Ultraconserved Elements Research Articles

Circumscription of the Ganaspis brasiliensis (Ihering, 1905) species complex (Hymenoptera, Figitidae), and the description of two new species parasitizing the spotted wing drosophila, Drosophila suzukii Matsumura, 1931 (Diptera, Drosophilidae)

Based on host specificity and distribution data, it has been hypothesized that Ganaspis brasiliensis (Ihering, 1905), a natural enemy of the horticultural pest spotted-wing drosophila, Drosophila suzukii Matsumura, 1931 (SWD), was composed of multiple, cryptic species. Parasitoid wasps assigned to the species name Ganaspis brasiliensis and Ganaspis cf. brasiliensis were investigated using a molecular dataset of ultra-conserved elements (UCEs) and morphology. We report strong evidence for the presence of cryptic species based on the combination of UCE data (1,379 UCE loci), host specificity, ovipositor morphology, and distribution data. We describe these new cryptic species as: Ganaspis lupinisp. nov., and Ganaspis kimorumsp. nov.Ganaspis lupini was formerly recognized as Ganaspis brasiliensis G3, and Ganaspis kimorum as Ganaspis brasiliensis G1. These two new species appear to be restricted to the temperate climates, whereas Ganaspis brasiliensis (formerly recognized as Ganaspis brasiliensis G5) has a more pan-tropical distribution. We investigated the characterization of the ovipositor clip of these species, and hypothesize that G. kimorum, which has a reduced ovipositor clip, has an advantage in ovipositing into fresh fruit, still on the host plant, while attacking SWD; as a corollary, G. brasiliensis and G. lupini, which both have a larger ovipositor clip, are better adapted to attacking hosts in softer, rotting fruit on the ground. As Ganaspis kimorum was authorized for release as a biological control agent against SWD under the name Ganaspis brasiliensis G1, the results here have direct impact on the field of biological control.

An ultraconserved element probe set for velvet worms (Onychophora)

Onychophora are cryptic, soil-dwelling invertebrates known for their biogeographic affinities, diversity of reproductive modes, close phylogenetic relationship to arthropods, and peculiar prey capture mechanism. The 216 valid species of Onychophora are grouped into two families – Peripatopsidae and Peripatidae – and apart from a few relationships among major lineages within these two families, a stable phylogenetic backbone for the phylum has yet to be resolved. This has hindered our understanding of onychophoran biogeographic patterns, evolutionary history, and systematics. Neopatida, the Neotropical clade of peripatids, has proved particularly difficult, with recalcitrant nodes and low resolution, potentially due to rapid radiation of the group during the Cretaceous. Previous studies have had to compromise between number of loci and number of taxa due to limitations of Sanger sequencing and phylotranscriptomics, respectively. Additionally, aspects of their genome size and structure have made molecular phylogenetics difficult and data matrices have been affected by missing data. To address these issues, we leveraged recent, published transcriptomes and the first high quality genome for the phylum and designed a high affinity ultraconserved element (UCE) probe set for Onychophora. This new probe set, consisting of ∼ 20,000 probes that target 1,465 loci across both families, has high locus recovery and phylogenetic utility. Phylogenetic analyses recovered the monophyly of major clades of Onychophora and revealed a novel lineage from the Neotropics that challenges our current understanding of onychophoran biogeographic endemicity. This new resource could drastically increase the power of molecular datasets and potentially allow access to genomic scale data from archival museum specimens to further tackle the issues exasperating onychophoran systematics.

Phylogeny and historical biogeography of the swallow family (Hirundinidae) inferred from comparisons of thousands of UCE loci

Swallows (Hirundinidae) are a globally distributed family of passerine birds that exhibit remarkable similarity in body shape but tremendous variation in plumage, sociality, nesting behavior, and migratory strategies. As a result, swallow species have become models for empirical behavioral ecology and evolutionary studies, and variation across the Hirundinidae presents an excellent opportunity for comparative analyses of trait evolution. Exploiting this potential requires a comprehensive and well-resolved phylogenetic tree of the family. To address this need, we estimated swallow phylogeny using genetic data from thousands of ultraconserved element (UCE) loci sampled from nearly all recognized swallow species. Maximum likelihood, coalescent-based, and Bayesian approaches yielded a well-resolved phylogenetic tree to the generic level, with minor disagreement among inferences at the species level, which likely reflect ongoing population genetic processes. The UCE data were particularly useful in helping to resolve deep nodes, which previously confounded phylogenetic reconstruction efforts. Divergence time estimates from the improved swallow tree support a Miocene origin of the family, roughly 13 million years ago, with subsequent diversification of major groups in the late Miocene and Pliocene. Our estimates of historical biogeography support the hypothesis that swallows originated in the Afrotropics and have subsequently expanded across the globe, with major in situ diversification in Africa and a secondary major radiation following colonization of the Neotropics. Initial examination of nesting and sociality indicates that the origin of mud nesting – a relatively rare nest construction phenotype in birds – was a major innovation coincident with the origin of a clade giving rise to over 40% of extant swallow diversity. In contrast, transitions between social and solitary nesting appear less important for explaining patterns of diversification among swallows.

Genome-wide Cas9-mediated screening of essential non-coding regulatory elements via libraries of paired single-guide RNAs

The functions of non-coding regulatory elements (NCREs), which constitute a major fraction of the human genome, have not been systematically studied. Here we report a method involving libraries of paired single-guide RNAs targeting both ends of an NCRE as a screening system for the Cas9-mediated deletion of thousands of NCREs genome-wide to study their functions in distinct biological contexts. By using K562 and 293T cell lines and human embryonic stem cells, we show that NCREs can have redundant functions, and that many ultra-conserved elements have silencer activity and play essential roles in cell growth and in cellular responses to drugs (notably, the ultra-conserved element PAX6_Tarzan may be critical for heart development, as removing it from human embryonic stem cells led to defects in cardiomyocyte differentiation). The high-throughput screen, which is compatible with single-cell sequencing, may allow for the identification of druggable NCREs.

Phylogeny and biogeography of harmochirine jumping spiders (Araneae: Salticidae)

We use ultraconserved elements (UCE) and Sanger data to study the phylogeny, age, and biogeographical history of harmochirine jumping spiders, a group that includes the species-rich genus Habronattus, whose remarkable courtship has made it the focus of studies of behaviour, sexual selection, and diversification. We recovered 1947 UCE loci from 43 harmochirine taxa and 4 outgroups, yielding a core dataset of 193 UCEs with at least 50 % occupancy. Concatenated likelihood and ASTRAL analyses confirmed the separation of harmochirines into two major clades, here designated the infratribes Harmochirita and Pellenita. Most are African or Eurasian with the notable exception of a clade of pellenites containing Habronattus and Pellenattus of the Americas and Havaika and Hivanua of the Pacific Islands. Biogeographical analysis using the DEC model favours a dispersal of the clade’s ancestor from Eurasia to the Americas, from which Havaika’s ancestor dispersed to Hawaii and Hivanua’s ancestor to the Marquesas Islands. Divergence time analysis on 32 loci with 85 % occupancy, calibrated by fossils and island age, dates the dispersal to the Americas at approximately 4 to 6 million years ago. The explosive radiation of Habronattus perhaps began only about 4 mya. The phylogeny clarifies both the evolution of sexual traits (e.g., the terminal apophyses was enlarged in Pellenes and not subsequently lost) and the taxonomy. Habronattus is confirmed as monophyletic. Pellenattus is raised to the status of genus, and 13 species moved into it as new combinations. Bianor stepposus Logunov, 1991 is transferred to Sibianor, and Pellenes bulawayoensis Wesołowska, 1999 is transferred to Neaetha. A molecular clock rate estimate for spider UCEs is presented and its utility to inform prior distributions is discussed.

UCE-derived mitochondrial phylogeny reveals pervasive mito-nuclear discordances in serotine bats (genus Eptesicus) and complex evolutionary history in Eptesicus (Histiotus)

Studies of evolution and biodiversity require solid understanding of species systematics revealed by molecular phylogeny using multilocus genomic data. Multilocus analyses, nevertheless, remain difficult in non-model taxa due to limited access to samples and molecular resources. To help overcome this limitation, ultra-conserved elements (UCEs) have been developed to generate large nuclear datasets and build more robust species phylogenies. Recently, MitoFinder pipeline was developed to further extract mitochondrial genes from the off-target sequences in UCE libraries to allow mito-nuclear comparison and increase the mitochondrial genomic database. Here we applied MitoFinder to published UCE datasets of serotine bats (genus Eptesicus) and focused on E. (Histiotus) whose evolutionary history is poorly understood. Our results showed extensive mito-nuclear discordances in the divergence of major clades in Eptesicus and within E. (Histiotus), indicating potential incomplete lineage sorting and historical mitochondrial introgression within and across subgenera. Moreover, we collected several new samples of E. (Histiotus), including the first molecular data of the recently described E. (H) diaphanopterus, and combined available published sequences to generate the most taxa-complete mitochondrial phylogeny of E. (Histiotus) bats. Results supported the early divergence of E. (H.) magellanicus and the species status of E. (H.) diaphanopterus. In addition, we found strong evidence of cryptic diversity, with potentially new taxa in Peru, Uruguay, and Brazil, which needs to be evaluated in future studies using complementary data. Our study enriched the sequence database of serotine bats and shed light on the hidden diversity and complex evolutionary history of E. (Histiotus).

Mitogenome architecture supports the non-monophyly of the cosmopolitan parasitoid wasp subfamily Doryctinae (Hymenoptera: Braconidae) recovered by nuclear and mitochondrial phylogenomics.

Mitochondrial DNA gene organisation is an important source of phylogenetic information for various metazoan taxa at different evolutionary timescales, though this has not been broadly tested for all insect groups nor within a phylogenetic context. The cosmopolitan subfamily Doryctinae is a highly diverse group of braconid wasps mainly represented by ectoparasitoids of xylophagous beetle larvae. Previous molecular studies based on Sanger and genome-wide (ultraconserved elements, UCE; and mitochondrial genomes) sequence data have recovered a non-monophyletic Doryctinae, though the relationships involved have always been weakly supported. We characterised doryctine mitogenomes and conducted separate phylogenetic analyses based on mitogenome and UCE sequence data of ~100 representative doryctine genera to assess the monophyly and higher-level classification of the subfamily. We identified rearrangements of mitochondrial transfer RNAs (tRNAs) that support a non-monophyletic Doryctinae consisting of two separate non-related clades with strong geographic structure ('New World' and 'Old World' clades). This geographic structure was also consistently supported by the phylogenetic analyses preformed with mitogenome and UCE sequence data. These results highlight the utility of the mitogenome gene rearrangements as a potential source of phylogenetic information at different evolutionary timescales.

The Rediscovery of a Relict Unlocks the First Global Phylogeny of Whip Spiders (Amblypygi).

Asymmetrical rates of cladogenesis and extinction abound in the tree of life, resulting in numerous minute clades that are dwarfed by larger sister groups. Such taxa are commonly regarded as phylogenetic relicts or "living fossils" when they exhibit an ancient first appearance in the fossil record and prolonged external morphological stasis, particularly in comparison to their more diversified sister groups. Due to their special status, various phylogenetic relicts tend to be well-studied and prioritized for conservation. A notable exception to this trend is found within Amblypygi ("whip spiders"), a visually striking order of functionally hexapodous arachnids that are notable for their antenniform first walking leg pair (the eponymous "whips"). Paleoamblypygi, the putative sister group to the remaining Amblypygi, is known from Late Carboniferous and Eocene deposits but is survived by a single living species, Paracharon caecusHansen (1921), that was last collected in 1899. Due to the absence of genomic sequence-grade tissue for this vital taxon, there is no global molecular phylogeny for Amblypygi to date, nor a fossil-calibrated estimation of divergences within the group. Here, we report a previously unknown species of Paleoamblypygi from a cave site in Colombia. Capitalizing upon this discovery, we generated the first molecular phylogeny of Amblypygi, integrating ultraconserved element sequencing with legacy Sanger datasets and including described extant genera. To quantify the impact of sampling Paleoamblypygi on divergence time estimation, we performed in silico experiments with pruning of Paracharon. We demonstrate that the omission of relicts has a significant impact on the accuracy of node dating approaches that outweighs the impact of excluding ingroup fossils, which bears upon the ancestral range reconstruction for the group. Our results underscore the imperative for biodiversity discovery efforts in elucidating the phylogenetic relationships of "dark taxa," and especially phylogenetic relicts in tropical and subtropical habitats. The lack of reciprocal monophyly for Charontidae and Charinidae leads us to subsume them into one family, Charontidae, new synonymy.

Skimming genomes for systematics and DNA barcodes of corals.

Numerous genomic methods developed over the past two decades have enabled the discovery and extraction of orthologous loci to help resolve phylogenetic relationships across various taxa and scales. Genome skimming (or low-coverage genome sequencing) is a promising method to not only extract high-copy loci but also 100s to 1000s of phylogenetically informative nuclear loci (e.g., ultraconserved elements [UCEs] and exons) from contemporary and museum samples. The subphylum Anthozoa, including important ecosystem engineers (e.g., stony corals, black corals, anemones, and octocorals) in the marine environment, is in critical need of phylogenetic resolution and thus might benefit from a genome-skimming approach. We conducted genome skimming on 242 anthozoan corals collected from 1886 to 2022. Using existing target-capture baitsets, we bioinformatically obtained UCEs and exons from the genome-skimming data and incorporated them with data from previously published target-capture studies. The mean number of UCE and exon loci extracted from the genome skimming data was 1837 ± 662 SD for octocorals and 1379 ± 476 SD loci for hexacorals. Phylogenetic relationships were well resolved within each class. A mean of 1422 ± 720 loci was obtained from the historical specimens, with 1253 loci recovered from the oldest specimen collected in 1886. We also obtained partial to whole mitogenomes and nuclear rRNA genes from >95% of samples. Bioinformatically pulling UCEs, exons, mitochondrial genomes, and nuclear rRNA genes from genome skimming data is a viable and low-cost option for phylogenetic studies. This approach can be used to review and support taxonomic revisions and reconstruct evolutionary histories, including historical museum and type specimens.

Redefining Ormyridae (Hymenoptera, Chalcidoidea) with establishment of subfamilies and description of new genera

AbstractThe circumscription of the family Ormyridae (Hymenoptera: Chalcidoidea) is revised after phylogenetic analysis based on ultra‐conserved elements (UCEs) and comparative morphological assessment of the chalcid ‘Gall Clade’. Six genera are treated in the family, including two new genera, Halleriaphagus van Noort and Burks, gen. nov., and Ouma Mitroiu, gen. nov. One genus, Eubeckerella Narendran, is re‐assigned to the family, and Ormyrulus Bouček is synonymised with Ormyrus Westwood, syn. nov., resulting in the new combination Ormyrus gibbus (Bouček), comb. nov. The six genera are classified in three subfamilies, two of which are newly described, Asparagobiinae van Noort, Burks, Mitroiu and Rasplus, subfam. nov., and Hemadinae van Noort, Burks, Mitroiu and Rasplus, subfam. nov. Halleriaphagus is established for the newly described type species Halleriaphagus phagolucida van Noort and Burks, sp. nov., and Ouma is erected for O. daleskeyae Mitroiu, sp. nov., and O. emazantsi Mitroiu, sp. nov. Asparagobius is revised with description of Asparagobius bouceki van Noort, sp. nov., and Asparagobius copelandi Rasplus and van Noort, sp. nov. Asparagobius and Halleriaphagus are classified in Asparagobiinae, Hemadas in Hemadinae and Eubeckerella, Ormyrus and Ouma in Ormyrinae. The molecular support defining the ormyrid clade is corroborated by the proposed morphological synapomorphy of a foliaceous prepectus overlying the tegula base. Identification keys to the genera of Ormyridae and to the species of Asparagobius and Ouma are provided. Online Lucid identification keys and images of all the species treated herein are available at: http://www.waspweb.org.Zoobank Registration: LSID urn:lsid:zoobank.org:pub:8811695B-EE57-4C18-A6B6-E63D267E2373.

A phylogenomic approach to a taxonomic revision: a combination, new synonymies, and a description of two new species within the camel spider genus Chanbria Muma, 1951 (Solifugae: Eremobatidae)

Abstract This study summarizes the taxonomic treatment of the camel spider genus Chanbria Muma, 1951. Taking an integrative taxonomic approach incorporating phylogenomic, morphological, and geographical information, the genus is herein revised. Of the four species currently placed in the genus, two are retained: Chanbria regalis Muma, 1951 and Chanbria serpentinus Muma, 1951. Eremochelis plicatus (Muma, 1962) is transferred to this genus because it is consistently recovered in a clade with Chanbria based on several phylogenetic analyses using hundreds of loci recovered from ultraconserved element data. In this study, we re-analyse previously acquired genomic data to assess former species hypotheses and identify new morphological synapomorphies that support the monophyly of Chanbria. The genetic data support the synonymization of Chanbria rectus Muma, 1962 syn. nov. with C. regalis. Furthermore, we synonymize Chanbria tehachapianus Muma, 1962 syn. nov. with C. regalis because C. tehachapianus was erected based on limited morphological information and lack of geographical separation between other populations of C. regalis. Two new species, Chanbria brookharti sp. nov. and Chanbria mapemes sp. nov., are described. This brings the total number of species of Chanbria described to five recognized species: C. regalis, C. serpentinus, C. plicatus com. nov., C. brookharti sp. nov., and C. mapemes sp. nov.

Phylogenomics of Psammodynastes and Buhoma (Elapoidea: Serpentes), with the description of a new Asian snake family

Asian mock vipers of the genus Psammodynastes and African forest snakes of the genus Buhoma are two genera belonging to the snake superfamily Elapoidea. The phylogenetic placements of Psammodynastes and Buhoma within Elapoidea has been extremely unstable which has resulted in their uncertain and debated taxonomy. We used ultraconserved elements and traditional nuclear and mitochondrial markers to infer the phylogenetic relationships of these two genera with other elapoids. Psammodynastes, for which a reference genome has been sequenced, were found, with strong branch support, to be a relatively early diverging split within Elapoidea that is sister to a clade consisting of Elapidae, Micrelapidae and Lamprophiidae. Hence, we allocate Psammodynastes to its own family, Psammodynastidae new family. However, the phylogenetic position of Buhoma could not be resolved with a high degree of confidence. Attempts to identify the possible sources of conflict in the rapid radiation of elapoid snakes suggest that both hybridisation/introgression during the rapid diversification, including possible ghost introgression, as well as incomplete lineage sorting likely have had a confounding role. The usual practice of combining mitochondrial loci with nuclear genomic data appears to mislead phylogeny reconstructions in rapid radiation scenarios, especially in the absence of genome scale data.

Systematics and biogeography of a Sunda-Papuan snake lineage (Natricidae: Tropidonophis Jan 1863)

Abstract Sunda-Papuan keelback snakes (Serpentes: Natricidae: Tropidonophis Jan 1863) include 20 species distributed from the Philippines south-east through the Moluccas to New Guinea and Australia. Diversity of this insular snake lineage peaks on the island of New Guinea. Previous phylogenetic studies incorporating Tropidonophis have been limited to multi-locus Sanger-sequenced datasets with broad squamate or family-level focus. We used a targeted-sequence capture approach to sequence thousands of nuclear ultraconserved elements (UCEs) to construct the most comprehensive sequence-based phylogenetic hypothesis for this genus and estimate ancestral biogeography. Phylogenies indicate the genus is monophyletic given recent taxonomic reassignment of Rhabdophis spilogaster to Tropidonophis. All UCE phylogenies recovered a monophyletic Tropidonophis with reciprocally monophyletic Philippine and New Guinean clades. Divergence dating and ancestral range estimation suggest dispersal to New Guinea from the Philippines to have occurred during the Mid-Miocene via the Oceanic Arc Terranes. From Late Miocene into the Pliocene the genus experienced rapid diversification from orogeny of the New Guinean Central Cordillera from Oceanic Arc Terrane accretion on the northern boundary of the Sahul Shelf. Future collecting of missing taxa from the Moluccas and Indonesian Papua will better the understanding of non-volant faunal biogeography and diversification in this tectonically complex Pacific arena.

Untangling deep‐sea corals systematics: Description of a new family, Stephanocyathidae (Anthozoa, Scleractinia), through a genomic approach

AbstractOnce considered the most speciose mostly azooxanthellate scleractinian group, the family Caryophylliidae is found to be highly polyphyletic based on molecular data and is undergoing a process of systematic revision. High‐throughput sequencing methods coupled with morphological analyses have facilitated revision of several scleractinian lineages, including the aforementioned family. In previous studies that relied on few mitochondrial and nuclear markers, the caryophylliid genera Stephanocyathus and Vaughanella were phylogenetically recovered in separate clades from the lineage that includes the type genus of the family, Caryophyllia, which is considered the ‘true’ Caryophylliidae. To help untangle the relationship among taxa of this family, here a new deep‐sea scleractinian family (Stephanocyathidae Vaga, Cairns & Kitahara fam. nov.) is proposed based on phylogenomic reconstructions coupled with molecular features, specifically gene order, of the complete mitochondrial genome. Evolutionary reconstructions were based on both mitochondrial and nuclear ultraconserved elements (UCEs) and exon loci data sets and confirmed the divergent position of the genera Stephanocyathus and Vaughanella. The new family shows a specific gene transposition in the mitochondrial genome, not present in the ‘true’ caryophylliid lineage, but instead already observed for the species Paraconotrochus antarcticus, recovered as sister taxon of the here proposed new family. Although its phylogenetic position is unknown, the genus Ericiocyathus is also added to the new family, based on macromorphological similarities. This study represents a step forward in our understanding of deep‐sea corals relationships and provide further information (e.g., mitochondrial gene order) that will aid in future efforts of assessing the systematic of caryophylliid lineages.

Revalidation of the jumping spider genus Cheliceroides Żabka, 1985 based on molecular and morphological data (Araneae, Salticidae).

The monotypic genus Cheliceroides Żabka, 1985 is revalidated based on both molecular sequence data (ultra-conserved elements and protein coding genes of mitochondrial genomes) and morphological evidence. Our molecular phylogenetic analyses show that Cheliceroides is not closely related to Colopsus Simon, 1902, not even in the same tribe, and a comparative morphological study also demonstrates significant differences in the genital structures (i.e. in the shape of embolus, and with or without pocket on epigynum) of the two genera. Therefore, we remove Cheliceroides from the synonymy of Colopsus, and its generic status is revalidated.

Phylogenomics of the geometrid tribe Palyadini (Lepidoptera: Geometridae) reveals contrasting patterns of phylogenetic signal in wing colour characters.

Next generation sequencing techniques currently represent a practical and efficient way to infer robust evolutionary hypotheses. Palyadini is a small Neotropical tribe of geometrid moths composed of six genera that feature strikingly colourful wings. Here, we investigated patterns of evolution and amount of phylogenetic signal contained in various colour characters featured in the wings of members of this tribe by (i) inferring a robust phylogenetic hypothesis using ultraconserved elements (UCEs), and afterwards, (ii) mapping the morphological characters onto the molecular topology under a parsimonious ancestral character optimization. Our matrix, obtained with 60% completeness, includes 754 UCE loci and 73 taxa (64 ingroup, nine outgroup). Maximum likelihood and parsimony generated largely identical topologies with strongly supported nodes, except for one node inside the genus Opisthoxia. According to our topology, most wing colour characters are reconstructed as homoplastic, particularly at the tribe level, but five of the seven provide evidence supporting common ancestry at the genus level. Our results emphasize, once again, that no character system is infallible, and that more research is necessary to take our understanding of the evolution of wing colour in moths to a level comparable with the knowledge we have for butterflies.

Enigmas no longer: using ultraconserved elements to place several unusual hawk taxa and address the non-monophyly of the genusAccipiter(Accipitriformes: Accipitridae)

AbstractHawks, eagles, and their relatives (Accipitriformes: Accipitridae) are a diverse and charismatic clade of modern birds, with many members that are instantly recognized by the general public. However, surprisingly little is known about the relationships among genera within Accipitridae, and several studies have suggested that some genera (in particular, the megadiverse genus Accipiter) are not monophyletic. Here, we combine a large new dataset obtained from ultraconserved elements, generated from whole genome sequencing of 134 species, with publicly available legacy markers (i.e. a suite of commonly sequenced mitochondrial and nuclear genes) to infer a well-supported, time-calibrated phylogeny of 237 extant or recently extinct species. Our densely sampled phylogeny, which includes 90% of recognized species, confirms the non-monophyly of Accipiter and provides a sufficient basis to revise the genus-level taxonomy, such that all genera in Accipitridae represent monophyletic groups.

Genome-wide data support recognition of an additional species of Neotropical river otter (Mammalia, Mustelidae, Lutrinae)

Abstract Cryptic biodiversity continues to be revealed worldwide, even in apparently well-known groups such as carnivorans. The Neotropical Otter (Lontra longicaudis) presents shape variation in its nose pad, a character that has been used to differentiate species in this group. Based on this, 3 subspecies are recognized: L. l. annectens (Mexico, Central America, and South America west of the Andes), L. l. enudris (Amazon and Orinoco basins), and L. l. longicaudis (Paraná basin and remaining distribution). Previous studies partially supported their distinctness based on mitochondrial DNA markers, morphometrics, and ecological niche modeling. We analyzed genome-wide nuclear markers (ultraconserved elements) of 29 L. longicaudis individuals across the species’ range to assess its population structure. Phylogenomic analysis recovered L. longicaudis as paraphyletic with robust support, with 1 clade comprising samples from Mexico and Colombia (trans-Andean populations) and another encompassing the remaining samples (cis-Andean populations), which grouped with 2 other South American species, L. felina and L. provocax. Principal component and admixture analyses strongly differentiated the 2 main L. longicaudis groups, and distinguished the Amazonian individuals from the remaining cis-Andean samples. Our results support the recognition of trans-Andean populations of L. longicaudis as a distinct otter species, which should be recognized as Lontra annectens.

Machine learning approaches delimit cryptic taxa in a previously intractable species complex

Cryptic species are not diagnosable via morphological criteria, but can be detected through analysis of DNA sequences. A number of methods have been developed for identifying species based on genetic data; however, these methods are prone to over-splitting taxa with extreme population structure, such as dispersal-limited organisms. Machine learning methodologies have the potential to overcome this challenge. Here, we apply such approaches, using a large dataset generated through hybrid target enrichment of ultraconserved elements (UCEs). Our study taxon is the Aoraki denticulata species complex, a lineage of extremely low-dispersal arachnids endemic to the South Island of Aotearoa New Zealand. This group of mite harvesters has been the subject of previous species delimitation studies using smaller datasets generated through Sanger sequencing and analytical approaches that rely on multispecies coalescent models and barcoding gap discovery. Those analyses yielded a number of putative cryptic species that seems unrealistic and extreme, based on what we know about species’ geographic ranges and genetic diversity in non-cryptic mite harvesters. We find that machine learning approaches, on the other hand, identify cryptic species with geographic ranges that are similar to those seen in other morphologically diagnosable mite harvesters in Aotearoa New Zealand’s South Island. We performed both unsupervised and supervised machine learning analyses, the latter with training data drawn either from animals broadly (vagile and non-vagile) or from a custom training dataset from dispersal-limited harvesters. We conclude that applying machine learning approaches to the analysis of UCE-derived genetic data is an effective method for delimiting species in complexes of low-vagility cryptic species, and that the incorporation of training data from biologically relevant analogues can be critically informative.

Phylogenomics of Afrotherian mammals and improved resolution of extant Paenungulata

Molecular investigations have gathered a diverse set of mammals—predominantly African natives like elephants, hyraxes, and aardvarks—into a clade known as Afrotheria. Nevertheless, the precise phylogenetic relationships among these species remain contentious. Here, we sourced orthologous markers and ultraconserved elements to discern the interordinal connections among Afrotherian mammals. Our phylogenetic analyses bolster the common origin of Afroinsectiphilia and Paenungulata, and propose Afrosoricida as the closer relative to Macroscelidea rather than Tubulidentata, while also challenging the notion of Sirenia and Hyracoidea as sister taxa. The approximately unbiased test and the gene concordance factor uniformly recognized the alliance of Proboscidea with Hyracoidea as the dominant topology within Paenungulata. Investigation into sites with extremly high phylogenetic signal unveiled their potential to intensify conflicts in the Paenungulata topology. Subsequent exploration suggested that incomplete lineage sorting was predominantly responsible for the observed contentious relationships, whereas introgression exerted a subsidiary influence. The divergence times estimated in our study hint at the Cretaceous-Paleogene (K-Pg) extinction event as a catalyst for Afrotherian diversification. Overall, our findings deliver a tentative but insightful overview of Afrotheria phylogeny and divergence, elucidating these relationships through the lens of phylogenomics.