Although Aspergillus fumigatus has been identified as a critical fungal pathogen by the World Health Organization, diagnosis of the various types of aspergilloses remains unsatisfactory. New biomarkers of disease and accessible test systems are needed. Moreover, new treatment concepts are required to address infections caused by this invasive pathogen. Siderophore production by A. fumigatus offers opportunities for both improved diagnosis and potential therapy. Here, we report the development of a competitive ELISA that detects ferrated triacetylfusarinine C (FeTAFC) using a recombinant monoclonal IgG [anti-FeTAFC]. The FeTAFC ELISA can detect FeTAFC in human urine, a matrix proposed to be especially suitable for disease diagnosis because of the non-invasive nature of specimen collection. In addition, a novel assay was developed to assess the in vitro inhibitory properties of the IgG [anti-FeTAFC] towards A. fumigatus mutant and wild-type growth under iron-limiting conditions. The growth of A. fumigatus ΔsidD, deficient in TAFC and precursor fusarinine C (FsC) biosynthesis, was significantly reduced (p = 0.0003) in the presence of the antibody. While the growth of A. fumigatus wild-type was less affected in the presence of the antibody, the ratio of secreted TAFC:FsC was increased due to increased conversion of FsC to TAFC-likely because of extracellular complexation of FeTAFC by the IgG [anti-FeTAFC]. This work shows the utility of the IgG [anti-FeTAFC] for the detection of A. fumigatus and initial evidence for limiting fungal growth by attenuation of FeTAFC uptake.
Mycofiltration uses saprophytic fungi immobilised on dead organic matter to treat contaminated water. This systematic review aimed to collate literature on mycofiltration, identify water sources subjected to mycofiltration, types of fungi employed, contaminants removed, and removal efficiencies (R%). Articles written in English between 1990 and 2023 were collected from various sources, screened based on inclusion criteria, and critically appraised. Metadata were extracted, and a narrative synthesis was conducted. Forty articles representing 156 studies passed appraisal, with 116 from journal articles, 24 from theses, and 16 from reports. Synthetic stormwater and real wastewater were the most frequently mycofiltered. Fungi of the Pleurotus genus were predominantly used in creating mycofilters. Organic contaminants removed included pharmaceuticals and pesticides, with R% between 60% and 100%. E. coli was the most studied microbial contaminant, and R% of 30%, 60%, and 90% were reported. Inorganic contaminants were mostly metals with R% above 60%. Overall, contaminant removal by mycofiltration varied, but the technology remained a promising tool. Research gaps observed included a lack of standardised methods for mycofilter preparation and design and little to no assessment of mycofilter saturation. Addressing gaps could aid in increasing mycofilter efficiency and reliable upscaling of mycofiltration.
Ceramidases hydrolyze ceramides to fatty acids and sphingolipids, but their role in fungal response to stress remains unclear. We investigated the function of neutral ceramidase (nCDase) response to aromatic fungicide (carvacrol, cuminaldehyde, and isoniazid) stress in Neurospora crassa. Comparative analysis of the wild-type strain, Δnc and OEnc showed that nCDase enhanced fungicide resistance through multiple mechanisms. nCDase improved β-1,3-glucan synthesis (30% increase), decreased membrane permeability, elevated superoxide dismutase and catalase activities, and promoted carotenoid accumulation (50%), which collectively improved stress tolerance. Δnc exhibited disruption of cellular integrity, altered fatty acid profiles (elevated oleic acid, reduced total fatty acids), and increased fungicide sensitivity. Collectively, these findings established that nCDase as a key regulator of cell wall dynamics, lipid homeostasis, and antioxidant defense, thereby facilitating fungal adaptation to abiotic stress. This study identified the role of nCDase in the response to aromatic fungicide stress and laid foundation for inhibiting pathogenic fungi in agricultural production and food preservation.
An isolate of Peniophora lycii was obtained from grapevine, and its interactions with several grapevine-associated fungi and the plant host were examined. The fungus was not able to infect intact leaves, but necrotized the margins of foliar disks and caused necrosis and white rot in woody tissues. In dual cultures, P. lycii and Aureobasidium pullulans showed mutual antagonism. Mycoparsitism of P. lycii was observed on epiphytic Botrytis cinerea, Alternaria sp., and endophytic Botryosphaeria dothidea interaction partners. In contrast, P. lycii showed trophic growth towards endophytic Phaeomoniella chlamydospora without any signs of harmful interactions. Dual inoculations of foliar disks with epiphytic fungi revealed no effects of fungal interactions on necrosis development by pathogens and verified mycoparasitic interactions in planta. Co-infection assays of cuttings with endophytic pathogen fungi showed cumulative effects of fungal interactions on wood symptom expression, with the exclusive contribution of P. lycii to white rot development. In addition to providing the first isolation of P. lycii from grapevine and the description of its mycoparasitic behavior, the present study suggests that the fungus may act as an opportunistic grapevine pathogen, probably as a secondary colonizer in trunk diseases. The observed dual host preference may allow trunk disease pathogens to initially feed on fungi, followed by damage to the grapevine. This may be in connection with the exceptionally long latency of these syndromes.
Drought and nitrogen deposition are major drivers of global change that can influence forest ecosystems and plant-microbe interactions, yet their combined effects on endophytic fungal communities and the roles of dark septate endophytes (DSE) remain unclear. In this study, we examined the diversity of culturable endophytic fungi in leaves and roots of Quercus dentata under different drought and nitrogen deposition treatments and evaluated the functional effects of representative DSE strains on host growth and physiology. A total of 1488 fungal isolates were obtained, revealing distinct organ-specific community patterns. Root-associated communities showed greater compositional stability across treatments, whereas leaf communities were more responsive to environmental variation. Severe drought reduced the dominance of several genera and promoted community restructuring, while nitrogen deposition had contrasting effects on α-diversity in leaves and roots. Beta diversity analyses indicated significant interaction effects between drought and nitrogen addition. Inoculation with four DSE strains produced strain-dependent effects on plant biomass, photosynthesis, water-use efficiency, physiological traits, and nutrient contents. These results indicate that drought and nitrogen deposition jointly influence endophytic fungal communities and that functional differences among DSE strains may affect host responses to combined stress.
Marine endophytic fungi inhabit the internal tissues of seaweed, seagrass, and mangroves without causing harm. These fungi are known to produce extracellular enzymes, including proteases and cellulases, which play crucial roles in various biological processes and have potential applications in diverse industrial sectors. This study aimed to screen the enzymatic potential of marine endophytic fungi, identify selected isolates, and characterize their enzyme activities. A total of 20 fungal isolates were obtained, comprising 16 isolates from seaweed, three from seagrass, and one from mangrove leaves, collected from the coastal areas of the Seribu Islands (Jakarta), Sukabumi (West Java), Nusa Dua (Bali), and the Buton Islands (Southeast Sulawesi). Screening results showed that 50% of the isolates exhibited proteolytic activity on skim milk agar, while 40% demonstrated cellulolytic activity on carboxymethylcellulose (CMC) agar. Two isolates with the highest clear zone indices for protease and cellulase activity were identified as Penicillium citrinum and Fomitopsis sp., with distinct morphological characteristics including velvety colonies and filamentous hyphal structures. The specific activities of the protease and cellulase were 5475.42 ± 2724.25 U/mg protein and 620.77 ± 607.71 U/mg protein, respectively, indicating high catalytic potential.
This paper systematically reviews the research progress on the physiological functions, gene cloning, classification basis, and expression regulation mechanisms of laccase in edible and medicinal fungi. Laccase is a copper-containing polyphenol oxidase widely distributed in these fungi, characterized by broad-spectrum substrate catalytic activity and redox properties. It plays a central role in lignin degradation, pigment synthesis, and environmental pollutant treatment. Regarding gene cloning, researchers have successfully isolated and identified laccase genes from multiple species using techniques such as transcriptome sequencing, RACE amplification, and gene knockout. Expression regulation studies have revealed that laccase genes exhibit stage-specific expression patterns during mycelial growth, fruiting body development, and lignin degradation. In recent years, breakthroughs in genomics, transcriptomics, and gene editing technologies have greatly advanced research into the cloning, classification, and regulatory mechanisms of laccase genes. This article systematically reviews the diversity, clonal classification, and regulatory mechanisms of these genes, aiming to provide a reference for further research and industrial development of laccase in edible and medicinal fungi.
Fungi are the main causative agents of plant diseases and are responsible for substantial and recurrent damage to agricultural systems. Their activity causes significant reductions in crop productivity and food quality, ultimately contributing to plant deterioration and economic losses. It is estimated that phytopathogenic fungi can compromise up to 30% of global agricultural production. To mitigate microbial deterioration, a wide range of control strategies have been employed, with chemical fungicides being one of the most widely used interventions. However, current approaches to fungal control are rapidly transforming owing to the growing prevalence of fungicide resistance, increasingly stringent regulatory frameworks governing chemical applications, and evolving market demands. Taken together, these factors impose new constraints and drive the development of more sustainable alternative options for effective food control. This review examines the diverse strategies used to control fungal diseases in plants, emphasizing advances in biocontrol agents and biofungicides, as well as emerging tools in the molecular biology, genomics, and biotechnology fields. The aim is to highlight recent developments and prospects that can be integrated into comprehensive disease-management approaches.
Phi Man Long Long Rak Cave, located in Mae Hong Son Province, northern Thailand, is a prehistoric burial site containing ancient wooden coffins that have undergone biodeterioration, likely due to fungal activity. Both culture-dependent and culture-independent approaches were employed to characterize fungal communities and assess their roles in wood degradation. Culture-dependent analysis identified five Aspergillus isolates from the wooden coffins, most of which produced cellulolytic and hemicellulolytic enzymes; some isolates also produced organic acids, indicating significant degradative potential. Culture-independent analysis revealed a community dominated by Aspergillus, together with additional taxa such as Penicillium and Ceriporia that were not detected by cultivation, highlighting greater community diversity and demonstrating the complementarity of the two methods. Functional prediction indicated a predominance of saprotrophic fungi. The presence of shared dominant taxa between soil and coffin-associated substrates suggests ecological connectivity at the soil-coffin interface, although the direction of dispersal cannot be determined from the present data. All tested fungicides inhibited fungal growth, with the highest efficacy observed in the formulation containing the highest proportion of active components. Taken together, these findings provide insights into fungal biodeterioration processes and inform conservation strategies.
Dark septate endophytes (DSE) and ericoid mycorrhizal fungi (ERMF) are employed to augment the abiotic stress resistance of fruits. However, their potential functions in enhancing the drought resistance of blueberry, an economically important fruit, remain unclear. Thus, this study aims to identify optimal inoculation combinations to enhance the drought resistance of blueberry seedlings. Specifically, the effects of single and dual inoculations with DSE (Cladosporium cladosporioides, D79) and ERMF (Oidiodendron citrinum, N12) on seedling physiology and metabolism were explored under varying drought conditions. The results showed that dual inoculation significantly improved leaf physiological characteristics. Under severe drought stress, the 1:2 DSE:ERMF ratio (D1N2) notably increased leaf relative water content (RWC) and reduced electrolyte leakage by up to 42.1% compared with the non-inoculated control. Dual inoculation also significantly decreased malondialdehyde (MDA) content, with the smallest increase observed in D1N2. Regarding antioxidant enzymes, dual inoculation sustained higher superoxide dismutase (SOD) activity under moderate drought and minimized the decline in SOD activity under severe drought (the lowest decrease was 36.4% in D1N2 versus 56.7% in CK). Moreover, the antioxidant losses under drought stress were reduced by upregulating various metabolic processes, especially the biosynthesis of phenylalanine, tyrosine, and tryptophan. A comprehensive evaluation suggested that inoculation with a 1:2 mixture of DSE and ERMF most effectively improved blueberry drought resistance, primarily by enhancing water and metabolite supply and stimulating the antioxidant defenses.
Alternaria blight, caused by fungi of the genus Alternaria, is one of the most common and damaging diseases affecting tomatoes, leading to significant yield losses. The intensive use of chemical fungicides faces the problems of pathogen resistance development and negative environmental impacts. This study investigated the possibility of using RNA interference technology based on exogenous double-stranded RNAs (dsRNAs) to protect tomatoes against the causal agent of early blight (EB), Alternaria alternata. Key genes of the pathogen A. alternata were selected as targets: Alt-a1 (a major allergen and virulence factor), TEF1a (translation elongation factor 1-alpha) and β-Tub (β-tubulin). Specific dsRNAs were synthesized in vitro and applied to tomato plants (Solanum lycopersicum L. cv. Micro-Tom) simultaneously with inoculation of A. alternata strain C7.24-T2-L-F1 spores. Visual assessment, measurement of chlorophyll A and B, and real-time quantitative PCR analysis showed that treatment with dsRNAs targeting the Alt-a1, TEF1a and β-Tub genes significantly suppressed infection development, reducing the amount of pathogen DNA in plant tissues by 7 to 27 times depending on the dsRNA type. The most effective was dsRNA to the Alt-a1 gene. Thus, the obtained results demonstrate the promise of spray-induced gene silencing (SIGS) as a strategy for protecting tomato plants against the pathogen A. alternata.
Mutations in an RNA-binding protein FUS are known to cause familial amyotrophic lateral sclerosis (ALS). Since this discovery, mutations in several other RNA-binding proteins (RBPs) have also been linked to ALS. Some of these ALS-associated RBPs have been shown to colocalize with ribonucleoprotein (RNP) granules such as stress granules and processing bodies (p-bodies). Increasing evidence has emerged supporting a hypothesis that the impaired clearance, inappropriate assembly, and dysregulation of RNP granules play a role in ALS. Through the genome-scale overexpression screening of a yeast model of FUS toxicity, we found that TAF15, a human RBP with a similar protein domain structure and belonging to the same FET protein family as FUS, suppresses FUS toxicity in yeast. The suppression by TAF15 is specific to FUS and not found in other yeast models of neurodegenerative disease-associated proteins. We showed that the RNA recognition motif (RRM) of TAF15 is required for its suppression of FUS toxicity. Furthermore, FUS and TAF15 physically interact, and the C-terminus of TAF15 is required for both the physical protein-protein interaction and its protection against FUS toxicity. Finally, while FUS induces and colocalizes with both stress granules and p-bodies, TAF15 only induces and colocalizes with p-bodies. Importantly, the co-expression of FUS and TAF15 induces more p-bodies than individually expressing each gene alone, and FUS toxicity is exacerbated in yeast that is deficient in p-body formation. Overall, our findings suggest a role of increased p-body formation in the suppression of FUS toxicity by TAF15.
The objective was to identify management strategies of IFI in critically ill patients through a Spanish national survey. A cross-sectional multicentre survey among ICU specialists, experienced in IFI, was performed (22 April-25 July 2024). The survey consisted of 13 questions with four closed answers. Sixty-three specialists from 51 hospitals of 16 regions completed the survey. 95% stated that, in high-risk patients with clinical suspicion of Pulmonary Aspergillosis (PA), galactomannan in BAL is performed to guide treatment. In the treatment of patients with PA and influenza, 86% declared that isavuconazole and liposomal amphotericin B are recommended treatments and in high suspicion of Aspergillus coinfection, 76% recommended empirical treatment waiting for microbiological confirmation. 90% declared that the use of Extracorporeal Membrane Oxygenation (ECMO) and Renal Replacement Therapies (RRT) could be associated with lower azole levels. Regarding intra-abdominal candidiasis, 78% that physiopathological changes in critically ill patients, reduce their entry into peritoneal fluid. The majority of the respondents agreed (>80%) on: In suspicion of PA, Galactomannan in BAL to guide treatment is mandatory; In case of aspergillosis and influenza, isavuconazole and liposomal amphotericin B are the recommended treatments; The use of ECMO and RRT could be associated with lower azole levels.
Samsoniella hepiali produces an array of pharmacologically valuable metabolites, but how environmental pH regulates its antioxidant system and lipid metabolism during submerged fermentation remains unclear. This study aimed to investigate the effects of different initial culture pH values (pH 4, 5, 6, and 7) on the antioxidant capacity and lipidomic metabolism of S. hepiali. The results demonstrated that at pH 5, the activities of peroxidase (POD) and superoxide dismutase (SOD), the contents of total phenolics (TP) and flavonoids, the scavenging rates of DPPH• and •OH, and the total antioxidant capacity all peaked. Conversely, the level of glutathione (GSH) reached its maximum at pH 6 (0.69 ± 0.014 μmol/g). Lipidomic analysis identified a total of 404 lipid molecular species, mainly TG, PE, and DG. Comparative analysis among pH 4 vs. pH 5, pH 6 vs. pH 5, and pH 7 vs. pH 5 revealed 27 core DALs belonging to 11 lipid subclasses, most of which were upregulated at pH 5. KEGG pathway enrichment analysis further revealed that sphingolipid metabolism was the sole core co-enriched pathway under different pH conditions. Particularly at pH 5, key signaling lipids, such as ceramides, underwent pronounced targeted accumulation. This study elucidates the molecular adaptation mechanisms of medicinal fungi in response to pH variation from a lipidomic perspective. It provides a basis for optimizing fermentation conditions to enhance antioxidant activity and functional lipid production.
The diagnosis of coccidioidomycosis is often achieved serologically by the detection of antibodies against fungal antigens. While several serologic tests are available for coccidioidomycosis, all of them are performed in a laboratory setting causing delays in diagnosis and therapeutic intervention. Point-of-care testing offers the ability to provide a shorter time to result by avoiding specimen send-out, minimizing processing steps, and employing expeditious immunochemical techniques. A preliminary trial of a rapid anti-coccidioidal antibody lateral flow assay (LFA) using fingerstick blood was performed on 22 patients with coccidioidomycosis at the point of care during outpatient clinic visits. Patients were tested longitudinally over the course of one year. An LFA reader was implemented to provide an objective result by quantifying the intensity of the test line band. There was close qualitative concordance observed between positive LFA results with send-out immunodiffusion (89.5%) and complement fixation (78.4%) standard of care clinical laboratory assays. Additionally, the relationship between LFA test line density values and traditional complement fixation antibody titers was assessed.
Candida albicans is a common opportunistic pathogen. Long-term use of azole antifungals faces challenges like resistance, necessitating novel agents. Tea tree oil (TTO), a natural broad-spectrum antimicrobial, shows promise, but its molecular mechanisms, particularly concerning novel cell death pathways, require clarification. This study comprehensively evaluated the antifungal mechanism of TTO against C. albicans using transcriptomics. Antifungal susceptibility assays were conducted to assess the effects of TTO and its components (4-terpineol, terpenes, and γ-pinene) on the growth of C. albicans hyphae and biofilms. Fluorescent labeling and biochemical analysis were employed to detect ferroptosis markers. Transcriptomic results indicate that TTO induces 423 differentially expressed genes and systematically inhibits the development of C. albicans hyphae through mechanisms such as oxidative stress, iron homeostasis disruption, disruption of cell wall integrity, and interference with ergosterol metabolism. Notably, the significant enrichment of redox enzyme activity and iron ion binding functions, along with changes in the glutathione metabolic pathway, suggest that ferroptosis may be involved in this process. Subsequent studies revealed that the compound 4-pinene most effectively inhibits the pathogenicity of C. albicans by suppressing its adhesion, hyphae formation, and biofilm formation, whereas terpinene induces the accumulation of reactive oxygen species (ROS) and increases lipid peroxidation in C. albicans; furthermore, following treatment with an iron-mediated apoptosis inhibitor, terpinene enhances the viability of the treated C. albicans cells.
Yam is a valuable traditional Chinese medicinal and nutritional crop that has gained significant popularity in recent years. However, the production of yam is severely impacted by brown rot caused by Fusarium solani, which affects both yield and quality. In this study, we isolated a promising biocontrol strain, designated AP6, from the rhizospheric soil of healthy yam. Strain AP6 exhibited strong antagonistic activity against F. solani, with an inhibition rate of 60.2%, and demonstrated broad-spectrum antagonistic activity. Based on morphological, physiological, biochemical characteristics, and whole-genome sequencing, this strain was identified as Bacillus velezensis. Strain AP6 can produce siderophores, amylase, protease, cellulase, and form biofilms. It can also change the hyphal morphology of F. solani, inhibit spore germination, reduce the viability of pathogens, and alleviate the disease severity of yam. Additionally, strain AP6 was shown to induce the accumulation of reactive oxygen species in yam, thereby enhancing the yam's defense responses. Field application trials demonstrated that a wettable powder formulation of strain AP6, when combined with commercial metalaxyl-M and fludioxonil, significantly reduced the incidence of brown rot in yam. These findings indicate that B.velezensis AP6 is a promising biocontrol agent, providing a practical and sustainable approach for integrated disease management in yam.
Twenty strains of Schizophyllum commune from the BIOTEC culture collection were selected for this study. S. commune is characterized by white to gray fan-shaped caps with lobed margins and distinctive split gills. Phylogenetic analysis of combined LSU rDNA and ITS rDNA sequences data using maximum parsimony placed the fungi in a strongly supported clade with S. commune. All strains were primarily screened for exopolysaccharide (EPS) and biomass production using potato dextrose broth (PDB) and peptone yeast glucose medium (PYGM) in 250 mL flasks shaken at 200 rpm for 7 days. The results revealed three strains with high EPS production, each exceeding 2.3 g/L, namely MMCR00487, MMCR00474 and MMCR00256. These strains were selected for media optimization using a Plackett-Burman design. Among them, MMCR00256 exhibited the highest EPS yield of 8.34 ± 1.47 g/L, followed by MMCR00487 and MMCR00474. Therefore, the strain MMCR00256 was further optimized by central composite design. The results revealed that the optimized medium for MMCR00256 increased the production of EPS to 10.39 ± 1.69 g/L, with a biomass yield of 26.28 ± 1.63 g/L (395 mg/g). The 5 L bioreactor optimization tested two inoculum types (mycelial and pellet) and two media (CCD and estimated) using strain MMCR00256. The mycelial inoculum grown in the estimated medium produced the highest EPS yield of 8.37 ± 0.26 g/L after 3 days, with 13.56 ± 2.94 g/L biomass. In conclusion, this study demonstrates that S. commune MMCR00256, when cultivated using the estimated medium and mycelial inoculum, can achieve enhanced exopolysaccharide production with improved efficiency, highlighting its significant potential for the development of efficient and scalable schizophyllan production processes at the industrial scale. Furthermore, this study provides essential insights into the cultivation and optimization of schizophyllan in S. commune.
Criolla grape varieties are native South American cultivars that represent an important reservoir of genetic and microbiological diversity. This study aimed to investigate the oenological potential of Saccharomyces and non-Saccharomyces yeasts isolated from three criolla grape varieties in order to support the future design of wine starters. Yeasts were isolated at different fermentation stages from four vineyards in Mendoza, Argentina. A total of 485 isolates were recovered and molecularly identified, revealing 12 species belonging to eight genera. Saccharomyces cerevisiae, Hanseniaspora guilliermondii and Hanseniaspora uvarum were the dominant species. Isolates were screened for H2S and acetic acid production, followed by physiological and enzymatic characterisation. Selected strains were further evaluated in small-scale fermentations to assess fermentative kinetics and metabolic performance. Significant variability was observed, particularly among non-Saccharomyces isolates, which generally exhibited lower ethanol yields and acetic acid production compared to S. cerevisiae. Several isolates of H. guilliermondii showed balanced fermentative behaviour and favourable metabolic and enzymatic profiles. Through the applied selection strategy, twelve strains emerged as promising wine starter candidates. These isolates combined a low production of undesirable metabolites, relevant enzymatic activities, and favourable fermentative performance. Overall, the results highlight the oenological potential of autochthonous yeasts as a resource for innovative winemaking strategies.
The functions of the putative bZIP (basic leucine zipper) transcription factors ZipV (Afu3g032301) and ZipZ (Afu2g14350) were investigated using wild-type, gene-deficient, and complemented strains of Aspergillus fumigatus. Deletion of zipV increased oxidative stress sensitivity and reduced in vivo virulence in a Galleria mellonella model, whereas complementation restored the wild-type phenotype. In contrast, deletion of zipZ resulted in no detectable phenotypic changes, even though transcription of both genes was modulated by oxidative stress. Phenotypic characterization of conidia, transcriptomic analyses of growing cultures and reverse-transcription quantitative real-time PCR of conidia-producing cultures suggested that ZipV regulates the development of certain conidial traits. The conidia of the ∆zipV mutant showed reduced heat stress tolerance, decreased catalase activity and delayed germination in comparison to the wild-type or the complemented strain. In the ∆zipV mutant, the transcription of catA encoding conidial catalase was impaired. This defect explains the reduced catalase activity and the oxidative stress sensitivity of the mutant and may contribute to its reduced virulence. The increased transcriptional activity of the alternative oxidase gene aoxA observed in the absence of ZipV suggests a compensatory response aimed at mitigating oxidative stress.