Morphological and phylogenetic evidence supports four new species and one new record of Entoloma (Entolomataceae, Agaricales) from Northeast China

Collybiopsis baiyunensis, a new species collected from Baiyun Mountain, Guangzhou, Southern China, is described based on phylogenetic analyses of combined ITS and nrLSU sequences and morphological evidence. It is characterized by its light brown to yellowish brown pileus with reddish orange to reddish brown disc and whitish margin, oblong to subcylindrical basidiospores, clavate to broadly clavate or spheropedunculate cheilocystidia, absence of pleurocystidia, and cylindrical or flexuose caulocystidia. Detailed morphological descriptions, phylogenetic analyses and comparisons with closely related species are presented.

A new species, Collybiopsis pakistanica, from Margalla Hills National Park, Pakistan, is described based on micro‐morphological and molecular phylogenetic analyses. This species is characterized by its milky white pileus, yellowish orange or light yellow to orange at central disc and margins, lacrymoid basidiospores, cheilocystidia with median construction, and broadly utriform and clavate to narrowly utriform pleurocystidia and epithelioid hymeniderm pileipellis. Its placement as a new taxon of Collybiopsis is confirmed by phylogenetic analyses of ITS and nrLSU sequences. Two new combinations, Collybiopsis ugandensis and Collybiopsis pleurocystidiata are also proposed as these species are transferred from Gymnopus.

Cortinarius represents not only the most extensive genus within the Agaricales but also constitutes a significant ectomycorrhizal lineage, a crucial ectomycorrhizal genus for maintaining ecosystem functionality. During a field survey in the Three Parallel Rivers Region, China, two specimens of Cortinarius were collected from an Abies forest. Based on morphological and phylogenetic analysis of ITS and nrLSU sequences, a novel species, Cortinarius orientiprivignipallens, is classified within the subgenus Telamonia, section Privignati. This species is characterized by its fibrillose pileus surface, which is greyish-white and becomes glabrous as it matures; the stipe is greyish-violet at the apex, while remnants of the universal veil appear brown. A description, illustration, phylogenetic analysis results, and comparison with allied taxa are provided. In addition, a key to the 20 new species of Cortinarius subgenus Telamonia published in China from 2019 to 2025 is also provided.

Panax notoginseng (Burk.) F. H. Chen is a perennial plant species in the family Araliaceae, and its roots and rhizome are precious materials for the production of traditional Chinese medicine. From April to June, 2018, new disease symptoms were detected on the mature leaves of 2- and 3-year-old Panax notoginseng (P. notoginseng) in Wenshan Autonomous Prefecture, Yunnan Province, China, and the disease incidence was about 10%-15% among the analyzed fields (3.6 ha, 23°49'46.99″ N, 104°06'12.99″ E, 1,631 m elevation). The diseased leaves had dark brown necrotic lesions (0.9-2.5 × 1.0-3.5 cm) and curled downward. As the disease progressed, the necrosis gradually spread along the vein to other leaf parts, eventually covering the whole leaf. In the late disease stage, the whole leaf was decayed and yellowed. For pathogen isolation, infected leaves (n=20) were surface sterilized in 1% sodium hypochlorite and washed with sterilized distilled water for 3 mins before being cut into smaller pieces (~1cm2), then placed onto potato dextrose agar (PDA) medium and incubated at 28°C under aseptic conditions for 3 days. The hypha around leaf discs were transferred onto the new PDA. A total of 20 colonies (SQ1~20) were obtained and purified by single spore culture for morphological characterization and molecular biological identification. The colonies of all isolates were nearly round, grayish white at the initial stage, and then turned to grayish brown. In addition, microscopic examination (100× magnification) of 20 isolates revealed dark, septate, and sparsely branched conidiophores as well as dark brown conidia with short conical beaks at their tip. Additionally, conidia (solitary or in short chains) were typically oval or club-shaped and had 0-2 longitudinal septa and 2-4 transverse septa (20-35 × 8-12 μm) (n = 50). Moreover, the conidia had a smooth or verrucose surface. Their morphological characteristics were similar to those descriptions given for members of section Alternaria by Lawrence et al. (2016). In order to further identify pathogenic species, genomic DNA was extracted from the colonies (SQ1~20) using a modified cetyltrimethylammonium bromide (CTAB) method (Loganathan et al. 2014). The sequences of internal transcribed spacer regions of ribosomal DNA (rDNA ITS) and partial RNA polymerase II second subunit gene (RPB2) were amplified by PCR using fungal universal primers ITS1/ITS4 (White et al. 1990) and fRPB2-5F/fRPB2-7cR (Liu et al. 1999), respectively. The DNA sequencing shows that ITS sequences from 20 isolates were totally same, and so did the RPB2 sequences (supplementary material). BLASTN analysis of NCBI database indicated that the RPB2 and ITS sequences have the highest nucleotide homology to A. Alternata ITS (MW008974) and RPB2 (LC132700), respectively. These two gene sequences were submitted to GenBank [Accession numbers ON075466 (ITS) and OP572232 (RPB2)]. Phylogenetic trees based on the combined ITS and RPB2 sequences were constructed by maximum parsimony method. The referenced ITS and RPB2 sequences of Alternaria were from three published articles (Rama et al. 2020; Sun et al. 2021; Wee et al. 2006). Phylogenetic analysis revealed that this isolate was clustered with A. alternata. Therefore, the morphology-based preliminary identification was verified by the phylogenetic analysis, and the isolate from diseased P. notoginseng leaves was A. alternata. To confirm its pathogenicity, the fungal isolate was assessed with 40 1-year-old healthy P. notoginseng plants in a greenhouse. Among them, the leaves of 20 of P. notoginseng plants were wounded using a sterile needle (seven or eight wounds per leaf) and then inoculated with 1mL conidial suspension (1 × 106 conidia/mL) prepared from 7-day-old fungal cultures grown on PDA medium. The inoculated plants were covered with plastic bags at 25°C for 24 h to maintain humidity, and then transferred to the greenhouse maintained at 28°C with a 16-h day/8-h night cycle and continuous misting. The other 20 control plants were only wounded and sprayed with sterile water. Typical necrotic lesions were detected on all of the inoculated P. notoginseng leaves cultivated in the greenhouse for 1 week post-inoculation. As the disease continued to develop, the necrotic lesions enlarged, and the infected leaves turned yellow and withered. These symptoms were similar to those observed on the naturally infected P. notoginseng. In contrast, the mock-inoculated control plants remained healthy. Furthermore, the fungus re-isolated from the infected P. notoginseng leaves in the pot experiment had similar morphological characteristics as the original strain. In addition, its genomic DNA was extracted for PCR analysis of ITS and RPB2 sequences, and the following DNA sequencing shows that the two DNA sequences were same as those of isolates SQ1~20, which confirmed that the re-isolated fungus was A. alternata. To the best of our knowledge, this is the first report of A. alternata causing a P. notoginseng leaf disease in China.

Astragalus phanothrix has long been a taxonomically controversial species due to conflicting synonymizations and section placements. Originally described under sect. Onobrychium by Bornmüller in 1915, it was later transferred to sect. Hypoglottidei and synonymized with A. saganlugensis. Recent treatments synonymized it with A. cancellatus within sect. Onobrychoidei, highlighting persistent taxonomic ambiguities arising from overlapping morphological characters and the lack of integrative analyses. This study reassessed the taxonomic status of A. phanothrix using morphological examinations and molecular phylogenetic analyses of ITS and matK sequences. Morphological comparisons revealed key diagnostic characters that distinguish A. phanothrix from its putative synonyms. Astragalus phanothrix differs from A. saganlugensis by its mixed black and white hairs on the calyx (compared to erect stems with sparser indumentum), and smaller ovate legumes (compared to larger, elliptic legumes). From A. cancellatus it is separated by the absence of bifurcate calyx hairs, its placement in sect. Hypoglottidei (versus sect. Onobrychoidei), and its predominantly white hairs on the peduncle (compared to mixed black and white hairs). Phylogenetic analyses placed A. phanothrix firmly within sect. Hypoglottidei, supporting its recognition as an independent species. These findings resolve the taxonomic doubts surrounding A. phanothrix, re-establishing it as a distinct species, and highlight the value of integrating morphological and molecular evidence to clarify complex taxonomic relationships in Astragalus.

Fungal diversity is rapidly expanding, with new species being discovered worldwide. Northern Thailand is considered a hotspot for fungal diversity. In this study, we collected a saprobic microfungus from decaying wood in Chiang Mai Province, Thailand. Morphological examination and phylogenetic analyses of ITS, LSU, and SSU sequences using maximum likelihood and Bayesian inference placed a new isolate within Kirschsteiniothelia. Kirschsteiniothelia chiangmaiensis sp. nov. is characterised by 8-spored, cylindrical-clavate asci with a small ocular chamber and 1-septate ascospores that are ellipsoid to fusiform, with distinct guttules and a smooth wall surrounded by a mucilaginous sheath. Kirschsteiniothelia chiangmaiensis differs from the type by the smaller ascomata, wider asci, and presence of pluri-guttulate, and diverges from K. thujina by smaller ascomata, ascospore, and the presence of mucilaginous sheath. Based on morphology and phylogeny evidence, we introduce K. chiangmaiensis as a new species.

An undescribed Colletotrichum species was isolated from leaf spots on Rubus idaeus (raspberry) in the summer of 2013 in the Jilin Province in northeast China. Based on morphology and ITS sequence data, it was identified as belonging to the Colletotrichum destructivum species complex. Phylogenetic analyses of combined ITS, GAPDH, ACT, CHS-1 and TUB2 sequence data revealed that the fungus represents a new species described here as Colletotrichum neorubicola sp. nov. Pathogenicity testing confirmed that this new species is pathogenic to R. idaeus.

Since the first formal description of the unicellular green algae Dunaliella salina, its presence in hypersaline environments worldwide and its physiological responses to different environmental conditions have been studied extensively. Moreover, due to massive carotenoid accumulation by some strains under specific growth conditions, its biotechnological applications have attracted a great deal of scientific interest. In this study, the phylogenetic relationship, growth, and carotenogenesis of a new strain of Dunaliella salina isolated from Maharlu salt lake in Shiraz (latitude 29.26°N, longitude 52.48°E), Iran were investigated. First, a phylogram based on neighbor-joining analysis of the nuclear rDNA ITS (ITS-1 + 5.8 rDNA + ITS-2) sequence data was constructed. The phylogenetic tree showed that the new isolate is part of a major clade containing several strains of D. salina and was designated as D. salina MSI-1. Then, the responses of the new isolate to the initial pH of the culture media and different concentrations of nitrate, NH4+, and citrate were examined. As with other strains of D. salina, growth and carotenogenesis were controlled by the levels of nitrate and NH4+ in the growth media. Low available nitrogen negatively affected growth but enhanced carotenoid accumulation. Insensitivity of carotenogenesis to citrate indicates a minor contribution of cytosolic IPP synthesis to the overall carotenoid production. Despite changes in the initial pH of the culture media over the experimental period, the initial pH had marked effects on the growth and carotenogenesis of the new isolate. These effects, together with the higher cell carotenoid content observed at pH 11.0, await further research. The results confirm that the analysis of the ITS sequences is a reliable basis for determination of the genetic relatedness among strains of the genus Dunaliella, and the search for strains with novel characteristics may have valuable biotechnological applications.

Four species of Psathyrella—P. subspadiceogrisea sp. nov., P. boreifasciculata, P. gordonii, P. senex were discovered in northeast China. P. subspadiceogrisea is described as new from Changbai Mountain; the other three species are recorded in China for the first time. The specimens were identified based on morphological characteristics and molecular analysis of ITS sequences. Detailed morphological descriptions, line drawings and photographs are presented.

ITS sequence analysis used for parent selection in Lilium lancifolium Thunb. cross-breeding

Analyses of ITS, ETS and matK sequences for genera of the subtribe Hinterhuberinae, including representatives of 17 subtribes from the tribe Astereae, rooted on Anthemideae, Calenduleae and Inuleae, provide no support for the monophyly of the Hinterhuberinae. Chiliotrichum and Celmisia groups, Olearia in part, Novenia, Oritrophium and Pteronia are in the basal group that also includes subtribe Felicinae. Madagaster, Remya, Olearia in part, Achnophora and Hinterhubera group lie in the Southern Hemisphere grade, with Asterinae, Bellidinae, Brachycominae, Grangeinae, Podocominae, Lagenophorinae and Baccharidinae. Monophyly of Asterinae, Grangeinae, Podocominae and Lagenophorinae is not supported. Instead South American representatives of Lagenophorinae, Grangeinae and Podocominae group with Baccharidinae and Hinterhubera group in the South American clade that is nested in the Southern Hemisphere grade. They are sister to the North American subtribes. Remya, Achnophora, Olearia in part and Australasian, European and African representatives of Asterinae, Brachycominae, Grangeinae, Podocominae and Lagenophorinae are grouped in a polytomy that is basal to the South American clade. Laennecia (Podocominae) is sister to Westoniella (Hinterhuberinae), and Myriactis panamensis (Lagenophorinae) and Plagiocheilus (Grangeinae) group with Archibaccharis (Baccharidinae). The Celmisia group is monophyletic. The radiate Diplostephium and Guynesomia do not group with other radiate genera from the Chiliotrichum group, but are in the Southern Hemisphere grade and close to the disciform Hinterhubera group. Transfer of Blakiella from Podocominae to Hinterhuberinae is supported by the three datasets. Madagaster is isolated from the other Hinterhuberinae in having basal position in the Southern Hemisphere grade. Evolution of dioecy in Baccharidinae and Aztecaster (Hinterhuberinae) evolved independently. Woody habit, epaleate receptacle, bisexual disk florets and ligulate peripheral florets are ancestral states in the tribe Astereae. Staminate disk florets and paleate receptacles are derived characters that evolved in several lineages independently. South America, southern Africa or New Zealand are proposed centers of origin for Astereae and for the Chiliotrichum and Celmisia groups. The Hinterhubera group evolved from the South American autochthonous elements, probably from the lower altitude plants during the uplift of the Andes and the development of páramo habitat.

Phylogenetic relationships among 85 species representing 35 genera in the grass tribe Andropogoneae were estimated from maximum parsimony and Bayesian analyses of nuclear ITS and chloroplast trnL–F DNA sequences. Ten of the 11 subtribes recognized by Clayton and Renvoize (1986) were sampled. Independent analyses of ITS and trnL–F yielded mostly congruent, though not well resolved, topologies. Arundinella is sister to Andropogoneae in the trnL–F phylogeny and is nested within the tribe in the ITS and combined data trees. Tristachya is sister to Andropogoneae + Arundinella in the ITS phylogeny. Four clades are common to the ITS and trnL–F phylogenies and the trees from the combined data set. Clade A consists of Andropogon, Diectomis, Hyparrhenia, Hyperthelia, and Schizachyrium. Within this clade, Andropogon distachyos, Hyparrhenia, and Hyperthelia form clade C. Clade B consists of Bothriochloa, Capillipedium, and Dichanthium, and clade D includes Chrysopogon and Vetiveria. Analysis of the combined data resulted in an unsupported larger clade comprising clades A and B plus Cymbopogon, and a sister clade of Heteropogon, Iseilema, and Themeda. This larger clade is similar to the core Andropogoneae clade previously reported (Spangler et al. 1999; Mathews et al. 2002). Based on our sample, which represents 41% of the tribe’s genera, most of Clayton and Renvoize’s (1986) subtribes are not monophyletic.

Human larva migrans is a zoonotic disease caused by larvae of various nematode species, with all previously confirmed pathogens originating from mammalian reservoirs. In 2020, a case series in Vietnam reported instances of cutaneous larva migrans caused by avian eyeworm larvae of the Oxyspirura genus. This nematode genus, belonging to the Thelaziidae family, includes 84 species that mainly parasitize the eyes of various bird species. They utilize different arthropod intermediate hosts, such as cockroaches, grasshoppers, and crickets, in their life cycle. Although the 18S sequences of the larvae were analyzed, the precise identification of the pathogen at the species level remains inconclusive. This study aims to identify the exact causative agent at the species level by analyzing larvae from a patient and adult eyeworms collected from chickens raised in his family using morphological and molecular methods. Molecular analysis of 18S rDNA, ITS2, and cox1 sequences revealed genetic identity between the larva from the patient and adult eyeworms from chickens, as well as with Oxyspirura mansoni from Thailand and Bangladesh. Additionally, morphological examinations further confirmed the adult chicken eyeworms as Oxyspirura mansoni. These findings confirm that O. mansoni larvae are responsible for human larva migrans, marking the first confirmation of nematode larvae from birds (chickens) as a pathogen causing an emerging neglected tropical disease in humans. Given the widespread distribution of this nematode, further research is crucial to investigate this neglected disease, not only in Vietnam but also in other regions. Understanding appropriate treatments and transmission routes is essential to prevent infections in both chickens and humans.

We revealed new deep-water species and cryptic speciation within freshwater sponges of the endemic family Lubomirskiidae (Porifera; Demospongiae; Spongillina) based on molecular and spicule morphology analyses of ITS and CO1 mtDNA. Lubomirskiidae contains a group of closely related species which are a dominant component of the benthos in Lake Baikal, the world's deepest and most ancient lake. Spicule morphology was similar between two Recent samples and species only known previously from fossils in Late Pliocene (3.2-2.8 mya) sediments. Despite the morphological similarity with the cosmopolitan family Spongillidae, molecular analysis of ITS sequences has reliably assigned these species to Lubomirskiidae. This not only indicates that species identification of freshwater fossil sponge spicules should be made with caution, but also suggests that the structure of megascleres may not be a reliable character for interpretations of paleoclimatic reconstructions for the Baikal region. Our results do not support the current classification of Lubomirskiidae into its morphologically defined genera and species, suggesting a strong discrepancy between molecular and morphological variation in Baikalian sponges. This present contribution is the first part of a study on the phylogenetic relationships of the Lake Baikal deep water sponge fauna.

Veluticeps microspora sp. nov. and V. ambigua new to Asia are described and illustrated based on morphological and molecular characters. V. microspora was collected from Yunnan Province, southwestern China, and is characterized by its special anatomical structure (without tomentum, cutis, and subiculum), simple septate hyphae, and small basidiospores. V. ambigua was collected from Jilin Province, northeastern China, and is reported in Asia for the first time. The molecular phylogeny of Veluticeps was preliminarily studied based on the analysis of ITS sequences from 18 species of Gloeophyllales. Two distinct clades of Veluticeps were formed in the phylogenetic tree. While V. microspora and V. ambigua clustered together to form a strongly supported clade, V. abietina, V. fimbriata, and V. berkeleyi grouped with Chaetodermella luna to form another clade.