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Grammatical gender in New Guinea is an often neglected area in typological research, even though it is extremely diverse. For example, in New Guinea, some languages have grammatical gender systems with two sex-based categories, more than four gender-indexing targets, and no gender marking on nouns, while some languages have grammatical gender systems with much more categories, which are only marginally sex-based. This paper infers the processes of development and change of grammatical gender in Torricelli languages from two perspectives. First, it synthesizes the available data in the existing literature and hypothesizes the evolutionary pathway of gender systems in Torricelli languages. Nineteen Torricelli languages are selected as a representative coverage of the 55 Torricelli languages listed in Glottolog within the limits of the available documentation. These languages are then coded based on 6 presence-absence features relating to gender marking on verbs, adjectives, nouns, numerals, pronouns, and demonstratives. Second, it conducts an analysis with phylogenetic comparative methods to provide a quantitative assessment of the evolutionary possibilities for gender systems in Torricelli languages. The preliminary results show that gender is likely marked at the root of Torricelli languages, with pronouns and verbs being at the core of the system. This is in agreement with trends reflecting the evolution of gender systems in languages across the world. [ABSTRACT FROM AUTHOR]

In this study, the complete mitochondrial genome of Triplophysa erythraea was determined for the first time. Results showed the mitogenome was 16 565 bp, including 2 ribosomal RNA genes, 22 transfer RNA genes, 13 protein-coding genes and 2 non-coding regions. The overall base composition of its mitochondrial genome was 28.9% A, 25.2% T, 28.7% C and 17.2% G. Phylogenetic tree revealed that Triplophysa erythraea had the closest relationship with Triplophysa xiangxiensis, which was also found in Xiangxi Tujia and Miao Nationality Autonomous Prefecture, Hunan province. In general, this study provided valuable information for conservation genetics analyses of Triplophysa erythraea and further displayed the evolution of species within the genus Triplophysa.

Abstract Background The phylum Nematoda is incredibly diverse and includes many parasites of humans, livestock, and plants. Peptide-activated G protein-coupled receptors (GPCRs) are central to the regulation of physiology and numerous behaviors, and they represent appealing pharmacological targets for parasite control. Efforts are ongoing to characterize the functions and define the ligands of nematode GPCRs, with already most peptide GPCRs known or predicted in Caenorhabditis elegans. However, comparative analyses of peptide GPCR conservation between C. elegans and other nematode species are limited, and many nematode GPCRs remain orphan. A phylum-wide perspective on peptide GPCR profiles will benefit functional and applied studies of nematode peptide GPCRs. Results We constructed a pan-phylum resource of C. elegans peptide GPCR orthologs in 125 nematode species using a semi-automated pipeline for analysis of predicted proteome datasets. The peptide GPCR profile varies between nematode species of different phylogenetic clades and multiple C. elegans peptide GPCRs have orthologs across the phylum Nematoda. We identified peptide ligands for two highly conserved orphan receptors, NPR-9 and NPR-16, that belong to the bilaterian galanin/allatostatin A (Gal/AstA) and somatostatin/allatostatin C (SST/AstC) receptor families. The AstA-like NLP-1 peptides activate NPR-9 in cultured cells and are cognate ligands of this receptor in vivo. In addition, we discovered an AstC-type peptide, NLP-99, that activates the AstC-type receptor NPR-16. In our pan-phylum resource, the phylum-wide representation of NPR-9 and NPR-16 resembles that of their cognate ligands more than those of allatostatin-like peptides that do not activate these receptors. Conclusions The repertoire of C. elegans peptide GPCR orthologs varies across phylogenetic clades and several peptide GPCRs show broad conservation in the phylum Nematoda. Our work functionally characterizes the conserved receptors NPR-9 and NPR-16 as the respective GPCRs for the AstA-like NLP-1 peptides and the AstC-related peptide NLP-99. NLP-1 and NLP-99 are widely conserved in nematodes and their representation matches that of their receptor in most species. These findings demonstrate the conservation of a functional Gal/AstA and SST/AstC signaling system in nematodes. Our dataset of C. elegans peptide GPCR orthologs also lays a foundation for further functional studies of peptide GPCRs in the widely diverse nematode phylum.

Abstract Bovine viral diarrhoea (BVD) is one of the most economically damaging livestock enzootic diseases in the world. BVD aetiological agents are three pestiviruses (BVDV-1, -2 and HoBi-like pestivirus), which exhibit high genetic diversity and complex transmission cycles. This considerably hampers the management of the disease, which is why eradication plans have been implemented in several countries. In France, a national plan has been in place since 2019. Our understanding of its impact on the distribution of BVDV genotypes is limited by the availability of French genetic data. Here, we conducted a molecular epidemiology study to refine our knowledge of BVDV genetic diversity in France, characterise its international relationships, and analyse national spatio-temporal genotypic distribution. We collated 1037 BVDV-positive samples throughout France between 2011 and 2023, with a greater sampling effort in two major cattle production areas. We developed a high-throughput sequencing protocol which we used to complete the 5’UTR genotyping of this collection. We show that two main BVDV-1 genotypes, 1e and 1b, account for 88% of genotyped sequences. We also identified seven other BVDV-1 genotypes occurring at low frequencies and three BVDV-2 samples (genotype 2c). Phylogenetic analyses indicate different worldwide distribution patterns between the two main BVDV-1 genotypes. Their relative frequencies present no major changes in France since the 1990s and few variations at the national scale. We also found some degree of local spatial structuring in western France. Overall, our results demonstrate the potential of large-scale sequence-based surveillance to monitor changes in the epidemiological situation of enzootic diseases.

Abstract The epidemiology and circulation patterns of various rhinovirus types within populations remains under-explored. We generated 803 VP4/VP2 gene sequences from rhinovirus-positive samples collected from acute respiratory illness (ARI) patients, including both in-patient and outpatient cases, between 1st January and 31st December 2014 from eleven surveillance sites across Kenya and used phylogenetics to characterise virus introductions and spread. RVs were detected throughout the year, with the highest detection rates observed from January to March and June to July. We detected a total of 114 of the 169 currently classified types. Our analysis revealed numerous virus introductions into Kenya characterized by local expansion and extinction, and extensive spatial mixing of types within the country due to the widespread transmission of the virus after an introduction. This work demonstrates that in a single year, the circulation of rhinovirus in Kenya was characterized by substantial genetic diversity, multiple introductions, and extensive geographical spread.

Abstract Background Diplomonads are anaerobic flagellates classified within Metamonada. They contain both host-associated commensals and parasites that reside in the intestinal tracts of animals, including humans (e.g., Giardia intestinalis), as well as free-living representatives that inhabit freshwater and marine anoxic sediments (e.g., Hexamita inflata). The evolutionary trajectories within this group are particularly unusual as the free-living taxa appear to be nested within a clade of host-associated species, suggesting a reversal from host-dependence to a secondarily free-living lifestyle. This is thought to be an exceedingly rare event as parasites often lose genes for metabolic pathways that are essential to a free-living life strategy, as they become increasingly reliant on their host for nutrients and metabolites. To revert to a free-living lifestyle would require the reconstruction of numerous metabolic pathways. All previous studies of diplomonad evolution suffered from either low taxon sampling, low gene sampling, or both, especially among free-living diplomonads, which has weakened the phylogenetic resolution and hindered evolutionary insights into this fascinating transition. Results We sequenced transcriptomes from 1 host-associated and 13 free-living diplomonad isolates; expanding the genome scale data sampling for diplomonads by roughly threefold. Phylogenomic analyses clearly show that free-living diplomonads form several branches nested within endobiotic species. Moreover, the phylogenetic distribution of genes related to an endobiotic lifestyle suggest their acquisition at the root of diplomonads, while traces of these genes have been identified in free-living diplomonads as well. Based on these results, we propose an evolutionary scenario of ancestral and derived lifestyle transitions across diplomonads. Conclusions Free-living taxa form several clades nested within endobiotic taxa in our phylogenomic analyses, implying multiple transitions between free-living and endobiotic lifestyles. The evolutionary history of numerous virulence factors corroborates the inference of an endobiotic ancestry of diplomonads, suggesting that there have been several reversals to a free-living lifestyle. Regaining host independence may have been facilitated by a subset of laterally transferred genes. We conclude that the extant diversity of diplomonads has evolved from a non-specialized endobiont, with some taxa becoming highly specialized parasites, others becoming free-living, and some becoming capable of both free-living and endobiotic lifestyles.

Abstract Chloroplast (cp) genome sequences have been extensively used for phylogenetic and evolutionary analyses, as many have been sequenced in recent years. Identification of Quercus is challenging because many species overlap phenotypically owing to interspecific hybridization, introgression, and incomplete lineage sorting. Therefore, we wanted to gain a better understanding of this genus at the level of the maternally inherited chloroplast genome. Here, we sequenced, assembled, and annotated the cp genomes of the threatened Quercus marlipoensis (160,995 bp) and Q. kingiana (161,167 bp), and mined these genomes for repeat sequences and codon usage bias. Comparative genomic analyses, phylogenomics, and selection pressure analysis were also performed in these two threatened species along with other species of Quercus. We found that the guanine and cytosine content of the two cp genomes were similar. All 131 annotated genes, including 86 protein-coding genes, 37 transfer RNA genes, and 8 ribosomal RNA genes, had the same order in the two species. A strong A/T bias was detected in the base composition of simple sequence repeats. Among the 59 synonymous codons, the codon usage pattern of the cp genomes in these two species was more inclined toward the A/U ending. Comparative genomic analyses indicated that the cp genomes of Quercus section Ilex are highly conserved. We detected eight highly variable regions that could be used as molecular markers for species identification. The cp genome structure was consistent and different within and among the sections of Quercus. The phylogenetic analysis showed that section Ilex was not monophyletic and was divided into two groups, which were respectively nested with section Cerris and section Cyclobalanopsis. The two threatened species sequenced in this study were grouped into the section Cyclobalanopsis. In conclusion, the analyses of cp genomes of Q. marlipoensis and Q. kingiana promote further study of the taxonomy, phylogeny and evolution of these two threatened species and Quercus.

ABSTRACT: Objectives: The emergence of carbapenem-resistant Klebsiella pneumoniae presents significant health challenges. Here, we present the structural genome sequence of an NDM-5–producing K. pneumoniae (HZKP2) in China. Methods: Antimicrobial susceptibility tests were conducted via broth microdilution. Whole-genome sequencing was performed for genomic analysis. Wzi and capsular polysaccharide (KL) were analysed using Kaptive. Resistance genes, virulence factors, and comparative genomics analyses were also conducted. Multilocus sequence typing (MLST), replicons type, and core genome MLST analysis were further conducted using BacWGSTdb server. Results: HZKP2 was resistant to cefepime, ceftazidime, ciprofloxacin, ciprofloxacin, meropenem, and ertapenem. It harboured fosA, blaSHV-187, oqxA, oqxB, sul1, dfrA1, tet(A), floR, aph(6)-Id, aph(3′')-Ib, sul2, blaCTX-M-55, and blaNDM-5. Based on the RAST results, 5563 genes that belonged to 398 subsystems were annotated. The complete genome sequence of HZKP2 was characterized as ST1, wzi 19, and KL19, 5 five contigs totalling 5 654 446 bp, including one chromosome and four plasmids. Further analysis found that blaNDM-5 was located in a 46 161 bp IncX3 plasmid (pHZKP2-3). The genetic structure of blaNDM-5 gene was ISKox3-IS26-bleMBL-blaNDM-5-IS5-ISAb125-IS3000. Further analysis revealed that insertion sequences mediated the dissemination of blaNDM-5 from other species of Enterobacterales. Phylogenetic analysis showed that the closest relative was from a human stool specimen in China, which differed by 53 core genome MLST alleles. Conclusions: Our study provides the first structural perspective of the ST1 K. pneumoniae isolate producing NDM-5 in China. These results could provide valuable insights into the genetic characteristics, antimicrobial resistance mechanisms, and transmission dynamics of carbapenem-resistant K. pneumoniae in clinical settings.

Divergence time estimation is crucial to provide temporal signals for dating biologically important events from species divergence to viral transmissions in space and time. With the advent of high-throughput sequencing, recent Bayesian phylogenetic studies have analyzed hundreds to thousands of sequences. Such large-scale analyses challenge divergence time reconstruction by requiring inference on highly correlated internal node heights that often become computationally infeasible. To overcome this limitation, we explore a ratio transformation that maps the original $N-1$ internal node heights into a space of one height parameter and $N-2$ ratio parameters. To make the analyses scalable, we develop a collection of linear-time algorithms to compute the gradient and Jacobian-associated terms of the log-likelihood with respect to these ratios. We then apply Hamiltonian Monte Carlo sampling with the ratio transform in a Bayesian framework to learn the divergence times in 4 pathogenic viruses (West Nile virus, rabies virus, Lassa virus, and Ebola virus) and the coralline red algae. Our method both resolves a mixing issue in the West Nile virus example and improves inference efficiency by at least 5-fold for the Lassa and rabies virus examples as well as for the algae example. Our method now also makes it computationally feasible to incorporate mixed-effects molecular clock models for the Ebola virus example, confirms the findings from the original study, and reveals clearer multimodal distributions of the divergence times of some clades of interest.

Phylogenetics has been foundational to SARS-CoV-2 research and public health policy, assisting in genomic surveillance, contact tracing, and assessing emergence and spread of new variants. However, phylogenetic analyses of SARS-CoV-2 have often relied on tools designed for de novo phylogenetic inference, in which all data are collected before any analysis is performed and the phylogeny is inferred once from scratch. SARS-CoV-2 data sets do not fit this mold. There are currently over 14 million sequenced SARS-CoV-2 genomes in online databases, with tens of thousands of new genomes added every day. Continuous data collection, combined with the public health relevance of SARS-CoV-2, invites an "online" approach to phylogenetics, in which new samples are added to existing phylogenetic trees every day. The extremely dense sampling of SARS-CoV-2 genomes also invites a comparison between likelihood and parsimony approaches to phylogenetic inference. Maximum likelihood (ML) and pseudo-ML methods may be more accurate when there are multiple changes at a single site on a single branch, but this accuracy comes at a large computational cost, and the dense sampling of SARS-CoV-2 genomes means that these instances will be extremely rare because each internal branch is expected to be extremely short. Therefore, it may be that approaches based on maximum parsimony (MP) are sufficiently accurate for reconstructing phylogenies of SARS-CoV-2, and their simplicity means that they can be applied to much larger data sets. Here, we evaluate the performance of de novo and online phylogenetic approaches, as well as ML, pseudo-ML, and MP frameworks for inferring large and dense SARS-CoV-2 phylogenies. Overall, we find that online phylogenetics produces similar phylogenetic trees to de novo analyses for SARS-CoV-2, and that MP optimization with UShER and matOptimize produces equivalent SARS-CoV-2 phylogenies to some of the most popular ML and pseudo-ML inference tools. MP optimization with UShER and matOptimize is thousands of times faster than presently available implementations of ML and online phylogenetics is faster than de novo inference. Our results therefore suggest that parsimony-based methods like UShER and matOptimize represent an accurate and more practical alternative to established ML implementations for large SARS-CoV-2 phylogenies and could be successfully applied to other similar data sets with particularly dense sampling and short branch lengths.

The antimalarial drug-resistance conundrum which threatens to reverse the great strides taken to curb the malaria scourge warrants an urgent need to find novel chemical scaffolds to serve as templates for the development of new antimalarial drugs. Plants represent a viable alternative source for the discovery of unique potential antiplasmodial chemical scaffolds. To expedite the discovery of new antiplasmodial compounds from plants, the aim of this study was to use phylogenetic analysis to identify higher plant orders and families that can be rationally prioritised for antimalarial drug discovery. We queried the PubMed database for publications documenting antiplasmodial properties of natural compounds isolated from higher plants. Thereafter, we manually collated compounds reported along with plant species of origin and relevant pharmacological data. We systematically assigned antiplasmodial-associated plant species into recognised families and orders, and then computed the resistance index, selectivity index and physicochemical properties of the compounds from each taxonomic group. Correlating the generated phylogenetic trees and the biological data of each clade allowed for the identification of 3 hot plant orders and families. The top 3 ranked plant orders were the (i) Caryophyllales, (ii) Buxales, and (iii) Chloranthales. The top 3 ranked plant families were the (i) Ancistrocladaceae, (ii) Simaroubaceae, and (iii) Buxaceae. The highly active natural compounds (IC50 ≤ 1 µM) isolated from these plant orders and families are structurally unique to the legacy antimalarial drugs. Our study was able to identify the most prolific taxa at order and family rank that we propose be prioritised in the search for potent, safe and drug-like antimalarial molecules.

BackgroundCOVID-19 pandemic mitigation measures, including travel restrictions, limited global circulation of influenza viruses. In Australia, travel bans for non-residents and quarantine requirements for returned travellers were eased in November 2021, providing pathways for influenza viruses to be re-introduced.AimWe aimed to describe the epidemiological and virological characteristics of the re-emergence of influenza in Victoria, Australia to inform public health interventions.MethodsFrom 1 November 2021 to 30 April 2022, we conducted an epidemiological study analysing case notification data from the Victorian Department of Health to describe case demographics, interviewed the first 200 cases to establish probable routes of virus reintroduction and examined phylogenetic and antigenic data to understand virus diversity and susceptibility to current vaccines.ResultsOverall, 1,598 notifications and 1,064 positive specimens were analysed. The majority of cases (61.4%) occurred in the 15-34 years age group. Interviews revealed a higher incidence of international travel exposure during the first month of case detections, and high levels of transmission in university residential colleges were associated with return to campus. Influenza A(H3N2) was the predominant subtype, with a single lineage predominating despite multiple importations.ConclusionEnhanced testing for respiratory viruses during the COVID-19 pandemic provided a more complete picture of influenza virus transmission compared with previous seasons. Returned international travellers were important drivers of influenza reemergence, as were young adults, a group whose role has previously been under-recognised in the establishment of seasonal influenza epidemics. Targeting interventions, including vaccination, to these groups could reduce future influenza transmission.

Turfgrass is a crop used extensively in athletic fields and golf courses in Maryland. A soil sample collected in July 2023 from an athletic field in Baltimore County, Maryland, part of a turfgrass nematode survey, contained Belonolaimus longicaudatus. In the southeastern United States, B. longicaudatus is an economically important pathogen of warm season turfgrass. The density was four individuals/100 cm3 of soil, and no visual symptoms were observed in the bermudagrass field. Morphological features and morphometrics of males and females were consistent with B. longicaudatus and placed the Maryland population in a subclade that was geographically represented by populations from north and west Florida, Texas, and South Carolina. Sequencing of the internal transcribed spacer region ITS1 and ITS2 and 28S large ribosomal subunit D2-23 expansion region confirmed the species' identity. Phylogenetic trees and parsimony network analysis placed the Maryland isolate in a large grouping of B. longicaudatus populations including those from Alabama, Delaware, Florida, Indiana, Mississippi, South Carolina, and Texas. To our knowledge, this is the first report of B. longicaudatus in Maryland.

Abstract Background Emerging zoonotic diseases arise from cross-species transmission events between wild or domesticated animals and humans, with bats being one of the major reservoirs of zoonotic viruses. Viral metagenomics has led to the discovery of many viruses, but efforts have mainly been focused on some areas of the world and on certain viral families. Methods We set out to describe full-length genomes of new picorna-like viruses by collecting feces from hundreds of bats captured in different regions of Spain. Viral sequences were obtained by high-throughput Illumina sequencing and analyzed phylogenetically to classify them in the context of known viruses. Linear discriminant analysis (LDA) was performed to infer likely hosts based on genome composition. Results We found five complete or nearly complete genomes belonging to the family Picornaviridae, including a new species of the subfamily Ensavirinae. LDA suggested that these were true vertebrate viruses, rather than viruses from the bat diet. Some of these viruses were related to picornaviruses previously found in other bat species from distant geographical regions. We also found a calhevirus genome that most likely belongs to a proposed new family within the order Picornavirales, and for which genome composition analysis suggested a plant host. Conclusions Our findings describe new picorna-like viral species and variants circulating in the Iberian Peninsula, illustrate the wide geographical distribution and interspecies transmissibility of picornaviruses, and suggest new hosts for calheviruses.

Abstract Background Pseudomonas juntendi is a newly identified opportunistic pathogen, of which we have limited understanding. P. juntendi strains are often multidrug resistant, which complicates clinical management of infection. Methods A strain of Pseudomonas juntendi (strain L4326) isolated from feces was characterized by MALDI-TOF-MS and Average Nucleotide Identity BLAST. This strain was further subject to whole-genome sequencing and Maximum Likelihood phylogenetic analysis. The strain was phenotypically characterized by antimicrobial susceptibility testing and conjugation assays. Results We have isolated the novel P. juntendi strain L4236, which was multidrug resistant, but retained sensitivity to amikacin. L4236 harbored a megaplasmid that encoded bla OXA−1 and a novel bla IMP−1 resistance gene variant. P. juntendi strain L4236 was phylogenetically related to P. juntendi strain SAMN30525517. Conclusion A rare P. juntendi strain was isolated from human feces in southern China with a megaplasmid coharboring bla IMP−1−like and bla OXA−1. Antimicrobial selection pressures may have driven acquisition of drug-resistance gene mutations and carriage of the megaplasmid.

Abstract Background LIM (Lineage-11 (LIN-11), Insulin-1 (ISL-1), and Mechanotransduction-3 (MEC-3)) genes belong to a family that hold ubiquitous properties contributing to organ, seed, and pollen development as well as developmental and cellular responses to biotic and abiotic stresses. Lettuce (Lactuca sativa) is a highly consumed vegetable crop susceptible heat stress. High temperatures limit lettuce’s overall yield, quality and marketability. Lettuce LIM genes have not been identified and their role in response to high temperatures is not known. Aiming to identify potential new targets for thermoresilience, we searched for LIM genes in lettuce and compared them with orthologous of several dicotyledons and monocotyledons plant species. Results We identified fourteen lettuce LIM genes distributed into eight different subgroups using a genome-wide analysis strategy. Three belonging to DAR (DA means “large” in Chinese) class I, two DAR class II, one in the WLIM1, two in the WLIM2, one in the PLIM1, two in the PLIM2 class, one ßLIM and two δLIMs. No DAR-like were identified in any of the species analyzed including lettuce. Interestingly, unlike other gene families in lettuce which underwent large genome tandem duplications, LIM genes did not increase in number compared to other plant species. The response to heat stress induced a dynamic transcriptional response on LsLIM genes. All heat stress regimes, including night stress, day stress and day and night stress were largely responsible for changes in LIM transcriptional expression. Conclusions Our global analysis at the genome level provides a detailed identification of LIM genes in lettuce and other dicotyledonous and monocotyledonous plant species. Gene structure, physical and chemical properties as well as chromosomal location and Cis-regulatory element analysis together with our gene expression analysis under different temperature regimes identified LsWLIM1, LsWLIM2b, LsDAR3 and LsDAR5 as candidate genes that could be used by breeding programs aiming to produce lettuce varieties able to withstand high temperatures.

Rhodiola, belonging to the Crassulaceae family, is a perennial herbaceous plant genus. There are about 90 Rhodiola species worldwide, some of which have been reported to have medicinal properties. Rhodiola sachalinensis is a perennial medicinal herb within this genus and, in the present study, its chloroplast genome was sequenced, assembled, annotated and compared with 24 other Rhodiola species. The results obtained show that the chloroplast genome of R. sachalinensis is 151 595 bp long and has a CG content of 37.7%. The inverted repeats (IR) region of the Rhodiola chloroplast genome is the most conserved region, with the main differences being observed in the ycf1 and ndhF genes at the IRb‐small single copy boundary, and rps19 and trnH genes at the IRa‐large single copy boundary. Phylogenetic analysis showed that Rhodiola species form two major clades, and species with recorded medicinal properties, clustered together in one branch except for R. dumulosa. Within the genus, R. sachalinensis is most closely related to Rhodiola rosea, although comparative analyses showed that only R. sachalinensis and Rhodiola subopposita contained the psbZ gene, which encodes a highly conserved protein subunit of the Photosystem II core complex. Overall, the present study contributes to the understanding of the chloroplast genome of Rhodiola species, and provides a theoretical basis for the study of their genetic diversity and possible use as medicinal plants.

Litter size plays an essential role in mammalian evolution and is one of the most important factors determining whether an organism is deemed to have a ‘slow’ or ‘fast’ life history strategy. Humans are distinct in being classified as having slow life history yet bearing singletons who have completed relatively less growth than other ape neonates. Previous work has proposed that the ancestral primate gave birth to singletons. However, primate litter size has not yet been contextualized within a broad phylogenetic assessment of mammalian life history. We performed a comprehensive investigation of primate litter size using life history data for 155 primate species, and litter size data for an additional 791 boreoeutherian mammals. Litter size and life history traits have strong phylogenetic signal in primates (Pagel’s lambda: 0.99, p < 0.001; Blomberg’s K: 0.6311. p < 0.001), and litter size is significantly negatively correlated with gestation length (p < 0.001). Our data support that the last common ancestors of both primates and Haplorhini gave birth to multiples (litter size 1.7 and 1.6, respectively). We also find that singleton-bearing pregnancies evolved convergently in multiple primate lineages, including tarsiers and other haplorhines. This study contributes significantly to our understanding of life history and litter size in mammals, and we emphasize the utility of a callitrichid model for investigating the evolution of human reproduction.

Abstract Background The deep (> 200 m) ocean floor is often considered to be a refugium of biodiversity; many benthic marine animals appear to share ancient common ancestry with nearshore and terrestrial relatives. Whether this pattern holds for vertebrates is obscured by a poor understanding of the evolutionary history of the oldest marine vertebrate clades. Hagfishes are jawless vertebrates that are either the living sister to all vertebrates or form a clade with lampreys, the only other surviving jawless fishes. Results We use the hagfish fossil record and molecular data for all recognized genera to construct a novel hypothesis for hagfish relationships and diversification. We find that crown hagfishes persisted through three mass extinctions after appearing in the Permian ~ 275 Ma, making them one of the oldest living vertebrate lineages. In contrast to most other deep marine vertebrates, we consistently infer a deep origin of continental slope occupation by hagfishes that dates to the Paleozoic. Conclusion Our results establish hagfishes as ancient members of demersal continental slope faunas and suggest a prolonged accumulation of deep sea jawless vertebrate biodiversity.

Abstract Background Fleas are one of the most common and pervasive ectoparasites worldwide, comprising at least 2500 valid species. They are vectors of several disease-causing agents, such as Yersinia pestis. Despite their significance, however, the molecular genetics, biology, and phylogenetics of fleas remain poorly understood. Methods We sequenced, assembled, and annotated the complete mitochondrial (mt) genome of the rodent flea Nosopsyllus laeviceps using next-generation sequencing technology. Then we combined the new mitogenome generated here with mt genomic data available for 23 other flea species to perform comparative mitogenomics, nucleotide diversity, and evolutionary rate analysis. Subsequently, the phylogenetic relationship within the order Siphonaptera was explored using the Bayesian inference (BI) and maximum likelihood (ML) methods based on concentrated data for 13 mt protein-coding genes. Results The complete mt genome of the rodent flea N. laeviceps was 16,533 base pairs (bp) in a circular DNA molecule, containing 37 typical genes (13 protein-coding genes, 22 transfer RNA [tRNA] genes, and two ribosomal RNA [rRNA] genes) with one large non-coding region (NCR). Comparative analysis among the order Siphonaptera showed a stable gene order with no gene arrangement, and high AT content (76.71–83.21%) with an apparent negative AT and GC skew except in three fleas Aviostivalius klossi bispiniformis, Leptopsylla segnis, and Neopsylla specialis. Moreover, we found robust evidence that the cytochrome c oxidase subunit 1 (cox1) gene was the most conserved protein-coding gene (Pi = 0.15, non-synonymous/synonymous [Ka/Ks] ratio = 0.13) of fleas. Phylogenomic analysis conducted using two methods revealed different topologies, but both results strongly indicated that (i) the families Ceratophyllidae and Leptopsyllidae were paraphyletic and were the closest to each other, and (ii) the family Ctenophthalmidae was paraphyletic. Conclusions In this study, we obtained a high-quality mt genome of the rodent flea N. laeviceps and performed comparative mitogenomics and phylogeny of the order Siphonaptera using the mt database. The results will enrich the mt genome data for fleas, lay a foundation for the phylogenetic analysis of fleas, and promote the evolutionary analysis of Siphonaptera. Graphical Abstract

Abstract Background Although RNA-seq data are traditionally used for quantifying gene expression levels, the same data could be useful in an integrated approach to compute genetic distances as well. Challenges to using mRNA sequences for computing genetic distances include the relatively high conservation of coding sequences and the presence of paralogous and, in some species, homeologous genes. Results We developed a new computational method, RNA-clique, for calculating genetic distances using assembled RNA-seq data and assessed the efficacy of the method using biological and simulated data. The method employs reciprocal BLASTn followed by graph-based filtering to ensure that only orthologous genes are compared. Each vertex in the graph constructed for filtering represents a gene in a specific sample under comparison, and an edge connects a pair of vertices if the genes they represent are best matches for each other in their respective samples. The distance computation is a function of the BLAST alignment statistics and the constructed graph and incorporates only those genes that are present in some complete connected component of this graph. As a biological testbed we used RNA-seq data of tall fescue (Lolium arundinaceum), an allohexaploid plant ( $$2n = 14\text { Gb}$$ 2 n = 14 Gb ), and bluehead wrasse (Thalassoma bifasciatum), a teleost fish. RNA-clique reliably distinguished individual tall fescue plants by genotype and distinguished bluehead wrasse RNA-seq samples by individual. In tests with simulated RNA-seq data, the ground truth phylogeny was accurately recovered from the computed distances. Moreover, tests of the algorithm parameters indicated that, even with stringent filtering for orthologs, sufficient sequence data were retained for the distance computations. Although comparisons with an alternative method revealed that RNA-clique has relatively high time and memory requirements, the comparisons also showed that RNA-clique’s results were at least as reliable as the alternative’s for tall fescue data and were much more reliable for the bluehead wrasse data. Conclusion Results of this work indicate that RNA-clique works well as a way of deriving genetic distances from RNA-seq data, thus providing a methodological integration of functional and genetic diversity studies.

Members of the bacterial genus Pantoea produce a variety of antimicrobial products that are effective against plant, animal, and human pathogens. To date, little is known about the distribution and evolutionary history of these clusters. We surveyed the public databases for the 12 currently known antibiotic biosynthetic gene clusters found across Pantoea strains to determine their distribution. We show that some clusters, namely pantocin B, PNP-3, and PNP-4 are found strictly in Pantoea, while agglomerin, andrimid, AGA, dapdiamide, herbicolin, PNP-1, PNP-2, PNP-5, and pantocin A, are more broadly distributed in distantly related genera within Vibrionaceae, Pectobacteriaceae, Yersiniaceae, Morganellaceae, and Hafniaceae. We evaluated the evolutionary history of these gene clusters relative to a cpn60-based species tree, considering the flanking regions of each cluster, %GC, and presence of mobile genetic elements, and identified potential occurrences of horizontal gene transfer. Lastly, we also describe the biosynthetic gene cluster of pantocin B in the strain Pantoea agglomerans Eh318 more than 20 years after this antibiotic was first described. [ABSTRACT FROM AUTHOR]

Background: Emerging zoonotic diseases arise from cross-species transmission events between wild or domesticated animals and humans, with bats being one of the major reservoirs of zoonotic viruses. Viral metagenomics has led to the discovery of many viruses, but efforts have mainly been focused on some areas of the world and on certain viral families. Methods: We set out to describe full-length genomes of new picorna-like viruses by collecting feces from hundreds of bats captured in different regions of Spain. Viral sequences were obtained by high-throughput Illumina sequencing and analyzed phylogenetically to classify them in the context of known viruses. Linear discriminant analysis (LDA) was performed to infer likely hosts based on genome composition. Results: We found five complete or nearly complete genomes belonging to the family Picornaviridae, including a new species of the subfamily Ensavirinae. LDA suggested that these were true vertebrate viruses, rather than viruses from the bat diet. Some of these viruses were related to picornaviruses previously found in other bat species from distant geographical regions. We also found a calhevirus genome that most likely belongs to a proposed new family within the order Picornavirales, and for which genome composition analysis suggested a plant host. Conclusions: Our findings describe new picorna-like viral species and variants circulating in the Iberian Peninsula, illustrate the wide geographical distribution and interspecies transmissibility of picornaviruses, and suggest new hosts for calheviruses. [ABSTRACT FROM AUTHOR]

Societal Impact Statement Summary As the second‐leading cause of mortality worldwide, cancer is a major focus of drug discovery research. Traditional plant knowledge can guide the search for undiscovered compounds, but the efficacy of this approach for cancer, a highly complex disease affecting diverse tissues, is unknown. We investigated the patterns underlying plant selection for cancer treatment globally, finding certain lineages are repeatedly targeted. While this indicates therapeutic value, their relatedness with plants used for unrelated ethnobotanical uses suggests that plants are probably selected to treat cancer‐associated symptoms, rather than addressing tumour growth. Careful re‐examination and scoring of ethnobotanical reports may make the prediction of lineages for drug discovery more informative. Cancer is a highly diverse disease and as the second‐leading cause of death worldwide is a focus of drug discovery research. Natural products have been shown to be a useful source of novel molecules for treating cancer. It is likely there are many plants with undiscovered molecules of therapeutic value, however identifying new leads from the vast diversity of plants is very challenging. Traditional knowledge might inform bioprospecting by predicting lineages of plants rich in therapeutically useful molecules. We characterise the phylogenetic diversity of plants used in traditional cancer management using a comprehensive genus‐level phylogeny of angiosperms, and a list of 597 genera used globally to treat different cancers. We phylogenetically predict which lineages may have elevated potential for drug discovery and assess the quality of the prediction. We demonstrate the independent and repeated targeting of specific lineages of plants by different peoples in different parts of the world. However, the lineages we report here as rich in plants used in traditional cancer management coincide with those for other ethnobotanical applications and contain few plants with proven anti‐cancer activity. That the same lineages are used to treat different cancers is suggestive of independent discovery of therapeutic value. However, it is likely that the traditional knowledge explored here is shaped by the selection of plants conferring milder effects for treating wider symptoms, such as tiredness or nausea, rather than for halting tumour growth. Accurate prediction of useful plant lineages for cancer management requires more nuanced information than is commonly provided in ethnobotanical records. [ABSTRACT FROM AUTHOR]

Constraint is a fundamental concept in evolutionary theory. Morphology and ecology both are limited by functional, historical and developmental factors to a subset of the theoretical range species could occupy. Cat-like carnivorans (Feliformia) offer a unique opportunity to investigate phenotypic constraint, as several feliform clades are purported to be limited to generalized ecomorphological roles, while others possess extremely specialized durophagous (bone-crushing) and sabretooth morphology. We investigated the evolutionary history of feliforms by considering their phylogeny, morphological disparity and rates of evolution. We recover results that show a mosaic pattern exists in the degree of morphological disparity per anatomical region per clade and ecology. Non-hypercarnivores, such as viverrids (civets and genets), Malagasy euplerids and lophocyonids (extinct hypocarnivores), have the greatest dental disparity, while hypercarnivores (felids, nimravids, many hyaenids) have the lowest dental disparity but highest cranial and mandibular disparity (excluding dentition). However, high disparity is not necessarily associated with high rates of evolution, but instead with ecological radiations. We reveal that relationships between specialization and disparity are not as simple as past research has concluded. Instead, morphological disparity results from an anatomical mosaic of evolution, where different ecologies correlate with and likely channel unique patterns/combinations of disparity per anatomical partition. [ABSTRACT FROM AUTHOR]

Aotearoa (New Zealand) is a biodiversity hotspot for temperate invertebrate taxa and home to high levels of endemicity. However, our knowledge of species‐level diversity and phylogeny of endemic New Zealand Onychophora (velvet worms) is at present limited. Here, we use mitochondrial cytochrome c oxidase subunit I (COI) barcoding to assess the extent of species diversity for the two velvet worm genera found in New Zealand, the ovoviviparous and endemic Peripatoides and the oviparous Ooperipatellus, found in Australia and New Zealand. Our results reveal that the estimated number of species of both genera in New Zealand is greater than currently described. We estimate there are between 13 and 67 species of Peripatoides and between 16 and 21 species of Ooperipatellus endemic to New Zealand. This is a stark increase from the two currently described New Zealand species of Ooperipatellus and previous work that has identified 10 species within Peripatoides. Our exploration of climatic variables shows that individuals of Ooperipatellus are predominantly found in wet, cool environments and Peripatoides are found across relatively drier, warmer habitats. We also generate ecological niche models to provide initial predictions of the distribution of climatically suitable habitats for each genus across New Zealand. [ABSTRACT FROM AUTHOR]

Background: LIM (Lineage-11 (LIN-11), Insulin-1 (ISL-1), and Mechanotransduction-3 (MEC-3)) genes belong to a family that hold ubiquitous properties contributing to organ, seed, and pollen development as well as developmental and cellular responses to biotic and abiotic stresses. Lettuce (Lactuca sativa) is a highly consumed vegetable crop susceptible heat stress. High temperatures limit lettuce's overall yield, quality and marketability. Lettuce LIM genes have not been identified and their role in response to high temperatures is not known. Aiming to identify potential new targets for thermoresilience, we searched for LIM genes in lettuce and compared them with orthologous of several dicotyledons and monocotyledons plant species. Results: We identified fourteen lettuce LIM genes distributed into eight different subgroups using a genome-wide analysis strategy. Three belonging to DAR (DA means "large" in Chinese) class I, two DAR class II, one in the WLIM1, two in the WLIM2, one in the PLIM1, two in the PLIM2 class, one ßLIM and two δLIMs. No DAR-like were identified in any of the species analyzed including lettuce. Interestingly, unlike other gene families in lettuce which underwent large genome tandem duplications, LIM genes did not increase in number compared to other plant species. The response to heat stress induced a dynamic transcriptional response on LsLIM genes. All heat stress regimes, including night stress, day stress and day and night stress were largely responsible for changes in LIM transcriptional expression. Conclusions: Our global analysis at the genome level provides a detailed identification of LIM genes in lettuce and other dicotyledonous and monocotyledonous plant species. Gene structure, physical and chemical properties as well as chromosomal location and Cis-regulatory element analysis together with our gene expression analysis under different temperature regimes identified LsWLIM1, LsWLIM2b, LsDAR3 and LsDAR5 as candidate genes that could be used by breeding programs aiming to produce lettuce varieties able to withstand high temperatures. [ABSTRACT FROM AUTHOR]

Background: Pseudomonas juntendi is a newly identified opportunistic pathogen, of which we have limited understanding. P. juntendi strains are often multidrug resistant, which complicates clinical management of infection. Methods: A strain of Pseudomonas juntendi (strain L4326) isolated from feces was characterized by MALDI-TOF-MS and Average Nucleotide Identity BLAST. This strain was further subject to whole-genome sequencing and Maximum Likelihood phylogenetic analysis. The strain was phenotypically characterized by antimicrobial susceptibility testing and conjugation assays. Results: We have isolated the novel P. juntendi strain L4236, which was multidrug resistant, but retained sensitivity to amikacin. L4236 harbored a megaplasmid that encoded blaOXA−1 and a novel blaIMP−1 resistance gene variant. P. juntendi strain L4236 was phylogenetically related to P. juntendi strain SAMN30525517. Conclusion: A rare P. juntendi strain was isolated from human feces in southern China with a megaplasmid coharboring blaIMP−1−like and blaOXA−1. Antimicrobial selection pressures may have driven acquisition of drug-resistance gene mutations and carriage of the megaplasmid. [ABSTRACT FROM AUTHOR]

Background: Pseudomonas juntendi is a newly identified opportunistic pathogen, of which we have limited understanding. P. juntendi strains are often multidrug resistant, which complicates clinical management of infection. Methods: A strain of Pseudomonas juntendi (strain L4326) isolated from feces was characterized by MALDI-TOF-MS and Average Nucleotide Identity BLAST. This strain was further subject to whole-genome sequencing and Maximum Likelihood phylogenetic analysis. The strain was phenotypically characterized by antimicrobial susceptibility testing and conjugation assays. Results: We have isolated the novel P. juntendi strain L4236, which was multidrug resistant, but retained sensitivity to amikacin. L4236 harbored a megaplasmid that encoded blaOXA−1 and a novel blaIMP−1 resistance gene variant. P. juntendi strain L4236 was phylogenetically related to P. juntendi strain SAMN30525517. Conclusion: A rare P. juntendi strain was isolated from human feces in southern China with a megaplasmid coharboring blaIMP−1−like and blaOXA−1. Antimicrobial selection pressures may have driven acquisition of drug-resistance gene mutations and carriage of the megaplasmid. [ABSTRACT FROM AUTHOR]

Seasonally tropical dry forests (SDTFs) in the American tropics are a highly diverse yet poorly understood and endangered ecosystem scattered from Northern Mexico to Southern Argentina. One floristic element of the STDFs is the genus Magoniella (Polygonaceae), which includes two liana species, M. laurifolia and M. obidensis, which have winged fruits and are distributed from Costa Rica to Southern Brazil. In a field expedition to the SDTFs of the Colombian Caribbean in 2015, morphologically distinctive individuals of Magoniella were found. In this study, we investigated the species boundaries within Magoniella and determined the phylogenetic position of these morphologically distinctive individuals in the tribe Triplaridae. We compiled morphological trait data across 19 specimens of both species and produced newly sequenced nuclear-plastid DNA data for M. obidensis. Morphometric analyses revealed significant differences in fruit length and perianth size among individuals from the Colombian Caribbean compared to M. obidensis and bract length when compared to M. laurifolia. Maximum likelihood analysis of non-conflicting nuclear and plastid datasets placed the Colombian Caribbean individuals as sister to M. obidensis with maximum statistical support. Additionally, pairwise sequence comparisons of the nuclear ribosomal ITS and the lfy2i loci consistently showed 15-point mutations (10 transitions, five transversions) and six 2 bp-long substitutions that differ between M. obidensis and the Colombian Caribbean individuals. Our morphological and molecular evidence thus suggests that the Colombian Caribbean individuals of Magoniella represent a divergent population from M. laurifolia and M. obidensis, which we describe and illustrate as a new species, M. chersina. Additionally, we provide nomenclatural updates for M. laurifolia and M. obidensis. This study highlights the power of combining morphological and molecular evidence in documenting and naming plant diversity. [ABSTRACT FROM AUTHOR]

A comprehensive and systematic examination of dengue virus (DENV) evolution is essential in Pakistan, where the virus poses a significant public health challenge due to its ability to adapt and evolve. To shed light on the intricate evolutionary patterns of all four DENV serotypes, we analyzed complete genome sequences (n = 43) and Envelope (E) gene sequences (n = 44) of all four DENV serotypes collected in Pakistan from 1994 to 2023, providing a holistic view of their genetic evolution. Our findings revealed that all four serotypes of DENV co-circulate in Pakistan with a close evolutionary relationship between DENV-1 and DENV-3. Among the genetically distinct serotypes DENV-2 and DENV-4, DENV-4 stands out as the most genetically different, while DENV-2 exhibits greater complexity due to the presence of multiple genotypes and the possibility of temporal fluctuations in genotype prevalence. Selective pressure analysis of the Envelope (E) gene revealed heterogeneity among sequences (n = 44), highlighting 46 codons in the genome experiencing selective pressure, characterized by a bias toward balancing selection, indicating genetic stability of the virus. Furthermore, our study suggested an intriguing evolutionary shift of DENV-4 toward the DENV-2 clade, potentially influenced by antibodies with cross-reactivity to multiple serotypes, providing a critical insight into the complex factors, shaping DENV evolution and contributing to the emergence of new serotypes. [ABSTRACT FROM AUTHOR]

Four species of the genus Wrangelia are presently known from the western Atlantic Ocean: W. argus, W. bicuspidata, W. penicillata, and W. gordoniae, with the first three historically being reported from Bermuda. Morphological and molecular barcode (COI‐5P) and phylogenetic analyses used in this study (SSU, LSU, rbcL) indicated eight species groupings of Wrangelia in Bermuda, excluding two of the historically recognized species, retaining only W. argus while adding seven new species, of which six are formally described. What had been historically reported as W. penicillata from Bermuda was shown to be distinct from Mediterranean Sea specimens (type locality) and was shown to be a mixture of W. hesperia sp. nov. and W. incrassata sp. nov. Along with these two, three other new species (W. laxa sp. nov., W. ryancraigii sp. nov., and W. secundiramea sp. nov.) have complete rhizoidal cortication tightly covering axial cells of indeterminate axes below the apices, distinguishing them from the two local incompletely corticated congeners W. argus and W. abscondita sp. nov., the latter a morphologically cryptic sister species with W. bicuspidata from the Caribbean Sea. Only one of the new species, W. ryancraigii, has thus far been observed in the mesophotic zone off the Bermuda platform, and it is morphologically cryptic with the euphotic zone's W. laxa. [ABSTRACT FROM AUTHOR]

Simple Summary: Parasitic flatworms of the family Cyclocoelidae parasitize various inner organs of birds, mainly in their respiratory and digestive systems. The taxonomic position of many cyclocoelid species remains controversial. In our study, we studied two species of trematodes from the family Cyclocoelidae (Harrahium obscurum and Morishitium dollfusi) using classical morphological and molecular phylogenetic approaches. We provided original drawings, morphological descriptions, and the results of a molecular phylogenetic analysis of these species of helminths from birds in Russia for the first time. Our work shows the morphological variability in individuals of both flatworm species. Since the variability of individual cyclocoelids may be rather high, morphological features alone are insufficient for their identification, and confirmation with the use of molecular data is required. The taxonomic status of many species of the family Cyclocoelidae is still unclear. Two species of cyclocoelids, Harrahium obscurum and Morishitium dollfusi, were collected from the air sacs of birds (Turdus merula and Tringa ochropus) inhabiting the Middle Volga region (European Russia). Here, we provide the first detailed morphological description of these cyclocoelids and combine it with the first molecular phylogenetic analysis of Cyclocoelidae from birds in Russia based on partial sequences of their 28s rDNA and coI mtDNA genes. Specimens of both flatworm species from different host individuals differ slightly in body shape and size, which probably reflects host-induced intraspecific variability. For the first time, we have shown that a stable morphological character such as the length of the vitelline fields in the studied digeneans is variable at the species level and cannot be used in their morphological diagnosis. [ABSTRACT FROM AUTHOR]

While conducting phylogenetic analyses of sequence-capture data of Australian members of Asteraceae tribe Astereae, we found that Elachanthus pusillus F.Muell., Isoetopsis graminifolia Turcz. and Kippistia suaedifolia F.Muell. formed a clade with Minuria. We, therefore, conducted an analysis focused on this clade, but with replicate samples of the three smaller genera, and confirmed our results. Kippistia had been synonymised under Minuria between 1876 and 1980, when it was reinstated. Elachanthus and Isoetopsis had not previously been suggested to be part of Minuria , and, indeed, Isoetopsis had been considered so morphologically divergent that even its tribal affiliations were long controversial. However, on closer examination, Elachanthus and Isoetopsis are strikingly similar to Minuria , especially in cypsela and pappus morphology. The evolution of both genera from a common ancestor in Minuria appears plausible in the light of their overall similarity to annual species of that genus such as M. annua , their multiple uniform rows of herbaceous phyllaries with scarious margins, a pappus of scales v. the tendency of Minuria to form pappi with scale-like bases, female outer and male inner florets, and a cypsela indumentum shared with some species of Minuria. We propose the following three new combinations under Minuria : M. pusilla , M. glabra and M. graminifolia. We conducted a phylogenetic analysis of Minuria and related genera and found that Elachanthus , Isoetopsis , and Kippistia phylogenetically nested in Minuria. Before 1980, Kippistia had long been considered part of Minuria. Morphological examination showed many similarities between Eleachanthus , Isoetopsis , and annual species of Minuria. We publish new combinations for four species, synonymising the three smaller genera. (Image credit: Alexander Schmidt-Lebuhn.) This article belongs to the collection Genomics for Australian Plants. [ABSTRACT FROM AUTHOR]

A new family, genus, and species record, Gayralia brasiliensis (Gayraliaceae, Ulotrichales, Chlorophyta) is reported in Singapore using chloroplast elongation factor Tu (tufA) and nuclear internal transcribed spacer (ITS) markers alongside gross and microscopic examination. This species was likely missed by previous surveys due to its limited local distribution, superficial similarity to the much more common Ulva, and its small size. Morphologically, G. brasiliensis in Singapore is more similar to G. oxysperma than to G. brasiliensis in Brazil. This study demonstrates the importance of integrative molecular and morphological analysis for characterising macroalgal diversity. [ABSTRACT FROM AUTHOR]

Gene duplication followed by nucleotide differentiation is one of the simplest mechanisms to develop new functions for genes. However, the evolutionary processes underlying the divergence of multigene families remain controversial. We used multigene families found within the diversity of toxic proteins in centipede venom to test two hypotheses related to venom evolution: the two-speed mode of venom evolution and the rapid accumulation of variation in exposed residues (RAVER) model. The two-speed mode of venom evolution proposes that different types of selection impact ancient and younger venomous lineages with negative selection being the predominant form in ancient lineages and positive selection being the dominant form in younger lineages. The RAVER hypothesis proposes that, instead of different types of selection acting on different ages of venomous lineages, the different types of selection will selectively contribute to amino acid variation based on whether the residue is exposed to the solvent where it can potentially interact directly with toxin targets. This hypothesis parallels the longstanding understanding of protein evolution that suggests that residues found within the structural or active regions of the protein will be under negative or purifying selection, and residues that do not form part of these areas will be more prone to positive selection. To test these two hypotheses, we compared the venom of 26 centipedes from the order Scolopendromorpha from six currently recognized species from across North America using both transcriptomics and proteomics. We first estimated their phylogenetic relationships and uncovered paraphyly among the genus Scolopendra and evidence for cryptic diversity among currently recognized species. Using our phylogeny, we then characterized the diverse venom components from across the identified clades using a combination of transcriptomics and proteomics. We conducted selection-based analyses in the context of predicted three-dimensional properties of the venom proteins and found support for both hypotheses. Consistent with the two-speed hypothesis, we found a prevalence of negative selection across all proteins. Consistent with the RAVER hypothesis, we found evidence of positive selection on solvent-exposed residues, with structural and less-exposed residues showing stronger signal for negative selection. Through the use of phylogenetics, transcriptomics, proteomics, and selection-based analyses, we were able to describe the evolution of venom from an ancient venomous lineage and support principles of protein evolution that directly relate to multigene family evolution. [ABSTRACT FROM AUTHOR]

Species of the genus Eubranchus Forbes, 1838 (Mollusca: Gastropoda: Nudibranchia) are common faunistic elements of boreal benthic ecosystems, associated with hydroid communities. Recent studies have suggested that the widely distributed trans-Arctic E. rupium (Møller, 1842) constitutes a complex of at least three candidate species, but the detailed taxonomy of the complex remains unresolved. The purpose of the present paper is to conduct an integrative taxonomic study including molecular genetic methods (a phylogenetic analysis using COI , 16S rRNA and histone H3 with application of species delimitation methods) and morphological study (light and scanning electron microscopy) of E. rupium and closely related species. The specific aims of this study were to establish the species boundaries, morphological variability, and the phylogeographic structure within this group. The phylogeographic analysis included a TCS -based network analysis, an analysis of molecular variance (AMOVA), divergence time estimations, and ancestral area reconstructions. We demonstrate that specimens initially identified as E. rupium included three distinctive species: the nominal E. rupium with an amphiboreal range, the new species Eubranchus novik sp. nov. from the Sea of Japan, for which a taxonomic description is provided in this paper, and Eubranchus sp. from the northern Kuril Islands, which requires the collection and study of additional material for formal description. Our results confirm the amphiboreal distribution of E. rupium , as no geographic structure was found across Pacific, Arctic and Atlantic populations, and the results of the AMOVA analysis showed no differences between groups of samples from different geographic regions. The divergence of the ' Eubrancus rupium species complex' is estimated from the late Miocene or the Miocene–Pliocene boundary to the late Pliocene. It is hypothesised that the most probable ancestral region for the Eubranchus rupium species complex is the north-western Pacific, and the subsequent speciation likely occurred due to dispersal followed by allopatric speciation. ZooBank: Species of the genus Eubranchus Forbes, 1838 are common in boreal benthic ecosystems. Recent studies suggest the trans-Arctic Eubranchus rupium (Møller, 1842) is a species complex. This paper conducts an integrative taxonomic study of Eubranchus rupium and related species, confirming its amphiboreal distribution. The divergence of the species complex is estimated from the late Miocene or the Miocene–Pliocene boundary to the late Pliocene with the north-western Pacific as the probable ancestral region, followed by dispersal and allopatric speciation. (Image credit: Darya Grishina.) [ABSTRACT FROM AUTHOR]

During the past two decades, the phylogenetic relationships and higher-level classification of the subfamily Rogadinae have received relevant contributions based on Sanger, mitogenome and genome-wide nuclear DNA sequence data. These studies have helped to update the circumscription and tribal classification of this subfamily, with six tribes currently recognised (Aleiodini, Betylobraconini, Clinocentrini, Rogadini, Stiropiini and Yeliconini). The tribal relationships within Rogadinae, however, are yet to be fully resolved, including the status of tribe Facitorini, previously regarded as betylobraconine, with respect to the members of Yeliconini. We conducted a phylogenomic analysis among the tribes of Rogadinae based on genomic ultraconserved element (UCE) data and extensive taxon sampling including three undescribed genera of uncertain tribal placement. Our almost fully supported estimate of phylogeny confirmed the basal position of Rogadini within the subfamily and a Facitorini clade (Yeliconini + Aleiodini) that led us to propose the former group as a valid rogadine tribe (Facitorini stat. res.). Stiropiini, however, was recovered for the first time as sister to the remaining rogadine tribes except Rogadini, and Clinocentrini as sister to a clade with Betylobraconini + the three undescribed genera. The relationships recovered and morphological examination of the material included led us to place the latter three new genera and recently described genus Gondwanocentrus within a new rogadine tribe, Gondwanocentrini Shimbori & Zaldívar-Riverón trib. nov. We described these genera (Ghibli Shimbori & Zaldívar-Riverón gen. nov., Racionais Shimbori & Zaldívar-Riverón gen. nov. and Soraya Shimbori gen. nov.) with two or three new species each (G. miyazakii Shimbori & Zaldívar-Riverón sp. nov., G. totoro Shimbori & Zaldívar-Riverón sp. nov., R. brunus Shimbori & Zaldívar-Riverón sp. nov., R. kaelejay Shimbori & Zaldívar-Riverón sp. nov., R. superstes Shimbori & Zaldívar-Riverón sp. nov., S. alencarae Shimbori sp. nov. and S. venus Shimbori & Zaldívar-Riverón sp. nov.). A new species of Facitorini, Jannya pasargadae Gadelha & Shimbori sp. nov., is also described. Our newly proposed classification expands the number of tribes and genera within Rogadinae to 8 and 66 respectively. ZooBank: Tribal relationships within the braconid subfamily Rogadinae are reconstructed based on nuclear UCE data and extensive taxon sampling. Our fully supported estimate of phylogeny and the morphological evidence led us to erect a new rogadine tribe, Gondwanocentrini Shimbori & Zaldívar-Riverón trib. nov. Facitorini was also confirmed as a separate rogadine tribe. Our updated classification expands the number of tribes and genera within Rogadinae to 8 and 66 respectively. (Image credit: Eduardo M. Shimbori.) [ABSTRACT FROM AUTHOR]

Chickpeas (Cicer arietinum L.) are an important legume crop known for their rich nutrient content, including proteins, carbohydrates, and minerals. Thus, they are enjoyed by people worldwide. In recent years, the production scale of chickpeas has been growing gradually. The planting area of chickpeas represents roughly 35–36% of the total planting area, and the output of the beans is roughly 47–48%. However, the growth and development process of chickpeas is limited by a number of factors, including high temperature, drought, salt stress, and so forth. In particular, high temperatures can reduce the germination rate, photosynthesis, seed setting rate, and filling rate of chickpeas, restricting seed germination, plant growth, and reproductive growth. These changes lead to a decrease in the yield and quality of the crop. Heat shock proteins (HSPs) are small proteins that play an important role in plant defense against abiotic stress. Therefore, in the present study, HSP20 gene family members were identified based on the whole-genome data of chickpeas, and their chromosomal positions, evolutionary relationships, promoter cis-acting elements, and tissue-specific expression patterns were predicted. Subsequently, qRT-PCR was used to detect and analyze the expression characteristics of HSP20 genes under different temperature stress conditions. Ultimately, we identified twenty-one HSP20 genes distributed on seven chromosomes, and their gene family members were found to be relatively conserved, belonging to ten subfamilies. We also found that CaHSP20 promoter regions have many cis-acting elements related to growth and development, hormones, and stress responses. In addition, under high-temperature stress, the relative expression of CaHSP20-17, CaHSP20-20, CaHSP20-7, CaHSP20-3, and CaHSP20-12 increased hundreds or even thousands of times as the temperature increased from 25 °C to 42 °C. Among them, excluding CaHSP20-5, the other five genes all contain 1-2 ABA cis-regulatory elements. This finding indicates that CaHSP20s are involved in the growth and development of chickpeas under heat stress, and the mechanisms of their responses to high-temperature stress may be related to hormone regulation. The results of the present study lay the foundation for exploring HSP20 gene family resources and the molecular mechanisms of heat resistance in chickpeas. Our results can also provide a theoretical basis for breeding high-temperature-resistant chickpea varieties and provide valuable information for the sustainable development of the global chickpea industry. [ABSTRACT FROM AUTHOR]

The evolution of animals and their gut symbionts is a complex phenomenon, obscured by lability and diversity. In social organisms, transmission of symbionts among relatives may yield systems with more stable associations. Here, we study the history of a social insect symbiosis involving cephalotine ants and their extracellular gut bacteria, which come predominantly from host‐specialized lineages. We perform multi‐locus phylogenetics for symbionts from nine bacterial orders, and map prior amplicon sequence data to lineage‐assigned symbiont genomes, studying distributions of rigorously defined symbionts across 20 host species. Based on monophyly and additional hypothesis testing, we estimate that these specialized gut bacteria belong to 18 distinct lineages, of which 15 have been successfully isolated and cultured. Several symbiont lineages showed evidence for domestication events that occurred later in cephalotine evolutionary history, and only one lineage was ubiquitously detected in all 20 host species and 48 colonies sampled with amplicon 16S rRNA sequencing. We found evidence for phylogenetically constrained distributions in four symbionts, suggesting historical or genetic impacts on community composition. Two lineages showed evidence for frequent intra‐lineage co‐infections, highlighting the potential for niche divergence after initial domestication. Nearly all symbionts showed evidence for occasional host switching, but four may, more often, co‐diversify with their hosts. Through our further assessment of symbiont localization and genomic functional profiles, we demonstrate distinct niches for symbionts with shared evolutionary histories, prompting further questions on the forces underlying the evolution of hosts and their gut microbiomes. [ABSTRACT FROM AUTHOR]

The objective of this paper is twofold. First, I present a framework called historical coherentism (Chang, 2004; Tal, 2016; Van fraassen 2008) and argue that it is the best epistemological framework available to tackle the problem of coordination, an epistemic conundrum that arises with every attempt to provide empirical content to scientific theories, models or statements. Second, I argue that the problem of coordination, which has so far been theorized only in the context of measurement practices (Reichenbach, 1927; Chang, 2001; Tal, 2012; Van fraassen 2008), can be generalized beyond the philosophy of measurement. Specifically, it will be shown that the problem is embodied in classificatory practices and that, consequently, historical coherentism is well suited to analyze these practices as well as metrological ones. As a case study, I look at a contemporary debate in phylogenetics, regarding the evolutionary origin of a newly identified archaeal phylum called Methanonatronarchaeia. Exploring this debate through the lens of historical coherentism provides a detailed understanding of the dynamics of the field and a foothold for critical analyses of the standard rationale used by practitioners. [ABSTRACT FROM AUTHOR]

The utility of the mitochondrial genomes (mitogenomes) in analyzing the evolutionary history of animals has been proven. Five deep-sea corals (Bathypathes sp.1, Bathypathes sp.2, Schizopathidae 1, Trissopathes sp., and Leiopathes sp.) were collected in the South China Sea (SCS). Initially, the structures and collinearity of the five deep-sea coral mitogenomes were analyzed. The gene arrangements in the five deep-sea coral mitogenomes were similar to those in the order Antipatharia, which evidenced their conservation throughout evolutionary history. Additionally, to elucidate the slow evolutionary rates in Hexacorallia mitogenomes, we conducted comprehensive analyses, including examining phylogenetic relationships, performing average nucleotide identity (ANI) analysis, and assessing GC-skew dissimilarity combining five deep-sea coral mitogenomes and 522 reference Hexacorallia mitogenomes. Phylogenetic analysis using 13 conserved proteins revealed that species clustered together at the order level, and they exhibited interspersed distributions at the family level. The ANI results revealed that species had significant similarities (identity > 85%) within the same order, while species from different orders showed notable differences (identity < 80%). The investigation of the Hexacorallia mitogenomes also highlighted that the GC-skew dissimilarity was highly significant at the order level, but not as pronounced at the family level. These results might be attributed to the slow evolution rate of Hexacorallia mitogenomes and provide evidence of mitogenomic diversity. Furthermore, divergence time analysis revealed older divergence times assessed via mitogenomes compared with nuclear data, shedding light on significant evolutionary events shaping distinct orders within Hexacorallia corals. Those findings provide new insights into understanding the slow evolutionary rates of deep-sea corals in all lineages of Hexacorallia using their mitogenomes. [ABSTRACT FROM AUTHOR]

Litylenchus crenatae mccannii is a foliar nematode that causes severe damage to beech, known as beech leaf disease (BLD). Previous studies have characterized and identified L. crenataein Japan and a subspecies L. crenatae mccannii in the USA. Litylenchus crenatae mccannii has been found to cause BLD in native and non-native beech hosts in North America. As little is known about the distribution and diversity of the nematode in Ontario, Canada, beech foliage was sampled throughout southcentral Ontario. Incidence and severity maps show that BLD spread from its first detections in 2017, in Elgin and Norfolk counties, to the west and northeast. A gradient in severity and incidence was observed with the highest ratings at the locations where it was originally found. The extent of the nematode's distribution is much broader than BLD, occurring throughout much of the range of American beech. Sequences of two barcoding loci for the L. crenatae mccannii isolates from 11 locations in Ontario showed 100% similarity within and among these locations. The genotype of L. crenatae mccannii found in Ontario is the same as that identified in the USA. This finding is consistent with the fact that 92% of beech imported into Canada is from the USA and suggests that the USA could be the origin of L. crenatae mccannii nematodes in Ontario or that both countries have imported beech trees from the same source country of BLD. This research provides motivation for further studies to clarify the sources of L. crenatae mccannii. [ABSTRACT FROM AUTHOR]

There are a staggering number of publicly available bacterial genome sequences (at writing, 2.0 million assemblies in NCBI's GenBank alone), and the deposition rate continues to increase. This wealth of data begs for phylogenetic analyses to place these sequences within an evolutionary context. A phylogenetic placement not only aids in taxonomic classification but informs the evolution of novel phenotypes, targets of selection, and horizontal gene transfer. Building trees from multi-gene codon alignments is a laborious task that requires bioinformatic expertise, rigorous curation of orthologs, and heavy computation. Compounding the problem is the lack of tools that can streamline these processes for building trees from large-scale genomic data. Here we present OrthoPhyl, which takes bacterial genome assemblies and reconstructs trees from whole genome codon alignments. The analysis pipeline can analyze an arbitrarily large number of input genomes (>1200 tested here) by identifying a diversity-spanning subset of assemblies and using these genomes to build gene models to infer orthologs in the full dataset. To illustrate the versatility of OrthoPhyl, we show three use cases: E. coli/Shigella, Brucella/Ochrobactrum and the order Rickettsiales. We compare trees generated with OrthoPhyl to trees generated with kSNP3 and GToTree along with published trees using alternative methods. We show that OrthoPhyl trees are consistent with other methods while incorporating more data, allowing for greater numbers of input genomes, and more flexibility of analysis. [ABSTRACT FROM AUTHOR]

The genus Eurytemora is a diverse group of copepods found in coastal, estuarine, brackish, and freshwater environments. The main research has focused on Eurytemora affinis (Poppe, 1880) and revealed it to be a species complex. Eurytemora velox (Liljeborg, 1853) has only recently been characterized in the Scheldt estuary but never within a global phylogenetic context. This study integrated nearly all European Eurytemora species sequences available, along with original ones from the Scheldt. A total of 351 sequences were analyzed using one mitochondrial gene (CO1) and one nuclear gene (nITS), with sequencing performed specifically on the CO1 gene. Phylogenetic reconstructions were performed using the Maximum Likelihood method, along with haplowebs and genetic diversity indices. A significant subdivision between six European species was observed. The monophyletic clade status of the E. affinis complex was confirmed alongside the existence of three geographically isolated lineages of the E. affinis species, the East Atlantic, the North Sea/English Channel, and the Baltic lineages, each exhibiting pronounced genetic differentiation. The population of E. velox from the Urals differs significantly from the European ones. These results provide both an overview of the genetic structure of the genus in geographic Europe as well as new insights on E. velox. [ABSTRACT FROM AUTHOR]

The Fusarium graminearum species complex (FGSC) is a worldwide phytopathogenic fungus of small grain cereals. Genetics and bioinformatics tools have been providing an efficient strategy for identifying FGSC. However, the potential reliability of tef1−α sequencing in FGSC members has not been well investigated. In this study, the tef1−α sequencing data of 246 FGSC members, one F. culmorum, and one F. solani isolate were subjected to distance-, character-, and PCA-based phylogenetic analysis. Linux terminals and the R programming language were used in phylogenetic analysis. The Unweighted Pair Group Method with Arithmetic Mean (UPGMA) and maximum likelihood methods produced relatively more homogenous F. graminearum sensu stricto (Fgss) and F. asiaticum isolates. Fgss and F. asiaticum isolates co-clustered in two separate sub-divisions in the ML and UPGMA methods, with significant differences in the Chi2 test (p < 0.05). PCA profiling revealed a low level of variation in FGSC members, with 99–99.5% percentages in axis 1. An increased number of taxa and isolates would be tested for tef1−α in future studies. To our knowledge, this is also the first study to combine phylogenetic methods with PCA tests for comprehensive characterization of FGSC members. [ABSTRACT FROM AUTHOR]

A formal cladistic analysis of the morphological features of taxa assigned to the Orthoseiraceae, including Phycavernosa, Guarreraea and species of the genus Orthoseira is developed for 35 taxa based on 16 characters along with two Melosira species and four biddulphioid species as ingroup taxa as well as Leptocylindrus danicus as the outgroup. A consensus tree of 266 843 most parsimonious topologies suggests that the Orthoseiraceae is monophyletic, and that Guarreraea plus all former species of Orthoseira represent a monophyletic group. This analysis of morphological data did not provide support for the members of the Orthoseiraceae being more closely related to melosiroid or biddulphioid diatoms. Relationships within the monophyletic orthoseiracean group are detailed. Due to the exclusion of the generitype of Orthoseira not possessing carinoportulae which are central tube processes, one of the diagnostic features of the former Orthoseiraceae, other taxon names are described and recognized herein. We recognize and describe a new order, the Liparogyrales, based on the genus Liparogyra for taxa that were previously assigned to Orthoseira. Two families, the Phycavernosaceae, comprising the genus Phycavernosa, and the Liparogyraceae, comprising Guarreraea and taxa formerly assigned to Orthoseira, are described. The genus Liparogyra is composed of three subgenera: the subgenus Liparogyra, the subgenus Porocyclia, and the subgenus Insularis which are diagnosed by synapomorphic features. Based on our analysis, the genus Stephanosira appears to lack synapomorphies. Liparogyra, Porocyclia and Stephanosira had previously been described as genera by Ehrenberg. Guarreraea and one newly proposed genus are also included in the Liparogyraceae. Several other lineages are aphyletic, due to them either being monotypic or without diagnostic features. Thus, a total of 29 species in five genera are recognized within the newly described order and two newly described families. Eighteen new combinations are proposed for the transfer of taxa from Orthoseira to other genera. Homoplasy amongst some of the character states is discussed. The diatom genus Orthoseira as typified by O. americana does not possess carinoportulae, requiring a new approach to the nomenclature and taxonomy of the group. A phylogenetic analysis of the group using morphological data was performed, demonstrating patterns of relationships of the species for this lineage. A new classification for this diatom lineage is presented. [ABSTRACT FROM AUTHOR]

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

The embedding problem of Markov matrices in Markov semigroups is a classic problem that regained a lot of impetus and activities through recent needs in phylogeny and population genetics. Here, we give an account for dimensions d ⩽ 4 , including a complete and simplified treatment of the case d = 3 , and derive the results in a systematic fashion, with an eye on the potential applications. Further, we reconsider the setup of the corresponding problem for time-inhomogeneous Markov chains, which is needed for real-world applications because transition rates need not be constant over time. Additional cases of this more general embedding occur for any d ⩾ 3 . We review the known case of d = 3 and describe the setting for future work on d = 4 . [ABSTRACT FROM AUTHOR]

Despite many studies conducted over a period of nearly two centuries since Reboul, who was the first to classify tulips as subgeneric level, there is still no consensus among researchers regarding the infrageneric classification of tulips. Nevertheless, in recent years, efforts to solve this taxonomic problem have been increasing. However, phylogenetic studies covering all species of a genus or at least with species representatives of all subgeneric taxonomy worldwide are very scarce. The main difference of the current study from previous studies is that it is a larger study focused on solving this problem by considering all perspectives together, covering many geographical areas and species in global distribution. For this purpose, we examined the phylogenetic relationships of the genus Tulipa with 82 accessions from 45 Tulipa species and six accessions from two Amana and three Erythronium species, and we positioned our results by discussing them in light of previous morphological, geographical, molecular and anatomical data. We investigated the phylogenetic relationships in the genus using DNA sequences from the internal transcribed spacer region of nuclear ribosomal DNA. The 45 species of Tulipa formed a monophyletic clade. Genera Amana and Erythronium were clustered as outgroups. In the genus Tulipa, two major clades were obtained; the first clade belonged to subgenus Orithyia. The second clade separated subgenus Tulipa and subgenera Eriostemones+Clusianae. Then, Eriostemones and Clusianae subgenera formed two separate clusters. Subgenus Eriostemones separated two main clusters; section Biflores and section Sylvestres. On the other hand, all previously proposed sections and series under the subgenus Tulipa were grouped into a single clade. This situation agrees with previous biosystematic data highlighting the morphological overlap of most of the sections and with previous molecular data showing that many sections are not monophyletic. Consequently, we propose a taxonomic concept consisting of four subgenera and two sections. [ABSTRACT FROM AUTHOR]