Ultraconserved elements-based systematics reveals evolutionary patterns of host-plant family shifts and phytophagy within the predominantly parasitoid braconid wasp subfamily Doryctinae.

Phytophagy has promoted species diversification in many insect groups, including Hymenoptera, one of the most diverse animal orders on Earth. In the predominantly parasitoid family Braconidae, an association with insect-induced, plant galls in angiosperms have been reported in three subfamilies, but in particular in the Doryctinae, where it has been recorded to occur in species of ten genera. Allorhogas Gahan is the most species-rich of these genera, with its species having different phytophagous strategies. Here we conducted a comprehensive phylogenomic study for the doryctine gall-associated genera, with an emphasis on Allorhogas, using ultraconserved elements (UCEs). Based on this estimate of phylogeny we: (1) evaluated their taxonomic composition, (2) estimated the timing of origin of the gall-associated clade and divergence of its main subclades, and (3) performed ancestral state reconstruction analyses for life history traits related to their host-plant association. Our phylogenetic hypothesis confirmed Allorhogas as polyphyletic, with most of its members being nested in a main clade composed of various subclades, each comprising species with a particular host-plant family and herbivorous feeding habit. The origin of gall-association was estimated to have occurred during the late Oligocene to early Miocene, with a subsequent diversification of subclades during the middle to late Miocene and Pliocene. Overlap in divergence timing appears to occur between some taxa and their host-associated plant lineages. Evolution of the feeding strategies in the group shows "inquilinism-feeding" as the likely ancestral state, with gall-formation in different plant organs and seed predation having independently evolved on multiple occasions.

Phylogenomics of braconid wasps (Hymenoptera, Braconidae) sheds light on classification and the evolution of parasitoid life history traits.

The parasitoid lifestyle is largely regarded as a key innovation that contributed to the evolutionary success and extreme species richness of the order Hymenoptera. Understanding the phylogenetic history of hyperdiverse parasitoid groups is a fundamental step in elucidating the evolution of biological traits linked to parasitoidism. We used a genomic-scale dataset based on ultra-conserved elements and the most comprehensive taxon sampling to date to estimate the evolutionary relationships of Braconidae, the second largest family of Hymenoptera. Based on our results, we propose Braconidae to comprise 41 extant subfamilies, confirmed a number of subfamilial placements and proposed subfamily-level taxonomic changes, notably the restoration of Trachypetinae stat. rev. and Masoninae stat. rev. as subfamilies of Braconidae, confirmation that Apozyx penyai Mason belongs in Braconidae placed in the subfamily Apozyginae and the recognition of Ichneutinae sensu stricto and Proteropinae as non-cyclostome subfamilies robustly supported in a phylogenetic context. The correlation between koinobiosis with endoparasitoidism and idiobiosis with ectoparasitoidism, long thought to be an important aspect in parasitoid life history, was formally tested and confirmed in a phylogenetic framework. Using ancestral reconstruction methods based on both parsimony and maximum likelihood, we suggest that the ancestor of the braconoid complex was a koinobiont endoparasitoid, as was that of the cyclostome sensu lato clade. Our results also provide strong evidence for one transition from endo- to ectoparasitoidism and three reversals back to endoparasitoidism within the cyclostome sensu stricto lineage. Transitions of koino- and idiobiosis were identical to those inferred for endo- versus ectoparasitoidism, except with one additional reversal back to koinobiosis in the small subfamily Rhysipolinae.

Genomic architecture of endogenous ichnoviruses reveals distinct evolutionary pathways leading to virus domestication in parasitic wasps.

BACKGROUND: Polydnaviruses (PDVs) are mutualistic endogenous viruses inoculated by some lineages of parasitoid wasps into their hosts, where they facilitate successful wasp development. PDVs include the ichnoviruses and bracoviruses that originate from independent viral acquisitions in ichneumonid and braconid wasps respectively. PDV genomes are fully incorporated into the wasp genomes and consist of (1) genes involved in viral particle production, which derive from the viral ancestor and are not encapsidated, and (2) proviral segments harboring virulence genes, which are packaged into the viral particle. To help elucidating the mechanisms that have facilitated viral domestication in ichneumonid wasps, we analyzed the structure of the viral insertions by sequencing the whole genome of two ichnovirus-carrying wasp species, Hyposoter didymator and Campoletis sonorensis. RESULTS: Assemblies with long scaffold sizes allowed us to unravel the organization of the endogenous ichnovirus and revealed considerable dispersion of the viral loci within the wasp genomes. Proviral segments contained species-specific sets of genes and occupied distinct genomic locations in the two ichneumonid wasps. In contrast, viral machinery genes were organized in clusters showing highly conserved gene content and order, with some loci located in collinear wasp genomic regions. This genomic architecture clearly differs from the organization of PDVs in braconid wasps, in which proviral segments are clustered and viral machinery elements are more dispersed. CONCLUSIONS: The contrasting structures of the two types of ichnovirus genomic elements are consistent with their different functions: proviral segments are vehicles for virulence proteins expected to adapt according to different host defense systems, whereas the genes involved in virus particle production in the wasp are likely more stable and may reflect ancestral viral architecture. The distinct genomic architectures seen in ichnoviruses versus bracoviruses reveal different evolutionary trajectories that have led to virus domestication in the two wasp lineages.

Running in circles in phylomorphospace: host environment constrains morphological diversification in parasitic wasps.

Understanding phenotypic diversification and the conditions that spur morphological novelty or constraint is a major theme in evolutionary biology. Unequal morphological diversity between sister clades can result from either differences in the rate of morphological change or in the ability of clades to explore novel phenotype ranges. We combine an existing phylogenetic framework with new phylogenomic data and geometric morphometrics to explore the relative roles of rate versus mode of morphological evolution for a hyperdiverse group: cryptine ichneumonid wasps. Data from genomic ultraconserved elements confirm that cryptines are divided into two large clades: one specialized in the use of hosts that are deeply concealed under hard substrates, and another with a much more diversified host range. Using a phylomorphospace approach, we show that both clades have experienced similar rates of morphological evolution. Nonetheless, the more specialized group is much more restricted in morphospace occupation, indicating that it repeatedly evolved morphological change through the same morphospace regions. This is in agreement with our prediction that host use imposes constraints in the morphospace available to lineages, and reinforces an important distinction between evolutionary stasis as opposed to a scenario of continual morphological change restricted to a certain range of morphotypes.

Testing the Dutilleul syndrome: host use drives the convergent evolution of multiple traits in parasitic wasps.

Common life-history aspects among independent lineages often result in the repeated evolution of suites of adaptive traits, or 'syndromes'. Such syndromes can be key avenues to understand relationships between morphological and ecological traits, but are rarely tested due to insufficient trait shift repetitions. We use a hyperdiverse lineage to investigate the evolution of a syndrome. Cryptine ichneumonid wasps that parasitize insects concealed in hard substrates display several traits that are putative adaptations to that end. Using a phylogenetic framework from a combined multigene molecular and morphological data set with 308 cryptine species, we tested whether these traits were part of a morphofunctional syndrome related to host use. Ancestral state estimations show multiple origins for six investigated traits, which are correlated to each other and to the use of deeply concealed hosts, suggesting adaptation. Putatively adaptive traits showed a much stronger link among themselves than with an assemblage of 49 other morphological traits. However, estimation of the order of evolution in adaptive traits showed no structured pattern. The results indicate that the challenge of attacking deeply concealed hosts induced the repeated evolution of a 'Dutilleul syndrome', named after the 'walker-through-walls' character from French literature. They also point towards a dynamic scenario in the evolution of complex functional systems. These findings highlight the power of morphology to illuminate poorly known aspects of natural history, and how hyperdiverse lineages can be used to understand the evolution of complex traits.

Phylogeny and historical biogeography of the paper wasp genus Polistes (Hymenoptera: Vespidae): implications for the overwintering hypothesis of social evolution.

The phylogeny of the paper wasp genus Polistes is investigated using morphological and behavioural characters, as well as molecular data from six genes (COI, 12S, 16S, 28S, H3, and EF1-α). The results are used to investigate the following evolutionary hypotheses about the genus: (i) that Polistes first evolved in Southeast Asia, (ii) that dispersal to the New World occurred only once, and (iii) that long-term monogyny evolved as an adaptation to overwintering in a temperate climate. Optimization of distribution records on the recovered tree does not allow unambiguous reconstruction of the ancestral area of Polistes. While the results indicate that Polistes dispersed into the New World from Asia, South America is recovered as the ancestral area for all New World Polistes: Nearctic species groups evolved multiple times from this South American stock. The final tree topology suggests strongly that the genus first arose in a tropical environment, refuting the idea of monogyny as an overwintering adaptation.

The Darwin wasp Camera thoracica (Szpligeti, 1916) (Ichneumonidae) as a natural enemy of the dreaded Brazilian wandering spider Phoneutria nigriventer (Keyserling, 1891) (Ctenidae).

Wandering spiders (genus Phoneutria) hold a prominent position as some of the worlds most medically significant venomous arachnids, especially in Brazil. In this study, we record and illustrate for the first time, the Darwin wasp Camera thoracica (Szpligeti, 1916) as a natural enemy of the ctenid Phoneutria nigriventer (Keyserling, 1891). Furthermore, we provide a description of the previously unknown male wasp, update and standardize the description of the female, and provide biological notes.

Phylogeny and revision of Messatoporus Cushman (Hymenoptera, Ichneumonidae, Cryptinae), with descriptions of sixty five new species.

The large New World genus Messatoporus Cushman is diagnosed, redefined, described, and its species revised. A total of 89 valid species are recognized, of which 65 are described as new. All previously known species are redescribed and illustrated. The following synonyms are proposed: M. jocosus (Provancher) and M. rufiventris Cushman under M. discoidalis (Cresson), syn. nov. and M. tricolor (Szépligeti) under M. variegatus (Szépligeti) syn. nov. The following new species are proposed: M. advenus, M. albiaterartus, M. allomeros, M. amarelus, M. amazonensis, M. amplithorax, M. andinus, M. anepomius, M. apiopharkis, M. argentinus, M. atlanticus, M. atrifoveus, M. aurantius, M. badius, M. basiflavus, M. bennetii, M. bicolor, M. bimaculatus, M. campanulatus, M. caxiuanensis, M. citreocephalus, M. complexifemur, M. concavivenus, M. convexus, M. copiosus, M. depressifrons, M. dialeipsis, M. dissidens, M. dominicanus, M. elektor, M. ellipsicavus, M. euryoikos, M. igneus, M. interceptus, M. keraiopetalus, M. laevilatus, M. latissulcus, M. longicaudis, M. longitergus, M. lordos, M. lunatus, M. maculiscus, M. matucanus, M. nigriangulatus, M. nigriscapus, M. nigriscutus, M. occidentalis, M. opacus, M. orientalis, M. paeneater, M. pallidus, M. paradoxus, M. paralissonotus, M. pleuriflavus, M. semialbiventris, M. semiaurantigaster, M. subalaris, M. tenuiorbis, M. tenuissimus, M. teutonicus, M. titans, M. unidentatus, M. unimaculatus, M. versicolor and M. xanthogaster spp. nov. As well, a lectotype is designated for M. townesi Alayo and Tzankov, and Messatoporus nigrispina (Cameron) is transferred to Prosthoporus Porter, comb. nov.. A key to species based on both sexes is also presented, and new distribution records and distribution maps are provided. The validity and relationships of the genus are investigated cladistically using 162 morphological characters for 27 species of Messatoporus and 105 outgroup taxa. Tree search was conducted using TNT under implied weighting, with values of 1­6 for the concavity constant (K). All analyses recovered Messatoporus as a monophyletic group, supported by 7­11 synapomorphies. The results also support the monophyly of the six studied genera of Osprynchotina, in a clade including also Dotocryptus Brèthes. Messatoporus is diagnosed by the combination of the following features: apicolateral corners of clypeus projected; mandible long and tapered, ventral tooth much shorter than dorsal one or indistinct; transverse sulcus at base of propodeum long and shallow; anterior margin of propodeum concave; posterior area of propodeum almost always transversely wrinkled; first metasomal spiracle placed approximately on midlength; median dorsal carina of first metasomal segment absent; ovipositor basally cylindrical, apically depressed, with ventral valve enclosing completely dorsal valve as a sheath. A second analysis including all species of the genus was performed to evaluate the phylogeny at the genus level. The character set for that analysis included 104 characters considered of phylogenetic importance within the genus, and used the same tree searching protocol as the first one. A distinct and gradual transition is detected, from species more similar to the remaining Osprynchotina to a significantly different morphotype with several convergences with the Gabuniina. These convergences are interpreted as adaptations to host location and substrate perforation, and may be related to use of more deeply concealed hosts.

Key innovations and the diversification of Hymenoptera.

The order Hymenoptera (wasps, ants, sawflies, and bees) represents one of the most diverse animal lineages, but whether specific key innovations have contributed to its diversification is still unknown. We assembled the largest time-calibrated phylogeny of Hymenoptera to date and investigated the origin and possible correlation of particular morphological and behavioral innovations with diversification in the order: the wasp waist of Apocrita; the stinger of Aculeata; parasitoidism, a specialized form of carnivory; and secondary phytophagy, a reversal to plant-feeding. Here, we show that parasitoidism has been the dominant strategy since the Late Triassic in Hymenoptera, but was not an immediate driver of diversification. Instead, transitions to secondary phytophagy (from parasitoidism) had a major influence on diversification rate in Hymenoptera. Support for the stinger and the wasp waist as key innovations remains equivocal, but these traits may have laid the anatomical and behavioral foundations for adaptations more directly associated with diversification.

A workflow for expanding DNA barcode reference libraries through 'museum harvesting' of natural history collections.

Natural history collections are the physical repositories of our knowledge on species, the entities of biodiversity. Making this knowledge accessible to society - through, for example, digitisation or the construction of a validated, global DNA barcode library - is of crucial importance. To this end, we developed and streamlined a workflow for 'museum harvesting' of authoritatively identified Diptera specimens from the Smithsonian Institution's National Museum of Natural History. Our detailed workflow includes both on-site and off-site processing through specimen selection, labelling, imaging, tissue sampling, databasing and DNA barcoding. This approach was tested by harvesting and DNA barcoding 941 voucher specimens, representing 32 families, 819 genera and 695 identified species collected from 100 countries. We recovered 867 sequences (> 0 base pairs) with a sequencing success of 88.8% (727 of 819 sequenced genera gained a barcode > 300 base pairs). While Sanger-based methods were more effective for recently-collected specimens, the methods employing next-generation sequencing recovered barcodes for specimens over a century old. The utility of the newly-generated reference barcodes is demonstrated by the subsequent taxonomic assignment of nearly 5000 specimen records in the Barcode of Life Data Systems.

Enhancing DNA barcode reference libraries by harvesting terrestrial arthropods at the Smithsonian's National Museum of Natural History.

The use of DNA barcoding has revolutionised biodiversity science, but its application depends on the existence of comprehensive and reliable reference libraries. For many poorly known taxa, such reference sequences are missing even at higher-level taxonomic scales. We harvested the collections of the Smithsonian's National Museum of Natural History (USNM) to generate DNA barcoding sequences for genera of terrestrial arthropods previously not recorded in one or more major public sequence databases. Our workflow used a mix of Sanger and Next-Generation Sequencing (NGS) approaches to maximise sequence recovery while ensuring affordable cost. In total, COI sequences were obtained for 5,686 specimens belonging to 3,737 determined species in 3,886 genera and 205 families distributed in 137 countries. Success rates varied widely according to collection data and focal taxon. NGS helped recover sequences of specimens that failed a previous run of Sanger sequencing. Success rates and the optimal balance between Sanger and NGS are the most important drivers to maximise output and minimise cost in future projects. The corresponding sequence and taxonomic data can be accessed through the Barcode of Life Data System, GenBank, the Global Biodiversity Information Facility, the Global Genome Biodiversity Network Data Portal and the NMNH data portal.

Revision of Fortipalpa Kasparyan & Ruíz-Cancino, (Ichneumonidae, Cryptinae).

Fortipalpa Kasparyan & Ruíz-Cancino, 2007 originally accommodates only its type species, F. yucatanica Kasparyan & Ruíz-Cancino, 2007. The genus can be distinguished from all other Neotropical Cryptini by having mandible with ventral tooth subequal to slightly longer than dorsal tooth; propodeum anteriorly smooth, posteriorly with transverse striation; T1 long and slender, without anterolateral tooth; and ovipositor moderately long and straight, with ventral valve without a lobe covering dorsal valve. The genus is revised, and the following new species are proposed, described, and illustrated: F. exelysae sp. nov., F. frida sp. nov., F. heredia sp. nov., F. panamensis sp. nov., F. pichincha sp. nov., F. sacha sp. nov., and F. shakira sp. nov. The male of F. exelysae sp. nov., the first for the genus, is also described and illustrated; other males remain unknown. The type species is redescribed and illustrated, including pictures of the paratype and a new record to Trinidad. Figures, distribution maps and a taxonomic key are provided for all valid species.

Many evolutionary roads led to virus domestication in ichneumonoid parasitoid wasps.

The investigation of endogenous viral elements (EVEs) has historically focused on only a few lineages of parasitoid wasps, with negative results consistently underreported. Recent studies show that multiple viral lineages were integrated in at least seven instances in Ichneumonoidea and may be much more widespread than previously thought. Increasingly affordable genomic and bioinformatic approaches have made it feasible to search for viral sequences within wasp genomes, opening an extremely promising research avenue. Advances in wasp phylogenetics have shed light on the evolutionary history of EVE integration, although many questions remain. Phylogenetic proximity can be used as a guide to facilitate targeted screening, to estimate the number and age of integration events and to identify taxa involved in major host switches.

Mitochondrial phylogenomics and mitogenome organization in the parasitoid wasp family Braconidae (Hymenoptera: Ichneumonoidea).

BACKGROUND: Mitochondrial (mt) nucleotide sequence data has been by far the most common tool employed to investigate evolutionary relationships. While often considered to be more useful for shallow evolutionary scales, mt genomes have been increasingly shown also to contain valuable phylogenetic information about deep relationships. Further, mt genome organization provides another important source of phylogenetic information and gene reorganizations which are known to be relatively frequent within the insect order Hymenoptera. Here we used a dense taxon sampling comprising 148 mt genomes (132 newly generated) collectively representing members of most of the currently recognised subfamilies of the parasitoid wasp family Braconidae, which is one of the largest radiations of hymenopterans. We employed this data to investigate the evolutionary relationships within the family and to assess the phylogenetic informativeness of previously known and newly discovered mt gene rearrangements. RESULTS: Most subfamilial relationships and their composition obtained were similar to those recovered in a previous phylogenomic study, such as the restoration of Trachypetinae and the recognition of Apozyginae and Proteropinae as valid braconid subfamilies. We confirmed and detected phylogenetic signal in previously known as well as novel mt gene rearrangements, including mt rearrangements within the cyclostome subfamilies Doryctinae and Rogadinae. CONCLUSIONS: Our results showed that both the mt genome DNA sequence data and gene organization contain valuable phylogenetic signal to elucidate the evolution within Braconidae at different taxonomic levels. This study serves as a basis for further investigation of mt gene rearrangements at different taxonomic scales within the family.

A new species, key and further redefinition of Nesolinoceras Ashmead (Hymenoptera, Ichneumonidae, Cryptinae).

A new species of Nesolinoceras Ashmead from the Dominican Republic is described and illustrated. A key to the species and a revised and expanded distribution map is presented. This is the third species recognized for the genus, and the first one confirmed to occur in high altitude (1100 m). When compared with the previously known species, a number of unique and shared features emerged, supporting a redefinition of the genus beyond that of Santos (2016): (1) the known inter-specific variability of 48% of the examined characters increased considerably, confirming their diagnostic value at the species-level, and (2) the stability of six features support them as additionally diagnostic for the genus: 2223 flagellomeres; mesoscutum subcircular, as long as wide; scuto-scutellar carina absent, axillary trough shallow, indistinct on scutellum; subalar ridge wide, somewhat ovoid, not keeled; crossvein 1cu-a arising basad of vein 1M+Cu by about 0.3 its length; and vein 2-M only slightly longer than vein 3-M. Furthermore, two important measurements for the diagnosis of Nesolinoceras now have new, expanded ranges: areolet 1.82.6 as long as pterostigma width, and areolet 0.71.0 as long as wide. The new taxon is readily recognizable by having the body mostly brown, fully infuscated wings, and the longest ovipositor of the species, among other diagnostic features. A new geographic record and in situ photo are also provided for N. laluzbrillante Santos.

A new species and southernmost record of Cestrus Townes (Hymenoptera, Ichneumonidae, Cryptinae).

Cestrus itatiensis sp. nov., from the Brazilian Atlantic Forest, is described and illustrated. This is the southernmost record for the genus and the first record in Brazil. The new species is characterized by having the body reddish brown; a narrow yellow stripe along eye margin on supra-clypeal area, reaching briefly the supra-antennal area; transverse carina of propodeum complete and stout; postpetiole and T2-8 progressively dark brown towards apical margin in a somewhat triangular pattern; and apex of dorsal valve of ovipositor with nine distinct teeth. Extensive sampling suggest this is a rare species.

Species delimitation, environmental cline and phylogeny for a new Neotropical genus of Cryptinae (Ichneumonidae).

A morphologically unusual Cryptini, Cryptoxenodon gen. nov. Supeleto, Santos & Aguiar, is described and illustrated, with a single species, C. metamorphus sp. nov. Supeleto, Santos & Aguiar, apparently occurring in two disjunct populations in northern and southeastern South America. The highly dimorphic female and male are described and illustrated. The phylogenetic relationships of the new genus are investigated using a matrix with 308 other species of Cryptini in 182 genera, based on 109 morphological characters and molecular data from seven loci. The analyses clearly support Cryptoxenodon gen. nov. as a distinct genus, closest to Debilos Townes and Diapetimorpha Viereck. Species limits and definition are investigated, but despite much morphological variation the analyses at the specimen level do not warrant the division of the studied populations into separate species. The considerable morphological variation is explored with principal component analyses of mixed features, and a new procedure is proposed for objective analysis of colors. The relationship of color and structural variation with altitude and latitude is demonstrated and discussed, representing an important case study for Ichneumonidae. Externally, Cryptoxenodon gen. nov. can be recognized mainly by its unusually large mandibles, but other diagnostic features include clypeus wide; sternaulus complete, distinct and crenulate throughout; areolet closed, about as long as pterostigma width; petiole anteriorly with distinct triangular projection on each side, spiracle near posterior 0.25; propodeum without posterior transverse carina; and propodeal apophyses conspicuously projected.

Discovery of two new Andean species of Scolomus (Townes Townes), with a key to all known species (Hymenoptera: Ichneumonidae: Metopiinae).

Scolomus Townes Townes is a widely distributed genus of the family Ichneumonidae, with most species occurring in the New World. Herein two new species from Chile are described and illustrated. Scolomus maculatus sp. nov., which is characterized by a large rhomboid areolet and very wide RS vein in the fore wing, resembling a petiole; head and pronotum green, mesoscutum yellow with dark brown spots on its lateral lobes and around the scutellum. Scolomus clypeatus sp. nov., which is characterized by its wide clypeus, 3.00× as wide as long, with a rectangular aspect; head, mesoscutum, postscutellum and pronotum entirely yellow. The first key to all known species of the genus is also presented.

Ichneumonid wasps of the subfamily Mesochorinae: new replacement names, combinations and an updated key to the World genera (Hymenoptera: Ichneumonidae).

Mesochorinae is a relatively small but widely distributed subfamily of Ichneumonidae, with most species occurring in the Neotropical Region. Currently, there are two classifications in use regarding mesochorine genera, causing numerous taxonomic conflicts. To resolve nomenclature conflicts, seven new replacement names and twelve new combinations are proposed for species now recognized as members of Mesochorus Gravenhorst. Herein the synonymy of Plectochorus Uchida and Stictopisthus Thomson  with the genus Mesochorus s. lat. is reaffirmed, supported by the following shared character states: transverse subantennal carina partially or completely developed; supraclypeal area evenly convex, without median protrusion; clypeus not separated from supraclypeal area by depression or groove; and hind wing without vein Cu1b. An updated key to the World genera of Mesochorinae is also presented.

Neither barriers nor refugia explain genetic structure in a major biogeographic break: phylogeography of praying mantises in the Brazilian Atlantic Forest.

The Atlantic Forest is one of the world's top biodiversity hotspots, but the diversification processes of its biota are still poorly known, with competing models attributing dominant roles to either Quaternary climatic changes or geographic barriers. Many studies identify the Doce river as a major phylogeographic break, but the reasons for this phenomenon are highly debated. Here we test the predictions of the refugial and barrier models for a common species of praying mantis, Miobantia fuscata, focusing in the areas immediately south and north of the Doce river. Our analyses show high intraspecific genetic diversity, deep coalescence times and no evidence for recent population expansion. Phylogeographic structure is inconsistent with a refugial hypothesis. Significant gene flow between northern and southern populations also conflicts with a strong role for geographic barriers. This study highlights the need for considering invertebrate taxa to infer recent landscape changes, and points towards a more complex picture of genetic diversification in the Atlantic Forest.