How the nucleolus recovers from acute proteostatic stress, particularly in multinucleate syncytia, remains poorly understood. In the highly polarized hyphae of the model filamentous fungus Magnaporthe oryzae, we uncover a novel stress-induced spatial quality control pathway that promotes the inheritance of rejuvenated nucleolar material during nuclear division. This pathway discriminates between newly formed and damaged nucleolar compartments, selectively partitioning and sequestering the latter. Our findings reveal a previously unrecognized mechanism for chaperone-mediated segregation of a membraneless nuclear organelle, extending principles of protein quality control to the unique demands of highly polarized syncytia.

This study reports the synthesis and characterization of fungal carbon dots (F-CDs) derived from filamentous microfungi belonging to the subphylum Pezizomycotina. Carbon dots were synthesized from a cell-free aqueous fungal homogenate obtained from the mycelial biomass of Phialomyces macrosporus, Penicillium spp., and Fusarium sp. The water-soluble mycelial fraction, containing intracellular metabolites, soluble proteins, and other low-molecular-weight compounds released upon cell disruption, was used directly as a carbon precursor in a hydrothermal synthesis. The resulting F-CDs exhibited intense green fluorescence with excitation-dependent emission, as confirmed by UV-Vis absorption and photoluminescence spectroscopy. Transmission electron microscopy revealed spherical nanoparticles with an average diameter of 3.9±1.1nm and crystalline domains with an interplanar spacing of 0.26nm. Zeta potential measurements indicated negatively charged surfaces (-15.6 to -18.4mV), suggesting good colloidal stability and potential for biological interactions. Fluorescence microscopy demonstrated efficient uptake of F-CDs by Aspergillus niger hyphae, resulting in bright green staining and indicating high cellular compatibility. These results establish filamentous microfungi as previously unexplored and versatile carbon precursors for the sustainable production of green-emitting carbon dots with promising applications in bioimaging.

Pathotype-specific roles of putative conidiation regulators in development, secondary metabolism, and pathogenicity in Fusarium fujikuroi.

Development and validation of a PCR-based detection assay for xerophilic Wallemia species informed by comparative mitogenomics.

Understanding metabolic processes of soil fungi is essential for elucidating their ecological roles in biogeochemical cycles and responses to emergent environmental stressors. Here, we demonstrate the potential of using stable isotope probing Raman (SIP-Raman) microspectroscopy in microfluidics technology-based soil chips to trace glucose metabolism rates and stress responses in laboratory grown filamentous soil fungus Psilocybe cf. subviscida. The time evolution of Raman spectral band intensities resulting from deuterated glucose uptake in the fungal hyphae allowed us to assess glucose metabolism rates. Under excess copper (Cu) stress, we observed suppression of both glucose metabolic activity and growth. In addition, reduced spectral signatures of intracellular cytochrome c further implied impaired mitochondrial function and potential onset of cell death. However, laser-induced radiation damage hampered repeated Raman measurements, including multispectral mapping, on individual hyphae, especially when exposed to the Cu stress. To overcome this, we employed stimulated Raman scattering (SRS) microscopy, which offers much higher sensitivity and mapping speeds, and therefore much lower radiation doses. This enabled localization of the uptaken glucose at the inner edges of the P. cf. subviscida hyphae and Cu-induced formation of putative vacuolar structures. While integration of this approach with soil chips requires future modifications to the chip design for increased optical transparency and ensured sterility, overall, our results demonstrate the potential of Raman-based microspectroscopy for spatially resolved, in situ analysis of fungal primary metabolism and stress physiology.

Dry bubble disease, attributed to the filamentous fungus Lecanicillium fungicola (Cordycipitaceae) results in huge yield losses in mushroom (Agaricus bisporus) cultivation worldwide. The possibilities for controlling the disease using commercial fungicides are highly limited, and therefore, there is an increasing demand for novel, alternative means of pest management. Our research objective was the comprehensive examination of viruses in the causal agents of dry bubble disease, which may open up an avenue for its virocontrol in the future. Out of 57 fungal isolates obtained from dry bubble-affected A. bisporus crops in various countries, 47 (82%) were confirmed by ITS (Internal Transcribed Spacer) sequence analysis as L. fungicola. In addition, different members of the genera Akanthomyces and Simplicillium (7 and 3 isolates, respectively), yet unknown to cause dry bubble symptoms, have also been detected. Cellulose column chromatography revealed the presence of double-stranded (ds) RNA in seven L. fungicola and three Akanthomyces sp. isolates, suggesting viral infection. The ten dsRNA-positive and eight randomly selected dsRNA-negative fungal strains were subjected to rRNA-depletion high-throughput RNA-sequencing analysis. The presence of seven new viruses representing four new species in the established families, Partitiviridae, Polymycoviridae, Botourmiaviridae and the narna-like virus group, and three previously established/proposed species in the families Chrysoviridae and "Mycovirgaviridae" were confirmed. The impact of the detected and identified viruses on their host fungi, and their potential applicability for virocontrol purposes will be examined in the future. This study provides the first detailed report on viruses of mushroom pathogenic fungi.

Reproducibility and standardization of methods are key approaches to microbiological research. However, laboratories differ widely in equipment, protocols, and operator expertise. These disparities are especially pronounced in fungal biology, since accurate spore counting and precise measurements of colony growth remain vulnerable to subjective bias and inconsistent technique. To standardize these foundational tasks, we developed two free, open-source image-analysis tools - SporeQuant and Orbisv2 - that automate hemocytometer spore counting and colony-area measurement, respectively. SporeQuant applies image thresholding and blob detection to identify individual spores and compute their concentration, while Orbisv2 segments colonies and calculates colony areas through thresholding and contour detection. Compared with manual analysis, SporeQuant and Orbis v2 markedly reduce analysis time by 63% and 76%, respectively, while achieving high measurement reliability, with MAE=0 for SporeQuant and R2>0.975 for Orbis v2. These tools reduce user-to-user variability, and achieve comparable or improved accuracy versus previously established methods. Furthermore, both tools can be accessed through a web page on both a computer or smartphone, requiring no specialized hardware, software, or prior knowledge. They provide accessible solutions for low-budget and high-throughput laboratories alike, enabling more consistent, scalable analysis of fungal growth and morphology in research and industrial settings.

Evaluation of Mitis Salivarius Agar for fungal growth in comparison with three standard fungal media.

Scedosporium apiospermum is a filamentous fungus increasingly implicated in infections across multiple organ systems, particularly in immunocompromised hosts. Due to its extensive intrinsic resistance patterns, it remains a challenging organism to treat. S. apiospermum is rarely implicated in skull base osteomyelitis and is associated with high morbidity and mortality in this clinical syndrome. We describe a case report of skull base osteomyelitis caused by S. apiospermum in a patient with poorly controlled diabetes mellitus, characterized by substantial diagnostic and therapeutic obstacles. Included in this article is a review of the existing literature on skull base osteomyelitis caused by this organism, highlighting common challenges in diagnosing and managing S.apiospermum . This case underscores the critical roles of early diagnosis and aggressive source control to optimize clinical outcomes.

SUMMARYTransporters are transmembrane proteins that mediate the selective translocation of metabolites, ions, and drugs across biological membranes. Their activity is essential for cellular communication, nutrition, detoxification, homeostasis, and responses to stress. Despite their fundamental biological and biomedical importance, particularly their involvement in genetic diseases and multidrug resistance in microbes and cancer cells, transporters remained relatively understudied until recently. This was largely due to the technical challenges of isolating and functionally characterizing these dynamic proteins, whose structure, function, and cellular expression continuously depends on specific interactions with membrane lipids. Nevertheless, since the mid-1960s, several transporters have been identified and extensively characterized at the genetic and physiological levels in model microorganisms, including fungi. In this review, we trace the 60-year research journey of UapA, a uric acid-xanthine transporter from the filamentous fungus Aspergillus nidulans (Ascomycota). The UapA story spans from early classical genetic analyses to the recent determination of its high-resolution structure. We describe the development of genetic, molecular, and cellular tools and how these enabled the functional dissection of UapA and were subsequently used for other A. nidulans transporters, including studies on cellular expression and turnover regulation. We additionally highlight up-to-date structural approaches that refined our knowledge on the transport mechanism, how substrate specificity is determined, and on membrane trafficking pathways underlying biogenesis and endocytosis of UapA and other transporters. The concepts developed through 60 years of persistent work on UapA have established important paradigms relevant to fungal physiology that have proven to be broadly applicable to understanding transporter-linked processes in other eukaryotic organisms.

The authors require two adjustments in the original manuscript [...]

Microbial interactions, particularly bacteria-fungus interactions, are research hotspots within microbial ecology and pathogenic biology. However, their underlying molecular mechanisms remain poorly understood, especially how bacterial pathogens recognize and exploit fungal signaling molecules for fungal predation. Here, we demonstrate that Pseudomonas aeruginosa employs an integrated tripartite farnesol-sensing system to detect and eliminate Candida albicans hyphae: The chemoreceptor PctA mediates directional migration toward hyphae; the type IV pilus sensor PilJ activates antifungal type III secretion system (T3SS) expression; and the quorum regulator PqsR monitors farnesol levels to coordinate virulence-metabolic switching. This mechanism enables bacteria to convert farnesol into their own signaling language (Pseudomonas quinolone signal) according to fungi status, thereby adaptively modulating their virulence expression and metabolism to cope with complex competitive microbial environments. Furthermore, bioinformatics analysis and functional validation confirm that the PctA-PilJ-PqsR triad is conserved across P. aeruginosa, suggesting that this interkingdom communication is widespread. In conclusion, this study reveals that P. aeruginosa orchestrates a targeted predation strategy against filamentous fungi by coordinating three interkingdom receptors, providing a theoretical foundation and potential molecular targets for understanding of interkingdom communication strategies among microorganisms and the development of signal molecule-based microbial prevention and control technologies.

Abstract Fungal antifungal proteins (AFPs) are promising biofungicides. PeAfpA and PeAfpB from Penicillium expansum show different activity profiles and potency, with PeAfpA being more active. Based on the PeAfpB solved structure, we had previously designed PeAfpB-PeAfpA chimeras that showed different properties. From these, we engineer here two additional variants, chPeAFPV6 and chPeAFPV7, that revealed novel aspects of the AFP structure, antifungal determinants and mechanism. chPeAFPV6, with a single E11K mutation in the loop L1 that is part of the γ-core motif, increased PeAfpB antifungal activity to that of PeAfpA against filamentous fungi but not yeasts, and promoted internalisation into Penicillium digitatum hyphae . However, changes in loop L3 of PeAfpB as in chPeAFPV7 abolished this increase, resulting in an inactive protein that still internalised. Overall, internalisation is neither sufficient nor essential for killing P. digitatum . Antifungal activity did not correlate with reactive oxygen species production, suggesting that oxidative burst is a fungal stress defence rather than a killing mechanism. Although cell permeabilisation was associated with antifungal activity, it does not seem to be a primary mode of action. Structural analysis showed interactions between the γ-core motif and loop L3, and suggests the importance of the conformation of the E7 residue of PeAfpB. Additionally, PeAfpA was identified as a protein able to penetrate Candida auris by a cell wall-dependent mechanism, and kill yeast cells. This study highlights the potential of the PeAfpB scaffold for engineering new-to-nature AFPs and provides novel insights into their modes of action, paving the way for future applications. Key points A single amino acid change in the γ-core of PeAfpB enhances antifungal potency Loop L3 of PeAfpB may block activity through interaction with the γ-core Antifungal activity does not correlate with ROS production

Heterologous protein secretion in filamentous fungi is often constrained by limitations in signal peptide recognition and intracellular trafficking. Aspergillus oryzae, a food-grade industrial fungus, has a robust native secretory system. However, its capacity for recombinant protein secretion remains suboptimal. Here, we developed a two-step, carrier-free engineering strategy to enhance protein secretion in A. oryzae. We identified endogenous signal peptides among highly secreted proteins using a green fluorescent protein (GFP) reporter. The oryzin signal peptide SPAoalp1 increased GFP secretion 5.50-fold compared with a no-signal-peptide control. We co-overexpressed Aosly1, a Sec1/Munc18 family protein that regulates soluble N-ethylmaleimide-sensitive factor attachment protein receptor-mediated vesicle trafficking, which, in combination with SPAoalp1, increased secretion approximately two-fold compared with SPAlp1 control and ten-fold with no-SP control. Applying the engineered platform for genetic improvement of heterologous bovine κ-casein increased secretion from 0.11 to 0.24 mg/L. Physiological optimization further increased secretion. The developed system provided initial evidence for secretion of a ~12 kDa band consistent with Aopafb transcription, with MIC90 values of 4.56-8.24% (v/v) against two Candida albicans strains and 4.68% (v/v) against Aspergillus niger. The system offers a modular framework for engineering fungal secretion and expands the utility of A. oryzae for recombinant protein production.

Lignocellulosic biomass represents a sustainable resource for enzyme production, yet indigenous microbial diversity remains underexplored in Algeria despite abundant agricultural residues. The present study investigates the potential of locally filamentous fungi isolated from Bejaia (Algeria) for lignocellulosic biomass valorization through cellulolytic enzyme production. A total of 201 native fungal strains were screened for cellulolytic potential, among which five showed high endoglucanase, exoglucanase, and β-glucosidase activities, with Aspergillus niger AZ193 identified as the most efficient strain. Cellulase production was carried out under solid-state fermentation using locally abundant lignocellulosic wastes. Date palm leaves resulted in the highest enzyme activities (201.78U/mL endoglucanase, 11.83U/mL exoglucanase, and 23.84U/mL β-glucosidase). The selected strain and substrate were further optimized using Response Surface Methodology, leading to a fivefold increase in endoglucanase activity (1000U/mL) and significant enhancement of other cellulases. The crude enzyme efficiently hydrolyzed pretreated date palm leaves, releasing up to 18mg/mL glucose within 72h. These findings highlight A. niger AZ193 as a robust cellulase producer and date palm leaves as an underutilized substrate for sustainable biotechnological applications. Further research should focus on enzyme purification, process scale-up, and integration into biorefinery applications for bioethanol production.

The review analyzes modern achievements in the field of biological processing of low- and intermediate-level liquid radioactive waste with an emphasis on the processes of biosorption of radionuclides by microorganisms, their syntrophic associations, as well as materials based on plant biomass. The aim of this work is to systematize data on the mechanisms of interaction of biological sorbents with fuel cycle radionuclides (U, Cs, Sr, Am, Pu, etc.), evaluation of their efficiency, and identification of promising directions for the development of treatment technologies. The results of studies over the past ten years are reviewed, including the use of living and heat-inactivated cultures of bacteria, microalgae, yeasts and filamentous fungi, artificial microbial consortia, as well as plant-derived wastes (straw, husks, wood residues) and products of their processing. A key role in the binding of actinides is played by phosphate groups of the cell wall and oxygen-containing functional groups of plant materials. Microbial associations and combined sorbents outperform monocultures in the efficiency of radionuclide removal due to the synergy of binding mechanisms. Factors influencing sorption capacity (pH, ionic strength, competing ions), conditions for biosorbent regeneration and subsequent immobilization of saturated biomass are analyzed. Biogeochemical processes in radioactive waste repositories are considered, where microbial activity can both contribute to the fixation of radionuclides (formation of biogenic mineral phases) and increase their mobility. The conclusion is made about the prospects of hybrid technological schemes combining biosorption with membrane methods and incorporation of spent biomass into geopolymer matrices, which ensures environmental safety and economic efficiency of liquid radioactive waste processing.

Aspergillus flavus is a globally distributed filamentous fungus of major agricultural and medical importance, capable of producing carcinogenic aflatoxins and forming two specialized developmental structures, conidia and sclerotia. While the molecular framework governing conidiation has been well characterized in Aspergillus nidulans, the corresponding mechanisms in A. flavus remain somewhat unelucidated. In this study, we identified and functionally characterized AfldrnA, a gene encoding a basic helix-loop-helix (bHLH) transcription factor. Targeted deletion of AfldrnA resulted in an aconidial phenotype accompanied by a significant increase in sclerotia formation, whereas complementation with the intact AfldrnA gene restored conidiation and reduced sclerotia development. Phenotypic assays revealed that the ΔAfldrnA mutant exhibited normal vegetative growth, unchanged antifungal susceptibility, and unaffected aflatoxin B1 production, indicating that AfldrnA primarily regulates developmental rather than metabolic differentiation. Additionally, observed differences between standard and dark incubation conditions suggest that AfldrnA may be involved in environmentally responsive regulation of fungal development. Overall, this study identifies AfldrnA as a pivotal transcriptional regulator essential for coordinating conidiation and sclerotia formation in A. flavus, providing new insights into the genetic and environmental regulation of fungal developmental programs.

Several species of saprophytic filamentous fungi are able of disrupting the life cycle of certain soil-born parasites that are of veterinary and agronomy importance, offering a promising sustainable control alternative. This study consisted of designing an experimental model, using catnip (Nepeta cataria) trays to simulate a vegetated environment for evaluating the parasiticidal activity of Mucor circinelloides, Trichoderma atrobrunneum, and Duddingtonia flagrans. Fungal spores were added to treated trays before adding feces with protozoan (Eimeria spp.), and gastrointestinal nematodes (roundworms, strongyles), and untreated-control water. No differences in plant growth or vigor, regardless of fungal presence, were observed, confirming the safety of these biological agents for vegetation. In the control trays, the viability of parasites ranged from 50% to 85%. In the treated trays, the viability of Eimeria and roundworms decreased by 40–100%, and the strongyle egg counts were reduced by 74% within 15 days. It is concluded that the vegetated tray model effectively simulates field conditions and provides a reliable platform for evaluating fungal efficacy against the free-living stages of parasites, offering a versatile tool for future research on soil-borne pathogens affecting animals and plants.

The control of fungal diseases in organic fruit production remains a major challenge due to the limited availability of authorized phytosanitary tools. This study evaluates the efficacy of two antagonistic yeasts - Metschnikowia pulcherrima L672 and Hanseniaspora uvarum L793 - in organic peach (Prunus persica) and plum (Prunus salicina) orchards over two consecutive seasons. Four treatments were compared: an untreated control, a standard organic management program, and foliar applications of each yeast strain. Microbiological monitoring revealed that Metschnikowia pulcherrima L672 demonstrated superior persistence and colonization, significantly reducing the abundance and colony size of filamentous fungi, including key pathogenic genera such as Alternaria, Cladosporium, and Penicillium spp. This biocontrol effect translated into improved fruit quality, notably a significant reduction in discarded plums at harvest, without compromising overall yield. In contrast, H. uvarum L793 exhibited limited persistence and a more transient antagonistic effect. These findings support the integration of Metschnikowia pulcherrima L672 as a promising and effective biocontrol agent for managing fungal diseases in organic stone fruit production systems. © 2026 The Author(s). Journal of the Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Petroleum hydrocarbon (PH) contamination in terrestrial environments is a serious global concern due to the toxicity and persistence of these compounds. Fungi are promising agents in bioremediation because of the ability to tolerate, degrade and metabolize complex and recalcitrant PHs. However, the use of other organisms, such as bacteria, remains more prevalent. Here, we evaluated the extent to which fungi have been investigated in PH bioremediation through a systematic review and meta-analysis following PRISMA guidelines. Among the 1,749 studies selected on bioremediation, only 348 included fungi, of which 97 met the inclusion criteria for quantitative analysis. Fungi were included in less studied (20% of articles) than bacteria and plants (63% and 31% respectively). However, our meta-analysis revealed that fungal treatments significantly reduced residual hydrocarbons by 50-60% in comparison with controls, with filamentous fungi (e.g., Trametes, Peniophora) showing the highest efficacy. Mycorrhizal fungi also demonstrated notable reduction (45-49%), while yeasts were less effective, particularly for polycyclic aromatic hydrocarbons (PAHs). We further identified that experimental variables, including treatment duration and the type of contaminated matrix, significantly influenced hydrocarbon reduction efficiency. Key limitations in the literature included a strong bias toward filamentous fungi and PAHs, as well as inconsistencies in experimental methodologies. Overall, our findings highlight the significant potential of fungi as bioremediation agents and emphasize the need to expand research on complex hydrocarbon mixtures (e.g., fuels) and underrepresented fungal groups, such as yeasts and mycorrhizal fungi, to better understand their role in reducing PH contamination in terrestrial ecosystems.