. In general, both CR2 sequences and microsatellites revealed genetic differentiation between immigrants from the Senkaku Islands (unringed birds) and chicks hatched in Tsubamezaki. These findings support the existence of two cryptic species. Each chick obtained from four ringed–unringed parent pairs at Hatsunezaki displayed a high proportion of alleles from just a single population. In contrast, some chicks in Tsubamezaki had a medium proportion of alleles from both populations. Breeding unringed subadult plumage birds, which were probable immigrants from the Senkaku Islands, were observed in Hatsunezaki but not in Tsubamezaki. Therefore, we propose that interspecific pairing occurred in the past but infrequently in recent generations on Torishima, suggesting historical reinforcement of reproductive isolation. Further microsatellite DNA studies of chicks from Hatsunezaki are required to confirm whether reinforcement of reproductive isolation is achieved. Alternatively, nearly complete pre-mating isolation between the two species was established in the past, but the scarcity of Senkaku-type birds in Torishima has facilitated hybridization.

Reproductive isolation via divergent genital morphology due to cascade reinforcement in Ohomopterus ground beetles.

Secondary contact between incipient species and selection against maladaptive hybridization can drive reinforcement between populations in contact and result in reproductive character displacement (RCD). Resultant divergence in mating traits within a species may generate downstream reproductive isolation between populations with displaced and non-displaced traits, referred to as the cascade reinforcement hypothesis. We examined this hypothesis using three allopatric populations of the ground beetle Carabus maiyasanus with a genital lock-and-key system. This species shows RCD in male and female genital morphologies in populations in contact with the sister species C. iwawakianus. In a reciprocal mating experiment using three allopatric populations with differences in male and female genital sizes, insemination failure increased as the difference in genital size increased. Based on the reproductive isolation index, insemination failure was the major postmating-prezygotic isolation barrier, at least in one population pair with comparable total isolation to those of other species pairs. By contrast, there was only incomplete premating isolation among populations. These results suggest that RCD in genital morphologies drives incipient allopatric speciation, supporting the cascade reinforcement hypothesis. These findings provide insight into the roles of interspecific interactions and subsequent trait diversification in speciation processes.

Ecological specialization and reproductive isolation among closely related sympatric ant-eating spiders.

Biological divergence results from several mechanisms. Defensive mechanisms, such as Batesian mimicry, can cause reproductive isolation via temporal segregation in foraging activity, particularly, in species that closely associate with their model. This seems to be the case of ant-eating spiders, which can be inaccurate Batesian mimics of their prey. Here, we focused on Zodarion nitidum, which has two forms occurring in sympatry, black and yellow. Given the expected noticeable impact of their colour differences on the spiders' interactions with their potential predators and prey, we investigated whether these morphotypes have diverged in other aspects of their biology. We measured the two morphotypes' phenotypic resemblance to a mimetic model, tested whether they were protected from predators, investigated their circadian activity, surveyed the prey they hunted, modelled their distributions, performed crossing experiments and estimated their degree of genetic differentiation. We found that the black morphotype is ant-like, resembling Messor ants, and it was not distinguishable from their ant models by four potential predators. In contrast, the yellow morphotype seems to use predator avoidance as a defensive strategy. Additionally, the two morphotypes differ in their circadian activity, the yellow morphotype being nocturnal and the black one being diurnal. The two morphotypes hunt and associate with different ant prey and possess marked differences in venom composition. Finally, crossing trials showed complete pre-mating isolation between the two morphotypes, but there was no evidence of genetic (mitochondrial data) or environmental niche differentiation. We conclude that the two morphotypes show evidence of a deep differentiation in morphological, behavioural, physiological and ecological traits that evolved together as part of the spider's diverging lifestyles.

Coexisting good neighbours: acoustic and calling microhabitat niche partitioning in two elusive syntopic species of balloon frogs, Uperodon systoma and U. globulosus (Anura: Microhylidae) and potential of individual vocal signatures

BackgroundMost amphibians use a repertoire of acoustic signals to propagate signals in social contexts. The description of these repertoires provides a key towards the understanding of the behaviour of individuals and the evolutionary functions of calls. Here, we assessed the variations in advertisement calls within and between two fossorial sympatric species, Uperodon systoma and Uperodon globulosus, that share their breeding season and breeding sites. For each species, we applied Beecher’s index of total information capacity (HS) for the individual vocal signature, determined the difference in call properties and demonstrated the segregation in the calling microhabitat niche between the two species.ResultsOur results demonstrated that the advertisement calls of U. systoma are pulsatile with a call rate of 3.00 ± 0.97 calls per second while those of U. globulosus are not pulsatile with a lower call rate of 0.53 ± 0.22 calls per second. For both species, the variations in call properties among individuals was higher than that within individual, a pattern consistent with that of other fossorial anurans. The body condition and air temperature did not significantly impact the call properties of either species. The outcome of the Beecher’s index (HS) showed that the calls of U. systoma can be used to identify 14 different individuals and the calls of U. globulosus can be used to identify 26 different individuals. The statistical analyses on the advertisement call of the two species showed a significant difference in the temporal properties as the call duration, and fall time and rise time were significantly different between the two species. Lastly, we successfully demonstrated that there is a clear segregation in calling site microhabitat between the two species, where U. globulosus calls floating close to the bank of the waterbody while U. systoma calls floating further away from the bank.ConclusionThis study highlights the potential for pre-mating isolation, character displacement and assortative mating in two syntopic fossorial anurans, leading to association between acoustic, calling microhabitat niche and body index divergence as important behavioural and ecological traits.