Delimiting Species Boundaries Research Articles

Current progress and future prospects for understanding genetic diversity of seed plants in China

AbstractGenetic diversity represents the fundamental basis of biological variation within species, and is therefore the ultimate representation of biodiversity. Despite that all forms of diversity being defined by genomic differences, genetic diversity has received relatively less attention compared with species and ecosystem diversity. Here, we review recent literature and conclude that progress in understanding genetic diversity of seed plants, is strongly associated with advances in sequencing technology. We here present case studies to illustrate the application of genetic diversity for tracing crop domestication and delimiting species boundaries. Understanding genetic diversity is particularly critical for the field of conservation biology, and there is a clear shift to population‐level genetic studies to understand rare species. We then document additional factors that potentially influence genetic diversity, including climate change, habitat fragmentation, and species invasion. Finally, we identify current research limitations and propose directions for future studies. We highlight the need to develop broad‐scale genetic diversity knowledge, combined with other aspects of diversity to improve biodiversity conservation outcomes. We conclude that populating global databases with genomic‐scale sequence data for all species is an urgent and achievable goal now.

Parallel evolution in an island archipelago revealed by genomic sequencing of Hipposideros leaf-nosed bats.

Body size is a key morphological attribute, often used to delimit species boundaries among closely related taxa. But body size can evolve in parallel, reaching similar final states despite independent evolutionary and geographic origins, leading to faulty assumptions of evolutionary history. Here we document parallel evolution in body size in the widely distributed leaf-nosed bat genus Hipposideros, which has misled both taxonomic and evolutionary inference. We sequenced reduced representation genomic loci and measured external morphological characters from three closely related species from the Solomon Islands archipelago, delimited by body size. Species tree reconstruction confirms the paraphyly of two morphologically designated species. The non-sister relationship between large-bodied H. dinops lineages found on different islands indicates large-bodied ecomorphs have evolved independently at least twice in the history of this radiation. A lack of evidence for gene flow between sympatric, closely related taxa suggests the rapid evolution of strong reproductive isolating barriers between morphologically distinct populations. Our results position Solomon Islands Hipposideros as a novel vertebrate system for studying the repeatability of parallel evolution under natural conditions. We conclude by offering testable hypotheses for how geography and ecology could be mediating the repeated evolution of large-bodied Hipposideros lineages in the Solomon Islands.

Population genomics and morphological data bridge the centuries of cyanobacterial taxonomy along the continuum of Microcoleus species

The filamentous cyanobacterium Microcoleus is among the most important global primary producers, especially in hot and cold desert ecosystems. This taxon represents a continuum consisting of a minimum of 12 distinct species with varying levels of gene flow and divergence. The notion of a species continuum is poorly understood in most lineages but is especially challenging in cyanobacteria. Here we show that genomic diversification of the Microcoleus continuum is reflected by morphological adaptation. We compiled a dataset of morphological data from 180 cultured strains and 300 whole genome sequences, including eight herbarium specimens and the type specimen of Microcoleus. We employed a combination of phylogenomic, population genomic, and population-level morphological data analyses to delimit species boundaries. Finally, we suggest that the shape of the filament apices may have an adaptive function to environmental conditions in the soil.

The Extraordinary Diversity of Merodon avidus Complex (Diptera: Syrphidae)-Adding New Areas, New Species and a New Molecular Marker.

In this paper, the Merodon avidus (Diptera, Syrphidae) species complex was revised, whereupon we discovered and described four new species for science: Merodon atroavidus Vujić, Radenković et Likov sp. nov., M. magnus Vujić, Kočiš Tubić et Ačanski sp. nov., M. nigroscutum Vujić, Radenković et Likov sp. nov. and M. pseudomoenium Vujić, Kočiš Tubić et Ačanski sp. nov. An integrative taxonomy approach was used to delimit species boundaries. Two molecular markers (the mitochondrial COI gene and nuclear 28S rRNA gene-newly analysed marker for the complex) and geometric morphometry of the wing shape, together with morphological data and distribution, successfully separated all species from the complex. The morphological variability of the analysed species is described and discussed and an illustrated diagnostic key for typical morpho-forms of species from the M. avidus complex is presented. A distribution map of all investigated species from the complex is provided. The level of endemicity of the M. avidus complex was discussed.

An integrative delimitation of the species' boundaries within one hornbeam species complex (Betulaceae: Carpinus)

AbstractBoth hybridization and intraspecific morphological variation across environmental gradients complicate species delineation. We aimed to discern both possibilities that may blur species boundaries in the Carpinus viminea–Carpinus laxiflora–Carpinus londoniana species complex. We conducted statistical analyses on 535 specimens encompassing the entire distribution of this species complex to identify phenotypic clusters. Additionally, we analyzed genetic divergence and probable hybridization between clusters using 76 individuals from 37 populations. Based on phenotypic and genetic clusters, we tentatively recognized four species: C. viminea, C. fargesii, C. laxiflora, and C. londoniana. Except for rare overlapping distributions between C. fargesii and C. londoniana, the redefined four species are mostly allopatric to each another based on their distributions. The morphological delimitation, species boundary and distribution of each species differ distinctly from past taxonomic treatments. For example, specimens previously identified under C. viminea, in fact, belong to three different species. Hybrids between C. fargesii and C. londoniana exhibit morphological traits similar to C. viminea, thereby contributing to difficulties in determining species boundaries and outlining species distributions. These findings suggest that local selection and geographical isolation may together have promoted both phenotypic and genetic divergences within this species complex. However, interspecific hybridization blurs species boundaries by producing hybrids with phenotypic similarity in addition to intraspecific variation. This study emphasizes the importance of statistical analyses of population‐level morphological and genetic variations across major distributional ranges for an integrative delimitation of species boundaries and the identification of hybridization and hybrids.

Molecular phylogeny and taxonomy of the genus Vernaya (Mammalia: Rodentia: Muridae) with the description of two new species.

The climbing mouse is a rare, small mammal listed as an endangered species on the China species red list. Molecular phylogenetic analyses and the evolutionary history of the genus remain unexplored because of the extreme difficulty in capturing individuals and their narrow distribution. Here, we collected 44 specimens, sequenced one mitochondrial and eight nuclear genes, and integrated morphological approaches to estimate phylogenetic relationships, delimit species boundaries, and explore evolutionary history. Molecular analyses and morphological results supported the validity of these four species. Here, we describe two new species, Vernaya meiguites sp. nov. and Vernaya nushanensis sp. nov., and recognize Vernaya foramena, previously considered a subspecies of Vernaya fulva, as a valid species. The estimated divergence time suggests that the climbing mouse began to diversify during the Pliocene (3.36 Ma).

Perspectives on the Grey Zone of Species Delimitation With Notes on Invertebrates in the Marine Environment

Defining species boundaries, or delimiting species, is a complex and often difficult task. Indeed, when such studies incorporate approaches that consider evolutionary mechanisms, there is much to be learned about species diversity and how the processes that play critical roles in speciation can impact species delineation. In 2021, a virtual workshop on species delimitation was held at the Smithsonian Institution National Museum of Natural History to train natural history scientists and taxonomists on the appropriate analytical tools that can be used to help delimit species when using molecular data. This perspective highlights some of the main themes discussed during that workshop while detailing three processes that can challenge any species delimitation study. Specifically, we discuss incomplete lineage sorting, gene flow, and population structure when delimiting species boundaries using molecular data. We highlight empirical studies and methodological approaches that have successfully met these challenges under various scenarios. Finally, we provide recommendations and considerations for undertaking species delimitation studies in a variety of taxa. To this end, we recommend that taxonomists fully embrace process-based species delimitation, which can provide important insights into speciation in their study systems. For those developing analytical approaches, we hope they consider incorporating less well-known taxa, such as marine invertebrates, into method testing. Marine invertebrates encompass many dark taxa across the tree of life yet represent the majority of animal phyla, many of which are vulnerable to extinction due to global ocean change. Thus, advancing species delimitation to address taxonomic revisions in these organisms will support conservation decisions on keystone ecosystems. Furthermore, the diversity of their life history strategies, the lack of obvious barriers to gene flow in the ocean environment, and their occurrence in isolated habitat patches can better inform our knowledge of speciation and the evolutionary processes that play a role in generating diversity in nature.

Deciphering phylogenetic relationships of and delimiting species boundaries within the controversial ciliate genus Conchophthirus using an integrative morpho-evo approach

The phylum Ciliophora (ciliates) comprises about 2600 symbiotic and over 5500 free-living species. The inclusion of symbiotic ciliates in phylogenetic analyses often challenges traditional classification frameworks due to their morphological adaptions to the symbiotic lifestyle. Conchophthirus is such a controversial obligate endocommensal genus whose affinities to other symbiotic and free-living scuticociliates are still poorly understood. Using uni- and multivariate morphometrics as well as 2D-based molecular and phylogenetic analyses, we attempted to test for the monophyly of Conchophthirus, study the boundaries of Conchophthirus species isolated from various bivalves at mesoscale, and reveal the phylogenetic relationships of Conchophthirus to other scuticociliates. Multidimensional analyses of morphometric and cell geometric data generated the same homogenous clusters, as did phylogenetic analyses based on 144 new sequences of two mitochondrial and five nuclear molecular markers. Conchophthirus is not closely related to ‘core’ scuticociliates represented by the orders Pleuronematida and Philasterida, as assumed in the past using morphological data. Nuclear and mitochondrial markers consistently showed the free-living Dexiotricha and the mouthless endosymbiotic Haptophrya to be the nearest relatives of Conchophthirus. These three highly morphologically and ecologically dissimilar genera represent an orphan clade from the early radiation of scuticociliates in molecular phylogenies.

Exploring the Diversity and Systematics of Phaeosphaeriaceae: Taxonomic Novelties from Ecologically Diverse Habitats and Their Phylogenetic Resolution.

The family Phaeosphaeriaceae is a diverse group of ascomycetous fungi that are commonly found in terrestrial habitats worldwide. In recent years, there has been increasing interest in the biodiversity of Phaeosphaeriaceae in China, particularly in Sichuan Province, which has not been fully explored for its high fungal diversity. In our study, we conducted extensive surveys in Sichuan Province to identify and describe new species of Ascomycota with diverse habitats. Here, we present a taxonomic revision of Phaeosphaeriaceae with taxonomic novelties from terrestrial habitats in Sichuan Province based on a multi-gene phylogenetic approach. Our study focuses on the description of four new species of Phaeosphaeriaceae, representing novel additions to the diversity of this fungal family. Using a combination of morphological and molecular data, we provide detailed descriptions of the new taxa and their placement within the family. Additionally, we discuss the phylogenetic relationships of these new taxa with other members of Phaeosphaeriaceae, providing insights into the correct taxonomic classification of the family. Our taxonomic revision contributes to understanding fungal diversity in China and provides a foundation for future studies investigating the taxonomy and ecological roles of Phaeosphaeriaceae fungi. Furthermore, our multi-gene phylogenetic approach provides increased resolution and accuracy in the delimitation of species boundaries within the family. Our study highlights the importance of continued exploration and taxonomic revision in order to fully understand the diversity and distribution of fungal species in China and beyond. New species:Paraloratospora sichuanensis, Phaeosphaeria chengduensis, P. sichuanensis, and Septoriella shoemakeri. New combinations:Paraloratospora breonadiae, P. fructigena, Septoriella ammophilicola, S. asparagicola, S. festucae, S. luzulae, and S. verrucispora. New names:Septoriella paradactylidis, and S. neomuriformis.

Widespread coexistence of genetically distinct morphotypes in theSatyrium longicaudacomplex (Orchidaceae)

AbstractSpecies-level taxonomy is traditionally based on herbarium collections that typically include few, or even single, representatives per site. This can lead to underestimation of diversity when there are sympatric populations of superficially similar plants belonging to different lineages. Satyrium longicauda (Orchidaceae) represents a taxonomic challenge for the delimitation of species boundaries due to the high degree of morphological variation detected within and among populations. Currently, just two varieties are accepted based mainly on length differences of the lateral sepal and nectar spur. However, there is extensive morphological variation within South African populations and evidence for several pollination ecotypes, indicating that this taxon represents an actively diverging species complex. Here, we evaluate intraspecific morphological variation through uni- and multivariate morphometrics and analyse internal transcribed spacer sequences for individuals sampled from 36 sites, including 14 sites where divergent morphotypes occur sympatrically. Morphometric analyses of 1802 individuals revealed the presence of eight morphotypes based on vegetative and floral characters. Up to six morphologically and genetically distinct morphotypes can coexist in sympatry. Morphological and genetic distances among populations were significantly correlated. Phylogenetic analyses of 120 accessions indicated that neither of the two varieties nor S. longicauda as a species is monophyletic, and provided evidence for the monophyly of some of the morphotypes including the newly described S. cernuiflorum. The presence of distinct morphological and genetic sympatric variants, which in several cases scale up to distinct evolutionary lineages, is consistent with the existence of different taxa according to morphological and biological species concepts. Our results therefore confirm that taxonomy based mainly on herbarium collections can grossly under-estimate actual diversity of disparate lineages, although further work is required to finalize taxonomic decisions. These findings have implications for efforts to estimate species diversity in groups that are in the process of diversifying and for conservation practice.

Phylogenetics of the Andean tree genusRuagea(Meliaceae): implications for taxonomy

AbstractRuagea (Meliaceae) comprises c. 15 species of small or medium-sized trees distributed in Central and South America, predominantly in montane rain forest and cloud forest of the Andes. The taxonomic position of Ruagea has changed several times in the past, and it was previously included in Guarea. Ruagea is currently accepted as a separate genus based on morphology and molecular phylogenetic analyses at the family level. However, its monophyly has not been accurately tested, and phylogenetic relationships among species are still largely unknown. In addition, delimitation of species boundaries is not straightforward due to morphological diversity within species. Here, we present the first phylogenetic reconstruction of Ruagea based on two nuclear ribosomal (ITS, ETS) and two plastid DNA regions (rps4-trnT, psbA-trnH). Our results show that Ruagea is monophyletic and composed of four main clades (A, B, C and D). Clade D, comprising the Ecuadorian endemic R. membranacea and a potentially new species from Colombia, is sister to the rest of the genus. Our results demonstrate that R. membranacea and R. insignis are monophyletic and provide support for the recognition of two new species with divergent morphologies. The delimitation of the remaining species remains unresolved as they are recovered as either poly- or paraphyletic. Our results highlight the complexity of the taxonomy of Ruagea, as found in other rainforest taxa, and the need for further detailed analyses to delimit these taxa. This study provides the first and most extensive phylogenetic analysis of the genus so far, representing a contribution towards understanding the evolutionary history of Ruagea.

Not out of the Mediterranean: Atlantic populations of the gorgonian Paramuricea clavata are a separate sister species under further lineage diversification.

The accurate delimitation of species boundaries in nonbilaterian marine taxa is notoriously difficult, with consequences for many studies in ecology and evolution. Anthozoans are a diverse group of key structural organisms worldwide, but the lack of reliable morphological characters and informative genetic markers hampers our ability to understand species diversification. We investigated population differentiation and species limits in Atlantic (Iberian Peninsula) and Mediterranean lineages of the octocoral genus Paramuricea previously identified as P.clavata. We used a diverse set of molecular markers (microsatellites, RNA-seq derived single-copy orthologues [SCO] and mt-mutS [mitochondrial barcode]) at 49 locations. Clear segregation of Atlantic and Mediterranean lineages was found with all markers. Species-tree estimations based on SCO strongly supported these two clades as distinct, recently diverged sister species with incomplete lineage sorting, P. cf. grayi and P.clavata, respectively. Furthermore, a second putative (or ongoing) speciation event was detected in the Atlantic between two P. cf. grayi color morphotypes (yellow and purple) using SCO and supported by microsatellites. While segregating P. cf. grayi lineages showed considerable geographic structure, dominating circalittoral communities in southern (yellow) and western (purple) Portugal, their occurrence in sympatry at some localities suggests a degree of reproductive isolation. Overall, our results show that previous molecular and morphological studies have underestimated species diversity in Paramuricea occurring in the Iberian Peninsula, which has important implications for conservation planning. Finally, our findings validate the usefulness of phylotranscriptomics for resolving evolutionary relationships in octocorals.

Diversity, distribution and extinction risk of native freshwater fishes of South Africa.

Extinction risk for 101 valid species and 18 unique genetic lineages of native freshwater fishes of South Africa was assessed in 2016 following the IUCN Red List criteria. An additional five species (three new species that were described and two species that were revalidated subsequent to the 2016 assessments) were assessed in the present study. A synthesis of the outcome of the assessments of the 106 valid species and 18 genetic lineages indicates that 45 (36%) of South Africa's freshwater fish taxa are threatened (7 Critically Endangered, 25 Endangered, 13 Vulnerable). Of the remaining taxa, 17 (14%) are listed as Near Threatened, 57 (46%) are Least Concern and five (4%) are Data Deficient. More than 60% of the endemic taxa are threatened. The Cape Fold Ecoregion has the highest proportion of threatened taxa (67%) due to the existence of a unique assemblage of narrow-range endemic species. Galaxias and Pseudobarbus have the highest number of highly threatened taxa as most of the species and lineages in these genera are classified as either CR or EN. Major threats to the native freshwater fishes of the country are invasive fish species, deterioration of water quality, impoundments and excessive water abstraction, land use changes and modification of riverine habitats. Immediate conservation efforts should focus on securing remnant populations of highly threatened taxa and preventing deterioration in threat status, because recovery is rare. Accurate delimitation of species boundaries, mapping their distribution ranges, improved knowledge of pressures and long-term monitoring of population trends need to be prioritised to generate credible data for the 2026 IUCN threat status assessments and designation of important fish areas as part of the National Freshwater Ecosystem Priority Areas (NFEPA) initiative.

An integrative approach reveals high species diversity in the primitively segmented spider genus

Accurate species delimitation is crucial for our understanding of evolution, biodiversity and conservation. However, morphology-based species delimitation alone appears to be prone to taxonomic errors and ineffective for taxa with high interspecific morphological homogeneity or intraspecific morphological variations, as is the case for mesothele and mygalomorph spiders. Combined molecular–morphology species delimitation has shown great potential to delimit species boundaries in such ancient lineages. In the present study, molecular and morphological evidence were integrated to delimit species of the primitively segmented spider genus Songthela Ono, 2000. The cytochrome c oxidase subunit I gene (COI) was sequenced for 192 novel specimens belonging to 12 putative morphospecies. The evolutionary relationships within Songthela and the 12-morphospecies hypothesis were tested in two steps – species discovery and species validation – using four single-locus species delimitation approaches. All species delimitation analyses supported the 12-species hypothesis. Phylogenetic analyses yielded three major clades in Songthela, which are consistent with morphology. Accordingly, we assigned 19 known and 11 new species (S. aokoulong, sp. nov., S. bispina, sp. nov., S. dapo, sp. nov., S. huayanxi, sp. nov., S. lianhe, sp. nov., S. lingshang, sp. nov., S. multidentata, sp. nov., S. tianmen, sp. nov., S. unispina, sp. nov., S. xiujian, sp. nov., S. zizhu, sp. nov.) of Songthela to three species-groups: the bispina-group, the multidentata-group and the unispina-group. Another new species, S. zimugang, sp. nov., is not included in any species groups, but forms a sister lineage to the bispina- and unispina-groups. These results elucidate a high species diversity of Songthela in a small area and demonstrate that integrating morphology with COI-based species delimitation is fast and cost-effective in delimiting species boundaries. http://zoobank.org/urn:lsid:zoobank.org:pub:AF0F5B31-AFAF-4861-9844-445AE8678B67

Multilocus species delimitation and phylogeny of the genus Calommata (Araneae, Atypidae) in southern China

AbstractSpecies delimitation is critical to our understanding of evolution, biodiversity, and conservation. However, delimiting species boundaries can be challenging, especially when intraspecific divergence is high (and interspecific divergence is low), and gene flow and incomplete lineage sorting are present. Mygalomorph spiders have become models for examining a variety of evolutionary and biogeographical questions. However, their extreme morphological homogeneity and high genetic structuring make delimiting mygalomorph species especially challenging. Integrative approaches to species delimitation, which combine multiple lines of evidence, have been used to infer the limits of many mygalomorph groups in an independent and reproducible way. Yet molecular species delimitation and phylogenetic relationships of the purseweb spider family Atypidae remain scarce. In this study, we explored the phylogeny and species delimitation of Calommata spiders from southern China using data sets including three mitochondrial (COI, 16S, and 12S) and two nuclear (28S, H3) markers. Our phylogenetic analyses yielded a well‐supported clade containing all southern Chinese Calommata and a sister relationship between C. signata and three new species within it. Based on the single‐locus and multilocus species delimitation analyses, we delimited four Calommata species from southern China: one known species (C. signata) and three new species (C. yuanjiangica sp. n., C. hangzhica sp. n. and C. jinggangica sp. n.). This study thus provides a framework for determining the limits of C. signata sensu lato species and calls for further study of systematics and evolutionary relationships within Calommata and other atypid lineages.

Phylogenomic Delimitation of Morphologically Cryptic Species in Globetrotting Nylanderia (Hymenoptera: Formicidae) Species Complexes

Abstract The ant genus Nylanderia Emery has a cosmopolitan distribution and includes 150 extant described species and subspecies, with potentially hundreds more undescribed. Global taxonomic revision has long been stalled by strong intra- and interspecific morphological variation, limited numbers of diagnostic characters, and dependence on infrequently collected male specimens for species description and identification. Taxonomy is further complicated by Nylanderia being one of the most frequently intercepted ant genera at ports of entry worldwide, and at least 15 globetrotting species have widespread and expanding ranges, making species-level diagnoses difficult. Three species complexes (‘bourbonica complex’, ‘fulva complex’, and ‘guatemalensis complex’) include globetrotting species. To elucidate the phylogenetic positions of these three complexes and delimit species boundaries within each, we used target enrichment of ultraconserved elements (UCEs) from 165 specimens representing 98 Nylanderia morphospecies worldwide. We also phased the UCEs, effectively doubling sample size and increasing population-level sampling. After recovering strong support for the monophyly of each complex, we extracted COI barcodes and SNPs from the UCE data and tested within-complex morphospecies hypotheses using three molecular delimitation methods (SODA, bPTP, and STACEY). This comparison revealed that most methods tended to over-split taxa, but results from STACEY were most consistent with our morphospecies hypotheses. Using these results, we recommend species boundaries that are conservative and most congruent across all methods. This work emphasizes the importance of integrative taxonomy for invasive species management, as globetrotting occurs independently across at least nine different lineages across Nylanderia.

Uncertainties and risks in delimiting species of Cambeva (Siluriformes: Trichomycteridae) with single-locus methods and geographically restricted data

Abstract Cambeva contains species with complex taxonomy or poorly delimitated in terms of morphology and geopraphic distribution. We conducted an extensive review of Cambeva populations from coastal drainages of Southern to Southeastern Brazil to evaluate species geographic limits with an integrative analysis including morphological and molecular data (COI). We test if two single-locus methods, Bayesian Poisson Tree Processes (bPTP) and Generalized Mixed Yule Coalescent (GMYC), are efficient to delimit species boundaries in Cambeva by the comparison with the diagnosable morphological units. Using GMYC, we also evaluated the combination of tree and molecular clock priors to reconstruct the input phylogeny and assessed how well the implemented model fitted our empirical data. Eleven species were identified using a morphological diagnosability criterion: Cambeva balios, C. barbosae, C. botuvera, C. cubataonis, C. davisi, C. guaraquessaba, C. iheringi, C. tupinamba, and C. zonata and two treated as undescribed species. In contrast with previous knowledge, many of them have wider distribution and high intraspecific variation. Species delimitation based on single-locus demonstrated incongruences between the methods and strongly differed from the morphological delimitation. These disagreements and the violation of the GMYC model suggest that a single-locus data is insufficient to delimit Cambeva species and the failure may be attributable to events of mitochondrial introgression and incomplete lineage sorting.

Species boundaries in the messy middle-A genome-scale validation of species delimitation in a recently diverged lineage of coastal fog desert lichen fungi.

Species delimitation among closely related species is challenging because traditional phenotype‐based approaches, for example, using morphology, ecological, or chemical characteristics, may not coincide with natural groupings. With the advent of high‐throughput sequencing, it has become increasingly cost‐effective to acquire genome‐scale data which can resolve previously ambiguous species boundaries. As the availability of genome‐scale data has increased, numerous species delimitation analyses, such as BPP and SNAPP+Bayes factor delimitation (BFD*), have been developed to delimit species boundaries. However, even empirical molecular species delimitation approaches can be biased by confounding evolutionary factors, for example, hybridization/introgression and incomplete lineage sorting, and computational limitations. Here, we investigate species boundaries and the potential for micro‐endemism in a lineage of lichen‐forming fungi, Niebla Rundel & Bowler, in the family Ramalinaceae by analyzing single‐locus and genome‐scale data consisting of (a) single‐locus species delimitation analysis using ASAP, (b) maximum likelihood‐based phylogenetic tree inference, (c) genome‐scale species delimitation models, e.g., BPP and SNAPP+BFD, and (d) species validation using the genealogical divergence index (gdi). We specifically use these methods to cross‐validate results between genome‐scale and single‐locus datasets, differently sampled subsets of genomic data and to control for population‐level genetic divergence. Our species delimitation models tend to support more speciose groupings that were inconsistent with traditional taxonomy, supporting a hypothesis of micro‐endemism, which may include morphologically cryptic species. However, the models did not converge on robust, consistent species delimitations. While the results of our analysis are somewhat ambiguous in terms of species boundaries, they provide a valuable perspective on how to use these empirical species delimitation methods in a nonmodel system. This study thus highlights the challenges inherent in delimiting species, particularly in groups such as Niebla, with complex, relatively recent phylogeographic histories.

New shallow water species of Caribbean Ircinia Nardo, 1833 (Porifera: Irciniidae)

Seven Ircinia morphospecies were collected from three sites in the Caribbean (Bocas del Toro, Panama; the Mesoamerican Barrier Reef, Belize; and the Florida Keys, United States of America). Previous research used an integrative taxonomic framework (genome-wide SNP sampling and microbiome profiling) to delimit species boundaries among these Ircinia. Here, we present morphological descriptions for these species, six of which are new to science (Ircinia lowi sp. nov., Ircinia bocatorensis sp. nov., Ircinia radix sp. nov., Ircinia laeviconulosa sp. nov., Ircinia vansoesti sp. nov., Ircinia ruetzleri sp. nov.) in addition to one species conferre (Ircinia cf. reteplana Topsent, 1923).

Coalescent-based species delimitation meets deep learning: Insights from a highly fragmented cactus system.

Delimiting species boundaries is a major goal in evolutionary biology. An increasing volume of literature has focused on the challenges of investigating cryptic diversity within complex evolutionary scenarios of speciation, including gene flow and demographic fluctuations. New methods based on model selection, such as approximate Bayesian computation, approximate likelihoods, and machine learning are promising tools arising in this field. Here, we introduce a framework for species delimitation using the multispecies coalescent model coupled with a deep learning algorithm based on convolutional neural networks (CNNs). We compared this strategy with a similar ABC approach. We applied both methods to test species boundary hypotheses based on current and previous taxonomic delimitations as well as genetic data (sequences from 41loci) in Pilosocereus aurisetus, a cactus species complex with a sky-island distribution and taxonomic uncertainty. To validate our method, we also applied the same strategy on data from widely accepted species from the genus Drosophila. The results show that our CNN approach has a high capacity to distinguish among the simulated species delimitation scenarios, with higher accuracy than ABC. For the cactus data set, a splitter hypothesis without gene flow showed the highest probability in both CNN and ABC approaches, a result agreeing with previous taxonomic classifications and in line with the sky-island distribution and low dispersal of P.aurisetus. Our results highlight the cryptic diversity within the P.aurisetus complex and show that CNNs are a promising approach for distinguishing complex evolutionary histories, even outperforming the accuracy of other model-based approaches such as ABC.