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Opportunistic infections caused by fungi, particularly those occurring in immunocompromised patients, are considered challenging worldwide. Therefore, a comprehensive understanding of pathogenic fungi is necessary. The present study reports the isolation of a strain of Apiotrichum cacaoliposimilis , which is difficult to detect using conventional clinical assays, from the sterile urine samples of a patient with a urinary tract infection. Sanger sequencing of the internal transcribed spacer regions confirmed the genus of the microbe, while whole-genome sequencing yielded the initial genome assembly of A. cacaoliposimilis . A total of 7,161 predicted proteincoding genes were mapped using multiple databases, including Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, non-redundant protein database, Pathogen-Host Interactions Database, and Comprehensive Antibiotic Resistance Database. The phenotypic data, biochemical reactions, and antimicrobial susceptibility analyses were conducted to reveal the metabolic properties, virulence, and drug resistance profile of the isolated A. cacaoliposimilis . The rank-sum test revealed the differences in the intergeneric distribution of the highly virulent genes UgeB and Pem2 . In addition, other genes exhibited significant overlap in terms of virulence factors with the clinical isolate Apiotrichum mycotoxinivorans GMU1709. Fortunately, similar to most fungi belonging to the Apiotrichum genus, the isolate investigated in the present study was also sensitive to the drug voriconazole (MIC = 0.06 μg/ml). In summary, the phylogenetic placement, potential pathogenic genes, drug sensitivity patterns, and morphological characteristics of the isolated A. cacaoliposimilis were determined precisely in the present study., (© 2024 Wei Wang et al., published by Sciendo.)

Dark Septate Endophytic (DSE) fungi can benefit plants by optimizing nutrient uptake, biosynthesis of phytohormones-like compounds, and stress relief such as toxic metals. The objective was to characterize in vitro 57 strains of the DSE Periconia macrospinosa isolated from sugarcane roots, indicating the most promising in solubilizing phosphate sources, growing in different metal and vinasse contents, and producing molecules related to indoleacetic acid (IAA). The strains were from the Embrapa Agrobiology Fungi Collection. Over 35% of the strains solubilize calcium phosphate, highlighting A356 and A155. No strain solubilizes aluminum phosphate. Three strains did not grow in the presence of Cd (A333, A334, A163), but A226, A332, and A423 stand out showing high growth with Cd. All strains grew in the presence of Cu, Zn, and Vinasse. A163, A164, and A328 were even stimulated by Cu addition. A331 and A335 showed a marked growth decrease with Zn. All strains are highly adapted to grow in vinasse presence. A257 and A226 produced the highest amount of IAA. The most biotechnological potential strains are indicated by presenting high growth under Cd, Cu, Zn, and vinasse stress, associated with a high tolerance index to these pollutants, further calcium phosphate solubilization, or IAA production.

The occurrence of mycothalli, symbioses between liverworts and fungi, is poorly documented in sub-Antarctica, and biogeographical patterns in Serendipita, the main fungal genus forming the symbiosis, remain understudied. Here, 83 specimens of 16 leafy liverwort species were sampled from sub-Antarctic South Georgia and were examined for mycothalli. Microscopy was used to enumerate fungal structures in liverwort tissues, and sequencing of fungal ribosomal DNA was used to determine the taxonomic and biogeographical affinities of the fungi. Stained hyphal coils, a defining feature of the symbiosis, were found to be frequent (>40% of stem length colonised) in Barbilophozia hatcheri, Cephaloziella varians and Lophoziopsis excisa. A single species of Serendipita, based on a 3% cut-off for ITS2 region sequence divergence, was a frequent colonist of these liverworts. A further 18 basidiomycete and ascomycete taxa colonised other liverwort species. The presence of the Serendipita species was positively associated with the occurrence of stained hyphal coils in stem epidermal cells. Phylogenetic analyses, incorporating worldwide accessions from leafy liverwort-associated Serendipita, showed that the same species, which also occurs in Chile, mainland Europe and on Svalbard, is apparently the sole symbiont of sub- and maritime Antarctic leafy liverworts, and indicated much higher species richness of the genus outside Antarctica., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

We describe a rare case of cutaneous phaeohyphomycosis by Veronaea botryosa in a renal transplant recipient from India. The patient presented with a nodule on the dorsum of right hand which resolved completely after 6 months of voriconazole therapy. The identity of the fungus was confirmed by sequencing internal transcribed spacer (ITS) region of rDNA. The case depicts its ubiquitous presence worldwide., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 SFMM. Published by Elsevier Masson SAS. All rights reserved.)

Leptosphaeria maculans is one of the major fungal pathogens on oilseed rape (Brassica napus), causing stem canker disease. The closely related Brassica species B. nigra, B. juncea, and B. carinata display extreme resistance toward stem canker. In this study, we demonstrate the nonhost status of B. carinata toward L. maculans in France through field experiments and inoculations performed in controlled conditions. A few isolates moderately adapted to B. carinata in controlled conditions were recovered in the field on B. nigra leaves, allowing us to investigate the unusual B. carinata-L. maculans interactions using molecular, macroscopic, and microscopic analyses. A cross between a L. maculans isolate adapted to B. napus and an isolate moderately adapted to B. carinata allowed the generation, in the lab, of recombinant L. maculans strains better adapted to B. carinata than the natural parental isolate obtained from B. nigra, and highlighted the polygenic determinism of the adaptation of L. maculans to B. carinata and B. napus. This biological material will allow further investigation of the molecular determinants of the adaptation of L. maculans to nonhost species and elucidate the genetic resistance basis of B. carinata., (© 2024 The Author(s). Environmental Microbiology Reports published by John Wiley & Sons Ltd.)

Lichens are composite organisms found throughout temperate terrestrial forests, with species-specific associations with industrial air pollution. Metagenomic analysis of lichen samples requires robust nucleic acid extraction methodology, a process that is challenging due to the protective cortex layers, high polysaccharide content, and the vast diversity of the internal microbiome. Our method includes physical lysis through garnet bead beating, chemical lysis using a sodium dodecyl sulfate buffer, phenol:chloroform:isoamyl alcohol extraction, and ethanol precipitation. The method was tested on three different lichen samples from two distinct species and yielded metagenomic DNA suitable for sequencing and PCR amplification. This procedure addresses the issues associated with DNA extraction from lichen using common laboratory equipment and reagents without the utilization of liquid nitrogen. This paper presents a cost-effective and accessible DNA extraction method for obtaining high-quality genetic material from dried and preserved lichen specimens., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Diplodia sapinea is the causal agent of Diplodia shoot blight, an emerging disease affecting pine forests worldwide. The range expansion of this pathogen in northern Europe has been suggested to be partially facilitated by recent warmer conditions. Although D. sapinea has been studied extensively, critical aspects of its infection biology and population structure remain unexplored. In this study, we developed nine simple sequence repeat (SSR) markers mined from D. sapinea genomes to assess the genetic diversity at higher resolution. Isolates from northern Spain, an area formerly regarded as having low genetic diversity and samples from a Californian population that was formerly regarded as clonal, were analysed in the study. In Spain, the nine SSR markers identified 56 genotypes in 285 samples. Isolates from symptomatic shoots, cones and asymptomatic tissues collected from different stands, suggested admixture between local populations. The same genotype tended to dominate within a single cone, and the same genotypes were usually found in both symptomatic and asymptomatic shoot tissues. The nine new SSR markers developed in this study revealed a high level of genetic diversity in both the northern Spanish and northern Californian populations than previously anticipated. Analyses using these nine SSR markers should contribute to a better understanding of the epidemiology, evolution and origin of D. sapinea, a pathogen that is gaining prominence in many parts of the world., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Lomentospora prolificans is an uncommon cause of invasive fungal disease, but it is associated with high mortality because it is difficult to treat. Most of severe cases are produced in immunossupressed patients, especially in those with neutropenia and/or hematological malignancies. Resistance to the majority of antifungal agents can be still observed. Here we report two cases of L. prolificans fungemia with different outcome, since in one of these patients treatment with one of the new antifungals could be applied. Both patients were treated with different antifungal drugs, but only the second one survived due to therapy with fosmanogepix®. The current treatment is still based on a combination of conventional antifungal drugs, although in much cases this strategy is not sufficient. The introduction of new promising antifungal agents such as fosmanogepix® and olorofim® may open new perspectives in the treatment of invasive infections caused by L. prolificans, as in our patient., Competing Interests: Declaration of competing interest The authors declare no competing interest, (Copyright © 2024 Elsevier Inc. All rights reserved.)

A novel enrofloxacin-degrading fungus was isolated from a rhizosphere sediment of the submerged macrophyte Vallisneria spiralis L.. The isolate, designated KC0924g, was identified as a member of the genus Humicola based on morphological characteristics and tandem conserved sequence analysis. The optimal temperature and pH for enrofloxacin degradation by strain KC0924g were 28 °C and 9.0, respectively. Under such condition, 98.2% of enrofloxacin with an initial concentration of 1 mg L -1 was degraded after 72 h of incubation, with nine possible degradation products identified. Four different metabolic pathways were proposed, which were initiated by cleavage of the piperazine moiety, hydroxylation of the aromatic ring, oxidative decarboxylation, or defluorination. In addition to enrofloxacin, strain KC0924g also degraded other fluoroquinolone antibiotics (ciprofloxacin, norfloxacin, and ofloxacin), malachite green (an illegal additive in aquaculture), and leucomalachite green. Pretreatment of cells of strain KC0924g with Cu 2+ accelerated ENR degradation. Furthermore, it was speculated that a flavin-dependent monooxygenase was involved in ENR degradation, based on the increased transcriptional levels of these two genes after Cu 2+ induction. This work enriches strain resources for enrofloxacin remediation and, more importantly, would facilitate studies on the molecular mechanism of ENR degradation with degradation-related transcriptome available., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: All authors have read the manuscript and agreed to its publication. Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Sclerotinia sclerotiorum, the necrotrophic cosmopolitan fungus, has become an emerging and re-emerging pathogen in the subtropical regions. Genetic diversity of 36 isolates of the fungus isolated from infected samples collected from the eastern and North eastern states was carried out using UP-PCR and SSR. Virulence variability was analysed based on four different measures. Among the eight UP-PCR primers and various combinations used, L-21, 3-2 and AA2M2-AS4 generated maximum number of fingerprints (13, 13 and 12, respectively) ranging from 100bp to 1kb. The isolates exhibited varied level of aggressiveness; majority (77.78%) were moderately virulent, 8.33% (22.22% of Assam and 6.67% of West Bengal) isolates were highly virulent, and 13.89% were less virulent. Several amplification products viz., 500bp generated by AA2M2-AS4, 150bp by AA2M2-L-21 and 100bp by L-21-3-2 were positively correlated with disease severity grading at 5% level of significance, whereas, 600bp band generated by AA2M2-3-2 was correlated at 1% level of significance. This indicates presence of these bands in highly virulent isolates. Out of the eight SSR primers, TATG9 did not generate any amplification and the isolates were divided into two major groups; the group II contained single isolate from Nagaland (NG4) indicating it to be genetically diverse from rest of the isolates. The subgroup A of the major group I was the largest and most diverse group with 11 members indicating genetic admixture within different geographic populations with different levels of similarity (70-100%). Genetic diversity based on SSR banding pattern showed highest value of Nei's gene diversity and Shannon's index of diversity (%pb = 61.11; h = 0.219; I = 0.330) for the Nagaland population with 9 members followed by West Bengal population with 15 members. Nei's genetic distance of all the tested populations was low, ranging from 0.0014 to 0.2350; however, genetic identity was high ranging from 0.7905 to 0.9986. The findings suggest that the pathogen populations of eastern and North eastern region were predominantly clonal with some evidence of infrequent out crossing., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Borah et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Introduction: Phaeohyphomycoses are uncommon and poorly understood opportunistic fungal infections, characterized by a wide spectrum of clinical manifestations ranging from localized skin lesions to disseminated disease. Most frequent genera are Alternaria, Cladophialophora, Exophiala or Curvularia. Less common ones, such as Verruconis gallopava, initially described as responsible of encephalitis of turkeys, pose significant challenges for diagnosis and treatment., Material and Methods: Following the description of a clinical case, we performed a comprehensive review of 48 cases of V. gallopava infection, a rarely reported species from 1986 to 2024., Results: Solid organ transplant recipients and patients with hematological malignancies are the population most at-risk. Clinical presentation is nonspecific but can be divided in two main entities, pulmonary and cerebral localizations. This later is associated with a mortality rate over 80% and was significantly more frequently reported in liver transplant recipients (p = 0.03). When tested, ß-D-glucans were positive in all cases. Antifungal susceptibility testing demonstrated low MICs for amphotericin B and all azoles but isavuconazole and fluconazole. Clinical outcomes support the use of amphotericin B, voriconazole, itraconazole and posaconazole as valid treatment options., Discussion: It is not known whether the cerebral cases are primary or secondary to pulmonary lesions. The indolent pulmonary lesions should prompt a complete work-up including biopsy with pathology and mycology expertise since the differential diagnosis is a cancer lesion., Competing Interests: Declarations. Conflict of interest: No conflict of interest to declare regarding this study for all authors., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Fungal endophytes play an important role in improving the health and productivity of native and cultivated plant species. Despite their ecological and industrial importance, few eucalypt species have been studied in terms of their endophyte communities. We examined the seasonal and spatial dynamics of fungal leaf endophytes in the model species, Eucalyptus crebra (narrow-leaved ironbark), using ITS-based amplicon sequencing. Alpha and beta diversity analyses revealed significantly higher species richness in summer compared to autumn and spring. Similarly, two-way ANOVA analysis showed significantly higher species diversity in summer compared to autumn (observed p < 0.001, Chao1 p < 0.005) and spring (observed p < 0.005, Chao1 p < 0.005). No difference in Shannon index was observed among different canopy levels across the season. Beta-diversity showed differences in fungal composition across the seasons and at various canopy levels based on unweighted UniFrac distance metric (PERMANOVA season p < 0.001, canopy p < 0.05), signifying distinct separation of fungi based on presence-absence. Ascomycota was the most abundant and diverse phylum and was present throughout the year. In contrast, Basidiomycota was only observed during cooler and drier seasons. Neofusicoccum was the most abundant genus, but distribution fluctuated significantly across the seasons. Pestalotiopsis and Neopestalotiopsis were most abundant in the low leaf canopy, whereas Pseudosydowia was most abundant in the high canopy. This study indicates that the diversity and abundance of endophytic fungi in the leaves of healthy E. crebra trees fluctuate seasonally and across canopy levels. The data generated can be used as a baseline for assessing and potentially modulating the health of E. crebra and other important Eucalyptus spp., Competing Interests: Declarations Competing Interests The authors declare no competing interests., (© 2024. The Author(s).)

A new species of Helicascus satunensis sp. nov. was collected on mature dead fruits of the Nypa palm in Satun Province, southern Thailand. Its morphological characteristics are similar to those of the genus Helicascus . Recently, a genus Helicascus with three species from marine habitats worldwide was studied. The morphology of this fungus was investigated and combined with multigene sequence analyzes of small subunit (SSU), large subunit (LSU), internal transcribed spacer (ITS) ribosomal DNA, translation elongation factor 1-alpha (TEF-1 α ) and RNA polymerase II (RPB2) genes. Morphologically, H. satunensis sp. nov. is characterized by semi-immersed, lenticular ascomata, multilocules, a bitunicate ascus and smooth, obovoid, dark brown ascospores that are one-septate and unequally two-celled. In addition, 3D visualization using synchrotron X-ray tomography was performed to investigate the interaction between fruiting body and substrata. Molecular phylogeny with multigene revealed that H. satunensis sp. nov. belongs to the family Morosphaeriaceae, order Pleosporales, class Dothideomycetes. Furthermore, H. satunensis sp. nov. forms a well-supported clade with Helicascus species described from marine habitats. Based on the unique morphological and molecular evidence, we propose this fungus, H. satunensis sp. nov., as a new species for Helicascus ., Competing Interests: The authors declare there are no competing interests., (©2024 Preedanon et al.)

Efficient early pathogen detection, before symptom apparition, is crucial for optimizing disease management. In barley, the fungal pathogen Pyrenophora teres is the causative agent of net blotch disease, which exists in two forms: P. teres f. sp. teres ( Ptt ), causing net-form of net blotch (NTNB), and P. teres f. sp. maculata ( Ptm ), responsible for spot-form of net blotch (STNB). In this study, we developed primers and a TaqMan probe to detect both Ptt and Ptm . A comprehensive k-mer based analysis was performed across a collection of P. teres genomes to identify the conserved regions that had potential as universal genetic markers. These regions were then analyzed for their prevalence and copy number across diverse Moroccan P. teres strains, using both a k-mer analysis for sequence identification and a phylogenetic assessment to establish genetic relatedness. The designed primer-probe set was successfully validated through qPCR, and early disease detection, prior to symptom development, was achieved using ddPCR. The k-mer analysis performed across the available P. teres genomes suggests the potential for these sequences to serve as universal markers for P. teres , transcending environmental variations.

A leaf spot disease affecting Curcuma kwangsiensis (Zingiberaceae) has been observed in Qinzhou City, Guangxi Province. Infected leaves exhibit yellow-brown spots that progressively expand and eventually lead to leaf death. Curvularia isolates were obtained from the diseased leaves with tissue isolation and single spore purification methods. To accurately identify these isolates, we analyzed their morphological characteristics and phylogenetic relationships using combinations of ITS, GAPDH, and EF-1α gene sequences. Phylogenetic analysis showed that the investigated strains formed a distinct clade separate from other recognized Curvularia species. Furthermore, the strains exhibited differences in conidiophore size and conidia shape/size. Based on phylogenetic studies, morphology, and pathogenicity tests, the pathogen was identified as a new species named Curvularia qinzhouensis. Optimal conditions for mycelial growth were observed at 30 °C and pH 8. The sensitivity of the pathogen to various phytochemicals was also examined. Honokiol, thymol, and citral demonstrated effective antifungal effects, with EC 50 values of 6.72 ± 1.75, 25.74 ± 4.30, and 54.24 ± 4.69 µg/ml, respectively. The present investigation provides the first report of leaf spot disease on C. kwangsiensis caused by C. qinzhouensis, and valuable insights for the prevention and control of this disease., (© 2024. The Author(s).)

Ascochyta blight caused by the ascomycete Ascochyta rabiei poses a major biotic threat to chickpea ( Cicer arietinum ) industries worldwide and incurs substantial costs to the Australian multimillion-dollar chickpea industry in both disease control and yield loss. The fungus was introduced to Australia in the 1970s from an unknown source population and, within a few decades, successfully established in all Australian agroecological chickpea-growing regions. Although genetically highly clonal, a broad range of phenotypic variation in terms of aggressiveness exists among the Australian A. rabiei isolates. More recently, highly aggressive isolates capable of causing severe disease symptoms on moderate to highly resistant chickpea cultivars have increased in frequency. To identify genetic loci potentially associated with A. rabiei aggressiveness on Australian chickpea cultivars, we performed deep genome sequencing of 230 isolates collected from a range of agroecological chickpea-growing regions between 2013 and 2020. Population genetic analyses using genome-wide SNP data identified three main clusters of genetically closely related isolates in Australia. Phylogenetic analyses showed that highly aggressive phenotypes developed multiple times independently throughout the phylogeny. The results point to a minor contribution of multiple genetic regions and most likely epigenomic variations to aggressiveness of A. rabiei isolates on Australian chickpea cultivars.

Aim. Melanized fungi were rarely studied for their antifungal resistance (AFR) or clinical outcome, despite rising incidence of melanized fungal ocular infections and AFR in general. We report the antifungal resistance patterns, clinical outcome and clinico-microbiological correlation in two commonly isolated melanized fungi from ocular infections, Curvularia lunata and Lasiodiplodia theobromae , at a tertiary eyecare centre in South India. Gap statement . Despite melanized fungi accounting for a significant proportion of ocular fungal infections in the Indian subcontinent, and despite there being a limited selection of effective antifungal agents available for these infections, the existing data and studies on these issues remain sparse. Therefore, this study aimed to investigate the prevalence of antifungal resistance in two of the most common melanized fungal pathogens in ocular infections, Curvularia lunata and Lasiodiplodia theobromae and correlate it with the treatment given and the clinical outcome in patients. Methodology. Electronic medical records provided the clinical data. Standard broth microdilution was performed for antifungal susceptibility testing (AFST) in 30 isolates (17 C . lunata and 13 L . theobromae ) for amphotericin B and natamycin (polyenes): voriconazole, ketoconazole, posaconazole, itraconazole and fluconazole (azoles) and caspofungin (echinocandin). Multidrug resistance (MDR) was defined as resistance to more than or equal to two classes of antifungals. DNA sequencing was performed for the isolates for species confirmation. The multivariate analysis was done for determining poor prognostic factors. Results. AFST showed highest susceptibility of study isolates for voriconazole (83.3% isolates), followed by natamycin (80%), fluconazole (80%), itraconazole (76.7%), ketoconazole (70%), posaconazole (66.7%), caspofungin (66.7%) and lastly amphotericin B (63.3%). All patients empirically received topical natamycin; additional oral ketoconazole/intraocular voriconazole was administered in select few. MDR was strongly associated with poor clinical outcome (multivariate analysis: P = 0.03, odds ratio = 7.8). All patients had microbial keratitis, one progressed to endophthalmitis. Additionally, therapeutic penetrating keratoplasty was required in 40% of cases. Globe salvage was possible in 80% patients, though good visual outcome was seen in only half of them. Both, anatomical and visual outcomes, were poor in 20% of patients. DNA sequencing showed C. lunata as the highest study species. Conclusion. C. lunata and L. theobromae showed varying in vitro antifungal susceptibility and clinical outcome in ocular infections. Voriconazole had significantly higher activity, while amphotericin B had lower activity in vitro for these melanized fungi. MDR isolates showed poorer clinical outcome.

Ectomycorrhizal fungi in the genus Tuber form hypogeous fruiting bodies called truffles. Many Tuber species are highly prized due to their edible and aromatic ascomata. Historically, there has been attention on cultivating and selling European truffle species, but there is growing interest in cultivating, wild-harvesting, and selling species of truffles endemic to North America. North America has many endemic Tuber species that remain undescribed, including some that have favorable culinary qualities. Here, we describe two such Tuber species from eastern North America. Maximum likelihood and Bayesian phylogenetic analyses of ITS (internal transcribed spacer), tef1 (translation elongation factor 1-alpha), and rpb2 (second largest subunit of RNA polymerase II) sequences were used to place these species within a phylogenetic context. We coupled these data with morphological analyses and volatile analyses based on gas chromatography-mass spectrometry. Tuber cumberlandense , sp. nov. (previously referred to as Tuber sp. 66), is a member of the Rufum clade that has been opportunistically harvested for commercial sale from T. melanosporum orchards across eastern North America. Tuber canirevelatum , sp. nov. belongs in the Macrosporum clade and thus far is only known from eastern Tennessee, USA. Both new species were discovered with the assistance of trained truffle dogs. The volatile profiles of T. canirevelatum and T. cumberlandense were measured in order to characterize aromas based on the chemical compounds produced by these fungi. Ascomata from both species were enriched in acetone, dimethyl sulfide, 1-(methylthio)-1-propene, and 1-(methylthio)propane. In this work, we celebrate and encourage the use of trained truffle-hunting dogs for fungal biodiversity discovery and research.

Morchella helvetica , sp. nov. ( Morchella sect. Distantes ) is a new species of true morels discovered in Switzerland. It is formally described in the present study using an integrative approach based on micro- and macromorphological characteristics, multilocus phylogenetics, and a brief description of its habitat. Molecular analyses clearly indicated that Morchella helvetica is a sister species to M. eximioides, M. angusticeps , and M. confusa . It can be distinguished by the two phylogenetic markers RNA polymerase II subunit 2 ( RPB2 ) and translation elongation factor-1 alpha (TEF1-α ). In addition, M. helvetica exhibits particular morphological features, notably the presence of pale hairs on the pileus, a mealy stipe, and darkening ridges when aging.

Background: Cladophialophora bantiana is one of the most virulent phaeohyphomycetes, typically causes non-angiogenic single (or sometimes multiple) cystic brain lesions, and has resulted in a mortality rate of up to 70%. Most C bantiana cases are described either in a series of isolated reports or in very small cohorts. The aim of this retrospective nation-based study was to share the data on C bantiana phaeohyphomycosis cases reported in France and French overseas territories over the past two decades to improve understanding of this disease., Methods: Patients with C bantiana infection were processed through the active surveillance programme of invasive fungal infections launched by the National Reference Center for Mycoses and Antifungals, Institut Pasteur (Paris, France), and the French Surveillance Network of Invasive Fungal Infections, which involved 29 hospitals from mainland France and overseas French territories. Only proven and probable cases of infection, according to the revised and updated consensus definitions from the European Organization for Research and Treatment of Cancer and Mycoses Study Group, were included in the study. Patients were diagnosed or confirmed, or both, using a polyphasic approach at the Institut Pasteur between 2002 and 2022. Patients were separated into two groups: those with CNS involvement and those with no CNS involvement. The primary outcome was the survival rate., Findings: A total of 23 patients with a C bantiana invasive infection were included during the study period (Jan 1, 2002, to Dec 31, 2022). The median age was 56 years in the CNS involvement group and 65 years in the non-CNS involvement group. Until 2021, the annual number of cases varied between zero and two, with six cases observed in 2022, the warmest year recorded in France since 1900. CNS involvement was observed in 15 (65%) patients, including three disseminated cases; skin and soft tissue involvement in seven (30%) patients and an isolated lung infection in one case. Diabetes was observed in five patients, and any immunodepression factor was observed in 14 (61%) of 23 patients. When considering only patients with CNS involvement, 9-month survival appeared higher in patients who underwent exeresis or large drainage (three [75%] of four patients vs three [27%] of 11 patients; p=0·24) and significantly higher in those treated for 2 or more weeks with triple antifungal therapy (liposomal amphotericin B plus posaconazole and flucytosine; seven [78%] of nine patients vs one [17%] of six patients; p=0·040). Two patients were treated with excision surgery alone (one patient with success, and the other patient lost to follow-up)., Interpretation: This study shows that the clinical presentations and underlying medical conditions of C bantiana infections are more diverse than previously described. It also emphasises a significant difference in mortality rate between those with and without CNS involvement. The prognosis improved when surgery was performed and triple antifungal therapy was administered. Such rare and devastating invasive fungal infections should be managed by a multidisciplinary team., Funding: Santé Publique France., Competing Interests: Declaration of interests We declare no competing interests., (Copyright © 2024. Published by Elsevier Ltd.)

An intense black pigmented halotolerant yeast GUBPC1, was obtained from the solar salterns of Nerul, Goa-India. The isolate could tolerate 0 to 20 % NaCl. FE-SEM analysis revealed its polymorphic nature, exhibiting oval cells at higher salt concentrations and filamentous spindle like shapes at lower concentrations. Initially, the cells appear oval, yeast like in shape but gradually after 15 days of incubation, it becomes elongated and undergoes budding, exhibiting various budding patterns, from single polar bud to bipolar buds with annellidic ring, to lateral buds and eventually forming filamentous hyphae. The intracellular black pigment was identified as melanin based on ultraviolet-visible spectroscopy analysis. The molecular identification of the culture showed closest similarity with Hortaea werneckii. Plant polymer-degrading enzymatic activities such as cellulase, laccase, chitinase, xylanase, pectinase, amylase and protease were exhibited by the isolate GUBPC1. To further understand and explore its biotechnological potential, we performed whole-genome sequencing and analysis. The obtained genome size was 26.93 Mb with 686 contigs and a GC content of 53.24 %. We identified 9383 protein-coding genes, and their functional annotation revealed the presence of 435 CAZyme genes and 16 functional genes involved in secondary metabolite synthesis, thus providing a basis for its potential value in various biotechnological applications., (Copyright © 2024 British Mycological Society. Published by Elsevier Ltd. All rights reserved.)

Boeremia was established to accommodate phoma-resembling fungi. Its species occur in terrestrial ecosystems as endophytes, saprobes and pathogens, except one species reported from a marine ecosystem. Boeremia species are characterized by hyaline, thin-walled, and aseptate (occasionally 1(-2)-septate) conidia that are variable in shape, and hyaline, straight or slightly curved, thick-walled, and 1-septate ascospores that are usually constricted at the septum. In the past, host associations were used to delimit Boeremia species. However, since Boeremia taxa have overlapping morphological characters and are cryptic, it renders taxonomic identification arduous. Therefore, the use of other approaches including multi-gene phylogenetic analyses are imperative. Recommended DNA markers for species delineation are the internal transcribed spacer (ITS, nuclear rDNA consisting of ITS1-5.8S-ITS2) and large subunit (28S, D1-D2 domains of nuclear 28S rDNA) loci, and the genes for actin (ACT1), beta-tubulin (TBB1), RNA polymerase 2 (RPB2) and translation elongation factor 1α (TEF1). Here, we applied morphological and molecular phylogenetic analyses to establish a new taxon (B. albae), and a new host and geographical record for B. maritima associated with leaf spots of Morus alba (Moraceae) in northern Thailand. By providing sequence data for three additional gene regions, our phylogenetic analyses impart a stable phylogenetic placement of the ex-type strain of B. maritima, as illustrated. This is the first study that reports Boeremia species from M. alba, and B. maritima from a terrestrial habitat., Competing Interests: Declaration of competing interest On behalf of all co-authors, I confirm that the submitted research is original and has not been published, nor is it currently under consideration for publication elsewhere. We have no conflicts of interest to disclose., (Copyright © 2024 British Mycological Society. Published by Elsevier Ltd. All rights reserved.)

Leaf blotch, caused by Zymoseptoria tritici, is a fungal disease that poses a severe threat to wheat production worldwide. Knowledge of virulence variability is crucial in choosing effective control measures. However, there have only been a few studies of the pathogenic variability and pathotypes within Ethiopian isolates. Hence, the objective of this study was to assess the virulence spectrum and variability of Z. tritici isolates. Forty-three isolates were tested for their virulence and pathotype against 7 wheat differential lines that have different resistance genes. A pathogenicity assay detected 41 differential line-specific virulent isolates among 301 interactions between a host and pathogen based on the percentage coverage of the leaf area by pycnidia. Some isolates were virulent against 50 %-60 % of the resistant genes, but most of them were virulent against some differential lines. Isolates such as EtA-11, EtSh-1, EtSh-2, EtSh-4, and EtA-19 expressed broad-spectrum virulence, highlighting that such isolates are useful for germplasm screening. The isolates were classified into 25 pathotypes, defined by their differential virulence responses. They were also assigned to two clusters according to their mean pycnidia percent. Pathotypes and principal component analysis detected 58.1 % and 62.2 % pathogenic diversity in Ethiopian isolates, respectively. The current findings provide information that breeders can use to identify and select more resistant varieties for farmers., Competing Interests: Declaration of Competing interest The authors state that there is no conflict of interest., (Copyright © 2024 British Mycological Society. Published by Elsevier Ltd. All rights reserved.)

Ambrosia beetles (Coleoptera: Curculionidae: Scolytinae) are small wood-boring insects that live in an obligate symbiosis with fungi, which serve as their primary food source. Beetles residing in the genus Euwallacea have evolved a unique association with a clade of Fusarium that falls within the aptly named Ambrosia Fusarium Clade (AFC). The discovery of the invasive polyphagous shot hole borer, E. fornicatus , in South Africa, has heightened awareness of ambrosia beetles and their symbionts in the country. In this study, we investigated the Fusarium symbionts of three species of Euwallacea in South Africa, with a specific focus on those associated with E. xanthopus . Isolations of Fusarium strains from both living and dissected beetles yielded nearly 100 isolates. Using multigene phylogenetic analyses, these isolates were identified as six different Fusarium species. Fusarium hypothenemi and F. euwallaceae have previously been reported from South Africa. Fusarium pseudensiforme and Fusarium AF-6 are new records for the country. The remaining two species are new to science and are described here as F. rufum sp. nov. and F. floriferum sp. nov. Targeted fungal isolation from specific beetle body parts revealed that the AFC species collected were typically associated with the dissected beetle heads and helped us identify the likely nutritional symbiont of E. xanthopus . This study highlights the understudied diversity of fungal associates of ambrosia beetles present in South Africa.

Tea (Camellia sinensis), a perennial evergreen shrub, is one of the most important cash crops in China. Tea leaves with symptoms of wilt disease was observed in Fengqing County, Lincang City, Yunnan Province, China. Large irregular jujube-red necrotic spots appeared on the leaves of tea plants, and the lesions with grayish white edge were accompanied by a certain degree of shrinkage. In the tea garden planting base, the natural disease incidence reached 40%-50 %, which significantly affects the yield of tea. One putative pathogen was isolated from three symptomatic tea plant leaves and was identified as Discosia brasiliensis using morphology and molecular phylogeny of multi-loci (ITS, LSU, tub, rpb2) sequence data. Using D. brasiliensis strains for artificial inoculation assay on the tea plant leaves, leaf atrophy symptom in leaves which is similar to those observed in the tea planting base, and the putative pathogen was re-isolated to fulfill Koch's postulates. This is the first report of wilt disease caused by Discosia brasiliensis in China., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Yong Wang reports financial support was provided by College of Agriculture, Guizhou University. Yan Li reports a relationship with College of Life Sciences, Guizhou University that includes: consulting or advisory. Jiaojiao Wu has patent pending to Yong Wang. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Soybean red leaf blotch (RLB), caused by the fungus Coniothyrium glycines , represents a foliar disease of soybean that is thus far restricted to Africa. The fungus is listed as a Select Agent by the Federal Select Agent Program because it could pose a severe threat to plant health were it to establish in the United States. Previous work uncovered tremendous molecular diversity at the internal transcribed spacer region, suggesting that there may be multiple species causing RLB. To determine whether multiple species cause RLB, we reconstructed the phylogeny of C. glycines and taxonomic allies using sequence data from four genes. We included 33 C. glycines isolates collected from six African countries and determined that all isolates form a well-supported, monophyletic lineage. Within this lineage there are at least six well-supported clades that largely correspond to geography, with one clade exclusively composed of isolates from Ethiopia, another exclusively composed of isolates from Uganda, and four composed of isolates from southern Africa. However, we did not detect any concordance for these clades between the four genes, indicating that all isolates included in this analysis are representative of a single species. Isolates in the Ethiopia clade are morphologically distinct from isolates in the other clades, as they produce larger sclerotia and smaller pycnida and more sclerotia in planta. Additionally, ancestral range estimations suggest that the C. glycines lineage emerged in southern Africa. These results show that there is significantly more genetic and morphological diversity than was initially suspected with this high-consequence fungal plant pathogen.

Dypsis lutescens, commonly known as areca palm, is a highly valued ornamental species due to its aesthetic value. However, the foliage is vulnerable to various pathogens, particularly those responsible for fungal leaf spot diseases. In October 2023, a severe incidence (93 %) of destructive leaf spots was recorded on Dypsis lutescens at the University of Agricultural Sciences, GKVK, Bangalore, and surrounding areas. The leaf spot symptoms manifested as frog-eye-like lesions, leading to complete leaf desiccation and significantly reducing the palms ornamental value. The pathogen exhibited the highest radial growth (90.00 mm) and prominent sporulation on oat meal agar, whereas Richard's synthetic agar resulted in the lowest radial growth (38.00 mm) with no sporulation. Morphological and multilocus phylogenetic analyses confirmed the pathogen as Bipolaris heliconiae. Pathogenicity tests fulfilled Koch's postulates, confirming that Bipolaris heliconiae is the causative agent of leaf spot disease in Dypsis lutescens in India. This novel finding underscores the emergence of a new disease and highlights the urgent need for effective management strategies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this research paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Bioherbicides are expected to be a supplement to integrated pest management, assisting in the control of problematic weed species. For instance, bioherbicides (Phoma and BioPhoma) were recently registered in Canada and the USA for the control of some perennial dicotyledonous weeds in lawns. These products are based on strains of the fungus Didymella macrostoma (syn. Phoma macrostoma ) that causes white tip disease (WTD) in Canada thistle ( Cirsium arvense ). In this study, WTD was reported for the first time in the Russian Federation. Analysis of the internal transcribed spacer (ITS) region of nuc rDNA and secondary metabolite profiling confirmed the identity of Russian WTD isolates to Canadian biocontrol strains identified as D. macrostoma . Multilocus phylogenetic analysis based on sequencing of the ITS region, partial large subunit nuc rDNA region (28S), RNA polymerase II second largest subunit gene ( rpb2 ), and partial β-tubulin gene ( tub2 ) has differentiated the WTD isolates from C. arvense and D. macrostoma isolates from other plant hosts. Based on phylogenetic, morphological, and chemotaxonomic features, these WTD isolates were described as a new species named Didymella baileyae , sp. nov. This study also demonstrated the low pathogenicity of the ex-type D. baileyae isolate VIZR 1.53 to C. arvense seedlings and its asymptomatic development in the leaves of aboveground shoots. The organic extracts from mycelium and culture filtrate of D. baileyae , as well as macrocidin A and macrocidin Z, displayed phytotoxicity both to C. arvense leaves and seedlings. Macrocidin A was only detected in the naturally infected leaf tissues of C. arvense showing WTD symptoms. Macrocidins A and Z demonstrated low antimicrobial and cytotoxic activities, exhibiting no entomotoxic properties. The data obtained within this study on the pathogenicity and metabolites of D. baileyae may be important for the rational evaluation of its prospects as a biocontrol agent.

Several members of the genus Peziza sensu stricto occur at the edge of melting snow. These nivicolous species have been widely reported in the Northern Hemisphere and are also known from Australia and New Zealand. We have used 16 specimens from North America and Australia to study morphology and to perform DNA sequencing. In sequence analyses, we have used ITS1 and ITS2 (internal transcribed spacers), 28S, RPB2 (RNA polymerase II gene), and two genes new to these studies, GAPDH (glyceraldehyde-3-phosphate dehydrogenase) and HSP90 (heat shock protein 90). Although not all regions are available for all samples, we have recognized the following species: Peziza heimii, P. nivalis , and P. nivis . Phylogenetic analyses were done using ITS alone; combined ITS1-5.8S-ITS2, 28S, and RPB2 ; ITS, and 28S, RPB2, GAPDH , and HSP90 . Even with this augmented set of genes and despite their widespread occurrence in North America, Europe, Australia, and New Zealand, we have not definitively distinguished species within this group. To assess these results, pairwise homoplasy index (PHI) analysis was employed. This showed evidence of recombination among the samples of P. nivalis and further supports the view of P. nivalis as a monophyletic cosmopolitan species. As part of this study, we also examined the variation in ITS copies in P. echinospora , for which a genome is available.

Purpose: We report two cases of fungal keratitis due to rare melanized fungal pathogens in ocular infection, one is the first case report of keratitis due to Pseudopithomyces maydicus and the second is keratitis due to rare fungal pathogen in ocular infections Phialophora chinensis ., Method: Conventional mycology during routine diagnostics helped in identifying these rare fungal isolates, following which we proceeded for the confirmation of identification by DNA sequencing and did in-vitro antifungal susceptibility test to understand their susceptibility pattern. The clinical information for these two patients were collected from hospital electronic medical records., Results: We discuss the clinical presentation, treatment given, and clinical outcome in these patients and correlate these with the conventional microbiology and sequencing techniques, which helped in identifying the pathogen and the in-vitro antifungal susceptibility of these rare isolates. We also do a brief literature review for these two rare fungal pathogens., Conclusions: Pseudopithomyces maydicus and Phialophora chinensis are rare causes of fungal keratitis due to melanized fungi. Both of this fungal keratitis respond well to medical therapy alone if diagnosed and treated early.

Barley grass ( Hordeum leporinum ), which often occurs in proximity to commercial barley ( H. vulgare ) cultivars, is an alternative host to Pyrenophora teres , an economically important pathogen causing net blotch in barley. This study is the first to report the sexual recombination of P. teres isolates collected from barley with those collected from barley grass. The sexual recombination between P. teres isolates from barley and barley grass was confirmed using a neighbor-net network and haploblock plots based on whole-genome sequencing of seven progeny isolates. Pathogenicity assays revealed that P. teres isolates from barley grass were not host specific and could infect both barley and barley grass, and the progeny isolates were virulent on commercially grown barley cultivars. Our results contradict previous population and pathogenicity studies of P. teres isolates obtained from barley and barley grass that have reported that the two populations are genetically distinct and host specific, suggesting that isolates collected from barley or barley grass could be two different entities. Despite the genetic divergence of P. teres isolates from barley and barley grass revealed through our phylogenomic analysis, there seems to be no complete host or reproductive separation between these populations. Therefore, there is a potential for generation of novel pathotypes through sexual recombination between P. tere s isolates associated with barley and barley grass, with a risk of increased impacts on commercial barley cultivars that do not carry resistance to these pathotypes., Competing Interests: The author(s) declare no conflict of interest.

Cotton root rot caused by Macrophomina phaseolina pose a significant threat to cotton production, leading to substantial yield and quality losses. Early and accurate diagnosis of this pathogen in soil is crucial for effective disease management. This study presents a pioneering investigation into the utilization of the nit gene encoding nitrilase for the development of a molecular diagnostic assay aimed at the rapid detection of M. phaseolina in field soils. The methodology involved the design and validation of primers targeting the Nit gene sequence, followed by the optimization of PCR conditions for efficient amplification. Leveraging state-of-the-art molecular techniques, the assay offers a novel protocol to accurately identify the presence of M. phaseolina in soil with high sensitivity and specificity. The specificity of the designed primers was confirmed through PCR amplification using DNA from M. phaseolina and other related fungi. Sensitivity tests demonstrated that the PCR assay reliably detected M. phaseolina DNA at concentrations as low as 1 ng. Furthermore, the performance of the diagnostic assay was rigorously evaluated using field soil samples with a known status of M. phaseolina infection, demonstrating its reliability and efficacy in real-world scenarios. This study introduces a novel molecular marker for the detection of M. phaseolina and offers a rapid and efficient means for screening M. phaseolina in large soil samples with minimal time and manpower., (© 2024 Wiley‐VCH GmbH.)

Chlorosplenium is a small genus comprising five species of inoperculate discomycetes in the order Helotiales (Leotiomycetes) often recognizable by their bright yellowish-green colors and gregarious growth on wood. In this study, we describe five new species- C. aotearoa, C. australiense, C. cusucoense, C. epimorsicum , and C. hawaiiense -based on a combination of recent fieldwork and examination of previously collected fungarium specimens. We use an integrative taxonomic approach to support the distinction of new species, incorporating morphology and DNA sequence data with biogeography. Macro- and micromorphological features of apothecia for all species and culture characteristics for four of the five new species are documented. A multilocus phylogeny based on nuc rDNA internal transcribed spacer region ITS1-5.8S-ITS2, partial large subunit nuc ribosomal DNA (28S nuc rDNA), and A-B regions of the largest subunit of RNA polymerase II ( RPB1 ) gene is presented. Additionally, we report Chlorosplenium chlora from Europe for the first time and expand our knowledge of the diversity and distributions of species in this genus in America, Australia, and New Zealand.

In walnut orchards, frequent symptoms of cankers and dieback (fruit blight, twig and branch cankers up to tree death) are caused by different agents, in particular by Botryosphaeriaceae, primarily Neofusicoccum parvum and Botryosphaeria dothidea. This study aimed at developing a sensitive, rapid, specific and internally controlled multiplex PCR assay for the detection of these species. The ability of the multiplex PCR, with an internal inhibition control (i.e. E. coli DNA), to specifically and successfully detect members of the two targeted species was validated using 11 different isolates for each fungal target (inclusivity) and 20 tree-associated fungal species different from B. dothidea or N. parvum (exclusivity). Applicability to plant materials was further investigated and showed an absence of amplification for asymptomatic husk or twig samples while the amplification profiles of symptomatic tissues ranged from no amplification to amplification of both species, in correlation with the observed fungal contamination level. In conclusion, we developed a rapid diagnostic tool for simultaneous detection of two major fungal pathogens of walnut. Although this protocol was tailored for walnut husks and twigs, applicability to any plant sample contaminated by these pathogens can be considered., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Hulless barley sheath rot is a spike disease caused by Dactylobotrys graminicola. In recent years, it has generally occurred in hulless barley growing areas in China, resulting in reduced hulless barley yields. In this study, primers and probes were designed based on conserved genome sequences, and a method was established using recombinant enzyme polymerase amplification-lateral flow burette (RPA-LFD) technology for the rapid diagnosis of sheath rot in hulless barley. The method can be successfully established in five minutes at a constant temperature of 39℃, and the results are consistent with those of normal PCR analysis. The method demonstrated high sensitivity, with a detection limit of 10 fg/µL. Furthermore, the rapid method was able to successfully detect D. graminicola in hulless barley during field incubation, which highlighted the significant advantage of the method in practical applications. In conclusion, the RPA method established in this study exhibited several advantageous characteristics, including high efficiency, simplicity, rapidity and practicality, which provide a theoretical basis for the early detection and prevention of D. graminicola., (© 2024. The Author(s).)

Chrysosporium-related fungi refers to an assemblage of fungi belonging to the Nannizziopsis, Ophidiomyces, and Paranannizziopsis genera. Chrysosporium-related fungi infection results in various skin lesions, such as necrosis and ulcers, in both captive and free-roaming reptiles. To update the prevalence of ophidiomycosis in Taiwan, which was first detected in 2019, we conducted a large-scale ecological survey of free-roaming native snakes with skin lesions in Taiwan. To the best of our knowledge, this is the first study on Chrysosporium-related fungi prevalence in Southeast Asia. Fungal samples collected from the skin lesions of snakes were cultured and subjected to morphological, histopathological, and molecular analyses. We examined 2382 free-roaming snakes representing 42 snake species; among them, 132 (5.54%) had skin lesions. Ten (0.42%) snakes, representing four species, tested positive for Ophidiomyces (five snakes, four species) or a possibly novel Nannizziopsis species (five snakes, three species). The infected snakes were generally healthy, with mild clinical signs. The low prevalence rate, mild clinical signs, and broad pathogen/host range suggest that Ophidiomyces and Nannizziopsis are endemic to Southeast Asia. The newly reported presence of Nannizziopsis in free-roaming snakes suggests the need for modifying the currently used surveillance strategy., (© The Author(s) 2024. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology.)

This is the first report of a cutaneous infection in an immunocompromised domestic cat caused by Setosphaeria turcica. The investigation encompasses an assessment of its virulence factors and susceptibility to antifungal drugs. The isolated strain originated from a domestic cat displaying cutaneous lesions that tested positive for feline leukemia virus (FeLV) infection. Identification procedures employed both microscopic and molecular techniques, with molecular identification relying on ITS DNA sequencing. Enzymatic assays targeting lipase, phospholipase, protease, and keratinase yielded negative results, suggesting the prevalence of alternative virulence mechanisms. Successful treatment of the infection was achieved with itraconazole, and susceptibility testing confirmed its sensitivity to azoles and polyene antifungal drugs., (Copyright © 2024 The Authors. Publicado por Elsevier España, S.L.U. All rights reserved.)

Eucalyptus spp. in plantations are negatively affected by canker and wilt diseases caused by several species of Ceratocystis, particularly those in the Latin American Clade (LAC). Ceratocystis eucalypticola and Ceratocystis manginecans are of particular concern where disease epidemics are reported globally, with recent outbreaks emerging in South African and Indonesian Eucalyptus plantations. Consequently, a rapid screening protocol is required for these pathogens. In this study, a high-resolution melting curve analysis (HRMA) was developed to detect C. eucalypticola and C. manginecans that bypasses time-consuming isolation and post-PCR procedures. Primers targeting a 172 bp region of the cerato-platanin (CP) gene were designed. Using these primers, the accuracy of HRMA to detect and distinguish between these two LAC species was assessed using pure fungal DNA, and DNA extracted directly from Eucalyptus samples naturally infected with C. eucalypticola. The assay accurately detected the presence of C. eucalypticola and C. manginecans and quantifies their DNA, both from cultures, and directly from wood samples. HRMA further differentiated these two species from all other tested LAC individuals. This assay was also able to detect the presence of all the tested LAC species and distinguish seven of these, including C. fimbriata, to species level. Ceratocystis polyconidia was the only non-LAC off-target species detected. Based on these results, the developed assay can be used to rapidly identify C. eucalypticola and C. manginecans directly from infected plant material or fungal cultures, with the potential to also screen for several other LAC species., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Dollar spot is a major fungal disease affecting turfgrass worldwide and can quickly destroy turfgrass swards. An assimilating probe-based loop-mediated isothermal amplification (LAMP) assay was developed to detect Clarireedia monteithiana and C. jacksonii , the causal agents of dollar spot within the continental United States. Five LAMP primers were designed to target the calmodulin gene with the addition of a 6-carboxyl-fluorescein florescent assimilating probe, and the temperature amplification was optimized for C. jacksonii and C. monteithiana identification. The minimum amount of purified DNA needed for detection was 0.05 ng μl -1 . Specificity assays against host DNA and other turfgrass pathogens were negative. Successful LAMP amplification was also observed for dollar spot-infected turfgrass field samples. Further, a DNA extraction technique via rapid heat-chill cycles and visualization of LAMP results via a florescent flashlight was developed and adapted for fast, simple, and reliable detection in 1.25 h. This assimilating probe-based LAMP assay has proved successful as a rapid, sensitive, and specific method for the detection of C. monteithiana and C. jacksonii in pure cultures and from symptomatic turfgrass leaves blades. The assay represents a promising technology to be used in the field for on-site, point-of-care pathogen detection., Competing Interests: The author(s) declare no conflict of interest.

Competing Interests: Financial disclosures/conflicts of interest: None reported.

Background: Sparse leaf patch (SLP) is one of the most significant diseases affecting seashore paspalum (Paspalum vaginatum Sw.), caused by Microdochium paspali. Fast and accurate detection of this pathogen is crucial for effective disease management. However, conventional culture-based methods are time-consuming and often compromised by the presence of other saprophytic or endophytic fungi., Results: In this study, we developed a real-time fluorescent quantitative (q)PCR method based on the internal transcribed spacer (ITS) region of the ribosomal RNA gene to rapidly detect and quantify M. paspali. The qPCR assay demonstrated the ability to detect all 12 tested isolates of M. paspali, with no cross-reactions observed when tested against 30 isolates of other fungal pathogens from turfgrass samples. The detection limit of the qPCR method was as low as 3.65 × 10 2 copies μL -1 of M. paspali genomic DNA, and the entire detection process could be completed within 1 h. The fluorescence signal was detectable in the leaf tissues of seashore paspalum without apparent disease symptoms as early as 24 h postinoculation with M. paspali. Moreover, the qPCR method successfully detected M. paspali in both asymptomatic and symptomatic turfgrass samples, including leaf, stem, root and rhizosphere soil, indicating that this assay can significantly enhance the detection of M. paspali., Conclusion: The study developed a rapid real-time qPCR assay for the detection of M. paspali causing SLP on seashore paspalum and in environmental samples, which has important implications for early warning and management of SLP. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

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)., (© 2024. The Author(s).)

Endophytic fungi have been shown to synthesize bioactive secondary metabolites, some of which promote plant growth through various mechanisms. In our previous study, endophytic fungi were isolated from mango trees ( Mangifera indica L.). The present study examined fifty endophytic fungal isolates for mineral solubilization activity, ammonia production, and siderophore production. It was shown that these isolates could produce phytohormones indole-3-acetic acid and gibberellic acid, as well as inhibit plant pathogens, specifically Colletotrichum gloeosporioides and Lasiodiplodia theobromae . The results showed that all the isolated fungal endophytes exhibited various activities. Based on the findings, two fungal endophytes- Aureobasidium pullulans CY.OS 13 and Aspergillus tamarii CY.OS 144-were selected for dual inoculation in chili plants under pot-scale conditions to investigate their potential to improve growth-related traits such as seed germination, shoot and root length, biomass, and chlorophyll content. Seed treated with A. pullulans CY.OS 13 and/or A. tamarii CY.OS 144 showed a significant ( p < 0.05) increase in seed germination and growth parameters of chili plants grown under pot-scale conditions. Particularly, chili plants whose seeds were injected with a combination of the two selected endophytic fungi showed the highest plant development traits. Therefore, the selected endophytic fungi have the potential to be used as biofertilizers, especially when combined. They could eventually replace chemical fertilizers because they are environmentally friendly, beneficial to humans, and can even promote sustainable agriculture.

Wilt and stem rot (WSR) is an emerging syndrome threatening cut lisianthus (Eustoma russellianum) production in Lam Dong province, Vietnam. The disease was observed in all 13 inspected commercial lisianthus greenhouses across major lisianthus cultivation areas in Lam Dong, including Da Lat, Lac Duong, Don Duong, and Duc Trong, with incidence increasing with plant age, ranging from 7.5 to 32.4%. Infected plants displayed stunting, wilting, stem rot and blight, and dieback, with predominance of wilt and stem rot. The disease showed polycyclic behavior, with symptoms shifting from random or scattered in young plants to clustered patterns after the initial flower cutting. Forty-one Fusaria-like fungal isolates recovered from diseased lisianthus plants were identified as Fusarium vanleeuwenii (28 isolates), Neocosmospora solani (11 isolates), and F. annulatum (2 isolates) based on morphological observations and phylogenetic analysis of the internal transcribed spacer (ITS) region and translation elongation factor 1-alpha (TEF-1α) genes. The composition of Fusaria species varied across sites, with F. vanleeuwenii being consistently present. Pathogenicity tests confirmed that isolates of F. vanleeuwenii Li-Fo9511, N. solani Li-Fs4311, and F. annulatum Li-Fp3051 caused typical stem rot in in-vitro assays. In-planta assays showed wilting in seedlings starting two weeks post-infection, with a remarkable increase in disease incidence and severity between five and six weeks, particularly for F. vanleeuwenii Li-Fo9511. The pathogens were re-isolated and morphologically confirmed, fulfilling Koch's postulates. This is the first report of F. vanleeuwenii, N. solani, and F. annulatum as pathogens of lisianthus WSR in Vietnam, highlighting the need for effective control strategies., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The cajuzinho do cerrado (Anacardium humile-Anacardiaceae), a shrub species native to Brazil, is harvested for multiple uses in food and medicine. Members of a harvesting community, near the municipality of Bonito de Minas, Minas Gerais state, Brazil reported characteristic symptoms of shoot blight and dieback reducing pseudofruit and seed production by this plant. This study aimed to identify the etiological agent of this disease. Two fungal isolates were obtained from symptomatic leaf samples and morphologically and molecularly characterized. The fungus was identified, based on morphological analyses, as a probable new species of Pseudoplagiostoma. Phylogenetic analyses based on a combination of DNA sequence data (nuc rDNA ITS1-5.8S-ITS2 region, tef1-α and tub2), confirmed this hypothesis. The isolates obtained were allocated to a distinct, well-supported clade (IB = 0.99, ML = 100%), placed as a unique lineage here proposed as a new species named Pseudoplagiostoma humilis. The pathogenicity test confirmed that this new species was the causal agent of shoot blight and dieback on A. humile. This is the fourteenth Pseudoplagiostoma species reported in the world and the third in Brazil., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Maple is an important ornamental plant in China. With the increasing use of maple trees in landscaping, a symptom of shoot dieback has been observed in Henan province, China., Results: In this study, 28 Diaporthe isolates were obtained from symptomatic shoots of maple trees between 2020 and 2023. Phylogenetic analyses based on five loci (ITS, TEF, CAL, HIS and TUB) coupled with morphology of 12 representative isolates identified three known species (D. eres, D. pescicola and D. spinosa) and one new species, namely D. pseudoacerina sp. nov. Koch's postulates confirmed that all these species were pathogenic. Additionally, D. pseudoacerina was able to infect China wingnut (Pterocarya stenoptera), pear (Pyrus sp.), and black locust (Robinia pseudoacacia). This study marks the first report of Diaporthe spinosa and D. pescicola pathogens infecting maple trees., Conclusions: These findings enhance the existing knowledge of the taxonomy and host diversity of Diaporthe species as, while also providing valuable information for managing of maple shoot dieback in Henan Province, China., (© 2024. The Author(s).)

The diversity of endophytes and their ecological relationships with the endangered conifer Araucaria angustifolia (a critically endangered species) are unrevealed. This study aimed to characterize the diversity of endophytic fungi associated with A. angustifolia. To this end, we analyzed 90 fragments from five individuals collected from a mixed localized fragment in Guarapuava-PR, Brazil. The total DNA of 61 morphotypes was extracted and the Internal Transcribed Spacer (ITS) region was amplified and sequenced. The sequence analysis allowed the identification of 37 genera belonging to the phylum Ascomycota and the classes Eurotiomycetes, Dothideomycetes, and Sordariomycetes, divided into 11 orders and 13 families. Most of the isolated fungi belonged to the Sordariomycetes class (40%) and to the Xylaria genus (14%), while Eurotiomycetes was the minority class within the community. Our results reveal the high endophytic richness supporting the life cycle of A. angustifolia and reinforce the necessity for the conservation of this conifer, as many genetic resources can be lost owing to its irrational exploration.

Fossils can unveil a long-vanished combination of character states that inform inferences about the timing and patterns of diversification of modern fungi. By examining the well-preserved stacked chained vesicular conidiophores developed in clusters from the basal stroma, we describe a new taxon of fossil Zygosporiaceae with a combination of characters unknown among extant taxa on compressed serrated-margined dicot leaf (cf. Fagaceae) recovered from the Siwalik sediments (Late Miocene; ca. 12-8 Ma) of Himachal Pradesh, western Himalaya. Based upon conidiophore morphology, our Siwalik fungal remains, similar to Zygosporium Mont. (Zygosporiaceae: Xylariales: Sordariomycetes), are recognized as a new fossil species, Z. stromaticum Kundu & Khan, sp. nov. Zygosporium stromaticum is the only known fossil anamorphic fungus that occurs on plant cuticles and has a cluster of stacked chained vesicular conidiophores arising from a poorly preserved basal stroma formed by irregular, thick-walled cells. Its combination of morphological characteristics is unknown in extant fungal taxa, so Z. stromaticum likely represents a new anamorphic foliicolous fungus that may now be extinct. This unique evidence may be essential for the calibration of divergence time estimations of fungal lineages.