Abstract Mantodea are among the most charismatic insects, and investigations of their evolutionary history interrogate some of their most compelling morphological traits. Despite this, studies of Mantodea systematics have often drastically changed historical perspectives about their evolutionary history, underscoring the lack of study on the group. The present account provides a comprehensive transcriptomic analysis with 51 species from 14 of the 16 established superfamilies that resolves several critical questions in mantodean phylogeny, including the relationship among the five major lineages, namely Chaeteessoidea, Mantoidoidea, Metallyticoidea, Amerimantodea and Cernomantodea. The inferred phylogeny highlights a well‐supported sister‐group relation between Chaeteessoidea and all remaining extant mantids. This placement of Chaeteessoidea implies that its unique thoracic musculature and foreleg morphology are indeed ancestral traits, which bridge cockroach‐like ancestors and derived Mantodea lineages. Our results are ambiguous about the position of Metallyticoidea in relation to Mantoidoidea, as we recover conflicting signals between the different analyses performed. Within Amerimantodea and Cernomantodea, we corroborate prior research by demonstrating the polyphyletic nature of traditional taxonomic groups and the significant role of morphological convergence in obscuring true evolutionary relationships. Divergence estimates place the split between cockroaches and praying mantises in the middle of the Permian period at ~274 ± 37 million years ago (mya), with crown‐Mantodea originating around 185 mya in the Jurassic, predating most previous estimates and suggesting a deep evolutionary history shaped by significant geological changes. In particular, the dated phylogeny and the distribution of Mantodea's major extant lineages provide additional evidence that vicariance associated with the breakup of Gondwana played a crucial role in the early diversification of the group. In summary, this study establishes a robust phylogenomic framework for Mantodea and clarifies long‐standing systematic uncertainties, providing a solid foundation for future research into praying mantises' evolution, biogeography and morphological diversification.
Complex diversification histories shaped by dynamic palaeogeographical processes, such as those associated with continental archipelagos, pose major challenges for delimiting taxonomic boundaries. The present study focuses on Dailognatha quadricollis, an insular group of darkling beetles with a complex taxonomic history that has presumably diversified non-adaptively across the Aegean archipelago. For the first time, the taxonomic status of the 12 subspecies assigned to D. quadricollis is revised under an integrative framework, utilizing single nucleotide polymorphism (SNP) data obtained from double digest restriction-site associated DNA (ddRAD) sequencing in combination with morphological characters and geographic information. Congruence among alternative species delimitation methods supports the subdivision of D. quadricollis into three species, resulting in the following taxonomic changes: Dailognatha quadricollis (Brull & eacute;) (= Tentyria laticollis Besser syn. nov.), Dailognatha carceli Solier stat. resurr. (= D. quadricollis anaphiana Koch syn. nov., = D. quadricollis sporadica Koch syn. nov.), Dailognatha rugata Solier stat. resurr. (= D. obtusangula Reitter syn. nov., = D. quadricollis montana Koch syn. nov., = D. quadricollis plicata Koch syn. nov., = D. quadricollis pseudobtusangula Koch syn. nov., = D. quadricollis rhodica Koch syn. nov., = D. quadricollis samosana Koch syn. nov.). A taxonomic key for the three delimited species is also provided, demonstrating the taxonomic utility of characters not previously studied within the group, including genital morphology and mesoventrite punctation. Overall, this study highlights the value of integrating genomic, morphological and geographical data when delimiting cryptic lineages across geographically fragmented systems and raises broader questions regarding the systematics of insular species complexes comprising multiple subspecies.
The systematic placement of the enigmatic South African moth Apoprogones hesperistis Hampson, which superficially resembles hesperiid skippers and sematurid moths, has long been controversial. Its rarity has hindered detailed morphological study, and the absence of fresh material has precluded molecular analyses. We investigated its phylogenetic position using non-destructive DNA extraction from the holotype collected in 1902, followed by museomic sequencing, and complemented these data with non-destructive micro-CT-based morphological examination. In a first analysis based on a Macroheterocera dataset including mitogenomes of 114 taxa, A. hesperistis was placed within Geometroidea as sister to an undescribed lineage from southern South America. A second, Geometroidea-focused phylogenomic analysis of 45 taxa confirmed this placement, recovering A. hesperistis within the lineage of geometroid families lacking tympanic hearing organs, again as sister to the same undescribed South American lineage. Together, these taxa formed a clade positioned between the Southeast Asian families Pseudobistonidae and Epicopeiidae. We describe a new genus, Ona Sihvonen, Hausmann & Brehm gen. n., with Ona australis Sihvonen, Hausmann & Brehm sp. n. from Tierra del Fuego, Argentina, as type species. Apoprogones and Ona are classified in the revived family Apoprogonidae, each representing a monotypic subfamily-Apoprogoninae and Onainae-to reflect their distinctiveness. We further show that Geometroidea comprises both tympanic and non-tympanic lineages, with 99.4% of described species possessing hearing organs. We hypothesize that the abdominal hearing organ, particularly the geometrid-specific structure called ansa, may represent a key innovation promoting diversification of the lineage post Cretaceous-Tertiary (K-T) boundary ca 66 Mya.
Ambrosia beetles farm fungi as a food source in tunnels within wood. Obligate fungus farming in ambrosia beetles has evolved more than 10 times independently and is primarily studied in the curculionid subfamilies Scolytinae and Platypodinae. In contrast, ambrosia beetles of the superfamily Lymexyloidea are sparsely studied, leaving the occurrence of fungus farming and its evolutionary history poorly understood. Larvae of the genus Elateroides Schaeffer cultivate mutualistic Alloascoidea Kurtzman & Robnett yeasts for nutrition within their tunnels in dead wood. These mutualistic fungi are vertically transmitted via specialized pouches, known as mycetangia, located at the ovipositor of adult females. This study aims to determine the origin of obligate fungus farming in Lymexyloidea. We reconstructed molecular phylogenies of the superfamily using two datasets: (1) a dataset with mitochondrial genome sequences from 47 taxa spanning the entire Lymexyloidea and (2) a phylogenomic dataset of 827 BUSCO orthologs from five representative lymexyloid taxa and four outgroups. We used micro-computed tomography (micro-CT) to detect mycetangia on the ovipositors of 13 extant lymexyloid taxa and on the extinct species Elateroides helmi sp. nov., preserved in Baltic amber. The discovery of E. helmi documents the presence of the Hylecoetinae in the Eocene. By integrating mycetangia presence/absence data with our phylogeny, we conclude that obligate fungiculture in the Lymexyloidea likely originated once within the subfamily Hylecoetinae.
A novel phylogenetic hypothesis for the subfamilies Dyopsinae and Thiacidinae is presented. For the first time, the type genus and species of Thiacidinae, Thiacidas postica Walker, is sequenced and analysed within an expanded dataset that includes a diverse array of representative Noctuidae. Utilizing a combination of phylogenomic data and traditional phylogenetic analyses based on legacy (Sanger) sequence data, the results strongly support its placement within Dyopsinae. Thiacidas postica and other Thiacidas Walker species are placed within the Ceroctena Guen & eacute;e clade (in the sense of Zahiri et al., 2013), forming a strongly supported sister relationship with the African genus Sciatta Walker. Consequently, the subfamily Thiacidinae is removed from synonymy with Pantheinae and reinstated with tribal status as Thiacidini (stat. nov.) within Dyopsinae. This tribe includes the Old World genera Amblyprora Bethune-Baker, Sciatta, Thiacidas and the Old World tropical Trisula Moore. Four additional tribes are recognized within Dyopsinae: Dyopsini, Desmolomini Schmidt & Zahiri, tribus nova, Momini (stat. rev.) and Litoprosopini (stat. rev.). The Neotropical genus Pachyplastis Felder is transferred from Erebidae to Dyopsinae: Desmolomini.
Here, we present a phylogenetic analysis of Curculionini based on 84 ingroup species from seven genera; 19 fig-associated species were newly sequenced for this study from material reared from approximately 110 fig trees representing 25 Ficus species, while the remaining 65 Curculionini species and all 61 outgroup taxa were obtained from previously published datasets on GenBank. Our analyses suggest that Curculionini originated in the early Palaeocene (ca. 65 Ma), with crown diversification beginning in the early Eocene, broadly coincident with the Eocene Climatic Optimum and the expansion of tropical Ficus (Moraceae). Although traditionally regarded as nut- and gall-feeding beetles, our results indicate that fig feeding was likely the ancestral host association within the tribe. At least three independent host shifts subsequently occurred towards other rosalean fruits, insect galls and hard seeds of Fagaceae and Juglandaceae. These shifts are temporally concentrated from the late Eocene onward, coinciding with global climatic cooling and the expansion of temperate broad-leaved forests. Biogeographic reconstruction supports an Afrotropical origin of Curculionini, followed by dispersal into the Oriental and western Palearctic regions. A single Beringian dispersal event gave rise to the Nearctic acorn-weevil radiation. Our analyses further reveal extensive polyphyly within Curculio and demonstrate that current subtribal classifications do not reflect evolutionary history. Taken together, our results are consistent with the hypothesis that morphological and physiological preadaptations for exploiting concealed and chemically defended plant tissues, interacting with major Cenozoic Earth-history events, shaped the global diversification of Curculionini.
This study evaluates the phylogenetic validity of traditionally defined species groups within the genus Eurytoma Illiger in the Palaearctic region using mitochondrial genome data, with a focus on genus- and species-group-level relationships. A total of 166 specimens were analysed, comprising 159 ingroup specimens and seven outgroups, covering 13 of the 18 historically recognized species groups. Ten groups were recovered within Eurytoma sensu stricto (s.s.), whereas several traditionally recognized species groups were polyphyletic or placed outside Eurytoma, thereby refining the generic limits of the genus. Notably, morphological traits used in species-group diagnosis, including the postgenal depression and ventral shelf structure, corresponded closely with the molecular phylogeny, whereas classifications based primarily on ecological similarity or putatively adaptive traits were not consistently supported. Ancestral state reconstruction revealed that entomophagy was ancestral across Eurytomidae, with a single transition to phytophagy inferred within Eurytoma s.s., corresponding to the fumipennis and amygdali groups. Multistate analyses further identified recurrent transitions among gall-associated parasitism, seed-feeding and coleopteran parasitism, including at least two independent origins of coleopteran parasitism within Eurytoma s.s. These results indicate that host-use is phylogenetically labile in Eurytoma and that convergent ecological strategies have evolved repeatedly across the genus. This study provides the first mitogenome-based phylogenetic test of historical species-group coherence in Eurytoma at the genus- and species-group levels. The revised framework clarifies long-standing ambiguity in the generic boundaries of Eurytoma and provides a practical phylogenetic and morphological reference for placing newly described species and previously unassigned taxa within species groups.
The genus Chelonus Panzer is one of the largest genera within the subfamily Cheloninae with 11 subgenera and more than 1000 valid species. However, the phylogenetic relationships of species within this genus and their coevolutionary relationships with endogenous bracoviriforms (BVfs) remain unclear. Here we provide the first phylogenomic analysis of 42 taxa representing all 11 subgenera of Chelonus and other species within the Cheloninae using 2380 loci of ultraconserved elements (UCEs). The results showed that the subgenus Arichelonus Viereck has an isolated position outside of the rest of Chelonus, while the other 10 subgenera formed a monophyletic group, the traditional genus Chelonus. Four subgenera within Chelonus, that is, Chelonus s. str., Megachelonus Baker, Microchelonus Sz & eacute;pligeti and Parachelonus Tobias are polyphyletic, suggesting that the traditional genus Chelonus could be further divided into 14 clades. Divergence time estimates indicated that the subgenus Arichelonus diverged approximately 35.5 million years ago, while most species within the genus Chelonus diversified <21.9 million years ago. Ancestral state reconstructions revealed morphological innovations of the subfamily, including metasomal curvature, which may be linked to host-use strategies. Cophylogenetic analyses demonstrated that a strong congruence of parasitoid-bracoviriform exists and confirmed that BVf nudivirus genes and virulence genes displayed different evolutionary trajectories. Altogether, our findings provide new insights into the evolutionary history of the genus Chelonus and substantiated the coevolutionary relationships between the parasitoids and their associated BVfs.
Recent Pleistocene radiations often generate cryptic diversity and semi-permeable species boundaries, challenging species delimitation and the reconstruction of evolutionary histories. Here, we investigate these processes in a species complex of Chorthippus grasshoppers from Greece using genome-wide ddRAD-seq data combined with geometric morphometrics. Phylogenomic analyses indicate that diversification occurred during the Middle Pleistocene and identify two major geographic clades separated by the Corinthian Gulf, followed by finer-scale divergence within the Peloponnese Peninsula. Most taxa form well-supported, reciprocally monophyletic lineages despite recent divergence, although several populations at range margins show signatures of historical hybridization, revealing semi-permeable species boundaries shaped by secondary contact. Introgression is geographically restricted and strongly asymmetric, occurring primarily from long-winged into short-winged taxa, consistent with differences in dispersal capacity. Species delimitation analyses largely support current taxonomy but also identify three previously unrecognized evolutionary lineages corresponding to geographically restricted populations, uncovering additional cryptic diversity. Morphological analyses reveal pronounced stasis in shape-related traits across lineages, with wing length emerging as the main axis of differentiation. Phylogenetic reconstructions indicate multiple independent reductions in wing size, highlighting repeated evolutionary transitions toward reduced dispersal. In at least one lineage, a pronounced geographic cline in wing length is mirrored by shallow phylogenetic structure, supporting an incipient evolutionary shift in dispersal capacity rather than phenotypic plasticity. Admixed populations consistently retain short-winged phenotypes, suggesting strong selection on dispersal-related traits despite genomic introgression. Overall, our results demonstrate how topographic complexity, Pleistocene climatic oscillations and dispersal-related morphology interact to shape diversification and maintain lineage integrity in a Mediterranean biodiversity hotspot.
Internal anatomical characters of Cicadellidae have rarely been incorporated into higher-level systematics, despite their potential phylogenetic signal. We compared the digestive, excretory (Malpighian tubules) and reproductive systems of 39 leafhopper species representing 12 cicadellid subfamilies, together with five outgroup taxa from related hemipteran lineages. Standardized dissections and light-microscope imaging were used to document organ architecture. Forty-two discrete characters were scored for phylogenetic inference using maximum parsimony using both equal and implied weighting (k = 9). Subfamilies differ significantly in the organization of salivary glands, the relative development of the filter chamber, conical segment and midgut loop, the morphology and pigmentation of Malpighian tubule segments, as well as the configuration of male and female reproductive systems (e.g., seminal vesicle encapsulation and connectivity; common oviduct dilation and accessory gland number/position). The resulting trees recover Cicadellidae as paraphyletic with Membracidae nested within it, and indicate non-monophyly of several traditionally recognized subfamilies (including Evacanthinae, Deltocephalinae and Eurymelinae). The results demonstrate that internal anatomical characters provide informative, complementary evidence for resolving relationships among major cicadellid lineages and will be useful for future systematic revisions.
This study presents the first total evidence dated phylogenetic analysis of the scorpion genus Urophonius, integrating 115 morphological characters and five molecular markers (28S, 18S, H3, 16S, COI). Our comprehensive phylogenetic framework provides novel insights into the genus' diversification timeline and evolutionary processes. Additionally, we described Urophonius andinus n. sp. from the central Chilean Andes, a high-altitude species found at 2400 m.s.a.l., representing the highest elevational record for the genus. This new species is placed within the granulatus species group, characterized by a spring-summer activity period. Este estudio representa el primer an & aacute;lisis filogen & eacute;tico de evidencia total del g & eacute;nero de escorpiones Urophonius, integrando 115 caracteres morfol & oacute;gicos y cinco marcadores moleculares (28S, 18S, H3, 16S, COI). Nuestro amplio marco filogen & eacute;tico brinda nuevas perspectivas sobre los tiempos y los procesos de diversificaci & oacute;n del g & eacute;nero. Adicionalmente, describimos a Urophonius andinus n. sp. de los Andes centrales de Chile, una especie de altura encontrada a unos 2400 m sobre el nivel del mar, lo que representa el r & eacute;cord altitudinal para el g & eacute;nero. Esta nueva especie forma parte del grupo de especies granulatus, que presenta un ciclo de actividad en primavera-verano.
Myrmeleontini is a widespread antlion tribe within the lacewing family Myrmeleontidae, well known for its larvae building cone-shaped pits in sandy soil to trap prey. The monophyly of the tribe is well supported, but the monophyly of many genera within the tribe as well as the definition of the genus Myrmeleon Linnaeus has long been questioned. Here, we present a phylogenomic analysis of Myrmeleontini with emphasis on the Eurasian fauna, using ultraconserved elements (UCE) data. Our results recovered Myrmeleon as a diverse grade, with Baliga Nav & aacute;s, Callistoleon Banks, Euroleon Esben-Petersen, Hagenomyia Banks and Megistoleon Nav & aacute;s deeply nested within it. Based on our sampling of type species of many genera (including those synonymized with Myrmeleon) and detailed morphological comparisons, we provide a new phylogeny-based classification of Myrmeleontini. Accordingly, Myrmeleon s. str. only refers to the formerly recognized M. formicarius group from Palaearctic region. The status of 12 genera that were previously considered as junior synonyms of Myrmeleon is restored (i.e., Banyaleon Zheng & Liu nom. n., Bordus Nav & aacute;s stat. rev., Callistoleon Banks stat. rev., Cocius Nav & aacute;s stat. rev., Enza Nav & aacute;s stat. rev., Macroleon Banks stat. rev., Moreyus Nav & aacute;s stat. rev., Morter Nav & aacute;s stat. rev., Neleon Nav & aacute;s stat. rev., Neseurus Nav & aacute;s stat. rev., Myrmeleodes Nav & aacute;s stat. rev. and Tafanerus Nav & aacute;s stat rev.), while Neohornius stat. rev., previously treated as a subgenus of Myrmeleon, is herein elevated to genus. Additionally, based on comprehensive sampling from the Oriental region, five new genera from this region are described, that is, Ancyroleon Zheng & Liu gen. n., Baligaptes Zheng & Liu gen. n. Melanobaliga Zheng & Liu gen. n., Orientaleon Zheng & Liu gen. n. and Sinobaliga Zheng & Liu gen. n.
Accurate species delimitation and identification are crucial for assessing biodiversity and conserving species. The Diachrysia cryptic moth complex, comprising D. chrysitis and D. stenochrysis, exhibits overlapping morphological traits, complicating species identification and rendering their status as distinct species debatable. We applied a target enrichment approach leveraging 1753 nuclear single-copy orthologs to clarify their relationships and test the power of this method in addressing complex taxonomic questions. Phylogenetic and population structure analyses revealed clear nuclear differentiation between the two taxa, contrasting with the variability in wing pattern observed within each species. We also detected rare cases where the mitochondrial DNA barcode region (COI) is shared between the species. This discordance between genetic and morphological variation suggests that wing pattern alone is often unreliable for species diagnosis. Our study advances understanding of the evolutionary and speciation history of Diachrysia and provides an efficient genomic model for addressing complex taxonomic questions of recently diverged sibling taxa.
In the absence of differential adaptation to ecological niche and the development of reproductive barriers, boundaries between coexisting species are expected to breakdown. The fruit fly species pair, Bactrocera tryoni and B. neohumeralis, have significant overlap in geographic range and host use, with mating time the only confirmed difference in their mating systems. Using a combination of ecological and genomic data, we tested the roles of competition and reproductive isolation on the maintenance of species boundaries in this pair. Genome-wide SNP analyses found strong genetic differentiation between the species with no evidence for hybridization. Most outlier SNPs were restricted to narrow regions towards the centromeres and telomeres of chromosomes, while high nucleotide diversity rates were observed in both species. Enrichment of annotation terms indicated an overabundance of genes with the 'abnormal neuroanatomy' term, but non-enrichment of terms associated with sleep and circadian rhythm. Field data found minimal evidence for ecological differentiation based on significant positive temporal, spatial and hostuse correlations between the species. However, B. tryoni appears less sensitive to low humidity than B. neohumeralis and used hosts which were not used by B. neohumeralis suggesting subtle microecological variation between species contributing to reproductive isolation. The simultaneous comparisons of molecular and ecological data in our study have provided a more nuanced understanding of how species boundaries have been maintained in this sibling species pair. Our analyses highlight the importance of genetic divergence for their maintenance in sympatry, but less for a role of competitive displacement or other ecological adaptation.
The ground beetles (Carabidae) are a highly species-rich lineage of the Coleoptera, with over half of their diversity concentrated in the similar to 20,000 described species in the subfamily Harpalinae sensu lato. As a presumed recent radiation lacking deeply distinct morphological divisions, their taxonomic classification has been challenging, while molecular studies remain limited in the number of genes and taxa sampled. Using similar to 450 mitochondrial genome sequences from across the Carabidae and the major biogeographic realms we investigate the tribal relationships in Harpalinae. Our phylogenetic analysis supports a revised system that broadly divides the harpalines into two major reciprocally monophyletic lineages, corresponding to a narrowly defined Harpalinae sensunovo and a distinct Lebiinae. Within Harpalinae, we recover well-supported subclades that mostly represent existing tribes (e.g., Harpalini, Pterostichini, Licinini, Platynini), while clades in Lebiinae required the recognition of three new or redefined clades: Lebiini, Agrini and Odacanthini. We also establish the polyphyletic status of the 'Truncatipennes' defined by truncated elytra and traditionally encompassing most 'lebiomorphs', which are split into the Lebiinae and at least two additional lineages, corresponding to the Dryptinae and Brachininae (bombardier beetles) branching below the Harpalinae + Lebiinae clade. The mitogenome data were extended to include similar to 7000 species of Carabidae by adding all available cytochrome c oxidase subunit I (COI) barcodes and other legacy sequences. The resulting phylogeny broadly concurs with the tribal boundaries defined by mitogenomes and provides a curated barcode reference library for species identification. The unprecedented scale of mitogenome sequencing, combined with dense taxon sampling of barcodes, resolves a particularly complex portion of the beetle tree-of-life.
Bibionomorpha is a large and diverse dipteran infraorder, of which its composition and family-level relationships have long been debated. In this study, we constructed a phylogenomic tree of Bibionomorpha using an extensive dataset of transcriptomic, genomic and anchored hybrid enrichment data. To further investigate the evolutionary timeline of the group, we also generated a fossil-calibrated time tree using data-driven calibration priors. Bibionomorpha, comprising Anisopodoidea, Bibionoidea, Scatopsoidea and Sciaroidea, is recovered as the sister group to Brachycera. Axymyiidae and Perissommatidae, which have been considered part of Bibionomorpha by some authorities, are instead recovered as sister to the Bibionomorpha + Brachycera. We discuss recalcitrant nodes within the infraorder, particularly regarding the placements of Bolitophilidae and Cecidomyiidae. Although tentative, phylogenetic network analysis suggests a possible reticulation event leading to Bolitophilidae, while mitochondrial data support a sex-specific hybridization event between ancestral Cecidomyiidae and Sciaridae + Diadocidiidae group. Time tree analyses suggest a Lower Triassic or deeper origin of Bibionomorpha, consistent with fossil evidence.
The cicada tribe Platypleurini, characterized by a remarkably flattened body, is distributed mainly in the Afrotropical and Oriental regions with a few species also occurring in the eastern Palearctic Region. Prior phylogenetic analyses revealed polyphyly among Oriental genera, and phylogeny and phylogeographical patterns of related 'sibling' species remain unclear. Here we investigated the phylogenetic relationships of representative platypleurines and population differentiation of two Oriental species using molecular data combined with genomic data of their obligate endosymbionts Karelsulcia and Hodgkinia. We revealed that platypleurines and Karelsulcia exhibit congruent phylogenies at both the species and population levels, and that Karelsulcia has undergone continuous genomic erosion, manifested as varying degrees of pseudogenization. The phylogeny of Hodgkinia also reflects that of platypleurines, and its genomes underwent further splitting and expansion concomitant with the diversification of host cicadas. The initial divergence of Platypleurini and that of the major Afrotropical and Asian lineages were ca. 44.58 Ma and ca. 37.27 Ma, respectively, corresponding to the initial collision between the Arabian Plate and the Eurasian Plate during the Late Eocene and Early Oligocene. We established Asianopleura gen. nov ., reassigned Eopycna coelestia to Neoplatypleura and described E. autumnalis sp. nov . Both E. repanda and N. coelestia comb. nov . diverged into distinct phylogroups during the Late Miocene and Early Pliocene, indicating that contemporaneous geological activities and climatic oscillations acted as pivotal drivers in the diversification of these mountain specialists. This study yields novel insights into ecological diversification of Platypleurini and co-evolutionary dynamics between cicadas and associated endosymbionts.
Typhlocybinae is one of the most diverse groups of leafhoppers, constituting an important component of phytophagous insect diversity. The traditional tribal-level phylogenetic relationships within this subfamily remain contentious, with differing hypotheses implying distinct evolutionary histories. This study contributes to resolving these controversies using phylogenomics. We newly sequenced low-coverage whole genomes for 54 species spanning the six classic tribes of Typhlocybinae. From these data, we extracted thousands of universal single-copy orthologs (USCOs) and ultraconserved elements (UCEs). Robust tribal-level phylogenies were reconstructed using multiple dataset matrices (USCO50, USCO70, USCO90, USCO_fna, ClipKIT_USCO70, UCE_fna) and tree-building strategies, including partitioned maximum likelihood with homogeneous models, unpartitioned heterogeneous mixture models and the multi-species coalescent (MSC) model. A particular focus was placed on elucidating the complex taxonomic status between Zyginellini and Typhlocybini, integrating molecular results with morphological evidence. Our findings indicate that the choice of molecular marker type and modelling methods can influence the inferred tribal-level relationships. Data filtering improves tribal-level support. The final analyses reveal that Zyginellini is not monophyletic but is intermixed with Typhlocybini, although some Zyginellini lineages appear to have originated earlier than the Old World Typhlocybini. The other four tribes of Typhlocybinae are all monophyletic. The tribal-level phylogenetic relationship is: ((Zyginellini_Typhlocybini) + (Dikraneurini + Erythroneurini)) + (Alebrini + Empoascini). Furthermore, integrating wing venation morphology with previous molecular evidence, we propose the Eualebrina subtribe nov. of Typhlocybini (sensu lato). This study provides unprecedented genomic-scale data for Typhlocybinae and offers a framework to address similar phylogenetic challenges in other organisms.
DNA barcoding, the use of a standard DNA fragment for species identification, has emerged as a major field of biodiversity research. The effectiveness of this approach rests on the premise that much less variation exists within species than between them. While exceptions occur, this has been demonstrated in many animal taxa where the COI gene is effective in species discrimination. Sawflies are an exception to this pattern because DNA barcodes often fail to distinguish congeneric species. Using high-throughput single-molecule DNA sequencing to recover COI sequences from thousands of sawflies, we found that single individuals often possess multiple, seemingly functional, full-length DNA barcodes (i.e., unrecognizable as nuclear pseudogenes)-a phenomenon not documented at similar prevalence in any animal taxon. While the evolutionary causes of multiple variants require further investigation, our observation is remarkable as it violates the one-barcode-one-specimen assumption. The presence of multiple variants of barcodes within individuals does not jeopardize the concept, but it introduces a complexity for species inventories based on metabarcoding. They will overestimate the species count when barcode-based operational species units are used as species proxies. Similarly, DNA barcode reference libraries must consider how best to deal with the high frequency of multiple intra-individual variants.
Aotearoa New Zealand is home to a remarkable number of endemic taxa, some of which have existed on the archipelago since before the breakup of Gondwana. The mite harvesters (suborder Cyphophthalmi), tiny non-spider arachnids that dwell in forest leaf litter and caves, are one such group. The mite harvester family Pettalidae Shear exhibits a classic Gondwanan distribution with notable diversity in Aotearoa, which is home to three pettalid genera. Our research focuses on the evolution of the most widespread and speciose Aotearoa pettalid genus, Rakaia Hirst, 1926. Through phylogenetic analysis, we provide a window into patterns of ancient diversification and infer historical biogeographic trends. We generated subgenomic data through target enrichment of ultraconserved elements (UCEs) using an Arachnida-specific probe set; the 50% and 75% taxon-occupancy matrix retrieved 848 and 585 loci, respectively. In addition to generating the first fully resolved phylogeny of Rakaia, we performed a molecular clock analysis and tested for shifts in diversification rates in order to explore the effect of geological events such as the Oligocene Drowning, the uplift of K & amacr; Tiritiri o te Moana, and forest habitat contraction and fragmentation during the Last Glacial Maximum.
