A Proteogenomic Approach for the Identification of Virulence Factors in Leishmania Parasites.

Identifying new genes involved in virulence and drug resistance may hold the key to a better understanding of parasitic diseases. The proteogenomic profiling of various Leishmania species, the causative agents of leishmaniasis, has identified several novel genes, N- and C-terminal extensions of proteins, and corrections of existing gene models. Various virulence factors (VFs) responsible for leishmaniasis have been previously annotated through a proteogenomic approach, including the C-terminal extension of heat shock protein 70 (HSP70). Furthermore, the diversity of VFs across Leishmania donovani, L. infantum, L. major, and L. mexicana was determined using phylogenetic analysis. Moreover, protein-protein interaction networks (PPINs) of VFs with HSPs aid in making significant biological interpretations. Overall, an integrated omics approach involving proteogenomics was used to identify and study the relationship among VFs with other interacting proteins, including HSPs. This chapter provides a step-by-step guide to the identification of new genes in Leishmania using a proteogenomic approach and their functional assignment using a bioinformatics-based approach.

Phylogenetic relationships of the Mangrove Hummingbird, "Amazilia" boucardi (Apodiformes: Trochilidae) of Costa Rica / Relaciones filogenéticas del Colibrí manglero "Amazilia" boucardi (Apodiformes: Trochilidae) de Costa Rica

Abstract Introduction: A recent revision of the generic classification of the Trochilidae based on DNA sequences revealed many inconsistencies with the current generic classification, largely based on plumage characters subject to homoplasy, especially in the Trochilini, the largest tribe. A thorough generic reorganization brought the classification into accord with the phylogeny, but due to lack of genetic data, two species remained unclassified. One of these was the Mangrove Hummingbird, "Amazilia" boucardi, endemic to Costa Rica and included in the IUCN red list of threatened species. Objective: To obtain molecular evidence to clarify the generic relationships of "A." boucardi. Methods: We isolated DNA from tissues of this species and amplified 4 nuclear and 4 mitochondrial fragments and compared these with homologous fragments from 56 species in the Trochilini, constructing phylogenetic trees with maximum likelihood and Bayesian methods. Results: Our phylogenetic analyses confirmed the placement of boucardi in the Trochilini and definitely excluded it from Amazilia but placed it with high confidence in the genus Chrysuronia Bonaparte, 1850, within which its closest relative is C. coeruleogularis, which also inhabits mangroves. Conclusions: Our genetic data based on nuclear and mitochondrial regions clearly indicate the relationship of A. boucardi and L. coeruleogularis. Moreover, it is also supported by their habitat distribution in the mangroves of the Pacific coast of Costa Rica and Western Panama. Therefore, we suggested to exclude A. boucardi as "incertae sedis".

Resumen Introducción: Una revisión reciente de la clasificación de la familia Trochilidae con base en secuencias de ADN demostró muchas incongruencias con la clasificación genérica previa, que había sido hecho con base en caracteres del plumaje muy sujetos a homoplasia, especialmente en la tribu más grande, Trochillini. Una reorganización de los géneros logró llevar su clasificación genérica a la concordancia con la filogenia, pero debido a la ausencia de datos genéticos, dos especies permanecieron sin clasificar. Una de estas fue el colibrí de manglar Amazilia boucardi, una especie endémica de Costa Rica, considerada como amenazada en la lista roja de la UICN. Objetivo: Obtener evidencia molecular para esclarecer las relaciones genéricas de A. boucardi. Métodos: Se aisló ADN de tejidos de esta especie y se amplificaron 4 fragmentos de ADN del núcleo y 5 de la mitocondria, y se compararon con fragmentos homólogos de 56 especies en la tribu Trochillini, generando árboles filogenéticos con métodos de máxima verosimilitud y bayesiano. Resultados: Los análisis filogénticos obtenidos confirmaron la ubicación de boucardi en Trochilini y definitivamente la excluyó del género Amazilia, pero la ubicó con un alto grado de confianza en el género Chrysuronia Bonaparte, 1850, dentro los cuales su pariente más cercano es C. coeruleogularis, que también habita manglares. Conclusiones: Nuestros datos genéticos basados en regiones nucleares y mitocondriales indican claramente la relación entre A. boucardi and L. coeruleogularis. Es más, lo anterior se sustenta por su distribución en los manglares de la costa Pacífica de Costa Rica y oeste de Panamá. Por lo tanto, sugerimos excluir a A. boucardi como "incertae sedis".

Refurbishing the marine parasitoid order Pirsoniales with newly (re)described marine and freshwater free-living predators.

Pirsoniales is a stramenopile order composed of marine parasitoids of diatoms with unique life cycle. Until recently, a single genus, Pirsonia, uniting six species, was known. The recent identification of new free-living eukaryotrophic Pirsoniales Pirsonia chemainus, Feodosia pseudopoda, and Koktebelia satura changed our understanding of this group as exclusively parasitic. However, their cell ultrastructure and feeding preferences were not fully studied due to the death of the cultures. In this study, we re-isolated some of these Pirsoniales and established six new strains exhibiting predatory behavior, including a first freshwater representative. This allowed us to describe five new genera and species, as well as to emend the diagnosis of the order Pirsoniales. The 18S rRNA gene phylogenetic analysis revealed the position of new strains within Pirsoniales and their relationships with parasitoid relatives and environmental sequence lineages. Feeding experiments on novel Pirsoniales strains using diverse algal prey showed that they were not able to form trophosomes and auxosomes. The ability of cell aggregation in Pirsoniales was observed for the first time. One of the studied strains contained intracellular gammaproteobacteria distantly related to Coxiella. Ultrastructural analyses revealed a more complex cytoskeleton structure in Pirsoniales than previously thought and supported the monophyly of Bigyromonadea and Pseudofungi.

Specific amino acid changes correlate with pathogenic flavobacteria.

Flavobacterium is a genus of microorganisms living in a variety of hosts and habitats across the globe. Some species are found in fish organs, and only a few, such as Flavobacterium psychrophilum and Flavobacterium columnare, cause severe disease and losses in fish farms. The evolution of flavobacteria that are pathogenic to fish is unknown, and the protein changes accountable for the selection of their colonization to fish have yet to be determined. A phylogenetic tree was constructed with the complete genomic sequences of 208 species of the Flavobacterium genus using 861 softcore genes. This phylogenetic analysis revealed clade CII comprising nine species, including five pathogenic species, and containing the most species that colonize fish. Thirteen specific amino acid changes were found to be conserved across 11 proteins within the CII clade compared with other clades, and these proteins were enriched in functions related to replication, recombination, and repair. Several of these proteins are known to be involved in pathogenicity and fitness adaptation in other bacteria. Some of the observed amino acid changes can be explained by preferential selection for certain codons and tRNA frequency. These results could help explain how species belonging to the CII clade adapt to fish environments.

Two names, one species: redescription and phylogenetic position of Schrankiana formosula Freitas, 1959 provides new insights into the evolutionary history of the Cosmocercidae.

Schrankiana are gastrointestinal parasites of anurans, distributed throughout Central and South America. Schrankiana formosula and Schrankiana inconspicata are some of the most commonly reported species parasitising anurans from Brazil, and the morphological differences between them are unclear. In the present study, we redescribed S. formosula based on a re-examination of type series and newly collected material from the frog Leptodactylus pentadactylus in the state of Amapá, Brazil. Additionally, we re-examined the type series of S. inconspicata, and propose it as a junior synonym of S. formosula. We provide detailed morphological and morphometric data with intraspecific variation analyses and new molecular data for S. formosula. In the present phylogeny, S. formosula formed a well-supported clade with Raillietnema sp. and Labeonema synodontisi. Based on molecular phylogenetic analyses and some morphological similarities, our findings support the hypothesis that Schrankiana is a member of the family Cosmocercidae, not Atractidae. Additionally, we provide the first ultrastructural descriptions of S. formosula, and establish the species' phylogenetic position from the Cosmocercidae.

Whole chloroplast genome sequence and phylogenetic analysis of Calanthe discolor (Orchidaceae).

The orchid Calanthe discolor, which has high ornamental and medicinal value, is mainly distributed in Zhejiang, Jiangsu, and southeast Hubei Provinces of China, as well as in Japan and the southern Korean peninsula. In this study, the whole chloroplast genome sequence of C. discolor was first assembled using high-throughput Illumina paired-end technology, providing data to evaluate the evolution of this species. The C. discolor chloroplast genome was158,286 bp long, including a large single-copy region of 87,095 bp, a small single-copy region of 18,407 bp, and two copies of a repeat region (26,392-bp each). The overall G + C content was 41.2%. A total of 133 genes were predicted from the genome, including 87 protein-coding genes, eight ribosomal RNAs, 38 transfer RNAs. Phylogenetic analysis indicated a close relationship between C. discolor and C. bicolor.

Molecular phylogenetic and estimation of evolutionary divergence and biogeography of the family Schizoparmaceae and allied families (Diaporthales, Ascomycota).

The Diaporthales includes 32 families, many of which are important plant pathogens, endophytes and saprobes, e.g., members of the families Pseudoplagiostomataceae, Pyrisporaceae and Schizoparmaceae. Nucleotide sequences derived from five genetic loci including: ITS, LSU, TEF1-α, TUB2 and RPB2 were used for Bayesian evolutionary analysis to determine divergence times and evolutionary relationships within the Schizoparmaceae. Molecular clock analyses revealed that the ancestor of Schizoparmaceae split during the Upper Cretaceous period approximately 75.7 Mya (95 % highest posterior density of 60.3-91.3 Mya). Reconstructing ancestral state in phylogenies (RASP) with using the Bayesian Binary Markov chain Monte Carlo (BBM) Method to reconstruct the historical biogeography for the family Schizoparmaceae indicated its most likely origin in Africa. Based on taxonomic and phylogenetic analyses, the Pseudoplagiostomataceae and Pyrisporaceae relationships were clarified and a total of four species described herein. For Pseudoplagiostomataceae, three new species and one known species that include, Pseudoplagiostoma fafuense sp. nov., Ps. ilicis sp. nov., Ps. sanmingense sp. nov. and Ps. bambusae are described and a key of Pseudoplagiostomataceae is provided. With respect to Pyrisporaceae, we considered Pseudoplagiostoma castaneae to be a synonym of Pyrispora castaneae. In addition, a new species of Schizoparmaceae, Coniella fujianensis sp. nov. is described and illustrated.

Museum specimens shedding light on the evolutionary history and cryptic diversity of the hedgehog family Erinaceidae.

The family Erinaceidae encompasses 27 extant species in two subfamilies: Erinaceinae, which includes spiny hedgehogs, and Galericinae, which comprises silky-furred gymnures and moonrats. Although they are commonly recognized by the general public, their phylogenetic history remains incompletely understood, and several species have never been included in any molecular analyses. Additionally, previous research suggested that the species diversity of Erinaceidae might be underestimated. In this study, we sequenced the mitochondrial genomes of 29 individuals representing 18 erinaceid species using 18 freshly collected tissue and 11 historical museum specimens. We also integrated previously published data for a concatenated analysis. We aimed to elucidate the evolutionary relationships within Erinaceidae, estimate divergence times, and uncover potential underestimated species diversity. Our data finely resolved intergeneric and interspecific relationships and presented the first molecular evidence for the phylogenetic position of Mesechinus wangi, Paraechinus micropus, and P. nudiventris. Our results revealed a sister relationship between Neotetracus and Neohylomys gymnures, as well as a sister relationship between Hemiechinus and Mesechinus, supporting previous hypotheses. Additionally, our findings provided a novel phylogenetic position for Paraechinus aethiopicus, placing it in a basal position within the genus. Furthermore, our study uncovered cryptic species diversity within Hylomys suillus as well as in Neotetracus sinensis, Atelerix albiventris, P. aethiopicus, and Hemiechinus auratus, most of which have been previously overlooked.

Calonectria eucalyptorum sp. nov., a new leaf blight pathogen of Eucalyptus from India.

In this study, Calonectria eucalyptorum sp. nov. is described from the blighted leaves of Eucalyptus in India using morphological and multi-locus phylogenetic analyses. The new species belongs to the Calonectria cylindrospora species complex, and its unique microscopic features and DNA sequence information enable clear separation from the 12 currently accepted species in this complex. Conidia of the new taxon are slightly longer than those of its phylogenetic neighbors. Additionally, this species produces central as well as lateral stipe extensions, which is a feature not known for the other members of the C. cylindrospora species complex. Analyses of the combined partial calmodulin, histone, translation elongation factor-1α, and ß-tubulin gene regions revealed a distinct phylogenetic position for C. eucalyptorum. Recombination analysis provided additional support for the new species hypothesis. Koch's postulates for the new taxon as a foliar pathogen of Eucalyptus were fulfilled. The discovery of novel and pathogenic Calonectria species is important because it sheds light on species diversity, potential threats, and disease control.

A new four-pored Amphisbaena Linnaeus, 1758 (Amphisbaenia, Amphisbaenidae) from the north of Espinhaço Mountain Range, Brazil.

A new species of Amphisbaena is described from the north of Espinhaço Mountain Range, municipality of Caetité, state of Bahia, Brazil. Amphisbaenaamethysta sp. nov. can be distinguished from its congeners by the following combination of characters: (1) snout convex in profile, slightly compressed not keeled; (2) pectoral scales arranged in regular annuli; (3) four precloacal pores; (4) distinct cephalic shields; (5) 185-199 dorsal half-annuli; (6) 13-16 caudal annuli; (7) conspicuous autotomic site between 4th-6th caudal annuli; (8) 16-21 dorsal and ventral segments at midbody; (9) 3/3 supralabials; (10) 3/3 infralabials; and (11) smooth and rounded tail tip. The new species is the 71st species of genus with four precloacal pores, and the 22nd species from the Caatinga morphoclimatic domain. The identification of Amphisbaenaamethysta sp. nov. indicates that the reptile fossorial fauna in the Espinhaço Mountain Range region is far from being completely known and that it may harbour a much greater diversity of endemic taxa.

Complete chloroplast genome sequence of Artemisia littoricola (Asteraceae) from Dokdo Island Korea: genome structure, phylogenetic analysis, and biogeography study.

The Asteraceae family, particularly the Artemisia genus, presents taxonomic challenges due to limited morphological characteristics and frequent natural hybridization. Molecular tools, such as chloroplast genome analysis, offer solutions for accurate species identification. In this study, we sequenced and annotated the chloroplast genome of Artemisia littoricola sourced from Dokdo Island, employing comparative analyses across six diverse Artemisia species. Our findings reveal conserved genome structures with variations in repeat sequences and junction boundaries. Notably, the chloroplast genome of A. littoricola spans 150,985 bp, consistent with other Artemisia species, and comprises 131 genes, including 86 protein-coding, 37 tRNA, and 8 rRNA genes. Among these genes, 16 possess a single intron, while clpP and ycf3 exhibit two introns each. Furthermore, 18 genes display duplicated copies within the IR regions. Moreover, the genome possesses 42 Simple Sequence Repeats (SSRs), predominantly abundant in A/T content and located within intergenic spacer regions. The analysis of codon usage revealed that the codons for leucine were the most frequent, with a preference for ending with A/U. While the chloroplast genome exhibited conservation overall, non-coding regions showed lower conservation compared to coding regions, with the Inverted Repeat (IR) region displaying higher conservation than single-copy regions. Phylogenetic analyses position A. littoricola within subgenus Dracunculus, indicating a close relationship with A. scoparia and A. desertorum. Additionally, biogeographic reconstructions suggest ancestral origins in East Asia, emphasizing Mongolia, China (North East and North Central and South Central China), and Korea. This study underscores the importance of chloroplast genomics in understanding Artemisia diversity and evolution, offering valuable insights into taxonomy, evolutionary patterns, and biogeographic history. These findings not only enhance our understanding of Artemisia's intricate biology but also contribute to conservation efforts and facilitate the development of molecular markers for further research and applications in medicine and agriculture.

Halostreptopolyspora alba gen. nov., sp. nov., a halophilic actinobacterium isolated from saline soil of Xinjiang, Northwest of China.

A Gram-stain-positive, aerobic, moderate halophilic actinobacterium, designated strain YIM 96095T, was isolated from a saline soil sample collected from Aiding Lake, Xinjiang, North-western China. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the isolate belonged to the family Nocardiopsidaceae, formed a distinct subclade, and was most closely related to Lipingzhangella halophila DSM 102030T and Allosalinactinospora lopnorensis DSM 45697T with sequence identity values of 95.8 and 95.1%, respectively. Optimal growth occurred at 37 °C, pH 7.0-8.0 and with 5-16% (w/v) NaCl, with well-developed, non-fragmented substrate mycelia and single-, double-, or triple-wrinkled spore(s) on the mature aerial hyphae. The chemical analysis presented meso-diaminopimelic acid as the diagnostic diamino acid of the cell-wall peptidoglycan, and glucose, galactose and rhamnose as the major whole-cell sugars, and iso-C15 : 0 and anteiso-C15 : 0 as the major fatty acids. The phospholipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, unidentified phospholipids and unidentified glycolipid. The menaquinones were MK-10(H8), MK-10(H6) and MK-9(H10). Its G+C content was 69.7 mol% in the determined genome sequence. Based on phenotypic, chemotaxonomic and phylogenetic characteristics, a novel genus and species named Halostreptopolyspora alba gen. nov., sp. nov. is proposed for isolate YIM 96095T (=KCTC 49266T=CGMCC 4.7636T).

Integrative morphological, cytogenetic and molecular characterization of the Andean climbing catfish Astroblepus mindoensis (Regan, 1916) (Siluriformes:Astroblepidae).

Astroblepus species, commonly known as Andean climbing catfish, exhibit a unique challenge in species delimitation, leading to ongoing taxonomic debates. Here we report data on Astroblepus mindoensis, a vulnerable species endemic to Ecuador, obtained by an integrative approach that includes cytogenetic analysis, molecular identification of the specimens, and recording of morphological and morphometric characters useful for species diagnosis. Thus, this study aimed to associate the karyotype data of the specimens analyzed with morphological and molecular characters, improving and expanding the existing taxonomic information, thus contributing to the systematics of the species. Our morphology results, unlike Regan's original description, which is brief and ambiguous, provide a more detailed morphometric and meristic description. Molecular phylogenetic reconstruction and genetic distance based on a fragment of the cytochrome c oxidase subunit I (COI) showed that our samples constitute a well-supported and monophyletic clade within the A. grixalvii species complex. The cytogenetic analysis identified distinct chromosomal markers, including a single cluster of major ribosomal genes (on chromosome pair 3) and of minor ribosomal genes (on chromosome pair 12) with their localization differing from those reported in other Astroblepus species analyzed. Additionally, the presence of a heteromorphic chromosome pair in males suggests the presence of an XX/XY sex-determination system that has not been identified in other congeneric species. Further investigation is necessary to determine if these chromosomes are associated with the accumulation of repeated sequences, as typically occurs with sex chromosomes, and to assess their presence in other species of the genus.

Museomics reveal origins of East African Pleophylla forest chafers and Miocene forest connectivity.

Here we present a nearly complete species-level phylogeny including 23 of the 25 known species of the forest-dwelling herbivorous scarab chafer beetle genus Pleophylla (Coleoptera: Scarabaeidae: Sericinae), based on the analysis of 950 nuclear genes (metazoan-level universal single-copy orthologs; mzl-USCOs). DNA sequences were obtained from freshly collected, ethanol-preserved samples and from dried museum specimens by target enrichment or genome shotgun sequencing. Alignment completeness of mzl-USCOs newly obtained here by target DNA enrichment of ethanol samples were very heterogenous and lower (29-62 %) than in Dietz et al. (2023a), while that of sequences recovered from dried samples was even lower (∼19 %). Alignment completeness of the sequences obtained from low coverage shotgun sequencing was highest (∼92 %), although the average coverage was much lower than for the target enrichment samples. We used the resulting phylogeny to reconstruct the historical biogeography of the group. To estimate a time-calibrated tree, we combined the mzl-USCO data of Pleophylla with a nucleotide alignment from an available transcriptomic dataset of Scarabaeoidea and used two different sets of secondary calibration points. Despite the problems associated with the capture rate of mzl-USCO sequences from museum specimens, we were able to infer a well-resolved phylogeny of the genus Pleophylla that also provided reliable estimates of the phylogenetic position of species for which we had little sequence data. Our study clearly identified South Africa as the geographic origin of Pleophylla. Timing and biogeographic history confirm a persistent fragmentation of forests since the Eocene. The occurrence of only one long-distance dispersal event from southern Africa to the Eastern African Arc even during the Miocene highlights the limited dispersal possibilities for these forest-adapted chafers, which do not seem to have had important northerly range expansions along hypothetical forest corridors during the Pleistocene.

Phylogenetic Analysis of Porcine Epidemic Diarrhea Virus (PEDV) during 2020-2022 and Isolation of a Variant Recombinant PEDV Strain.

Porcine epidemic diarrhea (PED) is an acute, highly contagious, and infectious disease caused by porcine epidemic diarrhea virus (PEDV). PEDV can affect pigs of all ages, with 50~100% mortality in neonatal piglets and substantial economic losses in the swine industry. In the present study, 347 fecal and intestinal samples were collected from seven regions in China during 2020-2022. A comprehensive molecular investigation of the spike (S) gene of PEDV strains was carried out, which included phylogenetic analysis of the obtained PEDV sequences. Epidemiological surveillance data indicate that the GIIc subgroup strains are widely distributed among pigs. A PEDV strain was successfully isolated from positive small intestine samples and identified through RT-PCR detection using specific N gene primers of PEDV, indirect immunofluorescence assay (IFA), TEM analysis, genome sequencing, and full-length S gene analysis, named PEDV/SC/2022. RDP and SimPlot analysis showed that the isolate originated from the recombination of PEDV/AH2012 and PEDV/AJ1102. In conclusion, our findings contribute to the current understanding of PEDV epidemiology and provide valuable information for the control of PED outbreaks in China.

Comparative Phylogenomic Study of Malaxidinae (Orchidaceae) Sheds Light on Plastome Evolution and Gene Divergence.

Malaxidinae is one of the most confusing groups in the Orchidaceae classification. Previous phylogenetic analyses have revealed that the relationships between the taxa in Malaxidinae have not yet been reliably established, using only a few plastome regions and nuclear ribosomal internal transcribed spacer (nrITS). In the present study, the complete plastomes of Oberonia integerrima and Crepidium purpureum were assembled using high-throughput sequencing. Combined with publicly available complete plastome data, this resulted in a dataset of 19 plastomes, including 17 species of Malaxidinae. The plastome features and phylogenetic relationships were compared and analyzed. The results showed the following: (1) Malaxidinae species plastomes possess the quadripartite structure of typical angiosperms, with sizes ranging from 142,996 to 158,787 bp and encoding from 125 to 133 genes. The ndh genes were lost or pseudogenized to varying degrees in six species. An unusual inversion was detected in the large single-copy region (LSC) of Oberonioides microtatantha. (2) Eight regions, including ycf1, matK, rps16, rpl32, ccsA-ndhD, clpP-psbB, trnFGAA-ndhJ, and trnSGCU-trnGUCC, were identified as mutational hotspots. (3) Based on complete plastomes, 68 protein-coding genes, and 51 intergenic regions, respectively, our phylogenetic analyses revealed the genus-level relationships in this subtribe with strong support. The Liparis was supported as non-monophyletic.

Morphological and molecular characterization of Paractinolaimus uljinensis n. sp. (Nematoda: Actinolaimidae) from Korea, with an updated compendium of the genus.

A new species of the genus Paractinolaimus isolated from the bark of a dead red pine tree was characterized using morphometric data and molecular DNA barcodes. Paractinolaimus uljinensis n. sp. was characterized by its medium sized body 2.50 to 2.98 mm long; lip region truncate, angular and offset by a depression; odontostyle 23.5 to 27.0 µm long; basal shield of pharynx present; vulval opening wide and longitudinal, positioned slightly anteriorly (V = 42.5-47.7); several advulval papillae; female tail long and filiform (324.0-435.0 µm long, c' = 10.1-14.2); a clearly visible copulatory hump; spicules 60.0 to 70.5 µm long; 12 to 15 (mostly 12-14) large contiguous ventromedian supplements, and male tail conoid to broadly rounded. The new species was morphologically compared with P. intermedius, P. sahandi, P. decraemerae, P. acutus, P. macrolaimus, and P. tuberculatus. The phylogenetic relationships among species were reconstructed using 18S- and 28S-rRNA gene sequences. The phylogenies showed well-supported sister relations of Paractinolaimus uljinensis n. sp. with P. sahandi, P. macrolaimus, and P. decraemerae. In addition, the ITS-rRNA gene sequences of Paractinolaimus uljinensis n. sp. were supplied, representing the first characterization of the gene for the genus.

The seventh species of the newt genus Tylototriton in Thailand: a new species (Urodela, Salamandridae) from Tak Province, northwestern Thailand.

A new species of the crocodile newt genus Tylototriton from Doi Soi Malai located at Mae Tuen Wildlife Sanctuary, Tak Province, northwestern Thailand is described based on molecular and morphological evidence, and named as Tylototritonsoimalai sp. nov. The new species is the seventh recorded species of the genus Tylototriton reported in Thailand. It differs morphologically from its congeners by a combination of the following morphological characteristics: head longer than wide; snout blunt or truncate; sagittal ridge on head narrow, short and distinct; dorsolateral bony ridges on head pronounced and rough; parotoids distinct; vertebral ridge prominent, wide and not segmented; 14-16 distinct, rounded and isolated rib nodules but posterior nodules connected; tips of fore- and hind limbs overlapping when adpressed along the body. The body background color is black, while the color markings are orange. Molecular analysis indicated that Tylototritonsoimalai sp. nov. is a distinct lineage and sister to T.uyenoi with a 4.1% genetic sequence divergence based on the mitochondrial NADH dehydrogenase subunit 2 gene. The new species is currently restricted to the hill evergreen forests of Doi Soi Malai. The implementation of a strategic plan is recommended to protect both the species and its habitat from anthropogenic activities.

Morphological and Phylogenetic Characterization of Trichomerium koreanum sp. nov. and Exophiala radicis Belonging to Chaetothyriales Fungi in Korea.

The order Chaetothyriales encompasses black yeasts and related fungi known for their diverse morphologies, asexual reproduction, and habitat preferences. In this study, we isolated two fungal strains belonging to Chaetothyriales, designated KNUF-23-13A and KNUF-22-020, from a longhorn beetle (Cyrtoclytus capra) and soil sample, respectively, in Korea. Phylogenetic analyses based on the concatenated nucleotide sequences of the internal transcribed spacer regions and partial sequences of the large subunit (LSU) rRNA, ß-tubulin (TUB2), and actin (ACT) genes revealed that the strains were grouped with strains of Trichomerium and Exophiala, respectively. For strain KNUF-23-13A, cultural and morphological differences were observed in the shape and size of mycelium between it and its phylogenetically closest species, Trichomerium lapideum CGMCC 3.17311T, supporting the novelty of this isolated species. In the case of strain KNUF-22-020, the cultural and morphological characteristics were similar to those previously reported for E. radicis P2854T. Based on morphological and phylogenetic analyses, we revealed KNUF-23-13A as a novel species, which we describe and name T. koreanum sp. nov., and KNUF-22-020 as Exophiala radicis, a previously unreported species in Korea.