ITS Analysis Research Articles

Phylogenetic and taxonomic evidence reveal Roussoella bambusicola sp. nov. (Pleosporales, Dothideomycetes) from Yunnan Province, China

During an investigation of saprobic microfungi associated with plants in Yunnan Province, China, an ascomycetous fungus collected from dead bamboo is proposed as a new species (Roussoella bambusicola) in Roussoellaceae based on morphological characteristics and phylogenetic analyses of combined ITS, LSU, tef1-α, and rpb2 sequence data. The new species is characterized by ampulliform or lageniform to subcylindrical, slightly curved, hyaline, smooth conidiogenous cells and oblong or ellipsoidal, aseptate, smooth to rough-walled conidia. This paper provides a description, illustrations, and a phylogenetic tree to show the placement of the new species.

Tubaria asiatica (Tubariaceae), a new species from Khyber Pakhtunkhwa, Pakistan

A new Tubaria (Tubariaceae) species, Tubaria asiatica is described and illustrated from district Swabi, Khyber Pakhtunkhwa Pakistan, based on morphological and phylogenetic analysis of ITS and 28S sequences. The most closely related species is Tubaria conspersa, but it differs from it due to its pale orange to orange‐tinged dark brown pileus adnate lamellae, dark brown–orange, cylindrical to subcylindrical stipe, oblong, ellipsoid, lacrymoid, rhomboid basidiospores, clavate cheilocystidia and basidia with long sterigmata. Morphological descriptions, colored photographs, microscopic structures and a comparative combined ITS‐28S‐based phylogenetic analysis is presented with reference to the newly described species.

Additions to Lentitheciaceae (Pleosporales, Dothideomycetes): Pseudosetoseptoria jiangxiensis sp. nov. from decaying culms of bamboo in China

In terms of morphology, ecology, and phylogeny, Lentitheciaceae species exhibit an enormous diversity. They have a cosmopolitan distribution worldwide, and lead diverse lifestyles. A novel species in Lentitheciaceae, Pseudosetoseptoria jiangxiensis occurring on decaying culms of bamboo in Jiangxi province, China is described and illustrated. Morphologically, Pseudosetoseptoria jiangxiensis has dark brown to black, semi-immersed, carbonaceous, pycnidial conidiomata and hyaline, obovoid, elliptic, or nearly spherical, aseptate conidia with minute guttules. Maximum likelihood and Bayesian inference analyses of combined ITS, LSU, SSU and tef1-α sequence data were used to clarify the phylogenetic affinities of the species.

A phylogeny of the Inocybe alienospora group (Agaricales) with emphasis on seven new species from China and emendation of sect. Leptocybe

ABSTRACT A multigene phylogeny of the Inocybe alienospora group is presented based on analyses of ITS, 28S, and rpb2 nucleotide data. Four major subclades and three additional independent lineages were identified in the I. alienospora group. Two species with superficial similarities with the I. alienospora group, viz, I. multicoronata and I. elata, however, were not nested within the I. alienospora group. Inocybe sect. Leptocybe is proposed to accommodate the I. alienospora group in a more formal classification system. As such, sect. Leptocybe now includes 12 documented species and seven new species described in this work. The seven new species include I. aprica sp. nov., I. casuarinoides sp. nov. I. haikouensis sp. nov., three species associated with Casuarina from tropical China; also I. heteromorpha, a species associated with fagaceous trees from tropical China; and I. aurescens sp. nov., I. juji sp. nov., and I. peppa sp. nov., three species associated with fagaceous trees from subtropical China. In addition, new geographical data for I. carpinicola and I. acutata are reported. The East Asian species I. acutata and the neotropical species I. lasseri were confirmed as members of sect. Leptocybe. A key to the 19 species in sect. Leptocybe is also provided.

Novel endophytic fungal species Pithoascus kurdistanensis producing morphine compounds

Papaver genus, commonly known as popies, is a valuable source of alkaloids used in medicine, including papaverine, morphine, codeine, and thebaine. We isolated six endophytic fungal isolates producing morphinan alkaloids from four Papaver species growing in Kurdistan Province, Iran. To do this, a 1:1 mixture of methanol and chloroform was used to extract fungal cultures. The contents of morphinan alkaloids in the extracts were subsequently determined using phase high-performance liquid chromatography. Among the morphinan alkaloid-producing fungal isolates, IRAN 4653C had the highest yield giving 23.06 (mg/g) morphine and 2.03 (mg/g) codeine when grown in potato dextrose liquid medium. The identity of this isolate was examined and recognized as a new fungal species named as Pithoascus kurdistanesis sp. nov. based on multi-gene phylogenetic analyses of ITS, TEF-1α, and TUB2 sequences data and morphological features. The morphinan-producing endophytic fungus and the isolated Pithoascus species from Papaver are being reported for the first time. Accordingly, this fungus shows promise as a new source of valuable compounds which is illustrated and introduced here as a new Microascaceae member belonging to Pithoascus from Kurdistan Province, Iran. Moreover, the morphinan productivity of P. kurdistanesis was further validated by gas chromatography-mass spectrometry (GC–MS).

Morphological characteristics and phylogenetic analyses revealed a new invertebrate-pathogenic fungus Akanthomyces bannaensis (Cordycipitaceae, Ascomycota), in China

The ubiquitous yet diverse associations between insects and fungi have long evolved. At the same time, the interactions between beetles, microbes, and hosts have not been well documented, especially for new fungi taxa. Two interesting specimens of Akanthomyces were found on adult moths of Dudusa sphingiformis in Yunnan Province, Southwest China. These specimens were identified as a new species based on morphology and phylogeny. Akanthomyces bannaensis sp. nov. is characterized by being determined to occur on an adult of Dudusa sphingiformis and synnemata arising from a moth body up to 5 cm long and 0.1–1.3 mm wide, unbranched, cylindrical to clavate with acute or blunt end. The morphological identification and phylogenetic analysis of combined ITS, nLSU, and RPB2 sequence data support A. bannaensis as a new species, in which A. bannaensis is sister to A. pyralidarum. A full description, illustrations and phylogenetic analysis results of the new species are provided.

Integrative description and histopathological lesions of Cyathostoma americana (Nematoda, Syngamidae) parasitizing a barn owl (Tyto alba) in Chile.

Syngamid worms (Nematoda: Syngamidae) parasitizing birds of prey are considered cosmopolitan, but the efforts to understand their biology and systematics are restricted to the Holarctic region. However, in the Neotropical region there is only one recent record with no data about its molecular characterization or its significance to the health of its hosts. Thus, this study aimed to identify through an integrative approach the Syngamid worms parasitizing a native owl, and to describe its pathological consequences. A total of 14 barn owls (Tyto alba) were dissected between 2015 and 2021, from which one bird was found to be parasitized by a high parasitic burden (i.e. 185 worms). Considering light microscopy and SEM, these nematodes were morphologically identified as Cyathostoma americana, and then supported through molecular analyses of nuclear loci ITS and LSU. The pathological changes were described as severe airsacculitis and pneumonia, which probably were the cause of death of the bird. This study represents the first effort to characterize the parasitism by this nematode in a nocturnal bird of prey from the Neotropics, with a lethal outcome. Additional studies are required from other geographical areas and Neotropical countries to better understand the dynamics of this parasite and their hosts, considering both diurnal and nocturnal birds of prey.

Morphological characteristics and phylogenetic analyses reveal Scopuloides yunnanensis (Polyporales, Basidiomycota), a new wood-decaying fungus from China

Wood-decaying fungi, as crucial decomposers in forest ecosystems, have been the subject of meticulous research. In this study, a new wood-decaying fungal species, Scopuloides yunnanensis, found in Yunnan, China, is proposed. Scopuloides yunnanensis was identified based on a comprehensive analysis of its membranaceous basidiomata, generative hyphae, lamprocystidia, projecting cystidia, and allantoid basidiospores. In addition to the morphology, phylogenetic analyses of ITS and nLSU rRNA markers were performed with Maximum Likelihood, Maximum Parsimony, and Bayesian inference methods, ensuring the robustness of our findings. The phylogenetic analysis showed that S. yunnanensis is sister to S. hydnoides. The new species’ description, illustrations, and results of the phylogenetic analysis are provided. In addition, an identification key to all Scopuloides species worldwide is provided.

Koliella bifissiva sp. nov (Chlorellaceae, Chlorophyta) and Analysis of Its Organelle Genomes.

Chlorellacean members are common in aquatic or subaerial habitats, and many of them have significant economic value. Taxonomic reports and organelle genome data for the Nannochloris clade, an important subgroup within this family, are limited, hindering the understanding and exploitation of this clade. In this study, a fusiform-celled strain, FACHB-3607, was isolated from a pond in China. Through examination of morphological characteristics and phylogenetic analyses of rbcL, 18S rDNA, and ITS, it was identified as a new species within the Nannochloris clade, named Koliella bifissiva sp. nov. In addition, this study provided a first insight into the organellar genomes of the genus Koliella. The K. bifissiva chloroplast had a 99.8 kb genome, and the mitochondrion had a 40.8 kb genome, which are moderate sizes within the Nannochloris clade. Phylogenomic analysis showed that K. bifissiva is most closely related to Nannochloris sp. "desiccata", followed by Marvania. In contrast, Picochlorum was the most distantly related species. The organelle genomes of the Nannochloris clade display dynamic evolution, reflected in variations in genome size, gene content and order, and selection pressure. This research enhances our knowledge of species diversity and evolutionary history in the Nannochloris clade.

Two new species of Nectriaceae (Hypocreales, Sordariomycetes) from Yunnan, China.

Nectriaceae is a highly diverse family, and members have a worldwide distribution, particularly in warm temperate to tropical regions. During the survey of fungal diversity in different habitats in Yunnan province, China, two new species isolated from soil and air respectively, namely Atractiumyunnanense sp. nov. and Nalanthamalaxishuangbannaensis sp. nov., were proposed based on morphological comparisons and the multi-gene phylogenetic analyses of combined ITS, LSU, rpb2, and tub2 sequence data. Phylogenetically, both species clustered in a monophyletic clade within Nectriaceae with strong support. A.yunnanense is characterized by synnematous conidiophores, pale olivaceous-green, clavate to oblong-ellipsoidal, multi-septate conidia, and pale olivaceous-green chlamydospores. N.xishuangbannaensis has acremonium-like or penicillium-like conidiophores and either obovate or ellipsoidal, cylindrical or fusiform conidia. Full descriptions, illustrations, and a phylogenetic tree showing the phylogenetic position of the two new species were provided.

Sex-specific responses of Taxus mairei to UV-B radiation involved altering the interactions between the microbiota assembly and host secondary metabolism

BackgroundTo adapt to constantly changing environments, ancient gymnosperms have coevolved with diverse endophytic fungi that are essential for the fitness and adaptability of the plant host. However, the effect of sex on plant-endophyte interactions in response to environmental stressors remains unknown. RNA-seq integrated with ITS analysis was applied to reveal the potential mechanisms underlying the sex-specific responses of Taxus mairei to ultraviolet (UV)-B radiation.ResultsEnrichment analysis suggested that sex influenced the expression of several genes related to the oxidation–reduction system, which might play potential roles in sex-mediated responses to UV-B radiations. ITS-seq analysis clarified the effects of UV-B radiation and sex on the composition of endophytic fungal communities. Sex influenced various secondary metabolic pathways, thereby providing chemicals for T. mairei host to produce attractants and/or inhibitors to filter microbial taxa. Analysis of fungal biomarkers suggested that UV-B radiation reduced the effect of sex on fungal communities. Moreover, Guignardia isolate #1 was purified to investigate the role of endophytic fungi in sex-mediated responses to UV-B radiation. Inoculation with spores produced by isolate #1 significantly altered various oxidation–reduction systems of the host by regulating the expression of APX2, GST7 NCED1, ZE1, CS1, and CM1.ConclusionThese results revealed the roles of endophytic fungi in sex-mediated responses to UV-B radiation and provided novel insights into the sex-specific responses of Taxus trees to environmental stressors.3pQjBcj-im_YZSujzFCGpbVideo

Three New Truffle Species (Tuber, Tuberaceae, Pezizales, and Ascomycota) from Yunnan, China, and Multigen Phylogenetic Arrangement within the Melanosporum Group.

Based on a multi-locus phylogeny of a combined dataset of ITS, LSU, tef1-α, and rpb2 and comprehensive morphological analyses, we describe three new species from the Melanosporum group of genus Tuber and synonymize T. pseudobrumale and T. melanoexcavatum. Phylogenetically, the three newly described species, T. yunnanense, T. melanoumbilicatum and T. microexcavatum, differ significantly in genetic distance from any previously known species. Morphologically, T. yunnanense is distinctly different from its closest phylogenetically related species, T. longispinosum, due to its long shuttle-shape spores (average the ratio of spore length to spore width for all spores (Qm) = 1.74). Tuber melanoumbilicatum differs from the other species in having a cavity and long shuttle-shaped spores (Qm = 1.65). Although T. microexcavatum sampled ascomata have relatively low maturity, they can be distinguished from its closely related species T. pseudobrumale by the ascomata size, surface warts, and spore number per asci; additionally, phylogenetic analysis supports it as a new species. In addition, molecular analysis from 22 newly collected specimens and Genebank data indicate that T. pseudobrumale and T. melanoexcavatum are clustered into a single well-supported clade (Bootstrap (BS) = 100, posterior probabilities (PP) = 1.0); and morphological characteristics do not differ. Therefore, based on the above evidence and publication dates, we conclude that T. melanoexcavatum is a synonym of T. pseudobrumale. By taking into account current knowledge and combining the molecular, multigene phylogenetic clade arrangement and morphological data, we propose that the Melanosporum group should be divided into four subgroups. Diagnostic morphological features and an identification key of all known species in the Melanosporum group are also included. Finally, we also provide some additions to the knowledge of the characterization of T. pseudobrumale, T. variabilisporum, and T. pseudohimalayense included in subgroup 1 of the Melanosporum group.

Natural Occurrence, Morpho‐Molecular Characteristics, and Pathogenicity of Fusarium spp. Associated With Chrysanthemum Wilt in Vietnam

ABSTRACTChrysanthemum Fusarium wilt (CFW) is a devastating soil‐borne disease affecting the cut chrysanthemum industry globally. In Vietnam, this disease has been occurring silently and is posing a risk of epidemic outbreaks in the southern provinces, especially Lam Dong. Results from a survey in 2023 showed that stunted growth, curved and small stems, dried‐rot roots, leaf drooping and green or yellow wilting of plants after transplanting or at their flower bud initiation were common symptoms observed across the main chrysanthemum‐growing farms in Lam Dong. The occurrence of CFW was recorded in relation with the host cultivar, irrigation method and fungicide‐spraying regime. Among the cultivars grown in Lam Dong, ‘Doa cu’ was the most susceptible to CFW. The use of sprinkler irrigation and infrequent application of fungicides were attributed to increase both disease incidence and severity. Thirty fungal isolates obtained from diseased plants were identified as Fusarium oxysporum and F. falciforme based on morphological features and molecular analysis of ITS and TEF‐1α genes. The detection frequency of these two species was almost equal, which shows their balanced role in the occurrence of CFW in Lam Dong. In‐planta pathogenicity assays of Fusarium isolates against plantlets grown from cuttings and invitro propagation showed a variation in pathogenicity and a specialisation of isolates for these planting materials. These results suggest that the profiles of population and aggressiveness of Fusarium isolates diverged with the type of planting material, and between the stock and the commercial farms.

Phylogenetic and taxonomic evidence reveal Punctulariopsis yunnanensis sp. nov. (Punctulariaceae, Basidiomycota) from southwest China

A new wood-rotting fungal species, Punctulariopsis yunnanensis, is proposed based on a combination of morphology and molecular data. The species is characterized by the resupinate basidiomata with a pinkish buff salmon hymenial surface, a monomitic system, generative hyphae with clamp connections, and subhymenial generative hyphae thick-walled, and the presence of dendrohyphidia, and broadly ellipsoid basidiospores measuring 7.8–12.8 × 6–9.8 µm. Phylogenetic analyses of the combined ITS and nLSU nrDNA gene regions demonstrated that P. yunnanensis formed an independent lineage and was sister to P. subglobispora with strong statistical support. A full description, illustrations and phylogenetic analysis results of the new species are provided.

The genus Peniophora (Russulales, Basidiomycota) from Patagonia revisited

The identity of Peniophora specimens from Patagonia is examined based on morphological, cultural, and biological features as well as phylogenetic analyses of nuclear ribosomal ITS and 28S sequences. Three new species are described from Patagonia: Peniophora patagonica, P. pitrae, and P. revoluta. In total, five species are known in the area, growing on endemic trees in Southern Argentina. Peniophora cinerea and P. rufomarginata are excluded from the Patagonian records, and the presence of P. incarnata and P. lycii is confirmed by molecular methods. A key to Peniophora species from Patagonia is presented.

Tricholyophyllum (Agaricales; Lyophyllaceae) an addition to the fungal flora of Pakistan with the description of a new species T. rubrum

Phylogenetic analysis of ITS and LSU sequence data and morphological characters support the recognition of Tricholyophyllum rubrum as a new species. The new species differs from the type species of the genus by its reddish brown pileus surface, the presence of pleurocystidia, and comparatively broader basidiospores. This is the first report of a Tricholyophyllum species from Pakistan and the second in the world.

A new species of Wongia (Papulosaceae, Sordariomycetes) from Taiwan

Wongia miscanthi sp. nov. from decaying culms of Miscanthus floridulus submerged in freshwater streams at Wufeng District, Taiwan, is described and illustrated, based on two collections. This new species was identified based on morphological comparison and multi-locus phylogenetic analyses of combined ITS, LSU, SSU, rpb2, and tef1a sequence dataset. Intraspecific variability between the two collections is discussed. A synopsis of Wongia species is compiled for comparison and a taxonomic key to the species is provided as an aid to their identification.

Living with bats: polyphasic approach reveals a new Fusarium species in Yunnan Province, China

ABSTRACT Bats are one of the most diverse mammal groups, with over 1400 species distributed across the planet, and they provide key ecological and economic services. In addition, bats provide an ideal habitat for fungal growth and harbour a high diversity of fungal species. Previous studies have described 423 fungal species associated with bats and bat carcasses. This paper describes a new fungal species, Fusarium rhinolophi, belonging to Nectriaceae, collected from bats in Mengla County, Xishuangbanna Dai Autonomous Prefecture, Yunnan Province, China. Phylogenetic analyses of the combined ITS, CAM, TEF-1α, TUB, RPB1, and RPB2 dataset confirmed that our fungus forms an independent lineage within the F. nisikadoi species complex. Thus, we describe our fungus as a new species of Fusarium based on morphology and molecular phylogeny. A description, phylogenetic tree showing species placement, and micrographs are provided.

Thyridium livistonae sp. nov. from Yunnan Province, China, with two new combinations

ABSTRACT An ascomycetous fungus was collected and isolated from dead leaves of Livistona chinensis in Lincang City, Yunnan Province, China, and morphology and multi-gene phylogenetic analyses of LSU, ITS, Actin, and β-tub were carried out to identify it. The morphology and multigene phylogeny revealed that this fungus is a distinct species of Thyridium. Therefore, this fungus is described and introduced as Thyridium livistonae sp. nov. In addition, two new combinations T. hipposidericola and T. xishuangbannaense are proposed based on morphology and phylogeny for Phialemoniopsis hipposidericola Karun., Tibpromma & X.F. Liu and P. xishuangbannaensis Karun., Tibpromma & X.F. Liu respectively. A detailed description, illustrations, and phylogenetic results are provided for the new species and two new combinations.

Fungal endophytes Fusarium solani SGGF14 and Alternaria tenuissima SGGF21 enhance the glycyrrhizin production by modulating its key biosynthetic genes in licorice (Glycyrrhiza glabra L.).

To identify promising fungal endophytes that are able to produce glycyrrhizin and enhance it in licorice and the mechanisms involved. Fifteen fungal endophytes were isolated from Glycyrrhiza glabra L. rhizomes among which SGGF14 and SGGF21 isolates were found to produce glycyrrhizin by 4.29 and 2.58µg g-1 dry weight in the first generation of their culture. These isolates were identified as Fusarium solani and Alternaria tenuissima, respectively, based on morphological characteristics and sequence analysis of internal transcribed spacer, TEF1, ATPase, and CAL regions. Subsequently, G. glabra plants were inoculated with these fungal isolates to examine their effect on glycyrrhizin production, plant growth parameters and the expression of key genes involved in glycyrrhizin pathway: SQS1, SQS2, bAS, CAS, LUS, CYP88D6, and CYP72A154. Endophytes were able to enhance glycyrrhizin content by 133%-171% in the plants. Natural control (NC) plants, harboring all natural endophytes, had better growth compared to SGGF14- and SGGF21-inoculated and endophyte-free (EF) plants. Expression of SQS1, SQS2, CYP88D6, and CYP72A154 was upregulated by inoculation with endophytes. LUS and CAS were downregulated after endophyte inoculation. Expression of bAS was higher in SGGF21-inoculated plants when compared with NC, EF, and SGGF14-inoculated plants. Two selected fungal endophytes of G. glabra can produce glycyrrhizin and enhance glycyrrhizin content in planta by modulating the expression of key genes in glycyrrhizin biosynthetic pathway.