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J. Fungi, Volume 12, Issue 2 (February 2026) – 80 articles

Cover Story (view full-size image): The anti-Candida albicans effect of cell-free supernatants (CFSs) from three different faecal isolates and one ATCC strain of Saccharomyces cerevisiae was assessed. The CFSs activity on C. albicans virulence traits demonstrates that CFSs significantly reduce C. albicans capacity to adhere and to form a biofilm and decrease the metabolic activity of biofilm-embedded fungal cells. The untargeted metabolomic analysis of the CFSs shows that compared to the reference strain, faecal isolates overexpress N-acetyl-DL-tryptophan and other molecules derived from its metabolism, L-leucine containing dipeptides, and the nucleotide inosine, suggesting that one or more of such molecules may impair C. albicans virulence, paving the way for a possible future employment of these CFSs as postbiotics. View this paper
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10 pages, 260 KB  
Opinion
Helminth Immune Modulation and Invasive Fungal Infections in Sub-Saharan Africa
by Luis Fonte, Yaxsier de Armas, Héctor R. Pérez-Gómez and Enrique J. Calderón
J. Fungi 2026, 12(2), 160; https://doi.org/10.3390/jof12020160 - 23 Feb 2026
Viewed by 908
Abstract
Sub-Saharan Africa, a region marked by enormous social and health inequalities, has the largest population infected with HIV and Mycobacterium tuberculosis, which are considered the main risk factors for fungal infections. At the same time, sub-Saharan Africa is the region of the [...] Read more.
Sub-Saharan Africa, a region marked by enormous social and health inequalities, has the largest population infected with HIV and Mycobacterium tuberculosis, which are considered the main risk factors for fungal infections. At the same time, sub-Saharan Africa is the region of the world with the highest rates of helminth infections, whose immunomodulatory effects impair the host’s immune responses to other microorganisms, including HIV and M. tuberculosis. Through this indirect way, helminth immune modulation could be another syndemic factor influencing the development of fungal infections. However, some epidemiological peculiarities of five fungal diseases in sub-Saharan Africa, which we analyze in this paper, suggest that the influence of helminth immune modulation on the development of fungal infections there could also be direct. In light of the knowledge of all those interactions, any healthcare and epidemiological approach to Invasive Fungal Infections in sub-Saharan Africa should be carried out from a syndemic perspective that takes into account the ways in which social environments contribute to the clustering of infections, the pathways through which infecting microorganisms could interact biologically in each individual, influencing the development and evolution of the disease in course, and the ways in which those interactions complicate diagnosis, treatment, and control. Full article
(This article belongs to the Special Issue Clinical and Epidemiological Study of Mycoses)
19 pages, 2091 KB  
Article
Evaluation of the Antifungal Potential of Different Photorhabdus Species Against Monilinia laxa and Colletotrichum fioriniae
by Emre Şen, Tímea Tóth, Szabolcs Ádám and Tamás Lakatos
J. Fungi 2026, 12(2), 159; https://doi.org/10.3390/jof12020159 - 22 Feb 2026
Viewed by 813
Abstract
Monilinia laxa and Colletotrichum fioriniae are major fungal pathogens causing brown rot and anthracnose in stone fruits and shell fruits, leading to significant economic losses. Chemical fungicides are widely applied but can result in resistance development, environmental contamination, and food safety concerns. Biological [...] Read more.
Monilinia laxa and Colletotrichum fioriniae are major fungal pathogens causing brown rot and anthracnose in stone fruits and shell fruits, leading to significant economic losses. Chemical fungicides are widely applied but can result in resistance development, environmental contamination, and food safety concerns. Biological control using entomopathogenic bacteria (EPB) of the genus Photorhabdus has emerged as an eco-friendly alternative. This study evaluated the in vitro antifungal activity of selected Photorhabdus species (P. kayaii 1723B, P. temperata 3017, P. cinerea 3086, P. laumondii 3196, and P. thracensis 3210) against M. laxa (M3) and C. fioriniae (VV081) using drop-to-drop confrontation and poisoned agar assays. Effects of fermentation time, preparation mode (original vs. centrifuged and filtered), and concentration (5, 10, 20%) were examined. Species-specific inhibition was observed, with Median Inhibition Index values indicated relatively higher antifungal activity for P. thracensis 3210 against M. laxa (0.718) and C. fioriniae (0.552), followed by P. cinerea 3086 (0.643 and 0.552) and P. kayaii 1723B (0.629 and 0.541). Fermentation time and preparation mode influenced antifungal activity in a strain-dependent manner, with longer fermentation periods and original culture preparations generally showing stronger inhibitory trends. Higher concentrations, especially 20%, were often associated with increased inhibition, although the magnitude of these effects varied among strain–pathogen combinations. Overall, these findings demonstrate that the strain- and pathogen-specific nature of antifungal responses in Photorhabdus, supporting their potential as components of targeted biological control strategies rather than uniform broad-spectrum agents. Full article
(This article belongs to the Section Fungal Pathogenesis and Disease Control)
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23 pages, 2992 KB  
Article
Enhanced Biocontrol of Root-Knot Nematodes Through Co-Cultivation of Clonostachys rosea and Bacillus velezensis: Proline-Driven Bacterial Fitness and Synergistic Metabolite Production
by Jie Zhang, Yajing Song, Manhong Sun, Jiangkuan Cui, Yuankai Chi, Mingcong Xia, Runhong Sun, Chao Wu, Qianqian Dong and Lirong Yang
J. Fungi 2026, 12(2), 158; https://doi.org/10.3390/jof12020158 - 22 Feb 2026
Viewed by 757
Abstract
The ascomycete fungus Clonostachys rosea is a promising biocontrol agent against root-knot nematodes. To develop a more effective and stable biocontrol strategy, we rationally constructed a co-culture system by partnering C. rosea with the plant growth-promoting bacterium Bacillus velezensis. Through systematic optimization [...] Read more.
The ascomycete fungus Clonostachys rosea is a promising biocontrol agent against root-knot nematodes. To develop a more effective and stable biocontrol strategy, we rationally constructed a co-culture system by partnering C. rosea with the plant growth-promoting bacterium Bacillus velezensis. Through systematic optimization of the medium and inoculation protocol, the co-culture demonstrated significantly enhanced performance, achieving 95.3% mortality of Meloidogyne incognita juveniles, a 78.0% increase in tomato shoot dry weight, and 69.2% disease control efficacy in pot trials. Metabolomic profiling indicated that the co-culture triggered a distinct metabolic profile compared to the respective monocultures. The enhanced efficacy was associated with the accumulation of two functional metabolite groups. First, the co-culture synergistically accumulated direct-effect compounds with reported nematicidal (e.g., daidzin, L-tryptophan) and plant-growth-promoting (e.g., isopentenyladenine, melatonin, and indole-3-propionic acid) activities. In parallel, L-proline emerged as a critical microbial interaction modulator. Targeted quantification showed a clear proline abundance gradient: highest in the C. rosea monoculture, intermediate in co-culture, and lowest in the B. velezensis monoculture. This gradient suggests that proline produced by C. rosea is likely utilized by B. velezensis, a finding further supported by the observation that proline enhanced bacterial biofilm formation and upregulated the matrix genes epsC and tasA. Accordingly, the co-culture itself formed significantly more robust biofilms. Thus, the enhanced biocontrol can be attributed to synergistic metabolite accumulation together with proline-mediated fitness gains in the bacterial partner, establishing a metabolic basis for rationally engineering microbial consortia. Full article
(This article belongs to the Section Fungi in Agriculture and Biotechnology)
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25 pages, 4997 KB  
Article
Regulation of ABC Transporters and Ergosterol Biosynthesis by the Transcription Factor FvADS-1 Controls Azole Resistance and Virulence in Fusarium verticillioides
by Yajing Yin, Hanxing Zhang, Zhenying Zhang, Mi Zhou, Shaojie Li and Chengcheng Hu
J. Fungi 2026, 12(2), 157; https://doi.org/10.3390/jof12020157 - 22 Feb 2026
Viewed by 682
Abstract
Fusarium verticillioides is a significant agricultural pathogen and an emerging causative agent of invasive fusariosis in clinical settings. Fusarium species frequently exhibit resistance to available antifungal agents, yet the molecular mechanisms underlying azole resistance remain poorly characterized. In this study, we identified the [...] Read more.
Fusarium verticillioides is a significant agricultural pathogen and an emerging causative agent of invasive fusariosis in clinical settings. Fusarium species frequently exhibit resistance to available antifungal agents, yet the molecular mechanisms underlying azole resistance remain poorly characterized. In this study, we identified the Zn(II)2Cys6 transcription factor FvADS-1 as a positive regulator of the azole stress response in F. verticillioides. The transcription of FvADS-1 was significantly induced by ketoconazole (KTC), and its deletion increased susceptibility to multiple azole compounds. Mechanistically, FvADS-1 positively regulates the KTC-induced expression of genes encoding ABC transporters and ergosterol biosynthesis enzymes, thereby modulating intracellular KTC accumulation and sterol homeostasis under azole stress. Furthermore, FvADS-1 positively regulates the transcriptional response of peroxisomal genes and contributes to fungal tolerance to oxidative stress. Notably, deletion of FvADS-1 attenuates the virulence of F. verticillioides on maize. The function of ADS-1 is evolutionarily conserved: heterologous expression of N. crassa ads-1 restored azole resistance in FvADS-1 deletion mutant, and the deletion of the F. oxysporum homolog FoADS-1 similarly increased azole susceptibility. Collectively, our study demonstrates that the conserved transcription factor ADS-1 plays a central role in regulating azole resistance and virulence in the pathogen F. verticillioides, offering new insights into antifungal resistance mechanisms in pathogenic filamentous fungi. Full article
(This article belongs to the Section Fungi in Agriculture and Biotechnology)
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17 pages, 3976 KB  
Article
Caspofungin Reshapes the Extracellular Vesicles Metabolome of Candidozyma (Candida) auris, Altering Amino Acid and Nucleotide Metabolism
by Vinicius Alves, Claire V. Mulholland, Daniel Zamith-Miranda, Susana Frases, Michael Berney and Joshua D. Nosanchuk
J. Fungi 2026, 12(2), 156; https://doi.org/10.3390/jof12020156 - 21 Feb 2026
Cited by 2 | Viewed by 992
Abstract
Candidozyma auris is an emerging multidrug-resistant fungal pathogen associated with severe invasive infections and high mortality, particularly in healthcare environments. Its rapid global expansion and resistance to multiple antifungal classes pose major challenges to treatment and containment. Extracellular vesicles (EVs) have recently been [...] Read more.
Candidozyma auris is an emerging multidrug-resistant fungal pathogen associated with severe invasive infections and high mortality, particularly in healthcare environments. Its rapid global expansion and resistance to multiple antifungal classes pose major challenges to treatment and containment. Extracellular vesicles (EVs) have recently been recognized as important mediators of fungal communication, virulence, and stress adaptation. Here, we examine how caspofungin, a frontline echinocandin, reshapes the EV metabolome of C. auris. Caspofungin exposure drives pronounced remodeling of EV size distributions, yielding a predominance of smaller, more uniform EVs alongside a minor population of larger subtypes. Metabolomic profiling of EVs revealed marked enrichment of metabolites involved in nucleotide salvage and recycling, along with altered amino acid abundances, including increases in amino acids associated with stress responses and redox regulation. These changes are consistent with altered nucleotide turnover and amino acid metabolism under antifungal stress. Importantly, these metabolic alterations reflect caspofungin-induced changes in cellular metabolism that are selectively exported via extracellular vesicles, rather than metabolic activity occurring within the vesicles themselves. Export of these metabolites via EVs may support population-level coordination, biofilm remodeling, and modulation of host immune responses, contributing to echinocandin tolerance. Together, our findings highlight nucleotide- and amino acid-associated metabolic features of EVs as informative readouts of caspofungin exposure and highlight the EV metabolome as a promising source of non-invasive biomarkers for monitoring drug exposure and resistance. This work advances understanding of C. auris adaptation under antifungal stress and reveals new opportunities for therapeutic and diagnostic innovation against this high-priority pathogen. Full article
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30 pages, 1490 KB  
Systematic Review
Unusual Sporotrichosis: A New Concept Proposal on the Unexpected Faces of Sporothrix spp. Infection
by Jayne Araújo da Silva, Adriany Lucas dos Santos, Júlia Andrade de Castro Rodrigues, Mariana de Paula Pires, Marcelo Cerilo-Filho, Gil Benard, José Rodrigo Santos Silva, Ricardo Luiz Dantas Machado, Jéssica Dornelas da Silva, Héctor Manuel Mora-Montes, Gutemberg Gomes Alves and Andréa Regina de Souza Baptista
J. Fungi 2026, 12(2), 155; https://doi.org/10.3390/jof12020155 - 21 Feb 2026
Viewed by 1501
Abstract
“Unusual sporotrichosis”, a concept proposed in this review, refers to severe, extracutaneous, or anatomically atypical manifestations of sporotrichosis occurring in immunocompetent hosts and represents an underrecognized clinical subset associated with important diagnostic and therapeutic challenges. This systematic review aimed to characterize unusual sporotrichosis [...] Read more.
“Unusual sporotrichosis”, a concept proposed in this review, refers to severe, extracutaneous, or anatomically atypical manifestations of sporotrichosis occurring in immunocompetent hosts and represents an underrecognized clinical subset associated with important diagnostic and therapeutic challenges. This systematic review aimed to characterize unusual sporotrichosis worldwide and to clarify its epidemiological, clinical, diagnostic, and therapeutic patterns. Following a registered protocol and PRISMA guidelines, PubMed, Scopus, and BVS/LILACS were searched up to November 2025 using a PICO-based strategy. Eligible studies included peer-reviewed case reports and case series with laboratory-confirmed sporotrichosis in patients without immunosuppression, diabetes mellitus, alcoholism, or other confounding comorbidities; classical lymphocutaneous and fixed cutaneous forms were excluded. From 922 records, 39 studies were included (13 case series and 26 case reports), yielding 55 cases reported between 1957 and 2024 across five world regions, mainly from the United States of America and Brazil. Adults aged 40–59 years (41.8%) and males (74.5%) predominated. Sapronotic transmission was most frequent (69.0%), although zoonotic transmission increased over time. Sporothrix schenckii/Sporothrix schenckii sensu stricto was the predominant species (87.3%). Osteoarticular (30.9%) and systemic (27.2%) forms were the most common presentations. Although cure was achieved in most cases (58.1%), sequelae were frequent (21.8%), and the worst prognosis—including most deaths—was observed in osteoarticular sporotrichosis. Unusual sporotrichosis is globally distributed and clinically distinct; therefore, early recognition and multimodal diagnostic and therapeutic strategies are essential to improve outcome. Full article
(This article belongs to the Special Issue Mycological Research in the Americas)
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18 pages, 2385 KB  
Article
Metagenomic and Genomic Analyses Reveal Prevalent Spread and Evolution of the Bat White-Nose Pathogen Pseudogymnoascus destructans in Western Canada
by Yue Wang, Chadabhorn Insuk, Cory Olson and Jianping Xu
J. Fungi 2026, 12(2), 154; https://doi.org/10.3390/jof12020154 - 21 Feb 2026
Viewed by 1272
Abstract
Bats play a crucial role in the ecosystem. However, North American bat populations have experienced a dramatic decline since 2006 due to white-nose syndrome, a disease caused by Pseudogymnoascus destructans (Pd). This fungus can invade and damage the skin on bat [...] Read more.
Bats play a crucial role in the ecosystem. However, North American bat populations have experienced a dramatic decline since 2006 due to white-nose syndrome, a disease caused by Pseudogymnoascus destructans (Pd). This fungus can invade and damage the skin on bat wings and muzzles during hibernation. Since 2021, Pd has been reported at selected sites in western Canada, the region with the highest bat diversity in Canada, eliciting urgent calls for action among diverse stakeholders. Here we analyze nine metagenomes of bat guanos and wing swabs and the genomes of five Pd strains from western Canada to investigate the distribution and diversity of Pd in this region. Pd was found in all nine metagenomic samples and the metagenome sequences enabled us to identify the associated bat species. Divergence time estimates of Pd based on whole-genome sequences suggest that Pd likely entered Alberta two to five years before its first official report. Furthermore, we found evidence of abundant gene copy number variations in this species. Together, our metagenomic and genomic analyses indicate that Pd is more prevalent than currently recognized and is evolving and diversifying. Continued surveillance with more comprehensive methods is needed to accurately track its spread and facilitate timely management of white-nose syndrome in North America. Full article
(This article belongs to the Section Fungal Genomics, Genetics and Molecular Biology)
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19 pages, 12218 KB  
Article
Taxonomy and Phylogeny of Amylocorticiales (Basidiomycota): Two New Genera, Six New Species, and Four New Combinations
by Yu-Qing Liu, Jing Ye, Si-Yi He, Yuan Yuan, Sen Liu, Yue Li, Man-Rong Huang, Ning Yang and Shuang-Hui He
J. Fungi 2026, 12(2), 153; https://doi.org/10.3390/jof12020153 - 20 Feb 2026
Viewed by 904
Abstract
Amylocorticiales forms a well-supported clade within Agaricomycetes, Basidiomycota, and most of the species have resupinate basidiomes and cause brown rot on wood. It is one of the smallest orders of the basidiomycetes, with the species diversity and phylogeny being understudied. In the present [...] Read more.
Amylocorticiales forms a well-supported clade within Agaricomycetes, Basidiomycota, and most of the species have resupinate basidiomes and cause brown rot on wood. It is one of the smallest orders of the basidiomycetes, with the species diversity and phylogeny being understudied. In the present study, we conduct phylogenetic analyses based on the concatenated ITS + nLSU sequence dataset of the order with an emphasis on the samples from southern China. As a result, ten new lineages were found. Combined with the morphological evidence, two new genera and six new species are described and illustrated, and four new combinations are proposed. Amylophanerochaete hainanense gen. et sp. nov. is closely related to Serpulomyces but differs in having smooth hymenophores with rhizomorphs and narrowly cylindrical to slightly sigmoid amyloid basidiospores. The new genus Pseudoathelia is proposed to accommodate Leptosporomyces linzhiense and Athelia septentrionalis, two athelioid species. Four new species, viz. Amylocorticium athelioides, A. bisporum, A. guangxiense, and A. luteolum, collected from southern China, formed distinct lineages within the Amylocorticium clade. Serpulomyces borealis, the only species of the genus, is proven to be a species complex, while one new species, S. subborealis, and two new combinations, S. rhizomorphus and S. yunnanensis, transferred from Ceraceomyces, are found in the lineage. An identification key to all the known genera is provided. Full article
(This article belongs to the Special Issue Diversity, Phylogeny and Ecology of Forest Fungi, 2nd Edition)
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19 pages, 1102 KB  
Review
Therapeutic Tools for Vulvovaginal Candidiasis: Current and Emerging Antifungal Agents
by Guillermo Quindós, Iker De-la-Pinta, Cristina Marcos-Arias, Nerea Jauregizar, Elena Sevillano, Lucila Madariaga and Elena Eraso
J. Fungi 2026, 12(2), 152; https://doi.org/10.3390/jof12020152 - 20 Feb 2026
Cited by 3 | Viewed by 1874
Abstract
Vulvovaginal candidiasis (VVC) represents a widespread gynaecological challenge, affecting approximately 75% of women at some point during their reproductive years, with a significant subset progressing to recurrent forms (RVVC). Classical azoles and polyenes remain the cornerstone of therapy. However, their clinical utility is [...] Read more.
Vulvovaginal candidiasis (VVC) represents a widespread gynaecological challenge, affecting approximately 75% of women at some point during their reproductive years, with a significant subset progressing to recurrent forms (RVVC). Classical azoles and polyenes remain the cornerstone of therapy. However, their clinical utility is undermined by the rise of azole-resistant non-Candida albicans species, the capacity of Candida to form biofilms, and a complex variety of host-related factors that complicate disease expression and therapeutic response. This narrative review provides a critical up-to-date examination of the therapeutic landscape, integrating current diagnostic algorithms with pharmacological strategies for both acute, recalcitrant and recurrent VVCs. We assess the efficacy and safety of established antifungal agents alongside the breakthrough introduction of novel drug classes, with a particular interest in the oral triterpenoid ibrexafungerp and the tetrazole oteseconazole, which offer new mechanisms of action for cases that fail to respond to standard regimens. Furthermore, we address the management of a special clinical scenarios, including pregnancy and lactation, and explore promising emerging innovative approaches such as mucoadhesive formulations, immunomodulatory approaches, and alternative non-antifungal therapies. Ultimately, this review aims to support clinical decision-making by balancing the accessibility and user-friendliness of conventional treatments with the targeted precision offered by modern antifungal agents. Full article
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14 pages, 1546 KB  
Article
Effects of Isaria cateniannulata on Enzyme Activities and Chitinase Genes in Tetranychus urticae
by Lingdi Gu, Xue Yang, Ying Ren, Kaiwei Tang, Can Liu, Weichen Yang, Qingfu Chen and Xiaona Zhang
J. Fungi 2026, 12(2), 151; https://doi.org/10.3390/jof12020151 - 19 Feb 2026
Viewed by 629
Abstract
Tetranychus urticae is a globally important economic pest mite. Isaria cateniannulata can infect the mite through its conidial penetration of the cuticle, ultimately leading to host mortality; however, the immune mechanisms involving enzyme activity systems and chitinase genes of T. urticae during this [...] Read more.
Tetranychus urticae is a globally important economic pest mite. Isaria cateniannulata can infect the mite through its conidial penetration of the cuticle, ultimately leading to host mortality; however, the immune mechanisms involving enzyme activity systems and chitinase genes of T. urticae during this process remain unclear. In this study, T. urticae infected with I. cateniannulata was used as experimental material to analyze changes in antioxidant and detoxification enzyme activities during infection. In addition, the expression patterns of six chitinase genes were analyzed, and significantly upregulated genes were selected for bioinformatics analysis and functional verification. The results showed that infection with I. cateniannulata enhanced the activity of defense-related enzymes in T. urticae, with Catalase (CAT) and mixed function oxidase (MFO) playing dominant roles. All six chitinase genes were activated, among which TuCHT7 and TuCHT12 were significantly upregulated at 24 h post-infection, and then TuCHT7 gradually declined. Whereas TuCHT12 maintained a sustained and stable induction pattern, TuCHT10 was suppressed, while the other genes exhibited transient expression. Feeding dsTuCHT12 suppressed the expression of this gene within 72 h, with significant suppression observed at 48 h. At this time, the expression levels of TuCHT8, TuCHT9 and TuCHT10 genes were activated, whereas TuCHT7 and TuCHT11 were suppressed. The combined application of dsCHT12 and I. cateniannulata increased the mortality of T. urticae by 3.19-fold and reduced egg production by 52.8%. This study preliminarily revealed the defense response mechanism of T. urticae against I. cateniannulata infection and provides a theoretical basis for eco-friendly pest control based on the combined application of RNAi and arthropodpathogenic fungi. Full article
(This article belongs to the Section Fungi in Agriculture and Biotechnology)
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26 pages, 2153 KB  
Review
Mining Genetically Encoded Biosensors from Filamentous Fungi
by Shuhui Guo, Shaozheng Song, Zhunzhun Liu, Yunjun Ge and Ye Chen
J. Fungi 2026, 12(2), 150; https://doi.org/10.3390/jof12020150 - 19 Feb 2026
Viewed by 1158
Abstract
Genetically encoded biosensors represent cutting-edge biosensors due to their capabilities in real-time monitoring and precise control in living cells. However, the development of eukaryotic genetically encoded biosensors for new analytes is constrained by the shortage of signal–receptor pairs. Bacterial biosensors have been transferred [...] Read more.
Genetically encoded biosensors represent cutting-edge biosensors due to their capabilities in real-time monitoring and precise control in living cells. However, the development of eukaryotic genetically encoded biosensors for new analytes is constrained by the shortage of signal–receptor pairs. Bacterial biosensors have been transferred to eukaryotes to expand the signal detection space, which has achieved remarkable success. However, due to the significant differences between eukaryotic and prokaryotic gene expression systems, optimizing bacterial biosensors has proven challenging. Successful cases indicate that developing orthogonal signal–receptor pairs directly from eukaryotic systems may offer a viable solution. Indeed, the potential of filamentous fungi—a highly diverse group of organisms that share conserved as well as specific signaling and metabolic pathways with yeast and mammalian cells—has been largely overlooked in biosensor development. In this review, we systematically examine biosensing systems in filamentous fungi, summarize their signal recognition receptors, signal transduction pathways, responsive transcription factors, and provide an overview of the biosensors and synthetic tools developed from them. Finally, we highlight the promise and challenges of biosensor development from filamentous fungi and discuss their potential applications. Full article
(This article belongs to the Section Fungal Genomics, Genetics and Molecular Biology)
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16 pages, 8308 KB  
Article
Transcriptomic and Physiological Profiling Reveals Metabolic Determinants and Key Regulatory Hubs of Fruiting Body Degeneration in Lentinula edodes
by Huiting Yang, Kun Liu, Jun Jiang, Xiaoya Song, Xinyan Lu, Jianfei Tan and Lingli Li
J. Fungi 2026, 12(2), 149; https://doi.org/10.3390/jof12020149 - 19 Feb 2026
Viewed by 778
Abstract
Frequent strain degeneration during subcultivation, characterized by impaired sporulation and fruiting body formation, represents a major constraint in fungal agricultural production. Our study systematically investigated two naturally degenerated Lentinula edodes strains classified as abortive (Abt: L808-13, L808-14) and malformed (Abn: L808-18) fruiting-body phenotypes, [...] Read more.
Frequent strain degeneration during subcultivation, characterized by impaired sporulation and fruiting body formation, represents a major constraint in fungal agricultural production. Our study systematically investigated two naturally degenerated Lentinula edodes strains classified as abortive (Abt: L808-13, L808-14) and malformed (Abn: L808-18) fruiting-body phenotypes, through comprehensive phenotypic characterization, enzymatic profiling, thermotolerance assessment, and transcriptomic analysis. While vegetative growth remained unaffected, degenerated strains exhibited premature hyphal knotting, significantly reduced thermotolerance, and Abn-specific suppression of carboxymethyl cellulase (CMCase) activity. Comparative transcriptomics revealed 1239 and 582 differentially expressed genes (DEGs) in Abt and Abn groups, respectively, accompanied by a global dysregulation in carbohydrate catabolism, phospholipid metabolism, and redox homeostasis. Furthermore, protein–protein interaction (PPI) networks and RT-qPCR data highlighted 12 core hub genes enriched in glycoside hydrolysis, cytochrome P450 signaling, and membrane lipid dynamics. These findings provide mechanistic insights into the molecular basis of fruiting body degeneration and establish a foundation for developing diagnostic indicators to screen for early-stage degeneration in industrial mushroom production. Full article
(This article belongs to the Section Fungal Cell Biology, Metabolism and Physiology)
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23 pages, 2719 KB  
Article
Screening and Validation of Interacting Proteins of Receptor-like Cytoplasmic Kinase OsRLCK118 Involved in Rice Blast Resistance
by Wenxiao Wang, Mingmin Wang, Ruiyu Wang, Shaojun Lin, Fenghuang Huang, Yidan Jin, Niqing He, Zhaoping Cheng, Qingshun Q. Li and Dewei Yang
J. Fungi 2026, 12(2), 148; https://doi.org/10.3390/jof12020148 - 19 Feb 2026
Viewed by 838
Abstract
Rice blast, caused by Magnaporthe oryzae, severely threatens global rice production. Although the receptor-like cytoplasmic kinase OsRLCK118 positively regulates rice immunity, its downstream signaling mechanism remains unknown. To systematically identify OsRLCK118-interacting proteins, we performed immunoprecipitation–mass spectrometry (IP-MS) and a yeast library screen, [...] Read more.
Rice blast, caused by Magnaporthe oryzae, severely threatens global rice production. Although the receptor-like cytoplasmic kinase OsRLCK118 positively regulates rice immunity, its downstream signaling mechanism remains unknown. To systematically identify OsRLCK118-interacting proteins, we performed immunoprecipitation–mass spectrometry (IP-MS) and a yeast library screen, yielding 781 and 287 candidates, respectively, with 35 overlapping hits. Among these, OsSAMS1, a known positive regulator of blast resistance, was selected for validation. Membrane yeast two-hybrid, split-luciferase complementation, and co-immunoprecipitation assays confirmed the physical interaction between OsRLCK118 and OsSAMS1. Furthermore, in vitro kinase assays showed that OsRLCK118 specifically phosphorylates OsSAMS1. These results uncover a novel signaling axis connecting pathogen recognition to ethylene biosynthesis via OsRLCK118-dependent phosphorylation of OsSAMS1, providing both mechanistic insight into rice immunity and potential genetic targets for resistance breeding. Full article
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17 pages, 2117 KB  
Article
Low-Intensity, Short-Duration Proton Irradiation Enhances Oxidative Stress Sensitivity of Aspergillus nidulans, with Transcriptomic Data Indicating Downregulation of Antioxidative Enzyme Genes
by Máté Szarka, Ildikó Vig, András Fenyvesi, Barnabás Cs. Gila, Károly Antal, Zita Szikszai, István Pócsi and Tamás Emri
J. Fungi 2026, 12(2), 147; https://doi.org/10.3390/jof12020147 - 19 Feb 2026
Cited by 1 | Viewed by 984
Abstract
Fungi regularly occur on spacecrafts, posing a serious risk to humans and equipment. In this study, we characterized how the model organism Aspergillus nidulans responds to low-intensity, short-duration proton irradiation designed to simulate a solar particle event, a common stress factor in space. [...] Read more.
Fungi regularly occur on spacecrafts, posing a serious risk to humans and equipment. In this study, we characterized how the model organism Aspergillus nidulans responds to low-intensity, short-duration proton irradiation designed to simulate a solar particle event, a common stress factor in space. The oxidative stress-sensitive ∆atfA mutant exhibited a lower survival rate than the wild-type strain. Pretreatment of the wild-type strain with menadione sodium bisulfite (MSB), which activates oxidative stress defense mechanisms, increased tolerance to proton beam radiation. These data are consistent with the idea that oxidative defense contributes to cellular responses to ionizing radiation. Unexpectedly, the applied radiation decreased the tolerance to MSB. To understand this unusual behavior, we compared the transcriptomes of the irradiated and non-irradiated mycelia. As expected, proton beam irradiation upregulated many genes involved in DNA repair but downregulated a large number of antioxidant enzyme genes. The downregulation of three key antioxidant genes—prxA (thioredoxin peroxidase), trxB (thioredoxin reductase), and gsh1 (γ-glutamylcysteine synthase)—was further confirmed by RT-qPCR analysis. One possible explanation is that, due to the rapid elimination of reactive oxygen species generated by water radiolysis, the effects of radiolysis-derived electrons could transiently dominate redox signaling. This shift may interfere with redox sensing in the fungus, resulting in reduced antioxidant gene expression and increased sensitivity to oxidative stress. Oxidative stress sensitivity caused by proton radiation may be the Achilles heel of cells that can survive this stress. Full article
(This article belongs to the Section Fungal Cell Biology, Metabolism and Physiology)
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19 pages, 1146 KB  
Review
Research Progress on the Interaction Mechanism Between Morchella and Mycoparasitic Fungi Causing Diseases and Their Biological Control: A Review
by Ruihua Zhao, Jiayi Xie, Pengfei Jin and Xiaolong He
J. Fungi 2026, 12(2), 146; https://doi.org/10.3390/jof12020146 - 17 Feb 2026
Cited by 2 | Viewed by 987
Abstract
Morchella is a highly valued edible and medicinal fungus with significant nutritional and economic value. In recent years, with the development of artificial cultivation techniques, the planting area of Morchella has been expanding continuously, while the incidence of fungal diseases has also increased [...] Read more.
Morchella is a highly valued edible and medicinal fungus with significant nutritional and economic value. In recent years, with the development of artificial cultivation techniques, the planting area of Morchella has been expanding continuously, while the incidence of fungal diseases has also increased sharply, seriously affecting its yield and quality and further restricting the development of the Morchella industry. To date, ten fungal diseases of Morchella have been reported, mainly including white mold, cobweb disease, pileus rot and fungal rot. The mycoparasitic fungi responsible for these diseases can infect Morchella by secreting proteins and secondary metabolites, while Morchella responds to fungal disease stress through genetic and metabolic regulation. Currently, biological control strategies for Morchella fungal diseases primarily rely on antagonistic microorganisms and natural products. This review summarizes the research progress on major fungal diseases of Morchella, including their causal fungi, pathogenic factors and infection mechanisms, host response mechanisms, and biological control. It also identifies existing research gaps and prospects for future research directions. Full article
(This article belongs to the Section Fungal Pathogenesis and Disease Control)
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18 pages, 2387 KB  
Article
Similiboletinus tomentopileatus gen. et sp. nov. from Tropical China, a New Boletoid Taxon Based on Morphological and Molecular Data
by Sipeng Jian, Xinjing Xu, Feng Gao, Yiwei Fang, Tianwei Yang, Jing Liu, Wenzhu Ai, Chunxia Zhang and Yanchun Li
J. Fungi 2026, 12(2), 145; https://doi.org/10.3390/jof12020145 - 16 Feb 2026
Viewed by 845
Abstract
Boletaceae, a family of profound ecological influence and economic value, has been the subject of prolific taxonomic research over the past decade, with new taxa continually being discovered. In this study, Similiboletinus tomentopileatus gen. et sp. nov., collected from a tropical region of [...] Read more.
Boletaceae, a family of profound ecological influence and economic value, has been the subject of prolific taxonomic research over the past decade, with new taxa continually being discovered. In this study, Similiboletinus tomentopileatus gen. et sp. nov., collected from a tropical region of China, is proposed based on integrative morphological and phylogenetic analyses. Phylogenetic analyses of a multigene dataset (LSU-rpb2-tef1-α-atp6) strongly support the position of Similiboletinus within the Suillelloideae. Morphologically, it is characterized by a tomentose and yellow–brown pileus, a subdecurrent hymenophore, unchanging context when injured, a boletoid hymenophoral trama, a cutis-type pileipellis, and broadly ellipsoid to ovoid basidiospores. The identification of Similiboletinus further underscores the striking and largely unexplored diversity of Boletaceae within tropical China. Full article
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17 pages, 4287 KB  
Article
Chromatin Remodeling Factor Arp8 Is Involved in the Regulation of Aflatoxin Biosynthesis and Fungal Pathogenicity of Aspergillus flavus
by Dongmei Ma, Yanfang Yao, Wenxin Luo, Penghui Wang, Dandan Wu, Yanling Yang, Kangfu Ye, Yuxin Hu, Ying Gao, Minghui Sun, Can Zhang, Lin Chen, Xixu Chen and Zhenhong Zhuang
J. Fungi 2026, 12(2), 144; https://doi.org/10.3390/jof12020144 - 15 Feb 2026
Viewed by 984
Abstract
Aspergillus flavus Link, 1809 is a pathogenic fungus widely present in the environment. It can infect plants and also acts as an opportunistic pathogen affecting humans and other animals. The aflatoxins (AFs), it produces, can cause cancers such as liver cancer. Therefore, in-depth [...] Read more.
Aspergillus flavus Link, 1809 is a pathogenic fungus widely present in the environment. It can infect plants and also acts as an opportunistic pathogen affecting humans and other animals. The aflatoxins (AFs), it produces, can cause cancers such as liver cancer. Therefore, in-depth research into the pathogenic mechanisms of A. flavus is crucial. Arp8 (Actin-like protein Arp8) is a unique subunit within the chromatin remodeling complex INO80, regulating processes including chromatin remodeling. However, the biological function of Arp8 in A. flavus remains unclear. This study constructed A. flavus arp8 knockout (Δarp8) and complementation (Com-arp8) strains via homologous recombination. Subsequent research revealed that following the deletion of arp8, A. flavus exhibits a reduction of approximately 51% in conidia production, complete abrogation of sclerotia formation, and significantly impairment of aflatoxin B1 (AFB1) biosynthesis. Crop grain colonization and Bombyx mori Linnaeus, 1758 infection models demonstrated that Arp8 plays a crucial role in A. flavus ability to infect hosts. Environmental stress experiments identified Arp8 as a vital factor for A. flavus in response to various environmental stresses. Quantitative RT-PCR (qRT-PCR) analysis indicated Arp8 achieves its biological functions through corresponding regulatory factors. This study elucidates the biological functions of Arp8 in A. flavus growth and development, pathogenicity, and aflatoxin synthesis, laying a foundation to illuminate the mechanisms of A. flavus pathogenicity and AFs production. Full article
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17 pages, 2525 KB  
Article
Cultivation of Inonotus hispidus on Jujube Wood Waste: Effects on Fruiting Body Biological Characteristics, Nutrients, Active Constituents and Metabolic Profiles
by Guangjie Zhang, Shuaichun Huang, Ying Zhang, Dongmei Wu, Yuan Cheng, Hong Li, Changtian Li and Yu Li
J. Fungi 2026, 12(2), 143; https://doi.org/10.3390/jof12020143 - 15 Feb 2026
Viewed by 800
Abstract
Inonotus hispidus is an important medicinal and edible fungus within the “Sanghuang” category, featuring a broad host range and rapid fruiting body growth. However, its wild resources are currently threatened by overharvesting. Simultaneously, large-scale jujube (Ziziphus jujuba) cultivation generates substantial pruning [...] Read more.
Inonotus hispidus is an important medicinal and edible fungus within the “Sanghuang” category, featuring a broad host range and rapid fruiting body growth. However, its wild resources are currently threatened by overharvesting. Simultaneously, large-scale jujube (Ziziphus jujuba) cultivation generates substantial pruning waste, often burned. This study explored the feasibility of using jujube wood as a cultivation substrate for I. hispidus. Three I. hispidus strains, Z1, Z2, and ZL, were cultivated on substrates with varying proportions of jujube wood replacing cottonseed hulls. The biological efficiency, nutritional components, active compounds, and free amino acid profiles of the resulting fruiting bodies were analyzed. Non-targeted metabolomics was used to investigate global metabolic changes. Results indicated that all strains successfully colonized the jujube-based substrates and produced fruiting bodies. Strain ZL exhibited the highest biological efficiency and the shortest growth period on the 48% jujube wood substrate, while others showed significantly increased triterpenoids and flavonoids content. Metabolomic analysis revealed substrate-dependent and strain-specific alterations in metabolic pathways, particularly in amino acid biosynthesis, the TCA cycle, and secondary metabolism. This study confirms jujube wood as a viable alternative substrate for the edible (ZL) and medicinal (Z1, Z2) cultivation of I. hispidus, providing a sustainable production method while establishing a valuable utilization pathway for jujube wood waste. Full article
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12 pages, 714 KB  
Review
Stress-Activated Protein Kinase Pathways as Potential Targets for the Development of New Antifungals
by Rebeca Alonso-Monge, José Pedro Guirao-Abad and Juan Carlos Argüelles
J. Fungi 2026, 12(2), 142; https://doi.org/10.3390/jof12020142 - 14 Feb 2026
Viewed by 888
Abstract
The World Health Organization WHO considers fungal infections as a significant global risk that necessitates the development of new therapies. The arsenal of antifungals is limited, and the eukaryotic organization of fungi makes it difficult to find selective antifungal targets. In the search [...] Read more.
The World Health Organization WHO considers fungal infections as a significant global risk that necessitates the development of new therapies. The arsenal of antifungals is limited, and the eukaryotic organization of fungi makes it difficult to find selective antifungal targets. In the search for potential targets for the design of new antifungals, the Stress-Activated Protein Kinase (SAPKs) pathways, and specifically, the two-component system, could be a plausible option since this upstream signaling system is absent in metazoans. SAPK pathways are involved in the response and adaptation to different environmental conditions. In pathogenic fungi, these signaling pathways are crucial for virulence, and some of them become activated in response to certain antifungals. Although further experimental evidence is required on the role of SAPKs in antifungal signaling and resistance, the possibility of impairing SAPK signaling by tagging the two-component system can be considered a useful strategy for implementing future antifungal therapies. In particular, the beneficial value of SAPK modulators combined with antifungal drugs should be a preferred line of research. In this review, we focused on the connection between the SAPK pathways and antifungal signaling in the four fungal priority pathogens, Cryptococcus neoformans, Candidozyma (formerly Candida) auris, Aspergillus fumigatus, and Candida albicans, defined by the WHO. Full article
(This article belongs to the Special Issue Advances in Antifungal Drugs, 2nd Edition)
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15 pages, 3260 KB  
Article
A Near-T2T Genome Assembly of Elsinoe fawcettii Provides Insights into Host Adaptation Driven by Cis-Regulatory Evolution
by Jiyu Su, Shujun Zhang, Qian Lu, Jie Yang, Cheng Zheng, Xiuxiu Li, Xiaofeng Chen, Hong Liu, Zonghua Wang and Hongli Hu
J. Fungi 2026, 12(2), 141; https://doi.org/10.3390/jof12020141 - 13 Feb 2026
Viewed by 837
Abstract
Elsinoe fawcettii is a devastating citrus pathogen worldwide, yet high-quality genomic resources are lacking, limiting insights into its adaptive mechanisms. Seventeen strains collected from 13 host species across 5 Chinese provinces were confirmed as E. fawcettii by multi-loci (ITS, rpb2, tef1-α [...] Read more.
Elsinoe fawcettii is a devastating citrus pathogen worldwide, yet high-quality genomic resources are lacking, limiting insights into its adaptive mechanisms. Seventeen strains collected from 13 host species across 5 Chinese provinces were confirmed as E. fawcettii by multi-loci (ITS, rpb2, tef1-α) phylogenetic and morphological analyses. A near-telomere-to-telomere (near-T2T) genome for representative strain FJ-Y-3 was constructed using integrated PacBio and Hi-C sequencing. The 24.40 Mb assembly was organized into 11 chromosomes with exceptional completeness (BUSCO: 97.1%) and continuity (scaffold N50: 2.18 Mb). Pan-genome analysis revealed a closed structure, with core genes representing 77.19% of the total, suggesting evolutionary adaptation through fine-regulation of conserved elements rather than extensive gene content variation. Accessory genes were significantly enriched in terpenoid/polyketide metabolism, cell surface remodeling, and xenobiotic degradation, underscoring metabolic plasticity. Whole-genome resequencing showed single-nucleotide polymorphisms as the dominant variant, with ~60% residing in regulatory regions, implicating cis-regulation as a key adaptive mechanism. This work provides a high-quality genome and multi-omics framework for E. fawcettii, establishing a crucial molecular foundation for understanding pathogen adaptation and developing sustainable disease management strategies. Full article
(This article belongs to the Section Fungal Genomics, Genetics and Molecular Biology)
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21 pages, 6567 KB  
Article
Comparative Metabolomics Reveals the Adaptive Strategy of White Auricularia cornea to Bamboo Substrate Variation
by Xianqi Shan, Fangjie Yao, Lixin Lu, Xiaoxu Ma, Ming Fang, Wei Liu, Jia Lu, Shengtao Qu, Zirui Zhao, Haimeng Zhao, Xu Sun and Zufa Zhou
J. Fungi 2026, 12(2), 140; https://doi.org/10.3390/jof12020140 - 13 Feb 2026
Viewed by 730
Abstract
To address the “fungus-forest conflict” in the edible mushroom industry and the challenge of resource utilization for bamboo substrate waste, this study focused on white Auricularia cornea, and cultivation systems were established with bamboo substrate replacing wood chips at ratios of 0%, [...] Read more.
To address the “fungus-forest conflict” in the edible mushroom industry and the challenge of resource utilization for bamboo substrate waste, this study focused on white Auricularia cornea, and cultivation systems were established with bamboo substrate replacing wood chips at ratios of 0%, 18%, 38%, 58%, and 78%. By integrating liquid chromatography–tandem mass spectrometry (LC-MS/MS) analysis with agronomic trait measurements, the study elucidated the metabolic adaptation mechanisms of the substrate. Results indicated that white A. cornea could grow normally in all bamboo substrates, with the 58% bamboo substrate replacement group (D_58) demonstrating the most optimal overall performance. The mycelial growth rate reached 3.55 ± 0.24 mm/d, and the growth period was the shortest (86.2 d), balancing growth efficiency with cost advantages. Metabolomics detected 3779 metabolites, primarily amino acids and their derivatives (42.2%) and organic acids (35.54%). Compared to the control group, each treatment group exhibited 104–528 upregulated and 192–630 downregulated differential metabolites, with 93 shared differential metabolites and numerous unique markers. KEGG pathway enrichment analysis revealed that varying bamboo substrate ratios shaped growth adaptation strategies by regulating core pathways such as nucleotide metabolism and ABC transporters. This study established the feasibility and optimal formulation of bamboo substrate substitution, elucidated the substrate–metabolite–phenotype linkage mechanism, and provided theoretical foundations and practical references for high-quality cultivation of white A. cornea and sustainable development through “substituting bamboo for wood” to reduce carbon emissions. Full article
(This article belongs to the Section Fungal Cell Biology, Metabolism and Physiology)
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2 pages, 151 KB  
Correction
Correction: Mourou et al. Diversity of Fungi Associated with Diseases of Cultivated Brassicaceae in Southern Italy. J. Fungi 2026, 12, 13
by Marwa Mourou, Maria Luisa Raimondo, Milan Spetik, Francesco Lops, Gaetana Ricciardi, Maria Grazia Morea, Ales Eichmeier and Antonia Carlucci
J. Fungi 2026, 12(2), 139; https://doi.org/10.3390/jof12020139 - 13 Feb 2026
Viewed by 477
Abstract
There was an error in the original publication [...] Full article
(This article belongs to the Section Fungal Evolution, Biodiversity and Systematics)
17 pages, 1034 KB  
Systematic Review
Can Artificial Intelligence Optimize the Early Diagnosis of Invasive Candidiasis? A Systematic Review and Meta-Analysis
by Hugo Almeida, Beatriz Rodríguez-Alonso, Montserrat Alonso-Sardón, Inmaculada Izquierdo, Ángela Romero-Alegría, Virginia Velasco-Tirado, Josué Pendones Ulerio, Javier Pardo Lledías and Moncef Belhassen-García
J. Fungi 2026, 12(2), 138; https://doi.org/10.3390/jof12020138 - 13 Feb 2026
Viewed by 866
Abstract
The early diagnosis of invasive candidiasis remains challenging in immunocompromised and other high-risk patients, prompting interest in artificial intelligence models for assisting clinical decision-making. We conducted a PROSPERO-registered systematic review and meta-analysis of artificial intelligence-based predictive models for the early identification of invasive [...] Read more.
The early diagnosis of invasive candidiasis remains challenging in immunocompromised and other high-risk patients, prompting interest in artificial intelligence models for assisting clinical decision-making. We conducted a PROSPERO-registered systematic review and meta-analysis of artificial intelligence-based predictive models for the early identification of invasive Candida infections. We searched multiple databases for studies reporting model performance in hospitalized immuno-compromised patients. Data on study characteristics, model details, validation strategy, and diagnostic accuracy were extracted. A bivariate random-effects meta-analysis was performed for candidemia prediction models with compatible data. Eight studies met inclusion criteria. Models were typically developed using retrospective hospital data with heterogeneous populations and predictors. Five candidemia studies provided threshold-based performance data for meta-analysis. Pooled sensitivity and specificity for candidemia prediction were 81.3% (95% confidence interval (CI) 72.9–87.6%) and 81.6% (95% CI 68.4–90.1%), respectively. Most models achieved high negative predictive values, whereas positive predictive values were modest, reflecting low event prevalence. The risk of bias was generally moderate to high (PROBAST), and the certainty of evidence was low (GRADE) due to study limitations and indirectness. AI models show promise for early candidemia identification with moderate diagnostic accuracy. They may be useful as decision-support tools, but further multicenter prospective validation is needed before routine clinical adoption. Full article
(This article belongs to the Section Fungal Pathogenesis and Disease Control)
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21 pages, 2370 KB  
Article
An Ammonium Transporter Gene Contributes to the Aggressiveness of the Dutch Elm Disease Pathogen Ophiostoma novo-ulmi
by Louis Bernier, Thais C. de Oliveira, Josée-Anne Majeau, Karine V. Plourde, Volker Jacobi, Philippe Tanguay, Paul Y. de la Bastide, Will E. Hintz, Ilga M. Porth, Josée Dufour, Pauline Hessenauer, Christine A. Roden, Cloé Laflamme and Lucie Varlet
J. Fungi 2026, 12(2), 137; https://doi.org/10.3390/jof12020137 - 13 Feb 2026
Viewed by 935
Abstract
Molecular mechanisms determining pathogenicity of the Dutch elm disease fungus Ophiostoma novo-ulmi are poorly understood. Prior identification of the pathogenicity locus pat1 prompted a chromosome walking approach to elucidate gene function in this region. Among 17 identified genes, ONUg0282 (amtA) was [...] Read more.
Molecular mechanisms determining pathogenicity of the Dutch elm disease fungus Ophiostoma novo-ulmi are poorly understood. Prior identification of the pathogenicity locus pat1 prompted a chromosome walking approach to elucidate gene function in this region. Among 17 identified genes, ONUg0282 (amtA) was predicted to encode a high-affinity ammonium transporter. In silico analyses confirmed the presence of four additional amt genes (amtB, amtC, amtD, and amtE) in both O. novo-ulmi and the less aggressive O. ulmi and that amtA and amtB belong to the Saccharomyces cerevisiae mep2 clade. The predicted amtA gene product showed features of Mep2-type transceptors, including amino acid residues corresponding to His-168 and His-318 in Escherichia coli AmtB protein, 11 transmembrane helices, and a conserved 22 amino acid motif immediately downstream of the last transmembrane helix. A knockdown amtA mutant with 25% residual expression was significantly less aggressive than wild-type O. novo-ulmi strain H327 when infecting Ulmus americana × U. parvifolia saplings. Predicted AmtA transporters from two CRISPR-Cas9 knockout mutants contained only five intact transmembrane helices. The ΔamtA mutants retained several wild-type phenotypic traits, including yeast–mycelium dimorphism, but were significantly less aggressive than H327 towards U. americana saplings. We concluded that ONUg0282 is an important determinant of aggressiveness in O. novo-ulmi. Full article
(This article belongs to the Section Fungal Pathogenesis and Disease Control)
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14 pages, 2873 KB  
Article
Genome-Wide Landscape of Position Effect Variegation in Pichia pastoris
by Xueyan Yao, Baosong Chen, Zhihui Zhang, Wenyan Gao, Qian Zhang, Yongpeng Yao, Yuanyuan Pan and Gang Liu
J. Fungi 2026, 12(2), 136; https://doi.org/10.3390/jof12020136 - 12 Feb 2026
Viewed by 768
Abstract
Exogenous genes are generally expressed by integration into the chromosomes of Pichia pastoris. However, systematic studies on the chromosomal position effect are lacking, and locations that are conducive to the high expression of foreign genes are rarely reported. In this study, a [...] Read more.
Exogenous genes are generally expressed by integration into the chromosomes of Pichia pastoris. However, systematic studies on the chromosomal position effect are lacking, and locations that are conducive to the high expression of foreign genes are rarely reported. In this study, a genomic random insertion mutagenesis library for P. pastoris was successfully constructed using the piggyBac (PB) transposon system. Through sequencing, the sequence TTAA was identified as the major recognition site of the PB transposon, which exhibited relatively high coverage on P. pastoris chromosomes, making it a valuable tool for studying position effect variegation in P. pastoris. Using the enhanced green fluorescent protein gene (eGFP) as a reporter, two libraries including low-expression positions and high-expression positions were obtained by flow cytometry. The low-expression sites were mainly located upstream of ORFs around the promoter region and downstream near the terminator region, while the high-expression sites were predominantly located at the gene interior. KEGG and GO analyses showed that genes in high-expression positions were significantly enriched in the ATP-dependent chromatin remodeling and histone binding pathways, and genes in low-expression positions were significantly enriched in the MAPK signaling pathway, autophagy, mitochondrial autophagy, ABC transporters, and the arginine synthesis pathway. This study has clarified the genome-wide landscape of position effect variegation in P. pastoris. Additionally, it has provided novel insights into high-throughput screening strategies for strains with high exogenous gene expression. Full article
(This article belongs to the Section Fungal Genomics, Genetics and Molecular Biology)
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8 pages, 1008 KB  
Technical Note
False-Positive IMMY® Serum Cryptococcal Antigen Lateral Flow Assay Results Due to Improper Use of Titration Buffer
by Asmus Tukundane, Wilber Bakka, Mercy Amanige, Dora Babirye, Timothy Mugabi, Abdu Musubire, David B. Meya, David R. Boulware, Brian Doherty, Richard Kwizera and Caleb P. Skipper
J. Fungi 2026, 12(2), 135; https://doi.org/10.3390/jof12020135 - 12 Feb 2026
Viewed by 1195
Abstract
The IMMY cryptococcal antigen (CrAg) lateral flow assay (LFA) is a reliable diagnostic tool for Cryptococcus detection, but false-positive results may arise from procedural or reagent-related errors, underscoring careful operation of the assay to ensure diagnostic accuracy and prevent unnecessary treatment. Two patients [...] Read more.
The IMMY cryptococcal antigen (CrAg) lateral flow assay (LFA) is a reliable diagnostic tool for Cryptococcus detection, but false-positive results may arise from procedural or reagent-related errors, underscoring careful operation of the assay to ensure diagnostic accuracy and prevent unnecessary treatment. Two patients who were initially reported as CrAg-positive by a peripheral laboratory were referred to Kiruddu Hospital in Kampala, Uganda, for clinical assessment and confirmatory testing. Repeat tests were conducted using specimen diluent following the manufacturer’s protocol, resulting in negative results. Semi-quantitative CrAg LFA testing and a series of control assays were performed to identify the source of error. We were able to consistently reproduce positive results when the titration diluent was inappropriately used instead of the specimen diluent. Serial dilutions confirmed persistent false positivity up to 1:80 when inappropriately using the titration diluent, while all dilutions that appropriately started with the specimen diluent remained negative. We hypothesize that the incorrect use of titration diluent instead of specimen diluent led to false-positive CrAg LFA results due to the absence of the blocking agent that neutralizes heterophilic antibodies. Procedural errors can lead to diagnostic misinterpretation and serious consequences in clinical management, emphasizing the importance of adherence to manufacturer’s instructions. Full article
(This article belongs to the Section Fungal Pathogenesis and Disease Control)
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21 pages, 8792 KB  
Article
Lipid Metabolism and Membrane Remodeling Drive Sclerotium Formation in Morchella eximia: Insights from Integrated Transcriptomics and Metabolomics
by Chunmou Wei, Jimeng Li, Zhongmei Mo, Wei Liu, Dan Zheng, Xueyan Chen, Fulin Li, Mingfeng Tai, Jiaxin Song, Changhua Gu and Qianqian Zhang
J. Fungi 2026, 12(2), 134; https://doi.org/10.3390/jof12020134 - 12 Feb 2026
Viewed by 897
Abstract
Sclerotium formation represents a critical transition phase in the life cycle of morel, shifting from vegetative growth to dormant structures. The capacity for sclerotium formation directly influences the yield and stability of artificial cultivation. To elucidate the molecular regulatory mechanisms underlying this process, [...] Read more.
Sclerotium formation represents a critical transition phase in the life cycle of morel, shifting from vegetative growth to dormant structures. The capacity for sclerotium formation directly influences the yield and stability of artificial cultivation. To elucidate the molecular regulatory mechanisms underlying this process, a combined transcriptomics and metabolomics approach was employed to analyze gene expression and metabolite dynamics during sclerotium development of Morchella eximia. A total of 2567 differentially expressed metabolites (DEMs) and 2314 differentially expressed genes (DEGs) were detected, primarily enriched in amino acid metabolism, lipid synthesis, and energy metabolism pathways. Amino acid metabolism facilitates protein synthesis and supplies carbon skeletons, while lipid metabolic networks, particularly de novo fatty acid synthesis from acetyl-CoA precursors, glycerophospholipid metabolism, sphingolipid metabolism, and unsaturated fatty acid biosynthesis, play a central role in sclerotium formation. A regulatory model was constructed, focusing on signal response, transcriptional regulation, nutrient transport and metabolism, morphology transition, lipid accumulation, and membrane system remodeling, demonstrating that lipids not only provide energy storage and membrane structural components for sclerotia but also mediate developmental transitions and environmental adaptation through signaling molecules and regulation of membrane properties. These findings systematically reveal the regulatory network governing morel sclerotium formation at the multi-omics level, with particular emphasis on the central role of lipid metabolism and membrane remodeling. The results offer a theoretical foundation for improving morel cultivation yield and stability through targeted metabolic regulation strategies. Full article
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16 pages, 7127 KB  
Article
An Efficient and Stable PEG-Mediated Transformation System for Medicinal Fungus Ophiocordyceps xuefengensis: Optimization and Functional Validation
by Xiaoting Feng, Xinyao Sheng, Jun Liu, Rongrong Zhou, Zhongxu Yang, Xiaojuan Tang and Shuihan Zhang
J. Fungi 2026, 12(2), 132; https://doi.org/10.3390/jof12020132 - 12 Feb 2026
Viewed by 817
Abstract
Ophiocordyceps xuefengensis is an important medicinal fungus with considerable pharmaceutical and economic value. However, its industrial and scientific utilization has been severely limited by the lack of an efficient genetic transformation system, largely due to limited genomic information and wild growth. In this [...] Read more.
Ophiocordyceps xuefengensis is an important medicinal fungus with considerable pharmaceutical and economic value. However, its industrial and scientific utilization has been severely limited by the lack of an efficient genetic transformation system, largely due to limited genomic information and wild growth. In this study, we established an efficient and stable plasmid transformation system within O. xuefengensis protoplasts mediated by PEG. To overcome low protoplast yield and transformation efficiency, key factors influencing protoplast preparation including enzyme composition and concentration, fungal age, and digestion conditions were systematically optimized. The optimal protocol involved digesting 4-day-old mycelia with a mixture of 1.5% lywallzyme 1 and 1.5% snailase at 34 °C and 130 rpm for 3.5 h, yielding at least 9.42 × 107 CFU/mL protoplasts. Protoplast regeneration was significantly enhanced in PY medium supplemented with 0.6 M mannitol. Under these optimized conditions, a transformation efficiency of 45.5% was achieved, with stable plasmid integration confirmed over four successive generations. Furthermore, the transformation system was successfully applied to functional gene characterization by driving exogenous gene expression using the endogenous gpd1 promoter. This study provides a foundational platform for functional gene analysis and paves the way for further elucidation of growth and development mechanisms and metabolic engineering in O. xuefengensis. Full article
(This article belongs to the Section Fungal Genomics, Genetics and Molecular Biology)
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17 pages, 913 KB  
Article
Development of Experimental Techniques to Study Blueberry Rust (Pucciniastrum minimum) Urediniospore Survival
by Karina Griffin and Jay M. Anderson
J. Fungi 2026, 12(2), 133; https://doi.org/10.3390/jof12020133 - 11 Feb 2026
Viewed by 880
Abstract
Blueberry rust disease is caused by the fungal pathogen Pucciniastrum minimum (syn. Thekopsora minima). Despite its importance as a plant pathogen, there are relatively few published studies on P. minimum. This study investigated and refined methodologies to cultivate and study this [...] Read more.
Blueberry rust disease is caused by the fungal pathogen Pucciniastrum minimum (syn. Thekopsora minima). Despite its importance as a plant pathogen, there are relatively few published studies on P. minimum. This study investigated and refined methodologies to cultivate and study this obligate parasite. P. minimum was successfully cultivated on detached blueberry leaves by misting leaves with water, followed by dusting with dry urediniospores. In vitro germination of urediniospores on water agar was achieved using a spore dusting technique, and germination rates were 70% higher compared to a spore suspension. Time after leaf detachment affected urediniospore germination and highlighted the importance of the processing time for replicability between experiments. Urediniospore viability could be evaluated by co-staining with fluorescein diacetate and propidium iodide, and the assessed viability was significantly higher than germination rates achieved in vitro. In detached leaf inoculations, leaves sourced from inside the glasshouse developed more rust than those from outside; this is discussed in the context of knowledge gaps on the infection process of P. minimum. This study resolves some key methodological issues involved with studying P. minimum rust urediniospores, and the general protocols we developed can be applied to other rust species for biological survival research. Full article
(This article belongs to the Special Issue Rust Fungi: From Systematics to Sustainable Management)
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19 pages, 596 KB  
Systematic Review
Breaking the Chain of Infection: A Systematic Review of Environmental Decontamination of Candidozyma auris (2017–2025)
by Aristotelis Papadimitriou, Lida-Paraskevi Drosopoulou, Maria Tseroni, Flora V. Kontopidou, Athanasios Tsakris and Georgia Vrioni
J. Fungi 2026, 12(2), 131; https://doi.org/10.3390/jof12020131 - 11 Feb 2026
Viewed by 1056
Abstract
Candidozyma auris is an emerging multidrug-resistant yeast that readily contaminates healthcare environments, persisting on dry surfaces and enabling transmission and difficult-to-control outbreaks. A systematic review of environmental hygiene interventions targeting C. auris was conducted, focusing on efficacy against planktonic cells and surface-associated biofilms [...] Read more.
Candidozyma auris is an emerging multidrug-resistant yeast that readily contaminates healthcare environments, persisting on dry surfaces and enabling transmission and difficult-to-control outbreaks. A systematic review of environmental hygiene interventions targeting C. auris was conducted, focusing on efficacy against planktonic cells and surface-associated biofilms (including dry-surface biofilms, DSB where available). PubMed and Scopus were searched for English-language records published from 1 January 2017 to 30 September 2025, and study selection followed PRISMA 2020. Thirty-six studies from nine countries met the inclusion criteria. These were predominantly laboratory efficacy evaluations using carrier/suspension or quantitative surface methods reporting log10 Colony Forming Unit (CFU) reductions; only seven studies assessed biofilm-associated C. auris. Across clades I–IV, chlorine-based disinfectants and oxidizing chemistries (hydrogen peroxide/peracetic acid formulations) most consistently achieved high-level reductions (often ≥ 5 log10 CFU) under label-relevant conditions. In contrast, products containing only quaternary ammonium compounds (QACs) frequently underperformed and demonstrated greater variability. No-touch methods, particularly 254 nm ultraviolet-C light (UV-C), provided meaningful adjunctive reductions, but were highly dependent on dose delivery and geometry, and evidence for ozone-based approaches was mixed. Limited data on C. auris DSBs suggest planktonic testing may overestimate real-world conditions and underscore the importance of endpoints, such as transfer prevention and regrowth suppression. Full article
(This article belongs to the Special Issue Advances and Innovations in Fungal Infections)
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