Molecular Species Delimitation Methods Research Articles

Bringing light into deep-sea biodiversity: a systematic revision and molecular phylogeny of the genus Scaphander Montfort, 1810 (Gastropoda: Cephalaspidea), with a focus on the Indo-Pacific

Abstract Scaphander is a genus of mostly deep-sea, soft-bottom gastropods distributed nearly worldwide. Its taxonomic history is complex, with 32 species currently accepted, most based on shells only. In this work, we revise the diversity and systematics of Scaphander, with a focus on the Indo-Pacific region, using a detailed morphological study and molecular phylogenetics. Conchological and anatomical characters, such as digestive and male reproductive systems were compared, and original descriptions and type material were investigated. Additionally, molecular species delimitation methods were used, such as Assemble Species by Automatic Partitioning (ASAP) based on the gene cytochrome c oxidase subunit I, and the multispecies coalescent method Species Tree And Classification Estimation, Yarely (STACEY) based on a dataset of five gene markers. Thirty-three Scaphander species were recognized worldwide, 10 still based only on shells. Five species are here described as new to science, namely Scaphander amygdalus sp. nov., Scaphander cornus sp. nov., Scaphander obnubilus sp. nov., Scaphander semicallus sp. nov. and Scaphander solomonensis sp. nov. The most diverse region is the Western Pacific (17 species), followed by the Atlantic (12 species) and the Indian Ocean (7 species).

Hidden diversity in eastern North America: The genus Ligidium (Oniscidea, Ligiidae) in the southern Appalachian Mountains

AbstractThe terrestrial isopod genus Ligidium includes 58 species from Europe, Asia, and North America. In Eastern North America four species are recognized: L. floridanum and L. mucronatum, known just from their type localities in Florida and Louisiana respectively, L. blueridgensis, endemic to the southern Appalachians, and L. elrodii, widespread from Georgia to Ontario. The genus shows a marked morphological conservatism, and species are differentiated mostly by small morphological differences; it is not always easy to determine if such variability represents inter‐ or intraspecific variation. Here, we explore the diversity of Ligidium from the southern Appalachian Mountains, exploring the congruence of morphologically defined groups with multilocus phylogenetic reconstructions and molecular species delimitation methods. We have studied a total of 130 specimens from 37 localities, mostly from the southern Appalachians, and analysed mtDNA (Cox1) and nuclear (28S, NaK) sequences. Morphologically, we recognized eight morphotypes, most of them assignable to current concepts of L. elrodii and L. blueridgensis. Phylogenetic analyses supported the evolutionary independence of all morphotypes, and suggest the existence of 8–9 species, including limited cryptic diversity. Single‐locus delimitation analyses based on mtDNA data suggest the existence of a much higher number of species than the multilocus analyses. The estimated age of the ancestors of sampled lineages indicates a long presence of the genus in eastern North America and old speciation events through the Miocene. Our results indicate a higher diversity than previously thought among the Ligidium populations present in the southern Appalachian Mountains, with several species to be described.

Application of Modern Approaches to Species Delimitation in Freshwater Erpobdellidae (Hirudinea: Arhynchobdellida) with a New Interpretation of Their Genera

The traditional taxonomy of freshwater invertebrates is a labor-intensive process requiring extensive knowledge and experience. In addition, this science is largely subjective, which makes its digitalization difficult. However, accurate species attribution is becoming increasingly important for environmental surveys and conservation efforts. In the 21st century, molecular biology methods have proven to be a successful tool for objectively determining biodiversity. Achieving objectivity in identifying the taxa of non-parasitic leeches of the family Erpobdellidae was the main goal of this study. To this end, various bioinformatic approaches to DNA sequence analysis have been tested. As a result, the phylogenetic diversity amounted to 47 species versus 29 morphospecies defined traditionally. The use of molecular species delimitation methods made it possible to identify seven complexes of morphologically hidden (cryptic) species and some morphological misidentifications, as well as to discover a new species from Eastern Siberia (Erpobdella sibirica) with a unique molecular diagnosis (DNA barcode). A pioneering attempt to apply the same approach to higher taxa showed that Erpobdellidae probably consists of seven genera, including the unambiguous elimination of the genus Motobdella. Obtaining quantitative and, therefore, objective data is an advantage of the molecular approach, which has proven to be effective in recognizing species and genera, at least in leeches.

Phylogenetic analyses of the leafhopper tribe Chiasmini Distant, 1908 and delimination of species of the genus Exitianus Ball, 1929 (Hemiptera: Cicadellidae: Deltocephalinae: Chiasmini) in China based on molecular data

Previous phylogenetic analyses of the grass-specialist leafhopper tribe Chiasmini have resolved relationships among genera but have included few representatives of individual genera. Here the phylogeny of 20 Chinese species belonging to 8 chiasmine genera was investigated by combining DNA sequence data from two mitochondrial genes (COI, 16S) and two nuclear genes (H3, 28S). In both maximum likelihood (ML) and Bayesian inference (BI) analyses, relationships among genera were largely consistent with prior analyses, with most members of the tribe placed into two sister clades: (Exitianus + Nephotettix) and the remaining five sampled genera. To examine morphology-based species definitions in the taxonomically difficult genus Exitianus Ball, 1929, one mitochondrial gene (COI) and one nuclear gene (ITS2) were used to infer the phylogenetic relationships and status of two common and widespread species and compare the performance of different molecular species-delimitation methods. These analyses divide the included populations into two well-supported clades corresponding to current morphological species concepts but some inconsistencies occurred under the jMOTU, ABGD and bPTP methods depending on the which gene and analytical parameter values were selected. Considering the variable results yielded by methods employing single loci, the BPP method, which combines data from multiple loci, may be more reliable in Exitianus.

Discovery of five new species of Allacta from Yunnan and Hainan, China (Blattodea, Pseudophyllodromiidae).

We examined new Allacta materials from Yunnan and Hainan Province, China, and discovered new species using both morphological and molecular species delimitation (ABGD) methods. Five new species are described: A.bifolium Li & Wang, sp. nov., A.hemiptera Li & Wang, sp. nov., A.lunulara Li & Wang, sp. nov., A.redacta Li & Wang, sp. nov., and A.unicaudata Li & Wang, sp. nov. All five species are placed under the hamifera species group. An updated key and checklist of Allacta species from China are provided.

Molecular and morphometric data provide evidence of intraspecific variation in shape and pigmentation pattern in Otocinclus cocama (Siluriformes: Loricariidae) across major river drainages.

Otocinclus cocama, a uniquely colored species of the loricariid catfish genus Otocinclus described solely from the type locality in the lower Ucayali River in northern Peru, is reported occurring in the Tigre River, a tributary to the Marañón River that drains a different section of the Andean Mountain range in the western Amazon. Both populations differ in the number of dark bars spanning the flanks of the body, and we investigated whether these morphotypes constitute distinct species. The body shapes of populations from the Tigre and Ucayali rivers were compared using geometric morphometrics. Although principal component analysis detected a broad overlap between populations, multivariate analysis of variance and linear driscriminat analysis revealed a subtle differentiation between the populations of the two hydrographic basins. Average body shape of the Ucayali River population tend to be slightly higher than that of the Tigre River, with the caudal peduncle stretched vertically in the Ucayali population. Multivariate regression of shape and centroid size revealed an allometric effect of 10.7% (p < 0.001), suggesting that the variation between Tigre and Ucayali populations was purely shape variation. Molecular data of coI, cytb, nd2, and 16S mitochondrial genes indicated a nucleotide diversity range from 0.001 to 0.003, and haplotypic diversity range from 0.600 ± 0.11 to 0.79 ± 0.07. The median-joining haplotype network for the concatenated matrix exhibited two divergent haplogroups related to the geographic area and separated by <10 mutational steps. The molecular species delimitation methods based on distance (automatic barcode gap discovery and assemble species by automatic partitioning) recovered two molecular lineages evolving independently, being one of the lineages formed by individuals from both populations. Tree-based methods (generalized mixed Yule coalescent and Bayesian implementation of the Poisson tree process) recovered similar topologies and supported single lineage recognition. Methods of molecular delimitation of species disclosed the high similarity between the two populations of Otocinclus cocama, further supported by the presence of old haplotypes common to both groups which could indicate that the populations still maintain gene flow. Although the morphological data reveal a subtle variation between both river basins, the molecular data suggest a weak population structuration based on hydrographic areas, but not different species lineages, therefore Otocinclus cocama is composed of a single lineage with two distinct morphotypes.

A systematic review of the Iberian springsnail subgenus Alzoniella (Navarriella) (Caenogastropoda: Hydrobiidae), with the description of a new potentially relict subfamily

Abstract The threatened springsnail subgenus Alzoniella (Navarriella) in the Iberian Peninsula has been suggested to be an old and relict lineage of the family Hydrobiidae. The subgenus is represented by two morphological species, both endemic to the Pyrenees and their southern foothills. We conducted phylogenetic analyses based on mitochondrial and nuclear gene fragments of topotypes and other populations, four molecular species delimitation methods, and morphological examinations to clarify the uncertain systematic position of the subgenus within the family, assess its species diversity, and understand the population genetic structure of the two geographically restricted species. Our phylogenetic results revealed that Alzoniella (Navarriella) is distantly related to all other species of Alzoniella, even belonging to an independent subfamily-level clade, for which we introduce the new genus Navarriella and the new subfamily Navarriellinae subfam. nov. Molecular methods and geometric morphometric analysis of shell shape identified a single species in the new genus. The significant phylogenetic distance from other hydrobiid taxa, narrow distribution, and limited gene flow among its populations (estimated from mitochondrial cytochrome c oxidase subunit I sequences) highlight Navarriella as an isolated lineage within the family that requires urgent conservation attention. Furthermore, our results cast a new light on the northern Iberian Mountains as a dispersal barrier for ancient spring lineages.

Species delimitation and molecular phylogeny of the grasshopper subfamily Gomphocerinae (Orthoptera: Acrididae) from Algeria based on mitochondrial and nuclear DNA markers

Gomphocerinae grasshoppers (Orthoptera: Acrididae) are generally characterised by their polymorphism and cryptic diversity, which can be confusing when relying on morphological identification alone. DNA taxonomy serves as a powerful molecular tool for species identification and biodiversity assessment. In the context of zoogeography and the biodiversity conservation of animal resources in ecosystems, DNA barcoding data for Algerian Gomphocerinae fauna remains limited in global databases, despite their agroeconomic and environmental importance. Therefore, in this study, we collected various species of this subfamily from the Biskra region in Algeria and conducted DNA barcoding analysis, employing different molecular species delimitation methods (the automatic barcode gap discovery, assemble species by automatic partitioning, single-threshold general mixed Yule coalescent model, Bayesian Poisson tree process and multi-rate Poisson tree process methods), as well as phylogenetic analyses (maximum-likelihood and Bayesian inference) based on two molecular markers (COI and ITS). The results of the 163 newly generated sequences demonstrated that DNA barcoding technology is highly efficient and valuable for species identification in the subfamily Gomphocerinae, showing strong congruence with morphological evidence for 12 species, including eight species that were sequenced for the first time. This study also reported a new record of the genus Stenohippus Uvarov, 1926 in Algeria, comprising two species. Additionally, taxonomic revision allowed the species Dociostaurus biskrensis Moussi & Petit, 2014, to be transferred to the genus Stenohippus. Molecular tree analyses revealed the phylogenetic positions of the newly sequenced species within the subfamily Gomphocerinae and provided insights into their evolutionary relationships. These new data serve as a starting point for future research in other geographical areas, enabling a better understanding of the biodiversity of this insect group in Algeria.

Phylogenetic structure and molecular species delimitation hint a complex evolutionary history in an Alpine endemic Niphargus clade (Crustacea, Amphipoda)

Subterranean fauna is an important contributor to the global fauna, but it is still understudied and a large part of its taxonomy is not yet resolved. One species complex with unresolved taxonomy is the groundwater amphipod Niphargus ruffoi, endemic to the Alpine chain. Here, we used new samples from across the Alpine arc to review the taxonomic status of the entire clade, including the species N. ruffoi and Niphargus arolaensis. We sequenced four genetic markers from the collected specimens, assessed the phylogenetic position of N. ruffoi within the genus, and studied the structure of this species complex using four molecular species delimitation methods. We tested for recombination using the alignments of the concatenated nuclear rDNA genes. The phylogenetic analyses revealed high support for the monophyly of the studied species complex, defining two lineages (i.e., N. arolaensis and N. ruffoi) within the clade. Molecular species delimitation methods suggested that N. arolaensis is a single species, while N. ruffoi should be considered as a species complex of three (using ITS) to eight (using COI) putative species. Moreover, we found a discrepancy between the different nuclear ribosomal DNA markers, indicating a possible recombination with fragments of 28S DNA of N. ruffoi s. lat. present in the genome of N. arolaensis. For the above-mentioned reasons, the internal phylogenetic structure of N. ruffoi s. lat. could not be fully resolved. Moreover, no clear morphological evidence supported the molecular species delimitation. Consequently, no taxonomic changes were proposed. We postulate that this complex scenario was influenced by Pleistocene climate oscillations with subsequent fragmentation events and secondary contacts, making this an interesting study system to investigate the evolution and biogeography of Alpine clades.

Integrative species delimitation helps to find the hidden diversity of the leaf-litter frog Ischnocnema manezinho (Garcia, 1996) (Anura, Brachycephalidae), endemic to the southern Atlantic Forest.

The delimitation of cryptic species is a challenge for biodiversity conservation. Anurans show high cryptic diversity levels, and molecular species delimitation methods could help identify putative new species. Additionally, species delimitation approaches can provide important results for cryptic species conservation, with integrative methods adding robustness to results. Ischnocnema manezinho was described from Santa Catarina Island (SCI), southern Brazil. More recently, some inventories indicated continental populations supposedly similar in morphology to it. If these records are confirmed as I. manezinho, it would likely change its endangered status on National Red List, removing the species from conservation agendas. We investigated the threatened frog Ischnocnema manezinho, to evaluate if the continental populations belong to this species or if they form an undescribed species complex. We used coalescent, distance, and allele-sharing-based species delimitation methods and integrative analyses of morphometric and bioacoustics traits to test evolutionary independence between I. manezinho from SCI, Arvoredo Island, and continental populations. Ischnocnema manezinho is restricted to Santa Catarina Island, while the five remaining lineages should be further investigated through a taxonomic review. Our results point to a small geographic range of Ischnocnema manezinho. Additionally, the species occurs in isolated fragments of forest in SCI surrounded by expanding urban areas, confirming its status as Endangered. Thus, the protection and monitoring of I. manezinho and the taxonomic description of the continental and Arvoredo Island candidate species should be priorities.

A new lizard species of the Liolaemus kingii group (Squamata: Liolaemidae) from northwestern Chubut province (Argentina).

We describe Liolaemus attenboroughi sp. nov., a lizard distributed in the northwestern Patagonian Steppe of Chubut province (Argentina) previously confused with L. kingii (Bell 1843). Recent studies based on molecular evidence supports its evolutionary independence. Here we provide a morphological diagnosis of this lineage, comparisons between three molecular species delimitation methods, and an updated phylogeny of the L. kingii group. Based on current knowledge of its distribution, this new species is allopatric with geographically close species of the L. kingii group.

Taxonomic status of Pseudophilautus conniffae (Anura: Rhacophoridae) an endemic shrub frog restricted to South–Western wet zone of Sri Lanka

Pseudophilautus conniffae from Lowland wet zone of Sri Lanka, was described as a new species in 2019. The validity of the new species was questionable and was often challenged as it shares strong morphological resemblance with P. limbus. Moreover, the phylogenetic placement of P. conniffae was unknown as no molecular data was available until now. Here, we generated 16S DNA sequences and re-examined the external morphological characters to assess its taxonomic distinctiveness. Pseudophilautus conniffae was recovered as being close to P. limbus with strong posterior probability and bootstrap support. The uncorrected pairwise genetic distance between P. conniffae and P. limbus was negligible, being less than 0.3% for the 16S gene fragment. Further two molecular species delimitation methods, ABGD and mPTP suggested that P. conniffae and P. limbus are a single operational taxonomic unit. The Principal Component Analysis of the morphometric characters also resulted in overlapping clusters. These results suggest that the newly described P. conniffae is not a valid species and we conclude that P. conniffae as a junior synonym of P. limbus.

Characterization of Hoplolaimus seinhorsti and Hoplolaimus pararobustus (Tylenchina: Hoplolaimidae) from banana, with phylogeny and species delineation in the genus Hoplolaimus.

The morphological and molecular characterisations of two lance nematode species isolated from the rhizosphere of banana, Hoplolaimus seinhorsti and H. pararobustus, are provided based on an integrative study that includes light and scanning electron microscopy, phylogenetic analysis and two tree-based molecular species delimitation methods (GMYC and bPTP). Nineteen new sequences were obtained, including 5 partial 18S rRNA, 6 D2-D3 of 28S rRNA, 1 ITS rRNA and 7 COI mtDNA (the first COI sequences of H. seinhorsti and H. pararobustus), and an updated morphological character comparison of 37 Hoplolaimus species is presented. The tree-based molecular species-delimitation approaches employed gave markedly differing results, and also showed remarkable discrepancies among the investigated genes, although the bPTP output was found to agree well with established morphological species delimitations. Both species-delimitation approaches did, however, provide the same output for the COI mtDNA sequences, and the COI mtDNA gene sequence was also found to correspond better to established morphological species. It is therefore recommended by this paper as representing the most suitable barcode marker for Hoplolaimus species identification. This integrative study also resulted in the corrective reassignment of 17 gene sequences that were previously unidentified or incorrectly classified, as well as concluding that H. pararobustus consists of two cryptic species.

Pseudophilautus pardus, a junior synonym of P. viridis (Amphibia: Anura: Rhacophoridae).

Pseudophilautus pardus, was first described in 2007, based on a single specimen collected in Sri Lanka during the 19th century. Its absence in recent surveys suggested that the species was extinct. The distinctive spotted dorsal coloration, together with other morphological features, was used to distinguish it from congeners. In 2013, we discovered a specimen resembling the holotype of P. pardus in Samanala Strict Nature Reserve. A DNA analysis based on the 16S rRNA mt-DNA locus recovered a low uncorrected pairwise genetic distance of 0.77-0.96% between the recent example of 'P. pardus' and a congener, P. viridis. Molecular species delimitation methods suggest P. pardus and P. virids to be a single operational taxonomic unit. Morphological analyses using the freshly collected specimen, together with numerous photographic records show P. viridis to be a highly polychromatic species within which the coloration observed in P. pardus too occurs, though rarely. We conclude that P. pardus is a junior synonym of P. viridis.

We don’t know the half of it: morphological and molecular evidence reveal dramatic underestimation of diversity in a key pollinator group (Nemestrinidae)

Nemestrinidae (tangle-veined flies) are important pollinators of numerous southern African plant species. Despite their known ecological importance, the family has received little taxonomic attention in recent years and the systematics of the group is poorly understood. In this study we aimed to assess the phylogenetic relationships and species diversity among three southern African nemestrinid genera from the Nemestrininae subfamily: Prosoeca, Moegistorhynchus and Stenobasipteron, with a specific focus on the largest among these, Prosoeca. We reconstructed a molecular phylogeny using both mitochondrial and nuclear (COI, 16S rRNA, 28S rRNA and CAD) DNA sequence data. Both morphology and molecular species delimitation methods (Automatic Barcode Gap Discovery and the Bayesian Poisson Tree Process) were used to estimate species diversity. The topology from the combined analysis places a monophyletic Moegistorhynchus as the sister group to a paraphyletic Prosoeca with Stenobasipteron nested inside Prosoeca. In all three genera, almost half of the putative species sampled did not match the concept of described species based on morphology. Analysis of phylogenetic diversity showed that undescribed putative species make a substantial contribution to the overall phylogenetic diversity among the sampled species. Comparisons among biogeographic regions suggested that diversity is concentrated in multiple biodiversity hotspots and biomes, particularly in Fynbos and Grassland biomes. The numerous undescribed species and paraphyly of Prosoeca both emphasise the need for increased taxonomic attention for this ecologically important group of flies in particular, and for southern African insect taxa in general.

Integrative taxonomy of the stalk‐eyed bug genus Chauliops (Heteroptera: Malcidae: Chauliopinae) reveals orogeny‐driven speciation

AbstractEconomically significant bean pests of the genus Chauliops are species rich in the areas surrounding the Qinghai–Tibet Plateau and provide an excellent system for speciation studies. Here, an integrative taxonomic approach, employing morphological analyses, population genetic methods, and multiple molecular species delimitation methods, was used to clarify the taxonomy of Chauliops in East and Southeast Asia. Four new species (Chauliops parahorizontalis Li &amp; Bu, sp. nov., Chauliops albida Li &amp; Bu, sp. nov., Chauliops bicoloripes Li &amp; Bu, sp. nov., and Chauliops paraconica Li &amp; Bu, sp. nov.) were described, which increases the number of Chauliops species in this area from six to 10; a key for Chauliops species is also provided. Phylogenetic analysis and divergence time estimation revealed that Chauliops was divided into four clades: Clade A (Chauliops bisontula + [Chauliops horizontalis + C. parahorizontalis sp. nov.]), Clade B (C. albida sp. nov. and C. bicoloripes sp. nov.), Clade C (Chauliops quaternaria and Chauliops zhengi), and Clade D (Chauliops fallax + [Chauliops conica + C. paraconica sp. nov.]). Two species diversification events of Chauliops estimated to have occurred 7–1 million years ago (Ma) and 25–13 Ma were detected. These speciation events were consistent with the two historical uplift events of the Qinghai–Tibet Plateau, suggesting that orogeny might have provided opportunities for the diversification of Chauliops species on the southeastern margin of the Qinghai–Tibet Plateau. Our findings show that population genetic analyses can be used to delimit related species and that orogeny is a key driver of species diversification on the southeastern margin of the Qinghai–Tibet Plateau.

Consolidation of Chloridium: new classification into eight sections with 37 species and reinstatement of the genera Gongromeriza and Psilobotrys.

Chloridium is a little-studied group of soil- and wood-inhabiting dematiaceous hyphomycetes that share a rare mode of phialidic conidiogenesis on multiple loci. The genus has historically been divided into three morphological sections, i.e. Chloridium, Gongromeriza, and Psilobotrys. Sexual morphs have been placed in the widely perceived genus Chaetosphaeria, but unlike their asexual counterparts, they show little or no morphological variation. Recent molecular studies have expanded the generic concept to include species defined by a new set of morphological characters, such as the collar-like hyphae, setae, discrete phialides, and penicillately branched conidiophores. The study is based on the consilience of molecular species delimitation methods, phylogenetic analyses, ancestral state reconstruction, morphological hypotheses, and global biogeographic analyses. The multilocus phylogeny demonstrated that the classic concept of Chloridium is polyphyletic, and the original sections are not congeneric. Therefore, we abolish the existing classification and propose to restore the generic status of Gongromeriza and Psilobotrys. We present a new generic concept and define Chloridium as a monophyletic, polythetic genus comprising 37 species distributed in eight sections. In addition, of the taxa earlier referred to Gongromeriza, two have been redisposed to the new genus Gongromerizella. Analysis of published metabarcoding data showed that Chloridium is a common soil fungus representing a significant (0.3 %) proportion of sequence reads in environmental samples deposited in the GlobalFungi database. The analysis also showed that they are typically associated with forest habitats, and their distribution is strongly influenced by climate, which is confirmed by our data on their ability to grow at different temperatures. We demonstrated that Chloridium forms species-specific ranges of distribution, which is rarely documented for microscopic soil fungi. Our study shows the feasibility of using the GlobalFungi database to study the biogeography and ecology of fungi. Taxonomic novelties: New genus: Gongromerizella Réblová; New sections: Chloridium section Cryptogonytrichum Réblová, Hern.-Restr., M. Kolařík & F. Sklenar, Chloridium section Gonytrichopsis Réblová, Hern.-Restr., M. Kolařík & F. Sklenar, Chloridium section Metachloridium Réblová, Hern.-Restr., M. Kolařík & F. Sklenar, Chloridium section Volubilia Réblová, Hern.-Restr., M. Kolařík & F. Sklenar; New species: Chloridium bellum Réblová & Hern.-Restr., Chloridium biforme Réblová & Hern.-Restr., Chloridium detriticola Réblová & Hern.-Restr., Chloridium gamsii Réblová & Hern.-Restr., Chloridium guttiferum Réblová & Hern.-Restr., Chloridium moratum Réblová & Hern.-Restr., Chloridium peruense Réblová & Hern.-Restr., Chloridium novae-zelandiae Réblová & Hern.-Restr., Chloridium elongatum Réblová & Hern.-Restr., Chloridium volubile Réblová & Hern.-Restr.; New varieties: Chloridium bellum var. luteum Réblová & Hern.-Restr., Chloridium detriticola var. effusum Réblová & Hern.-Restr., Chloridium chloridioides var. convolutum Réblová & Hern.-Restr.; New combinations: Chloridium section Gonytrichum (Nees & T. Nees) Réblová, Hern.-Restr., M. Kolařík & F. Sklenar, Chloridium section Mesobotrys (Sacc.) Réblová, Hern.-Restr., M. Kolařík & F. Sklenar, Chloridium section Pseudophialocephala (M.S. Calabon et al.) Réblová, Hern.-Restr., M. Kolařík & F. Sklenar, Chloridium simile (W. Gams & Hol.-Jech.) Réblová & Hern.-Restr., Chloridium chloridioides (W. Gams & Hol.-Jech.) Réblová & Hern.-Restr., Chloridium subglobosum (W. Gams & Hol.-Jech.) Réblová & Hern.-Restr., Chloridium fuscum (Corda) Réblová & Hern.-Restr., Chloridium ypsilosporum (Hol.-Jech.) Réblová & Hern.-Restr., Chloridium costaricense (G. Weber et al.) Réblová & Hern.-Restr., Chloridium cuneatum (N.G. Liu et al.) Réblová & Hern.-Restr., Fusichloridium cylindrosporum (W. Gams & Hol.-Jech.) Réblová, Gongromeriza myriocarpa (Fr.) Réblová, Gongromeriza pygmaea (P. Karst.) Réblová, Gongromerizella lignicola (F. Mangenot) Réblová, Gongromerizella pachytrachela (W. Gams & Hol.-Jech) Réblová, Gongromerizella pini (Crous & Akulov) Réblová; New name: Chloridium pellucidum Réblová & Hern.-Restr.; Epitypifications (basionyms): Chaetopsis fusca Corda, Gonytrichum caesium var. subglobosum W. Gams & Hol.-Jech.; Lectotypification (basionym): Gonytrichum caesium Nees & T. Nees. Citation: Réblová M, Hernández-Restrepo M, Sklenář F, Nekvindová J, Réblová K, Kolařík M (2022). Consolidation of Chloridium: new classification into eight sections with 37 species and reinstatement of the genera Gongromeriza and Psilobotrys. Studies in Mycology 103: 87-212. doi: 10.3114/sim.2022.103.04.

Turcogammarus aralensis (Uljanin, 1875), a relict pontogammarid amphipod crustacean from the Aralo-Caspian paleobasin: redescription, phylogenetic position and biogeography

The Ponto-Caspian region harbours the world's second most diverse lacustrine radiation of amphipod crustaceans. The taxonomic position of some of these species is unclear, due to incorrectly defined species limits, poor descriptions, and in some cases, poorly understood characters, such as armament of dorsal integuments. A pertinent example is Turcogammarus aralensis (Uljanin, 1875), which inhabits the Aral Sea and adjacent water bodies like the brackish lake Shalkar (Charkhal), lower reaches of the Syr Darya and Volga, and extends ca. 2500 km westwards in the Black Sea basin. This species has never been properly described, which has led to frequent taxonomic reshuffling. Herein, we provide a full re-description, ascertain its phylogenetic position using the cytochrome c oxidase subunit I (COI) marker, and review its geographical distribution. Our molecular analyses reveal T. aralensis as a sister species to the widespread Obesogammarus crassus (Sars, 1894), and its species status was further confirmed by three molecular species delimitation methods. Turcogammarus aralensis and O. crassus share many similarities such as the setation of the antennae and pereopods 3-4, and poorly setose posterior margin of pereopod 7 basis. Further study is needed to ascertain the validity of the genus Turcogammarus.

Phylogenetic analyses and species delimitation of Aconurella Ribaut (Hemiptera: Cicadellidae: Deltocephalinae: Chiasmini) in China based on molecular data.

The grassland leafhopper genus Aconurella is widespread in the Old World. Species of this genus are difficult to identify by traditional morphological characters but the morphology-based species classification in this genus has not previously been tested using molecular data. This study analysed DNA sequence data from two mitochondrial genes (COI, 16S) and one nuclear gene (ITS2) to infer the phylogenetic relationships and status of five previously recognized Aconurella species and compare the performance of different molecular species-delimitation methods using single and multiple loci. The analysis divided the included haplotypes into five well-supported subclades, most corresponding to existing morphology-based species concepts. However, different molecular species delimitation methods (jMOTU, ABGD, bPTP, GMYC and BPP) yielded somewhat different results, suggesting the presence of between 4 and 8 species, sometimes lumping the haplotypes of Aconurella diplachnis and Aconurella sibirica into a single species or recognizing multiple putative species within Aconurella prolixa. Considering the different results yielded by various methods employing single loci, the BPP method, which combines data from multiple loci, may be more reliable for delimiting species of Aconurella. Our results suggest that the morphological characters previously used to identify these species are reliable and adequately reflect boundaries between genetically distinct taxa.

Extremely divergent COI sequences within an amphipod species complex: A possible role for endosymbionts?

Some heritable endosymbionts can affect host mtDNA evolution in various ways. Amphipods host diverse endosymbionts, but whether their mtDNA has been influenced by these endosymbionts has yet to be considered. Here, we investigated the role of endosymbionts (microsporidians and Rickettsia) in explaining highly divergent COI sequences in Paracalliope fluviatilis species complex, the most common freshwater amphipods in New Zealand. We first contrasted phylogeographic patterns using COI, ITS, and 28S sequences. While molecular species delimitation methods based on 28S sequences supported 3–4 potential species (N, C, SA, and SB) among freshwater lineages, COI sequences supported 17–27 putative species reflecting high inter‐population divergence. The deep divergence between NC and S lineages (~20%; 28S) and the substitution saturation on the 3rd codon position of COI detected even within one lineage (SA) indicate a very high level of morphological stasis. Interestingly, individuals infected and uninfected by Rickettsia comprised divergent COI lineages in one of four populations tested, suggesting a potential influence of endosymbionts in mtDNA patterns. We propose several plausible explanations for divergent COI lineages, although they would need further testing with multiple lines of evidence. Lastly, due to common morphological stasis and the presence of endosymbionts, phylogeographic patterns of amphipods based on mtDNA should be interpreted with caution.