Molecular Systematic Studies Research Articles

Revealing the Genetic Differentiation of Rattus norvegicus (Berkenhout 1769) Populations by Analyzing Two Mitochondrial Markers

Rattus norvegicus (Brown rat) has a great importance for public health and economy because it lives in close association with human populations. However, molecular systematic studies on global populations of R. norvegicus are very few. In this study, sequences obtained from Europe, Asia, Africa, and America regions were analyzed using mitochondrial Cytochrome-b and Cytochrome oxidase-I gene regions and genetic differentiation levels between these populations were revealed. Accordingly, samples belonging to the studied populations did not split in Bayesian Inference trees and Median-joining networks; these samples also formed common haplotypes, and mean genetic distance and fixation index values were generally low. The results of the study showed that gene flow between these populations may be continuing due to the transportation activity by humans.

Two new species of Metacampanella (Agaricales, Marasmiaceae) from China and Mongolia.

Metacampanella is an important genus in the Marasmiaceae family. We collected specimens during our investigations in China and Mongolia. Through morphological and molecular phylogenetic analyses, we identified two new species of this genus: Metacampanellasubtricolor and Metacampanellacoprophila. In addition, we identified Metacampanellatricolor as a novel combination. Molecular systematic studies support these results. Illustrated descriptions, taxonomic discussions, and keys to the genus are provided.

Phylogenetic systematics of Vigna sensu stricto in the context of Physostigma and allies.

Vigna includes economically vital crops and wild species. Molecular systematic studies of Vigna species resulted in generic segregates of many New World (NW) species. However, limited Old World (OW) sampling left questions regarding inter- and intraspecific relationships in Vigna s.s. African species, including the putative sister genus Physostigma, were comprehensively sampled within the context of NW relatives. Maximum likelihood and Bayesian inference analyses of the chloroplast matK-trnK and nuclear ribosomal ITS/5.8 S (ITS) DNA regions were undertaken to resolve OW Vigna taxonomic questions. Divergence dates were estimated using BEAST to date key nodes in the phylogeny. Analyses of matK and ITS data supported five clades of Vigna s.s.: subg. Lasiospron, a reduced subg. Vigna, subg. Haydonia, subg. Ceratotropis, an enlarged subg. Plectrotropis, and a clade including V. kirkii and V. stenophylla. Genome size estimates of 601 Mb for V. kirkii are near the overall mean of the genus, whereas V. stenophylla had a larger genome (810 Mb), similar to some Vigna subg. Ceratotropis or Plectrotropis species. Former subg. Vigna is reduced to yellow- and blue-flowered species and subg. Plectrotropis is enlarged to mostly all white-, pink-, and purple-flowered species. The age of the split between NW and OW Vigna lineages is ~6-7 Myr. Genome size estimates cannot rule out a polyploid or hybrid origin for V. stenophylla, potentially involving extinct lineage ancestors of Vigna subg. Ceratotropis or Plectrotropis, as indicated by network and phylogenetic analyses. Taxonomic revisions are suggested based on these results.

Biogeography and phylogeny of the scavenging amphipod genus Valettietta (Amphipoda: Alicelloidea), with descriptions of two new species from the abyssal Pacific Ocean

Abstract Valettietta Lincoln & Thurston, 1983 (Amphipoda: Alicelloidea) is an infrequently sampled genus of scavenging amphipod, with a known bathymetric range from 17–5467 m encompassing a variety of habitats from anchialine caves to abyssal plains. Molecular systematics studies have uncovered cryptic speciation in specimens collected from the abyssal Pacific, highlighting uncertainty in the description of Valettietta anacantha (Birstein & Vinogradov, 1963). Here, we apply an integrative taxonomic approach and describe two new species, Valettietta trottarum sp. nov. and Valettietta synchlys sp. nov., collected at abyssal depths in the Clarion-Clipperton Zone, Pacific Ocean. Both species can be distinguished by characters of the gnathopods, uropod 3, and the inner plate of the maxilliped. Further, molecular phylogenetic analyses of two mitochondrial (16S rDNA and COI) and two nuclear (Histone 3 and 28S rRNA) regions found both new species to form well-supported clades and allowed us to re-identify previously published records based on genetic species delimitation. The biogeography of Valettietta is discussed in light of these re-evaluated records, and a new taxonomic key to the genus is provided. These new taxa highlight the strength of applying an integrated taxonomic approach to uncover biodiversity, which is critical in regions being explored for potential industrial purposes.

A new genus of soft coral (Octocorallia, Malacalcyonacea, Cladiellidae) and three new species from Indo-Pacific coral reefs.

Molecular systematic studies of the anthozoan class Octocorallia have revealed widespread incongruence between phylogenetic relationships and taxonomic classification at all levels of the Linnean hierarchy. Among the soft coral taxa in order Malacalcyonacea, the family Alcyoniidae and its type genus Alcyonium have both been recognised to be highly polyphyletic. A recent family-level revision of Octocorallia established a number of new families for genera formerly considered to belong to Alcyoniidae, but revision of Alcyonium is not yet complete. Previous molecular studies have supported the placement of Alcyoniumverseveldti (Benayahu, 1982) in family Cladiellidae rather than Alcyoniidae, phylogenetically distinct from the other three genera in that family. Here we describe a new genus, Ofwegenumgen. nov. to accommodate O.verseveldticomb. nov. and three new species of that genus, O.coronalucissp. nov., O.kloogisp. nov., and O.collisp. nov., bringing the total number of species in this genus to four. Ofwegenumgen. nov. is a rarely encountered genus so far known from only a few locations spanning the Indian and western Pacific Oceans. We present the morphological characters of each species and use molecular data from both DNA barcoding and target-enrichment of conserved elements to explore species boundaries and phylogenetic relationships within the genus.

Phylotranscriptomics supports numerous polyploidization events and phylogenetic relationships in Nicotiana.

Nicotiana L. (Solanaceae) is of great scientific and economic importance, and polyploidization has been pivotal in shaping this genus. Despite many previous studies on the Nicotiana phylogenetic relationship and hybridization, evidence from whole genome data is still lacking. In this study, we obtained 995 low-copy genes and plastid transcript fragments from the transcriptome datasets of 26 Nicotiana species, including all sections. We reconstructed the phylogenetic relationship and phylogenetic network of diploid species. The incongruence among gene trees showed that the formation of N. sylvestris involved incomplete lineage sorting. The nuclear-plastid discordance and nuclear introgression absence indicated that organelle capture from section Trigonophyllae was involved in forming section Petunioides. Furthermore, we analyzed the evolutionary origin of polyploid species and dated the time of hybridization events based on the analysis of PhyloNet, sequence similarity search, and phylogeny of subgenome approaches. Our results highly evidenced the hybrid origins of five polyploid sections, including sections Nicotiana, Repandae, Rusticae, Polydicliae, and Suaveolentes. Notably, we provide novel insights into the hybridization event of section Polydicliae and Suaveolentes. The section Polydicliae formed from a single hybridization event between maternal progenitor N. attenuata and paternal progenitor N. undulata; the N. sylvestris (paternal progenitor) and the N. glauca (maternal progenitor) were involved in the formation of section Suaveolentes. This study represents the first exploration of Nicotiana polyploidization events and phylogenetic relationships using the high-throughput RNA-seq approach. It will provide guidance for further studies in molecular systematics, population genetics, and ecological adaption studies in Nicotiana and other related species.

Phylogenomics and phylogeography of Menispermum (Menispermaceae).

Phylogenomics have been widely used to resolve ambiguous and controversial evolutionary relationships among plant species and genera, and the identification of unique indels in plastomes may even help to understand the evolution of some plant families. Menispermum L. (Menispermaceae) consists of three species, M. dauricum DC., M. canadense L., and M. mexicanum Rose, which are disjuncly distributed among East Asia, Eastern North America and Mexico. Taxonomists continue to debate whether M. mexicanum is a distinct species, a variety of M. dauricum, or simply a synonym of M. canadense. To date, no molecular systematics studies have included this doubtful species in phylogenetic analyses. In this study, we examined phylogenomics and phylogeography of Menispermum across its entire range using 29 whole plastomes of Menispermaceae and 18 ITS1&ITS2 sequences of Menispermeae. We reconstructed interspecific relationships of Menispermum and explored plastome evolution in Menispermaceae, revealing several genomic hotspot regions for the family. Phylogenetic and network analyses based on whole plastome and ITS1&ITS2 sequences show that Menispermum clusters into two clades with high support values, Clade A (M. dauricum) and Clade B (M. canadense + M. mexicanum). However, M. mexicanum is nested within M. canadense and, as a result, we support that M. mexicanum is a synonym of M. canadense. We also identified important molecular variations in the plastomes of Menispermaceae. Several indels and consequently premature terminations of genes occur in Menispermaceae. A total of 54 regions were identified as the most highly variable plastome regions, with nucleotide diversity (Pi) values > 0.05, including two coding genes (matK, ycf1), four introns (trnK intron, rpl16 intron, rps16 intron, ndhA intron), and 48 intergenic spacer (IGS) regions. Of these, four informative hotspot regions (trnH-psbA, ndhF-rpl32, trnK-rps16, and trnP-psaJ) should be especially useful for future studies of phylogeny, phylogeography and conservation genetics of Menispermaceae.

Taxonomy and Phylogeny of Peniophora Sensu Lato (Russulales, Basidiomycota).

Peniophora is an old corticioid genus, from which two small satellite genera, Dendrophora and Duportella, were derived based on morphological differences. Molecular systematic studies showed that they belong to Peniophoraceae, Russulales, but the inter- and intra-generic phylogenetic relationships are still unclear. Moreover, the species diversity of this group in subtropical and tropical Asia has not been sufficiently investigated and studied. In this study, we carried out an intensive taxonomic and phylogenetic study on Peniophora sensu lato based on analyses of concatenated ITS1-5.8S-ITS2 (ITS, Internal Transcribed Spacer) and 28S (D1-D2 domains of nuc 28S rDNA) sequence data of all available species worldwide. In the phylogenetic trees, species of Peniophora s. l. (sensu lato) including types of Peniophora s.s. (sensu stricto), Dendrophora and Duportella were interspersed within a strongly supported clade. It means that the morphological delimitations of the three genera are not reliable, and they should be regarded as a large genus. As a result, eight species of Duportella were transferred to Peniophora, although five of them have not been sequenced. Four new distinct lineages, corresponding to Peniophora cremicolor, P. major, P. shenghuae and P. vietnamensis spp. nov., were recovered in the trees. Peniophora taiwanensis is treated as a later synonym of P. malaiensis based on morphological and molecular evidence. Duportella renispora is excluded from Peniophoraceae, because BLAST results of the ITS and 28S sequences of the holotype showed that it is closely related to Amylostereum. Descriptions and illustrations are provided for the four new species, and an identification key is given for all 25 species of Peniophora in China. Our results indicated that the species diversity of the corticioid fungi in Asia is rich and deserves further study.

Secondary Structure Form of ITS2 Region: A Significant Labeling Tool at all Taxonomic Levels

The general purpose of the molecular systematic studies is to illuminate the ITS2 structure of the target populations, to determine its phylogenetic boundaries, and to clarify intra-species and inter-species relationships. Particularly, the internal transcribed spacer 2 (ITS2) region of nuclear ribosomal DNA is used in molecular systematics because of availability of conserved regions with its highly repeated in number in plant genomes. Addition to the primary sequences of ITS2, also secondary structure form of the region became a valuable feature in species divergence and became to use like a morphological character. In the current study to indicate the secondary structure form of the ITS2 region as a useful tool in systematics, different taxa from 22 genera were used. The DNA samples were collected in the field studies in 2021 and sequences were aligned using ClustalW and Kimura-2 parameter to calculate the genetic distances. Phylogenetic tree was also constructed with Maximum Likelihood method with the best suitable model at MEGA X software. The secondary structure predictions of species and ΔG (Gibbs) free energy calculations, the tools of both the ITS2 database and mFOLD web server were used. The results indicated that ITS2 secondary structure estimations represented the genetic differences visibly with its helices and motifs like a morphological character. Consequently, even if primary structure of the ITS2 region revealed as valuable marker in molecular systematic studies, also, all tested secondary structure forms of the region will be used as an ideal marker for taxonomic and phylogenetic reconstructions at all taxonomic levels.

A new species of the genus Podocerus from the Seto Inland Sea, Japan (Crustacea, Amphipoda, Podoceridae).

A new podocerid amphipod, Podocerussetouchiensis sp. nov., is described from the Etajima Island, the Seto Inland Sea, Japan. This new species differs from its congeners by the dorsal carination of pereonites and pleonites, and form of the antenna 1, gnathopods 1 and 2, uropods 1 and 2, and telson. Nucleotide sequence data of the mitochondrial cytochrome c subunit I (COI) from a paratype of Podocerussetouchiensis sp. nov. is provided for future molecular systematic studies.

Resolved and Redeemed: A New Fleck to the Evolutionary Divergence in the Genus Scomberomorus Lacepède, 1801 (Scombridae) With Cryptic Speciation

The genus Scomberomorus, with 18 nominal species, sustains a significant heterogeneous fishery throughout its range. The sole molecular systematic study of this genus concerned the species group S. regalis, which contains the new world taxa. The species diversity of Scomberomorus in the northern Indian Ocean has not been studied at the molecular level, often leading to misidentifications. Here, novel genetic data are provided that reconfigure species boundaries from the region. We used single and multilocus data (eight mitochondrial and three nuclear genes) to infer phylogenetic relationships, species delimitation, and the resurrection of a time-calibrated phylogenetic tree. Our aim was also to verify the hypothesis of geographical races in S. guttatus predicated on variable vertebral counts. Interestingly, all species delimitation analyses have recovered another highly cryptic species in the nominal S. guttatus previously believed to have an Indo-Pacific distribution. Scomberomorus guttatus (Bloch and Schneider, 1801) in the sensu stricto, is redeemed from its type locality based on genetic data and preliminary morphomeristic investigations and has a restricted distribution in the Bay of Bengal. The cryptic species Scomberomorus aff. guttatus which exhibits >10% genetic divergence from S. guttatus is resurrected here from the synonymy of the latter as Scomberomorus leopardus (Shaw, 1803). Widespread in the Indo-Pacific, this species contains two major molecular operational taxonomic units (MOTUs) with a divergence threshold of over 2% between them. Our analysis suggests that vertebral counts must be coupled with other features to identify the species/lineages in the nominal S. guttatus. The heterogeneity in the S. guttatus species group is discussed in relation to the ecological diversity of the region which facilitates larval recruitment and niche specialization. The results also revealed two allopatric putative species in S. commerson, found primarily in the Pacific and Indian Oceans. This study added genetic data from S. lineolatus and S. koreanus, not previously represented in the sequence repositories. Estimation of divergence time indicated that the Indo-West Pacific species group undergoes multiple diversification events besides the recent splits detected within S. leopardus.

Revalidation and redescription of Sarcocheilichthys sciistius (Abbott, 1901) (Cypriniformes: Cyprinidae) from the northern China.

Sarcocheilichthys sciistius was originally described by Abbott in 1901. It was considered as a synonym of S. nigripinnis by later researchers. However, some recent studies suggested that S. nigripinnis from the Haihe River Basin and Yellow River Basin were different to S. nigripinnis distributed in southern China, and that these populations show a closer relationship with the Heilongjiang (Amur) River endemic speciesS. czerskii and S. soldatovi. In this study, we examined one hundred and twenty-five specimens from nine sites of six river basins. Based on morphological and molecular systematic studies, we confirmed S. sciistius is a valid species, and S. czerskii, S. soldatovi were synonyms of S. sciistius. Sarcocheilichthys sciistius can be distinguished from S. nigripinnis by having more lateral-line scales (4042 vs. 3840), shorter lower lip and different pectoral and pelvic fins color. The phylogenetic trees reconstructed by Bayesian Inference and Maximum Likelihood based on the Cyt b gene also support S. sciistius as a distinct species.

DNA Sequence-Based Identification of Fusarium: A Work in Progress.

Accurate species-level identification of an etiological agent is crucial for disease diagnosis and management because knowing the agent's identity connects it with what is known about its host range, geographic distribution, and toxin production potential. This is particularly true in publishing peer-reviewed disease reports, where imprecise and/or incorrect identifications weaken the public knowledge base. This can be a daunting task for phytopathologists and other applied biologists that need to identify Fusarium in particular, because published and ongoing multilocus molecular systematic studies have highlighted several confounding issues. Paramount among these are: (i) this agriculturally and clinically important genus is currently estimated to comprise more than 400 phylogenetically distinct species (i.e., phylospecies), with more than 80% of these discovered within the past 25 years; (ii) approximately one-third of the phylospecies have not been formally described; (iii) morphology alone is inadequate to distinguish most of these species from one another; and (iv) the current rapid discovery of novel fusaria from pathogen surveys and accompanying impact on the taxonomic landscape is expected to continue well into the foreseeable future. To address the critical need for accurate pathogen identification, our research groups are focused on populating two web-accessible databases (FUSARIUM-ID v.3.0 and the nonredundant National Center for Biotechnology Information nucleotide collection that includes GenBank) with portions of three phylogenetically informative genes (i.e., TEF1, RPB1, and RPB2) that resolve at or near the species level in every Fusarium species. The objectives of this Special Report, and its companion in this issue (Torres-Cruz etal. 2022), are to provide a progress report on our efforts to populate these databases and to outline a set of best practices for DNA sequence-based identification of fusaria.

Ancient DNA extraction methods for herbarium specimens: When is it worth the effort?

PremiseHerbaria harbor a tremendous number of plant specimens that are rarely used for molecular systematic studies, largely due to the difficulty in extracting sufficient amounts of high‐quality DNA from the preserved plant material.MethodsWe compared the standard Qiagen DNeasy Plant Mini Kit and a specific protocol for extracting ancient DNA (aDNA) (the N‐phenacylthiazolium bromide and dithiothreitol [PTB–DTT] extraction method) from two different plant genera (Xanthium and Salix). The included herbarium materials covered about two centuries of plant collections. To analyze the success of DNA extraction using each method, a subset of samples was subjected to a standard library preparation as well as target‐enrichment approaches.ResultsThe PTB–DTT method produced a higher DNA yield of better quality than the Qiagen kit; however, extracts from the Qiagen kit over a certain DNA yield and quality threshold produced comparable sequencing results. The sequencing resulted in high proportions of endogenous reads. We were able to successfully sequence 200‐year‐old samples.DiscussionThis method comparison revealed that, for younger specimens, DNA extraction using a standard kit might be sufficient. For old and precious herbarium specimens, aDNA extraction methods are better suited to meet the requirements for next‐generation sequencing.

Phylogenetic Relationships of the Orchid Genus Coelogyne in Peninsular Malaysia Inferred from Morphological Characteristics and Internal Transcribed Spacer (ITS) Sequence Data

The phylogenetic relationships among the Peninsular Malaysian orchid genus Coelogyne were studied by morphological characteristics and sequence data of the internal transcribed region (ITS) from the nuclear ribosomal DNA (nrDNA). Coelogyne is a large genus of about 200 species distributed in pantropical areas from the Himalayas, Sri Lanka, India, Southern China and throughout South East Asia to Papua New Guinea. The widely accepted previous classification system was exclusively based on floral morphology. There were very few molecular systematic studies of Coelogyne done in Peninsular Malaysia thus far. In this study, 59 Coelogyne samples were collected throughout Peninsular Malaysia and 57 of them were identified to the species level. To study the phylogeny of this genus, morphological characters were utilized together with molecular evidences to generate the systematic hypotheses. Cluster analysis was performed using both the vegetative and floral characters. The results showed that three sections of Peninsular Malaysian Coelogyne, namely Longifoliae, Speciosae, and Fuliginosae were sister groups which were more closely related by forming one clade by itself. Another clade consisted of four other sections, namely Flaccidae, Coelogynae, Tomentosae, and Verrucosae. Molecular phylogenies obtained by using the Neighbour-Joining method showed the close relationship between the sections Tomentosae and Verrucosae, whereas usage of the Maximum Likelihood method demonstrated that three sections, namely Longifoliae, Speciosae, and Fuliginosae, were sister groups since they formed a single clade.

Complete mitochondrial genome of cylicocyclus auriculatus: molecular structure and phylogenetic analysis

Cylicocyclus spp. (Nematoda: Strongylida: Cyathostominae) are the common and important parasitic nematodes found in horses and donkeys worldwide. In this study, the complete mitochondrial genome of Cylicocyclus auriculatus Looss 1900, a representative member of this genus from the donkey in Southwest China was determined using the next-generation DNA sequencing technology. The genome was 13,851 bp in size and consisted of 36 genes including 12 protein-coding genes (atp6, cox1-3, cytb, nad1-6 and nad4L), 22 transfer RNA genes and two ribosomal RNA genes (rrnL and rrnS), as well as two non-coding regions. Phylogenetic analysis showed that C. auriculatus and Cylicocyclus insigne Boulenger 1917 were closely related, and then both grouped with other congeneric species and formed a monophyletic relationship with either species of Cyathostomum, Coronocyclus, Cyathostomum, Cylicostephanus or Cylicodontophorus, demonstrating their phylogenetic stability within Cyathostominae. These cumulative mitochondrial DNA data provide novel and useful genetic markers for molecular diagnostic, systematic and evolutionary biological studies of Cyathostominae nematodes.

Designation of the neotype of Triatomadimidiata (Latreille, 1811) (Hemiptera, Reduviidae, Triatominae), with full integrated redescription including mitogenome and nuclear ITS-2 sequences.

The taxonomic status of Triatomadimidiata (Latreille, 1811) is, by far, the most discussed within Triatominae. Molecular studies have recovered at least three independently evolving lineages in T.dimidiata across its range. The original description of T.dimidiata (as Reduviusdimidiatus) included few taxonomic characters, and no types were assigned. To define and describe the cryptic diversity within T.dimidiata sensu lato (s.l.), a neotype must be designated. For this purpose, all 199 specimens identified as T.dimidiata from the collections of the Smithsonian Institution – National Museum of Natural History and the American Museum of Natural History, ranging from Peru to Mexico, were studied. Only one specimen (from Tumbes, Peru) matched the combination of characters as listed in the original description, and it is herein formally designated as the neotype for T.dimidiata. The neotype is morphologically described and DNA sequences of its whole mitochondrial genome and the nuclear second internal transcribed spacer region (ITS2), commonly used in triatomine molecular systematics studies, are presented and compared to other publicly available sequences of T.dimidiata s.l. in GenBank. Our results suggest that T.dimidiata sensu stricto (s.s.) is somewhat rare and, therefore, unlikely to serve as a major vector of Chagas disease.

Systematic analysis of the genus Eriocaulon L. in India based on molecular and morphological evidence

The present study is the first-ever comprehensive molecular systematic study of Indian Eriocaulon species based on the critical appraisal of the freshly collected 552 accessions of 66 Eriocaulon spp. The genus Eriocaulon L. (Eriocaulaceae) is one of the most diverse genera of Eriocaulaceae and around one-quarter of the world’s representation is in India. The taxonomy of the genus has been challenging due to high intraspecific and low interspecific variation. With about 70% of Indian species endemic to the country, sparse representation in earlier phylogenetic studies did not provide phylogenetic resolution of Indian species. A critical analysis of morphology including microscopic seed surface characters of 55 species is presented here for the first time. Phylogenetic analyses based on ITS and trnL–F data yielded three major clades of Indian Eriocaulon species. Based on the morphological and molecular data, Eriocaulon baramaticum, E. govindiana, E. gulnarparianum, E. idukkianum, E. maharashtrense and E. pradeepii have been synonymized. Also, Eriocaulon rhodae has been reinstated. Molecular data exhibited little congruence with the earlier available taxonomic treatments for Indian species. Several morphological characters were mapped onto a phylogeny and none were found to characterize monophyletic groups.

Eurythenes atacamensis sp. nov. (Crustacea: Amphipoda) exhibits ontogenetic vertical stratification across abyssal and hadal depths in the Atacama Trench, eastern South Pacific Ocean

Eurythenes S.I. Smith in Scudder, 1882 (Crustacea: Amphipoda) are prevalent scavengers of the benthopelagic community from bathyal to hadal depths. While a well-studied genus, molecular systematic studies have uncovered cryptic speciation and multiple undescribed lineages. Here, we apply an integrative taxonomic approach and describe the tenth species, Eurythenes atacamensis sp. nov., based on specimens from the 2018 Atacamex and RV Sonne SO261 Expeditions to the southern sector of the Peru-Chile Trench, the Atacama Trench (24–⁠21°S). Eurythenes atacamensis sp. nov. is a large species, max. observed length 83.2 mm, possesses diagnostic features, including a short gnathopod 1 palm and a chelate gnathopod 2 palm, and a distinct genetic lineage based on a 16S rRNA and COI phylogeny. This species is a dominant bait-attending fauna with an extensive bathymetric range, spanning from 4974 to 8081 m. The RV Sonne SO261 specimens were recovered along a 10-station transect from abyssal to hadal depths and further examined for demographic and bathymetric-related patterns. Ontogenetic vertical stratification was evident across the trench axis, with only juveniles present at abyssal depths (4974–6025 m). Total length-depth analysis revealed that the size of females was unrelated to depth, whereas juveniles followed a sigmoidal relationship with a step-up in size at depths >7200 m. Thus, these bathymetric trends suggest that juveniles and females employ differing ecological strategies in subduction trench environments. This study highlights that even dominant and ecologically important species are still being discovered within the abyssal and hadal environments. Continued systematic expeditions will lead to an improved understanding of the eco-evolutionary drivers of speciation in the world’s largest ecosystem.

Genetically diverse yet morphologically conserved: Hidden diversity revealed among Bornean geckos (Gekkonidae: Cyrtodactylus )

The appreciation of cryptic biological diversity, and the pace at which it is recognized, has greatly increased with the use of molecular systematic techniques. The gekkonid genus Cyrtodactylus Gray, 1827 is one example of a group that has undergone a particularly rapid increase in recognized diversity due to molecular systematic studies. Many of these new species result from recognizing closely related but diagnosable lineages into sister taxa. Our study implements a multi-faceted approach to delimit cryptic Cyrtodactylus lineages on the Southeast Asian island of Borneo using morphological, ecological, and multilocus genetic data. We use multiple species delimitation models to assess species boundaries and identify clades that warrant further investigation. Unlike most morphologically cryptic species that have recently diverged, we find evidence of cryptic lineages being polyphyletic. Using multivariate statistical analyses, we show minimal phenotypic distinction between putative cryptic species within the C. pubisulcus complex. Despite not finding morphologically diagnostic characters, we demonstrate strong evidence for the specific recognition of C. hantu sp. nov. and C. miriensis sp. nov., which are currently considered conspecific with C. pubisulcus, from Sarawak, Malaysia. Our new concept for C. pubisulcus restricts the geographic range of the species to specific regions in western Sarawak, Malaysia, thus underscoring the need to conserve the limited remaining habitats of these species, as well as the considerable undescribed diversity across Borneo.