Journal of Fungi doi: 10.3390/jof12060413

Authors: Svetlana Vetrova Elena Kozar Irina Engalycheva Kseniya Mukhina Vera Chizhik Viktor Martynov

Fusarium fungi are known to infect table beet (Beta vulgaris subsp. vulgaris) plants at various stages of development worldwide. Fusarium root rot, which develops post-harvest during long-term storage, is of particular economic significance. In Russia, there is no up-to-date information about the species diversity of pathogens causing this disease of table beets, which determined the purpose of this study. A total of 28 Fusarium isolates were collected from affected beet roots grown in the Moscow region of the Russian Federation from 2018 to 2023 years. Molecular phylogeny based on the TEF-1α and RPB2 genes in combination with morphological characterization showed that five Fusarium species were involved in the pathogenesis of Fusarium root rot of table beet during storage: F. acuminatum (43% of the total number of isolates), F. avenaceum, F. campestre (FTSC); F. sporotrichioides (FSAMSC) and F. solani (FSSC). At the same time, the species F. acuminatum, F. campestre, and F. sporotrichioides were first discovered on beet root in the Russian Federation. Temperature sensitivity of the identified species was studied at 5 °C and 25 °C. According to the value of the cold sensitivity index (CTI) on the nutrient medium and native substrate, the isolates were distributed differently: F. campestre (0.32) > F. acuminatum (0.22) > F. avenaceum (0.21) > F. sporotrichioides (0.19) > F. solani (0.20) and F. acuminatum (0.32) > F. campestre (0.21) > F. solani (0.03) > F. avenaceum and F. sporotrichioides (0.01), respectively. This confirms the need to study the pathogenic properties of isolates on a natural substrate (host plant) under different temperature conditions. When infected with the dominant and most aggressive species F. acuminatum, there was a high variation in the size of the affected area, depending on the genotype of the lines, under both temperature conditions (Va = 2–8 mm3 at 5 °C and Va = 31–1760 mm3 at 25 °C). Therefore, this species can be considered to be the most objective differentiating factor in assessing the resistance of table beet roots to fusarium rot, which determines the need to include it in the breeding process for creating resistant varieties and hybrids for the Central region of Russia. The data obtained in this study are of great importance for developing strategies for managing Fusarium fungi associated with Fusarium rot of beet-root during storage. The research results will also be relevant for other vegetable crops that remain fresh for long periods of time or undergo vernalization in the case of seed production at low temperatures.

Journal of Fungi doi: 10.3390/jof12060412

Authors: Shuwen Liu Mian Liu Jian Liu Huali Li Yajiao Sun Mengyao Wang Hongliang Zhang Yunqiang Ma Junjia Lu

To explore efficient and sustainable biocontrol resources against anthracnose in Begonia benariensis, endophytic fungi were isolated from healthy host tissues and screened for antagonistic activity against Colletotrichum aotearoa SWBG5. Among 31 isolates, four showed strong inhibition, and the most potent strain, QYN6, exhibited an in vitro mycelial inhibition rate of 63.67%. Based on morphology and multi-gene phylogeny (ITS, TUB2, TEF-1α), QYN6 was identified as Nigrospora sphaerica. Mechanistic assays revealed that QYN6 secretes multiple cell wall-degrading enzymes (chitinase, β-1,3-glucanase, cellulase, protease) and displays hyperparasitism against the pathogen hyphae (entwining, deformation, swelling), acting synergistically to inhibit fungal growth. In greenhouse pot trials, QYN6 achieved a biocontrol efficacy of 48.91% against Begonia anthracnose. Additionally, QYN6 significantly activated host defense responses, increasing the activities of antioxidant enzymes (SOD, POD, PPO, CAT) and the contents of soluble protein and soluble sugar. Furthermore, QYN6 exhibited multiple plant growth-promoting traits, including IAA production, siderophore synthesis, and potassium solubilization. Inoculation with QYN6 markedly improved plant height, leaf number, root length, and biomass of B. benariensis. Overall, N. sphaerica QYN6 possesses dual biocontrol and growth-promoting potential, providing a promising microbial resource and theoretical basis for green management of Begonia anthracnose.

Journal of Fungi doi: 10.3390/jof12060411

Authors: Qianwen Xu Changlong Xie Dinghui Wang Xiaoxiao Zhu Wenfan Wei Xiaojia Niu Tianming Wang Hongchen Wang Daqiang Wu

The mitochondrial membrane protein UPS1, a conserved intermembrane space protein in Saccharomyces cerevisiae, possesses phosphatidic acid transfer activity and plays a positive regulatory role in processes such as cardiolipin metabolism and transport. The role of UPS1 protein in pathogenic fungi such as Candida albicans has not been explored, especially in relation to its influence on virulence factors like hyphal growth and biofilm formation, which are crucial for the pathogenicity of C. albicans. The research investigated the function of the UPS1 protein in C. albicans by using gene knockout techniques, analyzing mitochondrial function, and conducting tests for hyphal and biofilm development. The results revealed that deletion of the UPS1 gene leads to altered mitochondrial morphology, increased reactive oxygen species levels, and reduced intracellular ATP content, thereby causing severe growth defects in C. albicans. In addition, transcriptomic analysis indicated that loss of UPS1 significantly represses the expression of genes associated with hyphal growth and biofilm formation. Functional assays further confirmed that UPS1 deficiency markedly impairs cell adhesion capability, hyphal development, and biofilm formation of C. albicans. Notably, deletion of the UPS1 protein markedly reduces the susceptibility of C. albicans to membrane-targeted antifungal drugs. Finally, infection models using Galleria mellonella larvae and a murine vulvovaginal candidiasis model verified that UPS1 gene knockout attenuates the pathogenicity of C. albicans. In summary, our findings demonstrate that UPS1 protein modulates the pathogenicity of C. albicans by regulating mitochondrial function, hyphal growth, and biofilm formation.

Journal of Fungi doi: 10.3390/jof12060410

Authors: Hequn Li Hongjuan Yuan Bushra Iqbal Tianyu Wang Zhen Wang Huaiyuan Zhang

Autophagy is a central cellular process that recycles intracellular components and supplies precursors for biosynthesis. As a key regulator of autophagosome formation, autophagy-related protein 8 (ATG8) plays an essential role in macromolecular degradation and in the availability of lipid precursors. However, whether enhanced autophagic flux promotes lipid accumulation in oleaginous fungi remains unclear. In this study, atg8-1 and atg8-2 were homologously overexpressed in the oleaginous fungus Mucor circinelloides to evaluate their roles in lipid biosynthesis. The engineered strains McATG8-1T2 and McATG8-2T2 showed significantly increased total fatty acid (TFA) contents (32.9% and 32.5%), representing improvements of 15.0% and 13.7% compared with the control. γ-Linolenic acid levels were also elevated to 16.9% and 16.5%, relative increases of 25.2% and 22.0%, respectively. RT-qPCR analysis revealed coordinated upregulation of genes involved in autophagy, central carbon metabolism, lipid biosynthesis, and the pentose phosphate pathway. Ethanolamine supplementation further enhanced lipid accumulation, increasing TFA contents by 12.2–14.6%. In addition, inhibition of target of rapamycin complex 1 using rapamycin produced a strong synergistic effect with atg8 overexpression, leading to substantial lipid increases under nitrogen-limited and nitrogen-rich conditions. Collectively, these findings demonstrated that ATG8-mediated autophagy enhanced lipid accumulation and acted as a key determinant of lipid synthesis flux.

Journal of Fungi doi: 10.3390/jof12060409

Authors: Xinyi Zhang Zijia Peng Ruonan Jing Xinye Liu Tauseef Ullah Min Sheng Zhongdong Yu

White pine blister rust disease (WPBR), caused by Cronartium ribicola, ranks among the most destructive pathogens of five-needle pines. We developed a hydroxynaphthol blue (HNB)-based Loop-mediated isothermal amplification (LAMP) assay enabling rapid, visual detection of C. ribicola directly following DNA extraction. LAMP primers targeting the internal transcribed spacer (ITS) region were designed and validated through in silico comparison with related Cronartium species and in vitro testing against sympatric forest fungi. The optimized 25 μL reaction contained 8.0 mM Mg2+, 1.0 mM dNTPs, and an inner-to-outer primer ratio of 8:1, with amplification conducted at 62 °C for 40 min. Positive amplification produced a distinctive color transition from purple to sky blue, enabling visual interpretation without instrumentation. Under the tested conditions, the assay achieved a detection limit of 460 ± 3.2 fg/μL genomic DNA—a 10-fold improvement over conventional PCR in concentration-based sensitivity. Assay applicability was evaluated using 211 field-collected Pinus armandii samples sourced from China. Detection efficiency varied significantly across tissue types. Symptomatic bark exhibited a substantially higher positive detection rate (68.97%, 95% CI: 49.2–84.7%) compared to needles from symptomatic trees (18.75%, 95% CI: 4.1–45.7%). Among asymptomatic samples, 3.75% of bark samples tested positive for C. ribicola DNA, whereas all needle samples were negative. Geographically, positive detections clustered at several discrete sampling sites in southwestern China, predominantly at elevated elevations. The established LAMP-HNB assay provides a rapid, visually interpretable diagnostic tool for early detection and quarantine monitoring of WPBR following DNA extraction. Beyond its practical utility, this assay establishes valuable baseline data for targeted disease surveillance in the context of evolving climate conditions.

Journal of Fungi doi: 10.3390/jof12060408

Authors: Baojun Xu

Fungi represent one of the most diverse and functionally versatile groups of organisms on Earth, with profound impacts on human health, food security, industrial manufacturing, environmental remediation, and ecological sustainability [...]

Journal of Fungi doi: 10.3390/jof12060407

Authors: Ofelia Candolfi-Arballo Amanda Dávila-Lezama Erik Narváez-Hernández Manuel Ontiveros-Duries Jesús Manuel Soto-Reyes José Mauricio Galeana-Pizaña Nydia Alejandra Castillo-Martínez Laura Rosio Castañón-Olivares

Baja California is the second-highest state in Mexico for hospital discharges attributed to coccidioidomycosis (CM), yet epidemiological information on exposure patterns in affected communities remains limited. To estimate exposure to Coccidioides and assess its association with environmental factors, we conducted intradermal coccidioidin skin testing among 416 residents across nine regions of Baja California. We analyzed 24 environmental variables, including bioclimatic, topographic, and land use indicators. Overall, 31.9% of participants tested positive. Higher odds of exposure were observed in Valle de las Palmas and La Morita. Exploratory comparisons of environmental variables showed that, in unadjusted analyses, annual precipitation, precipitation during the wettest month, and elevation differed between high- and low-positivity localities. However, after applying the Benjamini–Hochberg false discovery rate correction, none of the evaluated continuous environmental variables remained statistically significant. These findings should therefore be interpreted as exploratory and hypothesis-generating rather than as evidence of an independently defined environmental profile. Overall, the results indicate heterogeneous exposure to Coccidioides across Baja California and suggest exploratory spatial variability in exposure across sampled localities. Because participants were recruited through nonprobability community-based sampling, these findings should be interpreted as exploratory rather than population-representative estimates.

Journal of Fungi doi: 10.3390/jof12060406

Authors: Narit Thaochan Chaninun Pornsuriya Thanunchanok Chairin Kodeeyah Thoawan Putarak Chomnunti Anurag Sunpapao

There was an error in the original publication regarding several quantitative statements in the Introduction section [...]

Journal of Fungi doi: 10.3390/jof12060405

Authors: Siyuan Yue Ya Li Peng Li Jing Zeng Junhui Nie Cheng Zhang Tong Wang Jianjun Guo Lin Yuan

Lignocellulosic biorefineries are limited by the high cost of cellulolytic enzymes. Consolidated bioprocessing (CBP), which integrates saccharification and fermentation in one step, offers a solution to this challenge. In this study, a cellulase-hyperproducing mutant of Talaromyces pinophilus, Y117, was generated from the parental strain TP117 via sequential ultraviolet irradiation and NTG (N-methyl-N′-nitro-N-nitrosoguanidine) mutagenesis. Enzymatic secretion and lignocellulose degradation capacities were evaluated, focusing on agricultural residues, particularly corncob. Y117’s performance was compared with TP117 and Trichoderma reesei Rut-C30 (TR30) under high-solids fermentation. Furthermore, the lactate dehydrogenase A (ldhA) gene from Rhizopus oryzae was heterologously expressed in Y117 to direct hydrolyzed sugars toward lactic acid (LA). Y117 exhibited significantly enhanced enzymatic secretion, achieving FPase activity of 8.9 IU/mL and a substrate utilization rate of 72.2% at 125 g/L corncob solids. Y117 outperformed TP117 and TR30 in cellulase, xylanase, and CMCase activities, as well as growth under high-solids fermentation conditions. In the LA fermentation process, Y117 produced 14.20 g/L LA, a notable improvement compared to TP117 (5.33 g/L) and TR30 (2.71 g/L). While LA productivity and yield currently remain below bacterial benchmarks, the unique CBP capability of Y117 provides a foundation for further metabolic engineering toward industrial viability. The engineered T. pinophilus Y117 demonstrates promising potential as a CBP platform for efficient straw-to-LA conversion, providing a sustainable approach for third-generation biobased materials production.

Journal of Fungi doi: 10.3390/jof12060404

Authors: Meng-Qian Chen Jun-Xi Liu Jia Ling Xi-Hui Du

The circadian clock is a widespread rhythmic phenomenon across organisms, characterized by distinct gene expression patterns and behaviors at specific times of the day. Extensive genetic studies in the model fungus Neurospora crassa have yielded critical insights into the components and molecular mechanisms of circadian oscillators. However, these understandings remain absent across fungal lineages, especially from edible mushrooms. Morels (Morchella spp.) are well-recognized edible ascomycetes of considerable economic value and are partially artificially cultivated, but their biological characteristics are poorly understood. Investigating the presence of their circadian clock components, as well as the molecular underpinnings of circadian rhythms, holds important biological implications. In this study, we firstly performed a genomic search for homologs of known circadian clock genes in Morchella sextelata. Homologs of seven circadian clock genes, including wc-1, wc-2, fwd-1, frh, frq, and two additional clock-controlled genes, were identified, indicating the components necessary for the operation of a FWC oscillator contained in M. sextelata. Then, using reverse transcription quantitative PCR (RT-qPCR), the expression profiles of these seven circadian clock-related genes and four mating-type genes were examined in RNA samples which were extracted from mycelia of MAT1-1, MAT1-2 and MAT1-1 × MAT1-2 co-culture/crossed condition during conidiation under in vitro cultivation across one day. The expression levels of seven circadian clock genes and four mating-type genes displayed similar time-of-day-specific rhythmic patterns, yet remained consistently distinct across the mating-type strains and their co-culture/crossed condition, indicating a potential correlation between circadian clock and mating-type loci. Collectively, these results suggest that M. sextelata harbors conserved circadian clock-related homologs and displays mating-type-associated temporal expression differences under the tested conidiation conditions, offering a novel perspective for exploring the potential link between clock-related regulation and mating-type background in the future.

Journal of Fungi doi: 10.3390/jof12060403

Authors: Maria Valentina Bedoya-Florez Ricardo A. Murcia-Galán Martha Viviana Roa-Cordero Sandra M. Leal-Pinto Juan David Puerta-Arias Yair Alvarez-Ricardo John J. Hurtado Tonny W. Naranjo

Candida albicans remains one of the leading causes of invasive fungal infections and is recognized as a critical-priority pathogen by the World Health Organization. The increasing emergence of resistance to azole antifungals such as fluconazole highlights the need for alternative therapeutic strategies. In this study, we evaluated the antifungal potential of a chromium(III)–triazole coordination complex (CrL1) against C. albicans. In vitro susceptibility testing showed that CrL1 exhibited notable antifungal activity against the fluconazole-resistant strain with low cytotoxicity in murine macrophages. To facilitate aqueous dispersion and enable in vivo administration, CrL1 was incorporated into an oil-in-water nanoemulsion (NE-CrL1). The antifungal activity of NE-CrL1 was evaluated in a murine model of invasive candidiasis. In mice infected with a fluconazole-resistant C. albicans strain, treatment with NE-CrL1 reduced renal fungal burden and was associated with attenuation of histopathological alterations and changes in local inflammatory responses. Although the present study has limitations, including the absence of mechanistic assays and additional physicochemical characterization, these results suggest in vivo antifungal activity of NE-CrL1 and warrant further preclinical evaluation against drug-resistant Candida infections.

Journal of Fungi doi: 10.3390/jof12060402

Authors: Isabella Burger Simon Leonhartsberger Kathrin Peikert Lukas Fourtis Polina Atanasova Lara T. S. Kramer Richard Fried Christian Stanetty Florian Rudroff Ruth Birner-Gruenberger Robert L. Mach Astrid R. Mach-Aigner Matthias Schittmayer Christian Zimmermann

The biosynthetic gene cluster (BGC) responsible for producing diaporthinic acid has remained genetically unassigned despite repeated isolation of this metabolite from several fungal species. In this study, we activated the dia BGC in Trichoderma reesei by overexpressing the cluster-associated zinc cluster protein DiaR1 to identify the BGC’s in vivo metabolic output and reconstruct the corresponding biosynthetic pathway. Metabolite production was analyzed by HPLC-MS/MS, and the major product was isolated and structurally confirmed by NMR spectroscopy. Individual genes of the dia cluster were deleted in the activated background to assess their functional roles, and transcript levels were quantified by RT-qPCR. Activation of the cluster resulted in the predominant accumulation of diaporthinic acid, accompanied by several related isocoumarin derivatives, while antibacterial and antifungal assays showed no detectable activity of diaporthinic acid under the tested conditions. Deletion analyses demonstrated that the polyketide synthase Dia1, the bifunctional halogenase/methyltransferase Dia5, and the FAD-dependent oxidoreductase Dia4 are essential for diaporthinic acid formation, whereas Dia2 and Dia3 are dispensable in vivo despite the previously proposed roles of their Aspergillus oryzae homologs based on in vitro studies. On the basis of intermediate accumulation patterns, we propose that Dia4 catalyzes the oxidation of dichlorodiaporthin to diaporthinic acid. Together, these results genetically link diaporthinic acid to the dia BGC and refine the previously proposed biosynthetic model derived from A. oryzae.

Journal of Fungi doi: 10.3390/jof12060401

Authors: Cong Liu Yinan Hao Siyuan Qi Jian Bai

Aspergillus nidulans, a model filamentous fungus endowed with well-established genetic tools and a repertoire of cryptic secondary metabolite biosynthetic gene clusters (BGCs), is extensively exploited as a microbial chassis for heterologous biosynthesis. Mining of its secondary metabolites facilitates the discovery of novel bioactive compounds and the development and application of chassis cells. In the course of heterologous expression of exogenous genes in A. nidulans, we unexpectedly observed the activation of cryptic host BGCs, which resulted in substantial alterations to its secondary metabolic profile. Four previously undescribed compounds (1–4), together with six known analogs (5–10), were isolated from three recombinant A. nidulans strains. Notably, compounds 1–3 are the first naturally occurring examples of diketopiperazine–isoindolinone hybrid alkaloids, while compound 4 is a previously unreported benzofuran carboxylic acid derivative. Their structures and absolute configurations were assigned by interpretation of a combination of spectroscopic data and electronic circular dichroism calculations. Compounds 4 and 5 exhibited potent DPPH radical scavenging activity (IC50, 6.01 and 7.00 μg·mL−1, respectively). This study uncovers a “metabolic perturbation” effect on the host metabolic network during heterologous expression and offers a new strategy for activating silent gene clusters and discovering novel natural products through genetic manipulation.

Journal of Fungi doi: 10.3390/jof12060400

Authors: Jia-Xue Liu Lin-Jiang Zhou Ya-Quan Zhu Hyang Burm Lee Hai-Sheng Yuan

Stipitate stereoid fungi are saprotrophic basidiomycetes characterized by a leathery basidiome, a central-to-lateral stipe and infundibuliform pilei. Although numerous species of stipitate stereoid fungi have been recorded worldwide, understanding of their phylogenetic relationships remains extremely limited, and research on this group of fungi in China is insufficient. In this study, specimens of the three stipitate stereoid genera, namely Podoscypha s. l., Cymatoderma s. l. and Stereopsis s. l., from southern China were investigated. Phylogenetic analyses of the internal transcribed spacer (ITS) regions and the large subunit of the nuclear ribosomal RNA gene (LSU) using maximum likelihood (ML) and Bayesian inference (BI) methods revealed that all three genera are polyphyletic. Consequently, Podoscypha s. s. and Cymatoderma s. s. were delimited, and Cladoderris—previously synonymized with Cymatoderma—was resurrected. Cladoderris is characterized by an imbricate basidiome, tomentose pilei and basidiospores typically shorter than 4 μm in length. Three new species, Podoscypha casiae, Stereopsis buccinata and Cladoderris perennis, were described and illustrated. The morphological distinctions and affinities between the new species and closely related taxa were discussed, the thresholds for the intraspecific and interspecific demarcation within the three genera in this study were provided, and identification keys for the species of each genus were presented.

Journal of Fungi doi: 10.3390/jof12060399

Authors: Sichen Liu James Townley Amir Seyedmousavi Joseph A. Frank

Invasive fungal infections pose significant clinical challenges, owing to limited antifungal efficacy and poor tissue drug penetration. This study investigated whether low-intensity pulsed ultrasound (LIPUS) could enhance the antifungal activity of sub-minimal inhibitory concentrations (sub-MICs) of amphotericin B (AmB) and micafungin (MFG) against two strains from two phylogenetically distinct Candida species: Candida albicans and Candida glabrata. Growth inhibition was assessed following LIPUS (15 min, 50% duty cycle, 1 W/cm2) alone or in combination with sub-MIC antifungals. Time-kill assays and adenylate kinase (AK) release evaluated the cell viability and membrane integrity, respectively. LIPUS transiently but markedly delayed C. albicans growth and enhanced the antifungal effects of both AmB and MFG at sub-MIC levels. The combination of LIPUS and ¼ MIC AmB reduced CFU counts by over 3 log at 12 h and induced higher AK release compared to controls, indicating membrane leakage. In contrast, Candida glabrata showed minimal susceptibility to LIPUS, with low AK release and mitigation of the fungistatic effects of sub-MIC MFG. Our study demonstrates that LIPUS can potentiate sub-MIC antifungals against C. albicans but may have the opposite effect in C. glabrata. This strain-specific variation in response to LIPUS underscores the need for further investigation before LIPUS can be considered as a treatment-enhancement strategy.

Journal of Fungi doi: 10.3390/jof12060398

Authors: Qingqing Liu Na Liu Jia Fu Yongting Bi Yunqi Xie Zhumei Jiang Bin Chen Shenghua Ying Zhenghong Zhao Yuejin Peng

Bombyx batryticatus is a traditional Chinese medicinal material derived from Bombyx mori infected by Beauveria bassiana; however, its formation mechanism remains poorly understood. This study compared infection processes in silkworms by two B. bassiana strains with markedly different virulence (highly virulent ZY027 and ARSEF2860). Integrated transcriptomic and proteomic analyses were employed to uncover, for the first time, the molecular basis of B. batryticatus formation at the systems biology level. The results demonstrated significant weight variations in B. batryticatus derived from different fungal strains. ZY027-induced stiff silkworms exhibited higher wet and dry weights than those infected by ARSEF2860. Large-scale gene reprogramming occurred in silkworm hemolymph post-infection, involving marked activation of Toll/Imd immune signaling pathways, ribosome biogenesis, and endoplasmic reticulum stress responses. A notable “uncoupling” between transcriptomic and proteomic profiles was identified, highlighting the critical role of post-translational regulation in host responses. The two strains triggered distinct metabolic reprogramming patterns: ZY027 notably suppressed oxidative phosphorylation and activated detoxification mechanisms, whereas ARSEF2860 presented characteristics of “immune–metabolic optimization.” These findings suggest that B. batryticatus formation involves complex fungus–silkworm molecular interactions in hemolymph, and that fungal strain characteristics are associated with significant differences in host molecular responses and product biomass. The study provides a theoretical foundation and innovative guidance for selecting strains with high B. batryticatus production potential and developing novel entomopathogenic fungal resources.

Journal of Fungi doi: 10.3390/jof12060397

Authors: Marko Gómez-Hernández Etelvina Gándara Eder Dorantes-Marín María Toledo-Garibaldi

Macromycetes are of great relevance to the functioning of terrestrial ecosystems, but habitat transformation can significantly alter the structure of macrofungal communities. Urbanization is regarded as a major threat to biological diversity; however, knowledge of its impact on macromycetes remains scarce. The present study aimed to assess diversity and distribution patterns of macrofungal species across an urbanization gradient in the Puebla–Tlaxcala Valley of Mexico and determine the effect of urbanization on macromycete communities. From May to October 2024, macromycetes were collected in four oak forests. Fungal specimens were classified based on their macromorphological and micromorphological characteristics. Topographic (1), microclimatic (4), vegetation structure (5), environmental (2), and urbanization (4) variables were included in the analyses. A total of 296 macrofungal species were recorded. Diversity has been shown to decline with increasing urbanization. Species composition shifted across the gradient, with the most urbanized sites showing higher turnover. The interplay of microclimate, vegetation structure, and urbanization was a key driver of the observed patterns, underscoring the sensitivity of macromycetes to urban environmental change. The findings highlight the importance of microclimatic buffering and habitat continuity for sustaining macrofungal communities within urban areas.

Journal of Fungi doi: 10.3390/jof12060395

Authors: Gaoyue Si Xifen Zhang Cheng Zhang Yaqian Li Xinhua Wang Ning Guo Jie Chen

Fusarium root rot (caused by Fusarium verticillioides) is a destructive soilborne disease in maize, significantly reducing crop yields. The root symbiotic fungi Trichoderma species have been confirmed as effective biocontrol microbes for Fusarium root rot; however, the mechanistic role of Trichoderma-induced endophytes in suppressing Fusarium root rot in maize remains unclear. This study found that Trichoderma harzianum T30 significantly reduced the abundance of pathogens by 48.9% and increased the abundance of potentially antagonistic Bacillus strains (33%) in the root endophytic bacterial community. In addition, the hyd1 gene in T. harzianum T30 induced a 7.5-fold upregulation of ZmOPR7 in maize roots compared to the Δhyd1 mutant treatment, a gene related to the jasmonic acid (JA) pathway. Further, several endophytic Bacillus strains were specifically induced by a hyd1-overexpressing strain, including B. amyloliquefaciens MX66, B. velezensis C9, and B. velezensis GAGAN3. Three endophytes significantly (p < 0.05) reduced Fusarium root rot incidence in maize by 46.6–55.0% and upregulated the expression of jasmonic acid/ethylene (JA/ET) pathway-related genes (ZmOPR7, ZmOPR8 and ZmEIL1) by 5.4-, 1.5-, and 4.6-fold, respectively, compared to untreated controls. Meanwhile, the Bacillus strain also improved maize plant growth. This study examined how overexpression of the T. harzianum elicitor gene hyd1 (in the OE-hyd1 strain) affects the colonization dynamics of beneficial endophytic bacteria in maize roots. Additionally, it further suggested the contribution of selected endophytic Bacillus strains in suppressing Fusarium root rot in maize.

Journal of Fungi doi: 10.3390/jof12060396

Authors: Jie Cheng Zhaoan Shao Zhenxia Zhu Shuohan Wang Donghui Gong Chengshuai Xu Chaobo Zhang Xiang Xiu Yongcheng Ding

Beauveria bassiana is one of the most widely used entomopathogenic fungi in insect pest management, and the need for rapid and reproducible quantification of fungal conidia to monitor process performance and to quality control products during biopesticide production is imperative. Conventional methodologies, such as hemocytometer counting and plate dilution assays, are time consuming, laborious and subject to significant operator-to-operator variability. Although optical methods have been increasingly explored for estimating fungal propagule concentrations, species-specific calibration, suspension stability, wavelength selection, and independent validation remain important for routine applications. In this study, we developed an agar–water-assisted UV–visible spectrophotometric calibration protocol for estimating conidial concentration using B. bassiana as a model entomopathogenic fungus. A 0.1% (w/v) agar–water suspension was used in order to get homogeneous, stable dispersions of conidia for optical measurements. Calibration of conidia concentration was accomplished through reliable optical density (OD) values measured at wavelengths 500 nm, 530 nm, 560 nm, 600 nm, and 650 nm. Linear correlations were observed across the tested wavelengths, with the highest goodness of fit for the model at 650 nm (R2 = 0.9907). The resulting regression equation, conidia concentration (×107 mL−1) = 4.184 × OD650—0.12450, has been independently verified with separate conidia batches, resulting in acceptable relative errors ranging from 13.78% and 18.98%. This agar–water-assisted OD650 calibration model provides a practical and species-specific tool for the standardization of conidial dosages in biopesticide research, facilitating the reliable evaluation and application of entomopathogenic fungi within integrated pest management systems.

Journal of Fungi doi: 10.3390/jof12060394

Authors: Shi-Xin Wang Jing Li Xin-Xin Zhang Han-Hong Xu Li-Fei Huang

Endophytic fungi contribute substantially to plant health, yet the diversity, community composition, and tissue distribution of culturable fungal endophytes in sweetpotato remain poorly characterized. In this study, endophytic fungi were isolated from roots, old stems, tender stems, petioles, and leaves of the scab-resistant leafy variety ‘Guangcaishu 16–19’, the scab-susceptible leafy variety ‘Guangcaishu No. 5’, and the tuberous variety ‘Guangshu 87’ were identified based on morphological characteristics and ITS sequencing. ITS sequencing identified 492 fungal isolates belonging to 63 putative taxa in 31 genera. Colletotrichum was the dominant genus in the leafy varieties, whereas Chaetomium was dominant in the tuberous variety. The isolated endophytic fungi were widely distributed and tissue-specific, with genus-level distribution following the pattern “leaves and stems > roots”. Alpha diversity followed the order ‘Guangcaishu 16–19’ > ‘Guangcaishu No. 5’ > ‘Guangshu 87’. The fungal communities of the leafy varieties were the most similar, and their root-associated communities exhibited higher alpha diversity than those of the tuberous variety. In addition, scab-resistant varieties exhibited higher endophytic fungal diversity. Overall, endophytic fungal communities in sweetpotato exhibited high alpha diversity, and significant differences in community structure were observed among variety tissues. These findings provide culturable fungal resources for future screening of beneficial endophytic fungi, bioactive metabolites, and potential biocontrol agents.

Journal of Fungi doi: 10.3390/jof12060393

Authors: Jie Yuan Wenna Zhao Xiaoqing Wu Minghui Xu Cheng Ji Cong Xu Fei Chen Yongchun Zhang Jidong Wang

Arbuscular mycorrhizal fungi (AMF) can improve plant performance, but how they coordinately influence root metabolism and associated bacterial communities in sweet potato remains unclear. Here, a pot experiment was conducted to investigate the effects of Glomus intraradices inoculation on sweet potato seedlings by integrating analyses of rhizosphere soil properties, plant growth and nutrient uptake, root metabolomics, and rhizosphere and endophytic bacterial communities using 16S rRNA gene sequencing with FAPROTAX-based functional prediction. AMF inoculation significantly increased whole-plant fresh and dry biomass, potassium concentration and accumulation, and the accumulation of starch and water-soluble carbohydrates, while no significant effects were observed on dry matter rate or plant nitrogen and phosphorus concentration. In the rhizosphere, AMF reduced soil electrical conductivity and increased organic matter content without significantly affecting pH, alkali-hydrolyzable nitrogen, available phosphorus, or available potassium. Root metabolomic profiling identified 289 differential metabolites, with enrichment of phenylpropanoid biosynthesis, glycerophospholipid metabolism, porphyrin metabolism, and nucleotide metabolism, together with broad up-regulation of lipid-related metabolites. Bacterial communities showed strong compartment specificity, with the root endosphere displaying lower alpha diversity than the rhizosphere. Higher rhizosphere bacterial Shannon diversity was observed in the AMF treatment, together with compartment-dependent shifts in bacterial community composition; enrichment of endophytic taxa such as Devosia and Niastella was detected following AMF inoculation. Functional prediction further suggested niche differentiation between rhizosphere and endophytic bacteria, together with AMF-associated shifts in carbon- and nitrogen-related functions. Overall, these results suggest that G. intraradices inoculation is associated with enhanced sweet potato growth and enhanced potassium and carbohydrate accumulation in association with coordinated changes in rhizosphere conditions, root metabolism, and bacterial community assembly.

Journal of Fungi doi: 10.3390/jof12060392

Authors: Luis Angel Morales-Mora Ignacio Eduardo Maldonado-Mendoza Soley Berenice Nava-Galicia Omar Romero-Arenas Analilia Arroyo-Becerra Miguel Angel Villalobos-López Diana Verónica Cortés-Espinosa Martha D. Bibbins-Martínez

Wild maize (teosinte) has been reported to be less susceptible to biotic and abiotic stresses than its modern relative, corn. The composition of the teosinte root microbiome may be linked to traits such as drought tolerance and pest resistance. Trichoderma spp. are ubiquitous saprotrophic fungi found in the plant rhizosphere, enhancing host plant growth and crop productivity while alleviating biotic and abiotic stresses. The present study identified ten Trichoderma fungal isolates associated with the rhizosphere microbiome of teosinte (Zea mays spp. mexicana) and performed in vitro screening to assess both their multi-trait plant growth-promoting activities and their biological control potential against the phytopathogens Aspergillus flavus and Fusarium verticillioides. Additionally, interaction tests were conducted to evaluate the phytostimulant effect of Trichoderma spp. on maize (Zea mays) seed germination. Taxonomic and phylogenetic analysis identified five different Trichoderma species: T. rifaii (TA and TH); T. azevedoi (TB and TI); T. afroharzianum (TE); T. hamatum (TF and TG); and Trichoderma sp. (aff. bannaense) (TC, TD, and TJ). Partial least squares discriminant analysis revealed the isolates TF, TG, and TJ to have the highest potential for use as biocontrol and biostimulant agents. The present study is the first to examine Trichoderma species associated with the teosinte microbiome, and the results suggest that Trichoderma isolates are a potential sustainable alternative for improving maize cultivation.

Journal of Fungi doi: 10.3390/jof12060391

Authors: Ekaterina S. Skolotneva Vasiliy N. Kelbin Margarita A. Rozova Evsey Kosman

For the first time, a race survey of Puccinia graminis f. sp. tritici (Pgt) population was conducted on Triticum durum in the Altai region of Western Siberia, Russia. A total of 34 single-pustule isolates with different virulence phenotypes were identified on durum wheat (Triticum durum) in 2025 and compared with Pgt from bread wheat (Triticum aestivum). The UPGMA-based clustering separated Pgt isolates into two distinct groups, suggesting the host-driven differentiation that was further proven using population genetics tools. The pathogen isolates from durum showed a wider range of virulence complexity, higher variability, and greater average singularity. Virulence frequencies of Pgt on T. durum and T. aestivum differed markedly for Sr6, Sr7b, Sr9e, Sr17+13, and several other genes, while Sr24 and Sr31 remained effective independently of the pathogen origin. Two races, PKCSF and NFMSF, were detected on both the hosts, indicating a shared pathogen gene pool between bread and durum wheat. Even assuming host-specific divergence of Pgt in the Altai region, there is a need for the deployment of the same resistance genes into both T. aestivum and T. durum cultivars to prevent an outbreak of stem rust in the event of favorable conditions for inoculum exchange between crops.

Journal of Fungi doi: 10.3390/jof12060390

Authors: Andrea Matthews Duy P. Le Sharon Hamill Jirah Villajuan Donald M. Gardiner Elizabeth A. B. Aitken Andrew Chen

Ginger (Zingiber officinale Roscoe) is a high-value horticultural crop widely cultivated for its culinary and medicinal applications, yet its production is increasingly constrained by soil-borne diseases. Among these, Fusarium yellows, caused by Fusarium oxysporum f. sp. zingiberi (Foz), is one of the most damaging constraints in ginger-growing regions around the world, leading to progressive yellowing, vascular blockage, and decline in rhizome quality. Members of the Fusarium oxysporum species complex are known to include both pathogenic and non-pathogenic lineages that often co-occur within the same host and environment, complicating disease diagnosis and epidemiological understanding. In this study, we examined Fusarium-like isolates recovered from both symptomatic and symptomless ginger plants within Southeast Queensland, the major ginger production region in Australia. We then investigated the genetic diversity, effector gene content, and pathogenic potential of these isolates. Comparative analyses revealed two genetically and functionally distinct groups: a clonal Foz lineage consistently associated with Fusarium yellows symptoms and characterised by a conserved set of Secreted In Xylem (SIX) effector genes (SIX7, SIX9, SIX10, and SIX12) and a diverse set of F. oxysporum isolates lacking these effectors. The conserved presence and co-localisation of SIX7, SIX10, and SIX12 within a 5 kb region on a 1.4 Mb contig in the Foz lineage is consistent with the retention of a stable lineage-specific effector module, likely associated with accessory genomic regions that may contribute to host specificity and pathogenicity in the Fusarium oxysporum species complex. Pathogenicity assays confirmed that only the Foz lineage induced disease, whereas non-Foz isolates caused no visible symptoms despite limited colonisation of host tissues. These findings highlight the coexistence of pathogenic and endophytic Fusarium lineages within ginger production systems and support the use of effector-based markers for improved detection and disease management.

Journal of Fungi doi: 10.3390/jof12060389

Authors: Yixuan Wang Wenjun Xu Shiting Qiu Siya Ying Ka Hong Wong Tianpeng Yin Siwen Yuan Kun Feng

15,16-Dihydrotanshinone I (DHT) is a prominent lipophilic diterpenoid from Salvia miltiorrhiza with significant pharmacological potential, though its therapeutic application is limited by poor aqueous solubility. In this study, a microbial biotransformation strategy using Ganoderma lingzhi, known for its wide variety of enzyme, was employed to diversify the chemical structure of DHT and improve its bioactivity profile. Through systematic screening and optimization of fermentation conditions, seven transformation products were isolated and characterized. Among these, five are reported as novel compounds: 17-hydroxy-salvianone (A), 18,19-dihydroxy-danshinspiroketallactone (B-2), epi-18,19-hydroxy-danshinspiroketallactone (B-3), 20-hydroxy-salvianone (C), and 19-hydroxy-danshinspiroketallactone (D). Biological evaluations demonstrated that these derivatives possess multi-target therapeutic potential, including moderate cytotoxic effects against 4T1 and A549 cancer cell lines, alongside anti-inflammatory and neuroprotective activities. However, no significant antibacterial activity was observed for any of the derivatives against six common pathogens. Specifically, compound A significantly inhibited nitric oxide (NO) production in LPS-stimulated RAW 264.7 cells, while B-3 protected SH-SY5Y cells against H2O2-induced oxidative stress. Transcriptomic profiling of the biotransformation process identified 2221 differentially expressed genes (DEGs), showing significant enrichment in cytochrome P450-mediated metabolism and oxidative stress response pathways, which were further validated by qPCR. These results establish G. lingzhi as an efficient biocatalyst for the structural modification of tanshinones and provide a library of novel DHT derivatives for drug discovery.

Journal of Fungi doi: 10.3390/jof12060388

Authors: Karla Jenifer Juárez-Cruz Alfredo Jiménez-Pérez Federico Castrejón-Ayala Gabriela Trejo-Tapia Lauro Soto-Rojas Alma Rosa Solano-Báez Guillermo Márquez-Licona

Root and crown rot of strawberry is an emerging disease of concern in Mexico. Like other soil-borne diseases, it has spread widely due to the vegetative propagation of this crop. This study aimed to evaluate the in vitro antagonistic activity of four native Trichoderma strains and one strain of Arcopilus cupreus against a population of Neopestalotiopsis rosae, as a potential biological control alternative. Thirty-five commercial strawberry fields across the three main strawberry-producing states were sampled, yielding 103 fungal isolates. Nearly 70% of the recovered isolates belonged to the Neopestalotiopsis genus. Sixteen representative isolates were identified as N. rosae based on morphology and multilocus sequence phylogeny (ITS, β-tub, tef1-α) and confirmed as pathogenic through pathogenicity tests. Four native T. afroharzianum isolates recovered from the rhizosphere of healthy strawberry plants were identified by multilocus sequence analysis (rpb2, tef1-α). All T. afroharzianum isolates exhibited antagonistic activity in dual-culture assays, inhibiting mycelial growth by 71–73%, surpassing the effectiveness of the commercial fungicide cyprodinil + fludioxonil (average inhibition 50%). In contrast, the A. cupreus isolate recovered from a native medicinal plant showed an average inhibition of 38%. These results highlight native T. afroharzianum as a promising sustainable alternative for integrated management of strawberry root and crown rot in Mexico.

Journal of Fungi doi: 10.3390/jof12060387

Authors: Eva Duborská Zeinab Zamani Bence Farkas Marek Bujdoš Hana Vojtková

This study investigates the accumulation, volatilization, and metabolic effects of two iodine species (iodide and iodate) in selected filamentous fungi. Six environmentally relevant fungal strains were cultivated under controlled conditions, and iodine distribution between biomass, culture medium, and volatilized fraction was quantified using ICP-MS. Additionally, changes in extracellular metabolite production were determined by capillary isotachophoresis. The results revealed pronounced species-specific differences in iodine transformation. Fusarium poae exhibited the highest accumulation capacity, reaching up to 409.5 mg·kg−1 dry weight, and showed the most efficient overall iodine uptake from the cultivation medium. In contrast, Alternaria tenuissima and Trichoderma viride demonstrated elevated volatilization rates, particularly under iodate treatment, indicating distinct transformation pathways. Exposure to iodine species also induced significant changes in extracellular organic acids production. Increased levels of acetate, succinate, and sorbate suggest that iodine affects fungal metabolic activity, likely through stress-related shifts in central carbon metabolism and redox balance. These findings demonstrate that filamentous fungi employ diverse strategies for iodine transformation, including accumulation, volatilization, and metabolic adaptation. The results highlight the important role of fungi in regulating iodine speciation and mobility and provide new insights into their contribution to the terrestrial iodine cycle.

Journal of Fungi doi: 10.3390/jof12060386

Authors: Wen-Xiu Liu Jing Zhu Ning Wang Heng Zhao Xiao-Yong Liu Zhi-Dong Zhang

Mucoralean fungi are mostly saprotrophic. During a fungal investigation of soil in Guangdong and Anhui provinces of China, five new species of Mucorales were discovered, namely Cunninghamella brevispora sp. nov., C. geminata sp. nov., Mucor chlamydosporiferus sp. nov., M. citrinus sp. nov., and M. magnisporus sp. nov. The identification is based on morphological characteristics, as well as molecular phylogenetics of the internal transcribed spacer (ITS), large subunit ribosomal RNA gene (LSU rDNA), translation elongation factor 1-alpha gene (TEF1α), and RNA polymerase II largest subunit gene (RPB1). Cunninghamella brevispora sp. nov. is sister to C. guizhouensis, and is distinguished by short sporangiophores. Cunninghamella geminata sp. nov. is sister to C. subclavata; rhizoids are absent in the former but well-developed in the latter. Mucor chlamydosporiferus sp. nov. is closely related to M. prayagensis, and is characterized by abundant chlamydospores. Mucor citrinus sp. nov. is closely related to M. paraorantomantidis, and is differentiated by pale yellow sporangiospores. Mucor magnisporus sp. nov. is sister to M. merdicola, and is discriminated by large sporangiospores. To date, with the addition of the five new species described herein, the total number of accepted species in the genus Cunninghamella and Mucor has increased to 49 and 163, respectively.

Journal of Fungi doi: 10.3390/jof12060385

Authors: Wen-Jing Wu Jin-Chao Yang Hong-Lin Zhang Qian Xu Guang-Rong Zhou Bo Xu Yan-Chun Li

Termitomyces is an economically and ecologically important genus mainly distributed in the southwestern and southern regions of China. We performed phylogenetic analyses of this genus using Maximum Likelihood and Bayesian analyses based on the nuclear ribosomal internal transcribed spacer (ITS), the small subunit of mitochondrial DNA (mtSSU), and the large subunit of nuclear ribosomal DNA (nrLSU) in this study. Three new species, namely T. albus, T. apicoannulatus, and T. pseudointermedius, and one new record to China, T. pakistanensis, were described and illustrated based on morphological and molecular evidence. Detailed morphological descriptions, colour photographs, line drawings of microstructures, and comparisons with phylogenetically and morphologically related species are provided.

Journal of Fungi doi: 10.3390/jof12060384

Authors: Lianghui Jiang Kunfa Gan Jinlan Xie Zhanghong Mo Qiang Liang Xing Huang Qian Nong Li Lin Changning Li

Sugarcane is a globally important crop, widely cultivated for sugar production and bioenergy. However, leaf spot disease leads to a reduction in its quality and yield. In this study, pathogen identification, biological characteristic analysis, and screening of antagonistic bacteria against the causal pathogens were done as a basis for epidemic prediction and green control of sugarcane leaf spot disease. The causal pathogens of sugarcane leaf spot disease were identified as Epicoccum latusicollum El532 and Fusarium sacchari Fs64, respectively, based on morphological characteristics, multi-gene phylogenetic analysis (ITS, TUB2, and RPB2 for El532; ITS, TEF1α, and RPB2 for Fs64), and pathogenicity tests. Biological characterization revealed that both pathogens exhibited optimal mycelial growth at 25 °C and under continuous darkness. However, light-dark cycles inhibited their growth. The optimal pH ranges for both isolates were 6–9 and 5–10, respectively. Maltose was the optimal carbon source for El532, whereas maltose, lactose, and starch were optimal for Fs64. Yeast extract served as the optimal nitrogen source for both. Isolation and screening of bacterial strains from healthy sugarcane roots, leaves, and rhizosphere soil yielded 13 antagonistic bacterial strains. Among them, six strains exhibited inhibition rates exceeding 57% against both pathogens. Bacillus subtilis A5 exhibited the highest antagonistic activity (68.85% against El532, 71.69% against Fs64), underscoring its potential as a promising biocontrol candidate. These findings provide a scientific basis for the diagnosis and management of sugarcane leaf spot disease.

Journal of Fungi doi: 10.3390/jof12060383

Authors: Valentina Daniela Sisu Anda Băicuș

Invasive candidiasis is a significant concern in healthcare environments, and awareness of these infections has increased in recent years. A growing number of risk factors, the ability of some Candida species to progress from colonization to tissue invasion, and their capacity to adhere to and survive on abiotic surfaces have all contributed to the spread of invasive candidiasis. The primary goal in cases of invasive candidiasis is to diagnose it as promptly as possible, as any delay can delay antifungal treatment. This review concentrates on clinical syndromes reunited under the definition of invasive candidiasis and the current diagnostic methods. Risk factor assessment is another major topic of this narrative review and recent updates are included. Research stage biomarkers are also explored and partial results are mentioned as there are continuous efforts to search for new tools for a more accurate prediction or an earlier identification of IC.

Journal of Fungi doi: 10.3390/jof12060382

Authors: Cihan Aslı Yunus Korkom Daniel Carrillo Ibrahim Cakmak

Fungi of the genus Trichoderma have attracted attention because of their potential activity against phytophagous mites; however, information regarding their non-target effects on predatory mites remains limited. This study evaluated the effects of spore suspensions and secondary metabolites of Trichoderma afroharzianum Tr132 and Trichoderma virens Tvr2 on three predatory mite species widely used in biological control programs: Phytoseiulus persimilis, Neoseiulus californicus, and Amblyseius swirskii. Predator egg hatchability and adult mortality were assessed under laboratory conditions. Spore suspension treatments did not significantly affect egg hatchability, which remained high (97–99%) across all predator species. In contrast, secondary metabolites slightly reduced egg hatchability to 94–96%, compared with 99.5% in the control. Exposure to spore suspensions caused moderate mortality in adult predatory mites, increasing from 10 to 13% at 3 days post-application (dpa) to 15–19% at 6 dpa. Secondary metabolites produced higher mortality that increased over time, reaching 9–11% at 1 dpa, 17–18% at 3 dpa, and up to 22–25% at 6 dpa. Mortality responses were similar among predator species. Overall, Trichoderma applications had minimal effects on predator egg hatchability but caused moderate mortality in adult predatory mites, particularly following exposure to secondary metabolites. These findings highlight the importance of evaluating the compatibility of Trichoderma-based products with beneficial predatory mites before their integration into IPM programs.

Journal of Fungi doi: 10.3390/jof12060381

Authors: Yuling Cui Fangjie Yao Xiaoxu Ma Tingting Liu Xu Sun Ming Fang Lixin Lu Youmin Zhang Yinghao Li Xinming Chen Xinyue Xu

For the first time, a regulatory network linking melanin, genes, pathways, and edible quality was constructed for 138 Auricularia cornea strains sourced domestically and internationally. This marks the inaugural study of A. cornea spanning from cellular to physical-mechanical properties. Correlation analysis between melanin and edible quality traits (hardness, springiness, cohesiveness, gumminess, chewiness, and resilience) revealed that hardness, cohesiveness, and gumminess increased with rising melanin content, while springiness correspondingly decreased. Genome-wide association analysis identified 15,597,589 SNP loci. A total of 39 genes related to food quality were annotated, including one melanin-related lacquer enzyme gene, ACW004924. Real-time quantitative PCR validation of key genes identified for melanin and edible quality traits revealed results consistent with those from correlation analysis. The lacquer enzyme genes ACW004736, ACW006232, which regulate melanin synthesis, and the tyrosinase genes ACW001451, ACW002443, and ACW001003 were also identified in edible quality traits. These genes perform similar functions in GO-enriched metabolic processes, catalytic activity, and cellular structural complexes, as well as in KEGG-enriched pathways such as carbon metabolism and polysaccharide synthesis. They catalyze melanin synthesis and promote interactions between melanin and cell wall polysaccharides, chitin, and structural proteins, thereby stabilizing the cellular scaffold structure, jointly mediating the effect of melanin on the edible quality of A. cornea. The results supplement the downstream regulatory chain of catalytic enzymes and edible quality in the γ-L-glutaminyl-3,4-dihydroxybenzene (GDHB) pigment synthesis pathway, and form an information network of melanin synthesis, cell wall structure optimization, and edible quality regulation.

Journal of Fungi doi: 10.3390/jof12050380

Authors: Nantana Mills Natasha Mills Nakarin Suwannarach Nuttapol Noirungsee Jaturong Kumla Sahutchai Inwongwan Rujipas Yongsawas Chanon Saksunwiriya Varis Domethong Rasmi Shoocongdej Terd Disayathanoowat

Phi Man Long Long Rak Cave, located in Mae Hong Son Province, northern Thailand, is a prehistoric burial site containing ancient wooden coffins that have undergone biodeterioration, likely due to fungal activity. Both culture-dependent and culture-independent approaches were employed to characterize fungal communities and assess their roles in wood degradation. Culture-dependent analysis identified five Aspergillus isolates from the wooden coffins, most of which produced cellulolytic and hemicellulolytic enzymes; some isolates also produced organic acids, indicating significant degradative potential. Culture-independent analysis revealed a community dominated by Aspergillus, together with additional taxa such as Penicillium and Ceriporia that were not detected by cultivation, highlighting greater community diversity and demonstrating the complementarity of the two methods. Functional prediction indicated a predominance of saprotrophic fungi. The presence of shared dominant taxa between soil and coffin-associated substrates suggests ecological connectivity at the soil–coffin interface, although the direction of dispersal cannot be determined from the present data. All tested fungicides inhibited fungal growth, with the highest efficacy observed in the formulation containing the highest proportion of active components. Taken together, these findings provide insights into fungal biodeterioration processes and inform conservation strategies.

Journal of Fungi doi: 10.3390/jof12050379

Authors: Rok Tomazin Tadeja Matos

Aspergillosis encompasses a heterogeneous spectrum of diseases caused by filamentous fungi of the genus Aspergillus, ranging from allergic airway disorders and chronic pulmonary infection to rapidly progressive invasive disease. Aspergillus fumigatus is the predominant pathogen worldwide, although other species, including Aspergillus flavus, Aspergillus terreus and cryptic species, contribute to morbidity and may exhibit intrinsic or acquired antifungal resistance. Early and accurate laboratory diagnosis is essential for timely treatment, appropriate antifungal selection, and stewardship. Traditional culture remains foundational, enabling confirmation of viable organisms, species-level identification, and antifungal susceptibility testing, but sensitivity is limited and turnaround times are prolonged. Non-culture approaches—including galactomannan, β-D-glucan, lateral flow assays, PCR, and next-generation sequencing—enhance diagnostic sensitivity, facilitate early detection, and allow identification of resistance-associated mutations. Optimal diagnostic performance is achieved through integrated, multimodal strategies combining laboratory tests with clinical and radiological findings. In invasive disease, concurrent use of biomarkers and molecular assays improves specificity and positive predictive value, while in allergic bronchopulmonary aspergillosis, immunological markers remain central. Future directions include standardised molecular protocols, novel antigenic and host-based biomarkers, and cost-effective, risk-adapted diagnostic algorithms to refine detection, guide therapy, and improve patient outcomes.

Journal of Fungi doi: 10.3390/jof12050378

Authors: Xiongjuan Huang Chengcheng Feng Xixi Ju Yuhui Huang Xiaofeng Chen Jiazuo Liang Xinglian Liu Zhendong Chen Rukui Huang

Bitter gourd (Momordica charantia L.) is an important vegetable and medicinal crop in tropical/subtropical regions, but suffers severe yield losses (even total failure) from Fusarium wilt caused by Fusarium oxysporum f. sp. momordicae (Fom). There is no specific detection system available to detect this pathogen, and the methods used for other pathogens exhibit cross-reactivity and require specialized equipment. Therefore, this study developed a loop-mediated isothermal amplification (LAMP) assay for early Fom diagnosis. Initially, five sets of LAMP primers targeting the conserved regions of Fom, located within the region amplified by the FOMM-SPF/SPR PCR primers, were tested for specificity and sensitivity. In this experiment, FoM-1-2 showed optimal specificity, identifying 44 Fom strains without cross-reactivity with 10 other non-Fom species after a 60 min incubation at 64 °C. A visual readout based on a fluorescent dye (green for positive, pale orange for negative) eliminated the need for gel electrophoresis and specialized instruments. The LAMP assay was 100-fold more sensitive than conventional PCR (detection limit: 5.6 pg/μL vs. 560 pg/μL). In inoculated seedlings, LAMP detected Fom in basal stems at four days post-inoculation and top leaves at six days, whereas conventional PCR yielded faint bands in the basal stem after eight days. Moreover, LAMP enabled non-destructive detection. Thus, the present study developed a rapid, specific, and sensitive visual LAMP assay, supporting early diagnosis of bitter gourd Fusarium wilt.

Journal of Fungi doi: 10.3390/jof12050377

Authors: Aron Osakina William J. Steinbach Praveen R. Juvvadi

Autophagy, also referred to as the “self-eating machinery”, is a crucial process where organisms maintain intracellular homeostasis through recycling or degrading non-essential and damaged cellular components. It is important in numerous biological functions such as cellular differentiation, aging, nutrient sensing, stress response, tissue homeostasis, immunity, and programmed cell death. Autophagy induction occurs with the formation of a double-layered membrane structure called “autophagosome”. The autophagosome wraps damaged organelles or proteins and transports them to the vacuole or lysosome for degradation. Autophagy is beneficial to organisms, and it should be optimally regulated because elevated or decreased levels are detrimental for survival. To date, more than 40 autophagy-related genes (ATGs) have been identified in the budding yeast Saccharomyces cerevisiae, with most having homologs in fungi and higher eukaryotes. Majority of the ATGs in industrial and pathogenic fungal species have been characterized and known to play vital roles in growth, development, and virulence. In this review we provide a comprehensive overview of ATGs in various fungal species and highlight how autophagy is regulated and controls various functions in plant, human, and industrial fungal species.

Journal of Fungi doi: 10.3390/jof12050376

Authors: Sanele Michelle Mnkandla Patricks Voua Otomo

Mycofiltration uses saprophytic fungi immobilised on dead organic matter to treat contaminated water. This systematic review aimed to collate literature on mycofiltration, identify water sources subjected to mycofiltration, types of fungi employed, contaminants removed, and removal efficiencies (R%). Articles written in English between 1990 and 2023 were collected from various sources, screened based on inclusion criteria, and critically appraised. Metadata were extracted, and a narrative synthesis was conducted. Forty articles representing 156 studies passed appraisal, with 116 from journal articles, 24 from theses, and 16 from reports. Synthetic stormwater and real wastewater were the most frequently mycofiltered. Fungi of the Pleurotus genus were predominantly used in creating mycofilters. Organic contaminants removed included pharmaceuticals and pesticides, with R% between 60% and 100%. E. coli was the most studied microbial contaminant, and R% of 30%, 60%, and 90% were reported. Inorganic contaminants were mostly metals with R% above 60%. Overall, contaminant removal by mycofiltration varied, but the technology remained a promising tool. Research gaps observed included a lack of standardised methods for mycofilter preparation and design and little to no assessment of mycofilter saturation. Addressing gaps could aid in increasing mycofilter efficiency and reliable upscaling of mycofiltration.

Journal of Fungi doi: 10.3390/jof12050375

Authors: Kun Yang Rongbo Wang Liguang Liu Kang An Jitao Liu Li Wang Jianwei Shan Chengchen Li Liang Qi Li Zheng Xiaobo Li

As a non-pathogenic oomycete, Pythium oligandrum possesses unique advantages, particularly in the context of being a biological control agent. With the increasing awareness of consumer consciousness, people are paying more attention to the use of environmentally friendly strategies in plant disease prevention and control. Pythium oligandrum is a type of biocontrol oomycete that can be developed as a biological control agent, and it does not have adverse effects on humans in the prevention and control of plant diseases. Consequently, there is increasing scientific interest in the beneficial plant–microbe interactions mediated by P. oligandrum. Currently, the main points of focus regarding the beneficial role of P. oligandrum in plant interactions are as follows: (i) P. oligandrum can activate plant defense responses and cause plants to produce resistance, thus protecting them from disease attacks; (ii) it is a strong mycoparasite that can coil around various oomycetes and fungi, directly killing pathogenic microorganisms; (iii) in addition, it can also promote plant growth. In this paper, we will discuss the aforementioned three main features in detail.

Journal of Fungi doi: 10.3390/jof12050374

Authors: Mirah Afiza Nurazizah Safrina Dyah Hardiningtyas Muhammad Arief Budiman Nurul Huda Abd Kadir Kustiariyah Tarman

Marine endophytic fungi inhabit the internal tissues of seaweed, seagrass, and mangroves without causing harm. These fungi are known to produce extracellular enzymes, including proteases and cellulases, which play crucial roles in various biological processes and have potential applications in diverse industrial sectors. This study aimed to screen the enzymatic potential of marine endophytic fungi, identify selected isolates, and characterize their enzyme activities. A total of 20 fungal isolates were obtained, comprising 16 isolates from seaweed, three from seagrass, and one from mangrove leaves, collected from the coastal areas of the Seribu Islands (Jakarta), Sukabumi (West Java), Nusa Dua (Bali), and the Buton Islands (Southeast Sulawesi). Screening results showed that 50% of the isolates exhibited proteolytic activity on skim milk agar, while 40% demonstrated cellulolytic activity on carboxymethylcellulose (CMC) agar. Two isolates with the highest clear zone indices for protease and cellulase activity were identified as Penicillium citrinum and Fomitopsis sp., with distinct morphological characteristics including velvety colonies and filamentous hyphal structures. The specific activities of the protease and cellulase were 5475.42 ± 2724.25 U/mg protein and 620.77 ± 607.71 U/mg protein, respectively, indicating high catalytic potential.

Journal of Fungi doi: 10.3390/jof12050373

Authors: Andrey R. Suprun Stanislava A. Vinogradova Alina A. Beresh Natalia S. Chopenko Alina A. Dneprovskaya Evgeniya V. Trubetskaya Artem Yu. Manyakhin Konstantin V. Kiselev

Alternaria blight, caused by fungi of the genus Alternaria, is one of the most common and damaging diseases affecting tomatoes, leading to significant yield losses. The intensive use of chemical fungicides faces the problems of pathogen resistance development and negative environmental impacts. This study investigated the possibility of using RNA interference technology based on exogenous double-stranded RNAs (dsRNAs) to protect tomatoes against the causal agent of early blight (EB), Alternaria alternata. Key genes of the pathogen A. alternata were selected as targets: Alt-a1 (a major allergen and virulence factor), TEF1a (translation elongation factor 1-alpha) and β-Tub (β-tubulin). Specific dsRNAs were synthesized in vitro and applied to tomato plants (Solanum lycopersicum L. cv. Micro-Tom) simultaneously with inoculation of A. alternata strain C7.24-T2-L-F1 spores. Visual assessment, measurement of chlorophyll A and B, and real-time quantitative PCR analysis showed that treatment with dsRNAs targeting the Alt-a1, TEF1a and β-Tub genes significantly suppressed infection development, reducing the amount of pathogen DNA in plant tissues by 7 to 27 times depending on the dsRNA type. The most effective was dsRNA to the Alt-a1 gene. Thus, the obtained results demonstrate the promise of spray-induced gene silencing (SIGS) as a strategy for protecting tomato plants against the pathogen A. alternata.

Journal of Fungi doi: 10.3390/jof12050372

Authors: Ofir Degani

Fungal pathogens constitute the largest group of plant disease agents, infecting crops through leaves, seeds, and soil, and causing substantial losses in agricultural productivity worldwide [...]

Journal of Fungi doi: 10.3390/jof12050371

Authors: Hua Zhao Chunyue Liu Xi Zhang Qingfeng Qiu Yangsheng Luo Xiwang Ke Biao Gu

Phytophthora infestans is the causal agent of late blight, one of the most destructive diseases of potato and tomato worldwide. Although qPCR-based methods are widely used to estimate pathogen biomass in infected tissues, methods for specifically assessing sporangial proliferation remain limited. In this study, we developed an RT-qPCR-based assay using PITG_13011, which encodes a predicted major facilitator superfamily transporter, as a sporangia-associated molecular marker in P. infestans. Among five candidate genes selected from transcriptomic data, PITG_13011 showed the strongest association with sporangia-associated samples in our validation assays. PITG_13011 transcripts were detectable from cDNA and genomic DNA derived from as few as 100 sporangia, and transcript abundance showed a strong positive correlation with sporangial number under controlled experimental conditions. In detached leaf inoculation assays, PITG_13011 transcript levels were associated with differences in sporangia-associated proliferation during infection. These results indicate that PITG_13011-based RT-qPCR can serve as a complementary molecular approach for estimating sporangia-associated proliferation of P. infestans in laboratory experiments. This method will be useful when sporangial production, rather than total pathogen biomass alone, is the parameter of interest.

Journal of Fungi doi: 10.3390/jof12050369

Authors: Yanqing Bi Guoshuai Zhang Xinyu Zhu Yumei Su W. G. Dilantha Fernando Xiaofeng Su Wenfang Guo Yue Li

Cotton Verticillium wilt (VW), caused by the soil-borne fungus Verticillium dahliae (V. dahliae), is a devastating disease that poses a serious threat to sustainable cotton production worldwide. Protein methylation plays a critical role in fungal adaptation to the host environment and manipulation of plant immunity. Protein arginine methyltransferases (PRMTs) are key enzymes catalyzing arginine methylation, yet their functions in V. dahliae pathogenicity remain largely unexplored. In this study, we identified VdPRMT4 in V. dahliae through homology-based screening. qRT-PCR analysis revealed that VdPRMT4 transcript levels were significantly increased during the early stages of V. dahliae infection in cotton. HIGS assays showed that silencing VdPRMT4 markedly alleviated cotton VW symptoms and reduced fungal biomass in cotton plants. Gene knockout and complementation experiments demonstrated that deletion of VdPRMT4 did not affect hyphal growth but significantly impaired sporulation capacity and severely attenuated pathogenicity on cotton. Transcriptomic analysis further indicated that loss of VdPRMT4 profoundly affected the metabolic pathways of V. dahliae, including protein processing in the endoplasmic reticulum, purine metabolism, and glycerolipid metabolism. Collectively, this study provides the first evidence that VdPRMT4 plays a critical role in stress adaptation and pathogenicity of V. dahliae, offering new insights into fungal pathogenesis and identifying potential targets for VW control.

Journal of Fungi doi: 10.3390/jof12050370

Authors: Juan Manuel Tovar-Pedraza Guillermo Estrada-Arroyo Rafael Macedo-Arzate Sami J. Michereff Kamila C. Correia Santos Gerardo Leyva-Mir José Antonio Mora-Aguilera Moisés Camacho-Tapia Guillermo Márquez-Licona Alma Rosa Solano-Báez

Mango (Mangifera indica L.) represents one of the most significant fruit crops cultivated across multiple regions of Mexico. In recent years, cases of stem-end rot and dieback have been observed in mango-producing areas. This research aimed to characterize the diversity of Lasiodiplodia species associated with these symptoms, determine their geographic distribution in five Mexican states, and evaluate their pathogenicity on mango fruits. During 2014, samples exhibiting dieback and stem-end rot symptoms were collected from 27 commercial orchards located in five states, resulting in the obtention of 87 Lasiodiplodia isolates. From these, 36 representative isolates were selected and identified through phylogenetic analyses (ITS, tef1-α, tub2), employing the Maximum Likelihood and Bayesian Inference approach. Eight Lasiodiplodia species were resolved: L. brasiliense, L. laeliocattleyae, L. subglobosa, L. theobromae, L. iraniensis, L. mexicanensis, L. hyalina, and L. pseudotheobromae. Among them, L. brasiliense, L. laeliocattleyae, L. subglobosa, L. iraniensis, L. mexicanensis, and L. hyalina are reported for the first time in association with mango tissues in Mexico. Pathogenicity tests conducted on detached mango fruits using the mycelial plug inoculation method demonstrated that all species were capable of inducing necrotic lesions. However, L. laeliocattleyae and L. brasiliense exhibited the highest levels of aggressiveness, while L. mexicanensis, L. hyalina, and L. pseudotheobromae were the least aggressive.

Journal of Fungi doi: 10.3390/jof12050368

Authors: Suzana Otašević Marija Trenkić Marko Stalević Marina Ranđelović Slavica Stojnev Milica Đorđević Jana Pešić Stanković Goran Koraćević Roberta Iatta

Exophiala endophthalmitis of exogenous origin is an exceptionally rare but severe ocular infection, characterized by diagnostic delays, limited therapeutic guidance, and frequently poor outcomes. Herein, we report one new case of an 80-year-old woman who presented with severe fungal keratitis progressing to endophthalmitis two years after an uncomplicated cataract surgery. The condition was initially misdiagnosed and treated with topical antibiotics and corticosteroids. By cultivation, microscopy, histopathological, and PCR analysis of the samples, Exophiala dermatitidis was identified as the causative agent. Despite targeted antifungal therapy with voriconazole, the disease rapidly progressed, resulting in corneal perforation and evisceration of the affected eye. The number of confirmed cases of this infection remains very limited. To address this gap, we conducted a structured review of all reported instances of exogenous Exophiala endophthalmitis, in which Exophiala dermatitidis emerged as the predominant causative species. Common predisposing factors included corneal barrier disruption, ocular surgery, diabetes mellitus, and corticosteroid use. Diagnostic confirmation was frequently delayed, and treatment outcomes varied. Amphotericin B-based regimens were associated with poor results, whereas voriconazole, particularly when combined with surgical intervention, demonstrated more favorable outcomes. Exogenous Exophiala endophthalmitis remains underrecognized, with limited evidence to guide management. This entity should be considered in postoperative or trauma-associated intraocular inflammation, and current evidence supports azole-based therapy combined with surgical intervention when indicated.

Journal of Fungi doi: 10.3390/jof12050367

Authors: Yan Tong Chuyu Tang Bing Jia Haoxu Tang Jinxuan Yan Yuling Li Xiuzhang Li

Samsoniella hepiali produces an array of pharmacologically valuable metabolites, but how environmental pH regulates its antioxidant system and lipid metabolism during submerged fermentation remains unclear. This study aimed to investigate the effects of different initial culture pH values (pH 4, 5, 6, and 7) on the antioxidant capacity and lipidomic metabolism of S. hepiali. The results demonstrated that at pH 5, the activities of peroxidase (POD) and superoxide dismutase (SOD), the contents of total phenolics (TP) and flavonoids, the scavenging rates of DPPH• and •OH, and the total antioxidant capacity all peaked. Conversely, the level of glutathione (GSH) reached its maximum at pH 6 (0.69 ± 0.014 μmol/g). Lipidomic analysis identified a total of 404 lipid molecular species, mainly TG, PE, and DG. Comparative analysis among pH 4 vs. pH 5, pH 6 vs. pH 5, and pH 7 vs. pH 5 revealed 27 core DALs belonging to 11 lipid subclasses, most of which were upregulated at pH 5. KEGG pathway enrichment analysis further revealed that sphingolipid metabolism was the sole core co-enriched pathway under different pH conditions. Particularly at pH 5, key signaling lipids, such as ceramides, underwent pronounced targeted accumulation. This study elucidates the molecular adaptation mechanisms of medicinal fungi in response to pH variation from a lipidomic perspective. It provides a basis for optimizing fermentation conditions to enhance antioxidant activity and functional lipid production.

Journal of Fungi doi: 10.3390/jof12050366

Authors: Kasun M. Thambugala Sanduni Dabare Asanthi Dhanusha Imalka Munaweera Dinushani A. Daranagama Sukanya Haituk Ratchadawan Cheewangkoon

Mycogenic nanomaterials, nanoparticles (NPs) biosynthesized through fungal enzymatic and metabolic activity, have emerged as a compelling alternative to chemically synthesized nanomaterials, offering fundamental biocompatibility, green production conditions, and biologically functional surface coatings. Fungi, acting as natural “nanofactories,” harness reductases, oxidoreductases, secreted proteins, and secondary metabolites to reduce metal ions into stable NPs under ambient conditions, simultaneously capping the particles with biomolecules that enhance colloidal stability, biocompatibility, and secondary biological activity. Unlike previous reviews that have addressed either biosynthesis mechanisms or applications in isolation, this review uniquely adopts a structured “Promise vs. Barrier” framework across six interconnected thematic pillars, offering the first comprehensive critical synthesis that simultaneously maps mechanistic frontiers, biodiversity gaps, and translational barriers within mycogenic nanotechnology. The present review critically examines both the extraordinary promise and the persistent barriers facing mycogenic nanotechnology across biosynthetic mechanisms, fungal biodiversity, nanomaterial portfolio expansion, biomedical applications, environmental and agricultural utility, and industrial scalability. We highlight how emerging multiomics approaches, integrating transcriptomics, proteomics, and metabolomics, are beginning to decode the molecular blueprints of fungal NP synthesis, while acknowledging that mechanistic knowledge gaps, limited genetic toolkits for non-model fungi, and the absence of standardized protocols continue to impede progress. The fungal kingdom represents a vast, underexplored reservoir of nanofactory potential, with fewer than 1% of known species evaluated to date; strategic bioprospecting using genome mining and machine learning is beginning to unlock this diversity. Mycogenic NPs demonstrate broad-spectrum antimicrobial activity against multidrug-resistant pathogens, selective anticancer activity, biosensing capacity, and applications in wound healing, sustainable agriculture, environmental remediation, and smart food packaging. However, critical deficits persist in clinical validation, long-term toxicity data, manufacturing reproducibility, and regulatory clarity. The review concludes with a tiered roadmap, spanning immediate mechanistic priorities through to long-term synthetic biology and AI-integrated commercialization, and calls for coordinated international action on standardization, reference material development, and harmonized regulatory frameworks to bridge the gap between laboratory promise and real-world application.

Journal of Fungi doi: 10.3390/jof12050365

Authors: Chengwei Liu Jianzhao Qi Xiuzhang Li

Fungi represent one of the most diverse groups of organisms on Earth, playing fundamental roles in ecosystems, agriculture, and biotechnology [...]

Journal of Fungi doi: 10.3390/jof12050364

Authors: Meng-Hsuan Lai Ni Tien Hsiao-Lien Yang Jun-He Huang Miin-Huey Lee Bang-Jau You

Ganoderma lucidum is a medicinal fungus widely utilized in traditional medicine and functional foods. Its primary bioactive constituents are ganoderic acids (GAs), a group of triterpenoid compounds. While chemical-induced apoptosis has previously been shown to enhance GAs production, this study investigates the role of physical stress in this regulatory pathway. We demonstrate that heat-induced apoptosis significantly increases GAs production in G. lucidum. To determine whether apoptosis directly regulates this process, we overexpressed the human anti-apoptotic gene Bcl-2 in G. lucidum, confirming expression via RT-PCR and Western blot analysis. Upon heat-induced apoptosis, these Bcl-2 overexpression mutants exhibited increased mycelial cell viability, accompanied by reduced metacaspase activity and, notably, decreased GAs production. Furthermore, we identified a Type I metacaspase gene in G. lucidum, Glmca1, which contains highly conserved catalytic domains common across fungal species. Silencing of Glmca1 followed by heat-induced apoptosis led to results similar to Bcl-2 overexpression: enhanced cell viability, suppressed metacaspase activity, and a significant reduction in GAs yield. These findings provide compelling evidence that apoptosis functions as a critical regulatory mechanism for secondary metabolite production in G. lucidum. Consequently, modulating apoptotic pathways through physical induction offers a promising strategy for enhancing the production of bioactive ingredients in medicinal fungi.

Journal of Fungi doi: 10.3390/jof12050363

Authors: Edwin Mondragon-Herrera Laydy Mitsu Mena-Chacon Santos T. Leiva-Espinoza Angel F. Huaman-Pilco

The use of entomopathogenic fungi as biological control agents has gained increasing relevance as a sustainable alternative to chemical insecticides in tropical agroecosystems. In this study, a naturally occurring isolate of Metarhizium brunneum infecting adults of Metamasius hemipterus was recovered from banana plantations in the Amazonas region, Peru, and evaluated for its potential as a biological control agent. Multilocus phylogenetic analysis based on tef1α, β-tubulin, rpb1, and rpb2 sequences confirmed its taxonomic identity within the M. brunneum clade. Physiological characterization revealed variability in growth and thermal response among isolates, while conidial production differed significantly depending on the substrate. Notably, isolate PM9 exhibited the highest conidial yield on rice substrate. Pathogenicity assays demonstrated high virulence against adult weevils, with an LC50 of 2.91 × 105 conidia·mL−1 and mortality exceeding 90% at the highest concentration tested. These findings indicate that isolate PM9 combines desirable physiological and pathogenic traits for biological control. The natural occurrence of this entomopathogen in banana systems suggests ecological adaptation to local conditions and supports its potential incorporation into integrated pest management strategies, although further field-based evaluation is required.

Journal of Fungi doi: 10.3390/jof12050362

Authors: Na Wu Xiaoxuan Du Chaowei Huang Xinru Xu Wenke Hu Suai Yin Xiaoxiao Ma Rong Shao Kyung-Min Kim Wei Xu

Morphological control in submerged fermentation is a well-established method for enhancing bioactive metabolite production in filamentous fungi. However, the molecular mechanisms linking morphology to fermentation efficiency remain insufficiently understood. In this study, supplementing 1.5% Tween 80 (P80) at the seed culture stage of Cordyceps militaris consistently induced the formation of compact, uniform mycelial pellets. This morphological induction at the seed stage enhanced fermentation performance, increasing exopolysaccharide (EPS) titer by 71.1% and reducing the production cycle by 24 h. Transcriptomic analysis revealed that pelletized cultures exhibited transcriptional patterns associated with MAPK signaling related to cell wall integrity and upregulation of genes involved in cell wall remodeling. Additionally, pelletized cultures displayed a reduced oxidative burden and were associated with enhanced antioxidant capacity. These findings link morphology induction to cell wall remodeling and oxidative stress defense, offering a potentially scalable strategy for industrial polysaccharide production in medicinal fungi.

Journal of Fungi doi: 10.3390/jof12050361

Authors: Sezer Akgöl Serpil Aylin Yaşar Fatih Kocabaş

Ubiquitination is a key post-translational modification regulating cellular signaling and innate immunity, and its reversal by deubiquitinases (DUBs) represents a critical mechanism exploited by pathogens for immune evasion. While ovarian tumor (OTU) domain-containing DUBs are well characterized in viral systems, their roles in fungal pathogens remain largely unexplored. In this study, we investigated two putative OTU domain-containing proteins derived from the plant pathogenic fungi Melampsora larici-populina (MlpOTU, EGG09943.1) and Taphrina deformans (TdOTU, CCG84064.1). Recombinant MlpOTU and TdOTU proteins were successfully expressed and purified from E. coli and exhibited high solubility and proper folding. Functional analyses in HEK293T cells demonstrated that both proteins significantly reduce global ubiquitination levels, confirming their deubiquitinase activity in vivo. Despite this shared enzymatic function, the two proteins displayed markedly distinct effects on host immune gene expression. MlpOTU selectively suppressed key antiviral effectors, most notably MX1, suggesting a targeted immune evasion strategy. In contrast, TdOTU induced robust upregulation of multiple immune-related genes, including type I interferons, indicating a divergent role. Neither MlpOTU nor TdOTU triggered robust apoptosis, supporting their role as modulators of host signaling rather than cytotoxic effectors. Collectively, these findings provide the first functional evidence that fungal OTU domain-containing proteins act as active deubiquitinases and reveal distinct strategies by which plant pathogens may manipulate host immune responses. This study establishes fungal OTU domains as promising targets for antifungal intervention and broadens our understanding of cross-kingdom evasion mechanisms.

Journal of Fungi doi: 10.3390/jof12050360

Authors: Samantha Cruz-López Emiret Analy Albavera-Ramírez Roberto Arenas Claudia Erika Fuentes-Venado Claudia Camelia Calzada-Mendoza Eunice D. Farfán-García Juan Castillo-Cruz Edwin Chávez-Gutiérrez Erick Martínez-Herrera Rodolfo Pinto-Almazán

Background/Objectives: Onychomycosis is a fungal nail infection that may present with severe, atypical, or treatment-resistant features in people living with HIV. Despite its clinical importance, evidence regarding its epidemiology, causative agents, and relationship with immune status remains limited. This systematic review aimed to evaluate the association between onychomycosis and HIV, focusing on prevalence, clinical characteristics, etiologic agents, and CD4+ T lymphocyte counts at diagnosis. Methods: A systematic review was conducted following PRISMA guidelines. MEDLINE/PubMed, SciELO, Scopus, and Scilit were searched for studies published between October 2015 and July 2025 in English and Spanish. Eligible studies included case reports, case series, and observational studies involving people with HIV and confirmed onychomycosis. Data extraction was performed independently, and findings were analyzed descriptively. Results: Thirty studies comprising 1296 patients were included; 306 had detailed clinical descriptions. Most cases were reported in the Americas (85.8%) and predominantly involved male patients. CD4+ counts were available in 123 individuals; 52% had <200 cells/µL, including 18 with <50 cells/µL. Trichophyton rubrum was the most frequently identified etiologic agent. Conclusions: Onychomycosis in HIV shows etiologic diversity and commonly affects patients with advanced immunosuppression, though it may also occur with partial immune preservation. Prospective standardized studies are needed.

Journal of Fungi doi: 10.3390/jof12050359

Authors: Pingliang Huang Ruifeng Yao Li Chen

Plant hormones and their synthetic analogueues offer sustainable alternatives for crop protection, yet the direct antifungal activity of strigolactone (SL) and its analogues against necrotrophic pathogens remain largely unexplored. Here, we screened eight phytohormones and related analogues for treatments of Botrytis cinerea and identified the SL analogue rac-GR24 (racemic GR24) as a concentration-dependent growth inhibitor active at low micromolar concentrations. Given the stereochemical complexity of SLs and their analogues, we evaluated multiple enantiopure isomers and found that ent-5DS and GR24ent-5DS, which differ in configuration from natural SLs, exhibited the strongest inhibitory activity. This stereospecific response was further validated using another filamentous fungus, Sclerotinia sclerotiorum, which displayed an identical susceptibility profile. Combinatorial treatments with enantiopure isomers and double-concentration rac-GR24 revealed that the antifungal effect of the racemate is primarily attributable to the GR24ent-5DS enantiomer, whereas the opposite enantiomer GR245DS is almost inactive. Collectively, our findings uncover a stereospecific response in fungal pathogens, demonstrating that B. cinerea and S. sclerotiorum respond to exogenous SL analogues in a chirally selective manner. This work establishes a stereochemically defined framework for developing enantioselective fungicidal agents with potential applications in sustainable agriculture.

Journal of Fungi doi: 10.3390/jof12050358

Authors: Joanna Burger Christian Jeitner Kelly Ng Robert T. Zappalorti John Bunnell Emile DeVito David Schneider David Burkett Michael Gochfeld

Ophidiomyces ophidiicola (O. ophidiicola) causes ophidiomycosis and has deleterious effects in some North American snakes. Studies have suggested that it is endemic in some species, but most studies have been conducted on individuals of unknown age, for only a season or two. This paper fills that gap by exploring prevalence of qPCR-confirmed ophidiomycosis in known-aged Northern pine snakes (Pituophis melanoleucus melanoleucus) during a six-year testing period, followed by two years of monitoring survival. Some individuals testing positive for O. ophidiicola lived for at least 8 years following initial infection, and their O. ophidiicola status could change from positive to negative and back again in successive years, while still seeming healthy. Approximately 85% were positive in at least one year, and overall positivity was 65% in the snakes. Detection frequency was 45% for ventral swabs, and only 23% for head swabs. Of 31 snakes found at least a year after first testing positive, 71% lived at least 3 years, and 23% lived 6 or more years. Females lived longer after testing positive than males, and more females changed from positive to negative than did males. These data help understand infections, recovery and re-infection in individuals, as well as survival of marked individuals, and have implications for endemism and long-term population viability of snake populations exposed to O. ophidiicola.

Journal of Fungi doi: 10.3390/jof12050357

Authors: Bo Tu Hui Chen Xu Zhang De-Xiang Tang Van-Minh Dao Chanhom Loinheuang Yao Wang

Gibellula (Cordycipitaceae, Hypocreales) represents a group of highly specialized obligate fungal pathogens restricted to spider hosts. Species delimitation was conducted using morphological characteristics in combination with multilocus phylogenetic analyses (nrSSU, ITS, nrLSU, tef1-α, rpb1, rpb2), and we recognized nine spider-associated Gibellula species from specimens collected in the Lancang–Mekong biodiversity hotspot (China, Laos, Thailand, and Vietnam). Among them, four are described as new to science: Gibellula longiconidiophora sp. nov., G. mekongensis sp. nov., G. ovorum sp. nov., and G. pseudopilosa sp. nov. The other five species represent new national distributional records: G. yunnanensis (new to Laos), G. pseudopigmentosa (new to Thailand), G. trimorpha (new to Vietnam), G. penicillioides (new to Laos), and G. scorpioides (new to China and Laos). Phylogenetic analyses resolved these taxa into well-supported lineages. Notably, G. ovorum is a rare example of a Gibellula species parasitizing spider egg sacs rather than adult spiders, revealing an unusual substrate shift. Morphological distinctions among the new species include differences in conidiophore length, synnematal development, conidial size, and sporulation patterns. Detailed descriptions, illustrations, and taxonomic comparisons are provided. This study significantly expands the known diversity and geographic distribution of Gibellula in the Lancang–Mekong region and underscores the importance of integrative taxonomy for uncovering hidden diversity in spider-pathogenic fungi.

Journal of Fungi doi: 10.3390/jof12050354

Authors: Yutao Zhou Jiahao Xu Chang Su Weina Wu Fengping Yi

Candida albicans is a common opportunistic pathogen. Long-term use of azole antifungals faces challenges like resistance, necessitating novel agents. Tea tree oil (TTO), a natural broad-spectrum antimicrobial, shows promise, but its molecular mechanisms, particularly concerning novel cell death pathways, require clarification. This study comprehensively evaluated the antifungal mechanism of TTO against C. albicans using transcriptomics. Antifungal susceptibility assays were conducted to assess the effects of TTO and its components (4-terpineol, terpenes, and γ-pinene) on the growth of C. albicans hyphae and biofilms. Fluorescent labeling and biochemical analysis were employed to detect ferroptosis markers. Transcriptomic results indicate that TTO induces 423 differentially expressed genes and systematically inhibits the development of C. albicans hyphae through mechanisms such as oxidative stress, iron homeostasis disruption, disruption of cell wall integrity, and interference with ergosterol metabolism. Notably, the significant enrichment of redox enzyme activity and iron ion binding functions, along with changes in the glutathione metabolic pathway, suggest that ferroptosis may be involved in this process. Subsequent studies revealed that the compound 4-pinene most effectively inhibits the pathogenicity of C. albicans by suppressing its adhesion, hyphae formation, and biofilm formation, whereas terpinene induces the accumulation of reactive oxygen species (ROS) and increases lipid peroxidation in C. albicans; furthermore, following treatment with an iron-mediated apoptosis inhibitor, terpinene enhances the viability of the treated C. albicans cells.

Journal of Fungi doi: 10.3390/jof12050356

Authors: Xiaolan Guo Jinbin Hu Yaqin Wang Lingda Zeng Dun Wang Yu Cao Delu Wang

Dark septate endophytes (DSE) and ericoid mycorrhizal fungi (ERMF) are employed to augment the abiotic stress resistance of fruits. However, their potential functions in enhancing the drought resistance of blueberry, an economically important fruit, remain unclear. Thus, this study aims to identify optimal inoculation combinations to enhance the drought resistance of blueberry seedlings. Specifically, the effects of single and dual inoculations with DSE (Cladosporium cladosporioides, D79) and ERMF (Oidiodendron citrinum, N12) on seedling physiology and metabolism were explored under varying drought conditions. The results showed that dual inoculation significantly improved leaf physiological characteristics. Under severe drought stress, the 1:2 DSE:ERMF ratio (D1N2) notably increased leaf relative water content (RWC) and reduced electrolyte leakage by up to 42.1% compared with the non-inoculated control. Dual inoculation also significantly decreased malondialdehyde (MDA) content, with the smallest increase observed in D1N2. Regarding antioxidant enzymes, dual inoculation sustained higher superoxide dismutase (SOD) activity under moderate drought and minimized the decline in SOD activity under severe drought (the lowest decrease was 36.4% in D1N2 versus 56.7% in CK). Moreover, the antioxidant losses under drought stress were reduced by upregulating various metabolic processes, especially the biosynthesis of phenylalanine, tyrosine, and tryptophan. A comprehensive evaluation suggested that inoculation with a 1:2 mixture of DSE and ERMF most effectively improved blueberry drought resistance, primarily by enhancing water and metabolite supply and stimulating the antioxidant defenses.

Journal of Fungi doi: 10.3390/jof12050355

Authors: Xian-Yan Zhou Tolgor Bau

Lepiota sect. Stenosporae is characterized by a trichodermal or cutis-like pileus covering and spurred basidiospores. Although macroscopic similarities among its members complicate field identification, species can be delimited by combining multi-locus (ITS, nrLSU, rpb2, and tef1-α) phylogenetic analyses with distinct micro-morphological features. Using this integrative approach, we investigated specimens of sect. Stenosporae collected from Northeast China. A total of 12 species were successfully delimited, including six species new to science (Lepiota dolichospora, L. hongshiensis, L. jilinensis, L. microstenospora, L. sinocastanea, and L. sirupa) and one new record for China (L. grangei). Comprehensive morphological descriptions and line-drawing illustrations of microscopic features are provided for all recognized taxa. These findings expand the known species diversity of Lepiota in China and contribute morphological and molecular data for further systematic studies of this fungal group.

Journal of Fungi doi: 10.3390/jof12050353

Authors: Jianyue Li Jien Xi Yuanwu Yang Wei Wang Chengti Xu Jinling Ma Xixi Yao Haodong Liu

Legume–grass mixtures are widely used to improve productivity and soil quality in alpine grasslands; however, their effects on soil fungal communities and the underlying mechanisms remain unclear. In this study, a two-factor field experiment (species richness (three, four, and five species) × legume–grass ratio (4:6, 3:7 and 2:8)) was conducted in an alpine artificial grassland on the Qinghai–Tibetan Plateau. Soil fungal communities were assessed using high-throughput sequencing combined with multivariate analyses. The results showed that Ascomycota, Mortierellomycota, and Basidiomycota were the dominant fungal phyla. Both species composition and seeding ratio significantly influenced fungal community structure and α-diversity primarily through indirect pathways mediated by plant community characteristics and soil properties. Plant height and soil total phosphorus (TP) were identified as key drivers of fungal α-diversity. Specifically, the four-species mixture (Z2) at a 3:7 legume–grass ratio resulted in relatively higher and more stable aboveground biomass and improved soil nutrient status, whereas increasing species richness to five species did not further enhance these benefits. Overall, our findings indicate that optimizing species composition and legume–grass ratio, rather than simply increasing species richness, is more effective for regulating soil fungal diversity and ecosystem function.

Journal of Fungi doi: 10.3390/jof12050352

Authors: Linhai Hong Qi Fan Nan Tao Peng Wang Ping Liu Jing Leng Chunxin Yao Qinghong Liu

This study presents a high-quality chromosome-level genome assembly of Morchella sextelata (54.64 Mb, 26 pseudochromosomes) and systematically characterizes its genomic and evolutionary features. Phylogenetic analysis indicates that M. sextelata diverged early within the Morchella genus (~14.2 million years ago) and underwent substantial genomic remodeling, with 1124 expanded and 1961 contracted gene families. Enrichment analysis of rapidly expanded gene families highlights two prominent functional themes: genes associated with small molecule/ion binding and secondary metabolite biosynthesis, and genes linked to the Fanconi anemia pathway and DNA repair/recombination. Notably, 56.96% of the COG-annotated M. sextelata-specific genes encode retrotransposon-related proteins, and this enrichment coincides with the expansion of DNA repair systems—a pattern reminiscent of the “transposon domestication” model. Functional genomic analyses further reveal that the glycoside hydrolase system is dominated by GH5, GH43, and GH3 families, suggesting a predicted capacity for plant cell wall polysaccharide degradation, while 12 biosynthetic gene clusters indicate genetic potential for terpenoid and non-ribosomal peptide biosynthesis. These findings provide a valuable genomic resource for M. sextelata and offer new insights into the role of transposable element mediated remodeling in fungal genome evolution.

Journal of Fungi doi: 10.3390/jof12050351

Authors: Thanawat Sridapan Chalisa Jaturapaktrarak Thidarat Rujirawat Wilasinee Konsue Pattarana Sae-Chew Chompoonek Yurayart Theerapong Krajaejun

Pythiosis is an emerging infectious disease caused by the oomycete Pythium insidiosum, affecting humans and animals primarily in subtropical and tropical regions. The pathogen is commonly found in swampy environments, and exposure can lead to diverse clinical manifestations. In humans, ocular and vascular infections predominate, whereas in animals, cutaneous/subcutaneous or gastrointestinal disease is more common. Medical therapy is frequently ineffective, and many patients require extensive surgical intervention. Advanced cases may progress to fatal outcomes. Therefore, early and accurate detection is critical for improving clinical management. This study evaluated a colorimetric loop-mediated isothermal amplification (c-LAMP) assay compared with an established multiplex PCR (m-PCR) assay for the detection of P. insidiosum in clinical specimens from animals with and without pythiosis. When tested on 47 frozen tissue samples, c-LAMP demonstrated superior diagnostic performance, with markedly greater sensitivity (83.9% vs. 41.9%), higher accuracy (78.7% vs. 61.7%), and a shorter turnaround time (65 vs. 180 min). However, c-LAMP yielded five false-positive results, likely due to nonspecific amplification or contamination. Improved sample-handling practices increased the specificity from 68.8% to 93.8%. In contrast, m-PCR showed perfect specificity (100.0%) but substantially lower sensitivity, resulting in a high false-negative rate. In conclusion, these preliminary findings suggest that c-LAMP is a promising rapid screening tool for suspected pythiosis, particularly in resource-limited settings. Nevertheless, confirmatory testing remains necessary for positive or equivocal c-LAMP results.

Journal of Fungi doi: 10.3390/jof12050350

Authors: Yanshu Zhao Xiaojia Zhang Yuxin Jiang Shuyuan Song Chuang Han Xiaodong Dai

This paper systematically reviews the research progress on the physiological functions, gene cloning, classification basis, and expression regulation mechanisms of laccase in edible and medicinal fungi. Laccase is a copper-containing polyphenol oxidase widely distributed in these fungi, characterized by broad-spectrum substrate catalytic activity and redox properties. It plays a central role in lignin degradation, pigment synthesis, and environmental pollutant treatment. Regarding gene cloning, researchers have successfully isolated and identified laccase genes from multiple species using techniques such as transcriptome sequencing, RACE amplification, and gene knockout. Expression regulation studies have revealed that laccase genes exhibit stage-specific expression patterns during mycelial growth, fruiting body development, and lignin degradation. In recent years, breakthroughs in genomics, transcriptomics, and gene editing technologies have greatly advanced research into the cloning, classification, and regulatory mechanisms of laccase genes. This article systematically reviews the diversity, clonal classification, and regulatory mechanisms of these genes, aiming to provide a reference for further research and industrial development of laccase in edible and medicinal fungi.

Journal of Fungi doi: 10.3390/jof12050349

Authors: Zhen Zhang Zhe Wang Qinyuan Xue Jiahui Yu Tailong Li Lan Wang Hongzu Feng

The occurrence of pear Valsa canker caused by Cytospora pyri poses a serious threat to the healthy and sustainable development of the Korla fragrant pear industry. To effectively control the disease, endophytic strains were isolated from the bark of Korla fragrant pear trees and screened for strong antagonistic activity against the pathogen. The selected strain was identified based on morphological characteristics and 16S rRNA phylogenetic analysis. Its biocontrol potential and functional traits were further evaluated, along with its growth-promoting ability assessed through in vitro tests and preliminary tomato pot experiments. Results showed that a total of 264 endophytic isolates were obtained from 200 pear tissue samples using dilution plating and tissue separation methods. Among them, strain FB-4 exhibited significant inhibition against C. pyri. Morphological observations and phylogenetic analysis identified the strain as Bacillus velezensis, named FB-4. The cell-free supernatant inhibited conidial germination and mycelial growth of the pathogen by 88.21% and 85.51%, respectively, and showed preventive and curative efficacies of 74.49% and 58.97% against pear Valsa canker. In vitro assays indicated that FB-4 could produce indole-3-acetic acid, protease, amylase, and cellulase, and demonstrated abilities to solubilize inorganic phosphate and synthesize siderophores. Inoculation with FB-4 bacterial suspension promoted the growth of tomato seedlings, with higher concentrations yielding more pronounced effects. In conclusion, strain FB-4 represents a dual-functional biocontrol agent with both antagonistic and plant-growth-promoting properties.

Journal of Fungi doi: 10.3390/jof12050348

Authors: Nikolett Molnár Dóra Szabó Adrienn Gomba-Tóth Ádám Novák Kálmán Zoltán Váczy Zoltán Karácsony

An isolate of Peniophora lycii was obtained from grapevine, and its interactions with several grapevine-associated fungi and the plant host were examined. The fungus was not able to infect intact leaves, but necrotized the margins of foliar disks and caused necrosis and white rot in woody tissues. In dual cultures, P. lycii and Aureobasidium pullulans showed mutual antagonism. Mycoparsitism of P. lycii was observed on epiphytic Botrytis cinerea, Alternaria sp., and endophytic Botryosphaeria dothidea interaction partners. In contrast, P. lycii showed trophic growth towards endophytic Phaeomoniella chlamydospora without any signs of harmful interactions. Dual inoculations of foliar disks with epiphytic fungi revealed no effects of fungal interactions on necrosis development by pathogens and verified mycoparasitic interactions in planta. Co-infection assays of cuttings with endophytic pathogen fungi showed cumulative effects of fungal interactions on wood symptom expression, with the exclusive contribution of P. lycii to white rot development. In addition to providing the first isolation of P. lycii from grapevine and the description of its mycoparasitic behavior, the present study suggests that the fungus may act as an opportunistic grapevine pathogen, probably as a secondary colonizer in trunk diseases. The observed dual host preference may allow trunk disease pathogens to initially feed on fungi, followed by damage to the grapevine. This may be in connection with the exceptionally long latency of these syndromes.

Journal of Fungi doi: 10.3390/jof12050347

Authors: Soma Araki Shunya Susukida Ken Miyazawa Toshitaka Kumagai Jikian Tokashiki Keietsu Abe

During fermentation in stirred-tank bioreactors (STBR), filamentous fungi are frequently exposed to hypoxic conditions. However, their responses, especially short-term ones (≤6 h), remain unclear. In this study, we performed a short-term multi-omics profiling in an Aspergillus oryzae hyphal dispersion mutant (AGΔ-GAGΔ) during a controlled transition to hypoxia (a decrease in dissolved oxygen (DO) from 10% to ≤1%) in a 4 L STBR. In transcriptome analysis, the genes encoding mitochondrial respiratory chain Complexes I–III were transiently downregulated at 1 h from DO depletion and were then upregulated, whereas those of Complex IV were upregulated immediately at the onset of hypoxia. In relation to this respiratory remodeling, we also observed an immediate induction of an alternative oxidase (AOX) gene. However, our metabolome data showed no significant change in the ATP level. This result could be explained by the upregulation of the glycolytic genes in hypoxic cultures. Fluorescence imaging revealed a transient increase in intracellular reactive oxygen species (ROS) in hypoxia, and metabolomics data revealed a decrease in the reduced glutathione/oxidized glutathione ratio in hypoxic cultures. Deletion of the AOX gene prolonged the ROS increase. Together, these data indicate that early hypoxia triggers a transient increase in oxidative stress, mitigated by antioxidant systems and mitochondrial respiratory rebalancing including an AOX-mediated bypass.

Journal of Fungi doi: 10.3390/jof12050346

Authors: Jefferson Brendon Almeida dos Reis Thayssa Monize Rosa de Oliveira Samia Gomes-da-Silva Maria Regina Silveira Sartori da Silva Fabyano Alvares Cardoso Lopes Alessandra Monteiro de Paula Nadson de Carvalho Pontes Helson Mario Martins do Vale

Understanding how agricultural soil management affects soil fungal communities is essential for assessing the resilience of biodiversity hotspots such as the Brazilian Cerrado. In this study, we characterized fungal community structure across three contrasting land-use systems within the same agricultural landscape: a native Cerrado remnant, a cover-cropping system, and a spatially isolated potato monoculture field. The soil’s chemical and enzymatic characteristics differed from one another and were clustered by area. However, the same pattern was not observed for the fungal community. Alpha-diversity indices did not differ significantly among sites, although native Cerrado soils showed slightly higher richness and evenness. Beta-diversity analyses based on Bray–Curtis and Jaccard distances, supported by NMDS, ANOSIM, beta-dispersion, and PERMANOVA, indicated no significant compositional differences among communities. Core-mycobiota analysis identified 157 shared ASVs, including genera such as Fusarium, Cladosporium, Chrysosporium, Trichoderma, and Clonostachys. As a preliminary assessment based on a limited spatial design and sequencing-based inference, these findings should be interpreted with caution. These results underscore the need for further research on the mechanisms driving fungal dispersal, edge effects, and the long-term impacts of agricultural land-use on fungal diversity and ecological integrity in the Cerrado.

Journal of Fungi doi: 10.3390/jof12050345

Authors: Yangtian Li Bowen Tang Jinchi Xie Yaohui Pan Taixin Yang Xiaoxiao Feng Bin Zhao Yingchao Liu

Yam is a valuable traditional Chinese medicinal and nutritional crop that has gained significant popularity in recent years. However, the production of yam is severely impacted by brown rot caused by Fusarium solani, which affects both yield and quality. In this study, we isolated a promising biocontrol strain, designated AP6, from the rhizospheric soil of healthy yam. Strain AP6 exhibited strong antagonistic activity against F. solani, with an inhibition rate of 60.2%, and demonstrated broad-spectrum antagonistic activity. Based on morphological, physiological, biochemical characteristics, and whole-genome sequencing, this strain was identified as Bacillus velezensis. Strain AP6 can produce siderophores, amylase, protease, cellulase, and form biofilms. It can also change the hyphal morphology of F. solani, inhibit spore germination, reduce the viability of pathogens, and alleviate the disease severity of yam. Additionally, strain AP6 was shown to induce the accumulation of reactive oxygen species in yam, thereby enhancing the yam’s defense responses. Field application trials demonstrated that a wettable powder formulation of strain AP6, when combined with commercial metalaxyl-M and fludioxonil, significantly reduced the incidence of brown rot in yam. These findings indicate that B.velezensis AP6 is a promising biocontrol agent, providing a practical and sustainable approach for integrated disease management in yam.

Journal of Fungi doi: 10.3390/jof12050344

Authors: Sehee I. Rim Matthew L. Duley Michael R. Hughes Jing Jing Marcia R. Lee

Members of Fusarium oxysporum species complex (FOSC) and Fusarium solani species complex (FSSC) are common etiologic agents of opportunistic mycoses. Their hyphae grow via apical extension followed by invasion of host epithelial surfaces, particularly corneal and epithelial tissues. Thigmotropism is the contact-sensing response of these growing hyphae to change the direction of their apical growth in response to changes in the topography of their contacted surfaces, such as clefts between adjacent host cells. Investigations of fungal thigmotropism among etiologic agents of opportunistic mycoses are sparse. In this study, we assessed thigmotropic capacities of 10 fusarial strains. Thigmotropic activity was quantified using a chemotaxicell system followed by scanning electron microscopy (SEM). Isolates belonging to the FSSC had significantly greater rates of thigmotropism than FOSC strains (p < 0.0001). In summary, this work offers a creative method to assess thigmotropism, a crucial aspect of fungal pathogenesis, and, secondly, demonstrates potent thigmotropic capacities of fusarial isolates belonging to the FSSC.

Journal of Fungi doi: 10.3390/jof12050343

Authors: Marlene Baptista Jean-Marc Daran Lucília Domingues

Isoprene, a highly volatile hydrocarbon with numerous industrial applications, has traditionally been produced from petrochemical sources through processes associated with significant environmental impacts. Microbial production of isoprene has emerged as a promising and more sustainable alternative. In this study, the potential of the non-conventional yeast Kluyveromyces marxianus to produce isoprene from a renewable feedstock was explored, contributing to the development of a more sustainable process. K. marxianus was engineered to produce isoprene from glucose through the expression of an isoprene synthase gene, and the gene copy number of this enzyme was found to significantly influence isoprene production. Furthermore, enhancing the supply of the isoprene precursors acetyl-CoA and dimethylallyl diphosphate (DMAPP) via engineering of the mevalonate pathway led to increased production. A higher headspace-to-culture ratio in sealed serum bottles also facilitated isoprene accumulation. Importantly, isoprene production was achieved from the cellulosic fraction of pretreated Eucalyptus globulus wood. To our knowledge, this is the first report of isoprene production in K. marxianus using a lignocellulosic feedstock, providing proof of concept for its potential in integrated processes based on sustainable substrates under stressful conditions.

Journal of Fungi doi: 10.3390/jof12050342

Authors: Marie Dwyer Rebecca A. Owens Claudia Garcia Revuelto Kieran G. Walshe Cathal M. Murphy Nicola M. Moloney Sean Doyle

Although Aspergillus fumigatus has been identified as a critical fungal pathogen by the World Health Organization, diagnosis of the various types of aspergilloses remains unsatisfactory. New biomarkers of disease and accessible test systems are needed. Moreover, new treatment concepts are required to address infections caused by this invasive pathogen. Siderophore production by A. fumigatus offers opportunities for both improved diagnosis and potential therapy. Here, we report the development of a competitive ELISA that detects ferrated triacetylfusarinine C (FeTAFC) using a recombinant monoclonal IgG [anti-FeTAFC]. The FeTAFC ELISA can detect FeTAFC in human urine, a matrix proposed to be especially suitable for disease diagnosis because of the non-invasive nature of specimen collection. In addition, a novel assay was developed to assess the in vitro inhibitory properties of the IgG [anti-FeTAFC] towards A. fumigatus mutant and wild-type growth under iron-limiting conditions. The growth of A. fumigatus ΔsidD, deficient in TAFC and precursor fusarinine C (FsC) biosynthesis, was significantly reduced (p = 0.0003) in the presence of the antibody. While the growth of A. fumigatus wild-type was less affected in the presence of the antibody, the ratio of secreted TAFC:FsC was increased due to increased conversion of FsC to TAFC—likely because of extracellular complexation of FeTAFC by the IgG [anti-FeTAFC]. This work shows the utility of the IgG [anti-FeTAFC] for the detection of A. fumigatus and initial evidence for limiting fungal growth by attenuation of FeTAFC uptake.

Journal of Fungi doi: 10.3390/jof12050341

Authors: Elliott Hayden Aicha Kebe Shuzhen Chen Abagail Chumley Chenyi Xia Widad El-Zein Quan Zhong Shulin Ju

Mutations in an RNA-binding protein FUS are known to cause familial amyotrophic lateral sclerosis (ALS). Since this discovery, mutations in several other RNA-binding proteins (RBPs) have also been linked to ALS. Some of these ALS-associated RBPs have been shown to colocalize with ribonucleoprotein (RNP) granules such as stress granules and processing bodies (p-bodies). Increasing evidence has emerged supporting a hypothesis that the impaired clearance, inappropriate assembly, and dysregulation of RNP granules play a role in ALS. Through the genome-scale overexpression screening of a yeast model of FUS toxicity, we found that TAF15, a human RBP with a similar protein domain structure and belonging to the same FET protein family as FUS, suppresses FUS toxicity in yeast. The suppression by TAF15 is specific to FUS and not found in other yeast models of neurodegenerative disease-associated proteins. We showed that the RNA recognition motif (RRM) of TAF15 is required for its suppression of FUS toxicity. Furthermore, FUS and TAF15 physically interact, and the C-terminus of TAF15 is required for both the physical protein–protein interaction and its protection against FUS toxicity. Finally, while FUS induces and colocalizes with both stress granules and p-bodies, TAF15 only induces and colocalizes with p-bodies. Importantly, the co-expression of FUS and TAF15 induces more p-bodies than individually expressing each gene alone, and FUS toxicity is exacerbated in yeast that is deficient in p-body formation. Overall, our findings suggest a role of increased p-body formation in the suppression of FUS toxicity by TAF15.

Journal of Fungi doi: 10.3390/jof12050340

Authors: Pengxu Chen Yingying Chen Lize Wang Ziyi Lan Xin Zheng Luoyuan Wang Xi Gan Sijia Zhang Yanxia Zhao

Ceramidases hydrolyze ceramides to fatty acids and sphingolipids, but their role in fungal response to stress remains unclear. We investigated the function of neutral ceramidase (nCDase) response to aromatic fungicide (carvacrol, cuminaldehyde, and isoniazid) stress in Neurospora crassa. Comparative analysis of the wild-type strain, Δnc and OEnc showed that nCDase enhanced fungicide resistance through multiple mechanisms. nCDase improved β-1,3-glucan synthesis (30% increase), decreased membrane permeability, elevated superoxide dismutase and catalase activities, and promoted carotenoid accumulation (50%), which collectively improved stress tolerance. Δnc exhibited disruption of cellular integrity, altered fatty acid profiles (elevated oleic acid, reduced total fatty acids), and increased fungicide sensitivity. Collectively, these findings established that nCDase as a key regulator of cell wall dynamics, lipid homeostasis, and antioxidant defense, thereby facilitating fungal adaptation to abiotic stress. This study identified the role of nCDase in the response to aromatic fungicide stress and laid foundation for inhibiting pathogenic fungi in agricultural production and food preservation.

Journal of Fungi doi: 10.3390/jof12050339

Authors: Rafael Zaragoza Ángel Estella Xavier Nuvials Mireya Robles-Plaza Araceli Casado-Gómez

Background: The objective was to identify management strategies of IFI in critically ill patients through a Spanish national survey. Methods: A cross-sectional multicentre survey among ICU specialists, experienced in IFI, was performed (22 April–25 July 2024). The survey consisted of 13 questions with four closed answers. Results: Sixty-three specialists from 51 hospitals of 16 regions completed the survey. 95% stated that, in high-risk patients with clinical suspicion of Pulmonary Aspergillosis (PA), galactomannan in BAL is performed to guide treatment. In the treatment of patients with PA and influenza, 86% declared that isavuconazole and liposomal amphotericin B are recommended treatments and in high suspicion of Aspergillus coinfection, 76% recommended empirical treatment waiting for microbiological confirmation. 90% declared that the use of Extracorporeal Membrane Oxygenation (ECMO) and Renal Replacement Therapies (RRT) could be associated with lower azole levels. Regarding intra-abdominal candidiasis, 78% that physiopathological changes in critically ill patients, reduce their entry into peritoneal fluid. Conclusions: The majority of the respondents agreed (>80%) on: In suspicion of PA, Galactomannan in BAL to guide treatment is mandatory; In case of aspergillosis and influenza, isavuconazole and liposomal amphotericin B are the recommended treatments; The use of ECMO and RRT could be associated with lower azole levels.

Journal of Fungi doi: 10.3390/jof12050338

Authors: Dong Li Chun Yin Wanying Zhao Youyang Wang Shoujian Zang Wenzhi Wang Youxiong Que Qibin Wu Weixiang Wang

Histone demethylases regulate epigenetic modifications and DNA damage repair in fungal pathogens, yet their specific functions in Magnaporthe oryzae remain poorly understood. This study identifies MoRph1, a JmjC domain-containing histone demethylase that interacts with the COMPASS complex. Targeted deletion of MoRph1 resulted in significantly reduced vegetative growth, impaired conidiation, and defective appressorium formation. The mutant displayed compromised appressorial turgor pressure due to delayed degradation of glycogen and lipid reserves, leading to inefficient host penetration and attenuated virulence on rice and barley. MoRph1 localized to the nucleus, and its absence caused increased nuclear abnormalities under DNA damage stress, suggesting impaired genome stability maintenance. Biochemical analysis confirmed that MoRph1 specifically demethylates histone H3 lysine 36 trimethylation. Transcriptome analysis revealed altered expression of genes associated with DNA replication, mismatch repair, and oxidative stress response. These results establish MoRph1 as a crucial epigenetic regulator coordinating fungal development, infection structure function, energy mobilization, and DNA damage repair. This study underscores the importance of chromatin-level regulation in fungal pathogenicity and provides a foundation for future evaluation of MoRph1 as a potential antifungal target.

Journal of Fungi doi: 10.3390/jof12050337

Authors: Kinga Edina Varga Zsigmond Benkő Károly Antal Brigitta Povazsanyecz Katalin Forgács István Pócsi Tamás Emri

The functions of the putative bZIP (basic leucine zipper) transcription factors ZipV (Afu3g032301) and ZipZ (Afu2g14350) were investigated using wild-type, gene-deficient, and complemented strains of Aspergillus fumigatus. Deletion of zipV increased oxidative stress sensitivity and reduced in vivo virulence in a Galleria mellonella model, whereas complementation restored the wild-type phenotype. In contrast, deletion of zipZ resulted in no detectable phenotypic changes, even though transcription of both genes was modulated by oxidative stress. Phenotypic characterization of conidia, transcriptomic analyses of growing cultures and reverse-transcription quantitative real-time PCR of conidia-producing cultures suggested that ZipV regulates the development of certain conidial traits. The conidia of the ∆zipV mutant showed reduced heat stress tolerance, decreased catalase activity and delayed germination in comparison to the wild-type or the complemented strain. In the ∆zipV mutant, the transcription of catA encoding conidial catalase was impaired. This defect explains the reduced catalase activity and the oxidative stress sensitivity of the mutant and may contribute to its reduced virulence. The increased transcriptional activity of the alternative oxidase gene aoxA observed in the absence of ZipV suggests a compensatory response aimed at mitigating oxidative stress.

Journal of Fungi doi: 10.3390/jof12050336

Authors: Dana L. Surwill Patricia Cruz Mark P. Buttner Jennifer R. Pharr Nancy Lough Theresa T. Roehr

Background: Fungal infections pose a significant threat to public health, with over 6.55 million cases and 2.55 million deaths annually. Exposure to fungal spores in indoor environments primarily occurs through inhalation or direct contact with surfaces. Monitoring is critical for early detection and prevention of outbreaks, yet routine airborne fungal testing is not universally mandated across healthcare settings. Methods: A systematic review of peer-reviewed articles from four databases was conducted to identify current airborne fungal monitoring guidelines and best practices for sample collection, culture media, incubation conditions, and results interpretation. Results: Eighteen articles met the inclusion criteria, and four studies discussed potential guidelines for acceptable airborne fungal levels in healthcare environments. Guidelines ranged from <1 CFU/m3 for HEPA-filtered environments to >1000 CFU/m3 for non-filtered areas. The most common fungi identified were Aspergillus, Penicillium, Alternaria, Cladosporium, and Rhizopus, with six WHO-listed critical fungal pathogens found. Impaction was the sole sampling method used, with most studies employing Sabouraud dextrose or malt extract agar with chloramphenicol, incubation for 2–7 days at 25–30 °C, and morphological identification. Conclusions: The need for globally recognized fungal monitoring standards is pressing. Without them, preventable fungal exposure will persist, risking severe, potentially fatal infections for patients and healthcare workers.

Journal of Fungi doi: 10.3390/jof12050334

Authors: Adiyadolgor Turbat Aruna Vigneshwari Thu Huynh Nomuun Oyunbat Apoorv Tiwari Pramod W. Ramteke Mónika Varga Tamás Papp Csaba Vágvölgyi András Szekeres

Indole-3-acetic acid (IAA) is a central auxin regulating plant growth and developmental plasticity, including cell division, elongation, vascular differentiation, and root architecture. Beyond plants, many fungi—including endophytic fungi that reside within plant tissues without visible disease symptoms—can synthesize IAA and thereby modulate plant performance and plant–fungus interactions. However, fungal IAA biosynthesis remains less resolved than bacterial or plant pathways, and mechanistic evidence is uneven across taxa. This review summarizes recent (2020–2025) advances in IAA-producing endophytic fungi, the main proposed biosynthetic routes and intermediates, and methodological considerations for attributing IAA production to the fungal partner. The reported associations of fungal IAA with beneficial interactions are then discussed, including growth promotion, stress tolerance, and interaction-mediated changes in the root system’s architecture, with emphasis on the concentration- and context-dependent nature of plant responses. Overall, fungal IAA is associated with diverse plant–fungus interactions, as discussed in this review.

Journal of Fungi doi: 10.3390/jof12050335

Authors: Qiaoming Zhang Wenjing Yang Caiyun Zhang Lirong Ren Na Bai Lin Zhang Chen He Minggui Gong

Cotton (Gossypium hirsutum L.) is a promising candidate for an Arsenic (As)-tolerant plant due to its low As accumulation in fibers. The individual arbuscular mycorrhizal fungi (AMF) inoculation or exogenous calcium (Ca2+) application is known to enhance heavy metal tolerance in higher plants; however, their synergistic mechanisms in alleviating As stress in cotton remain poorly understood. A three-factor pot experiment was conducted, including two levels of AMF (Funneliformis mosseae C.Walker & A.Schüßler) inoculation (non-inoculated/inoculated), As stress (0/100 mgAs5+·kg−1soil), and exogenous Ca2+ (CaCl2) application (0/20 mmol·L−1 CaCl2). AMF inoculation and Ca2+ application were investigated for their effects on cotton growth, root morphology, photosynthetic characteristics, osmotic regulators, antioxidant enzyme activities, and ion homeostasis under As stress. Results showed As stress significantly disrupted cotton growth (decreased plant height, shoot and root dry weight) and root morphology (reduced total root length, root area, and root fork number), photosynthetic capacity (reduced Pn, Ci, Fv/Fm, and ΦPSII), osmotic adjustment (decreased proline, soluble sugar and protein), antioxidant defense (inhibited SOD, POD, CAT activities), and K+/Ca2+ homeostasis (reduced concentration of K+ and Ca2+, and K+/Ca2+ ratio). Both AMF inoculation and Ca2+ application independently alleviated these adverse effects of As stress. At the same time, AMF symbiosis combined with exogenous Ca2+ was better than AMF inoculation or Ca2+ application alone in optimizing root architecture, improving stomatal function and photosynthetic efficiency, enhancing osmotic regulator accumulation and antioxidant enzyme activities, and restoring ion balance under As stress. Three-way ANOVA confirmed significant As×AMF×Ca2+ interactions on key parameters such as Pn and ΦPSII. In summary, both AMF inoculation and Ca2+ application synergistically enhanced cotton As tolerance through regulating growth, root morphology, photosynthetic characteristics, osmotic regulators, antioxidant enzyme activities, and ion homeostasis, demonstrating its potential for sustainable cotton cultivation in As-contaminated soils.

Journal of Fungi doi: 10.3390/jof12050333

Authors: Asanka Madhushan Paul W. J. Taylor Ahmed Mahmoud Ismail Jian-Kui Liu Sajeewa S. N. Maharachchikumbura

This study investigated Fusarium species associated with seven wild relatives of four economically important Rosaceae fruits in Sichuan Province, China, including wild strawberry (Fragaria sp. and Potentilla indica), wild raspberry (Rubus rosaefolius), wild cherry (Prunus sp., Maddenia sp. and Prunus leveilleana), and wild apple (Malus kansuensis). Based on multi-gene phylogenetic analyses and morphological characteristics, seven Fusarium species within the Fusarium tricinctum species complex (FTSC) were identified. Among these, four are described as new species (F. fragariae, F. potentillae, F. pruni and F. fructicola), while the remaining three represent new host records (F. avenaceum, F. diversisporum and F. paeoniae). In addition, phylogenetic and morphological evidence indicated that F. rosiradicicola is conspecific with F. diversisporum. Prioritizing the oldest epithet, we synonymized F. rosiradicicola under F. diversisporum. The pathogenicity of the isolates was evaluated on both their wild hosts and the corresponding cultivated fruits using detached, wound-inoculated assays. All tested isolates produced symptoms, showing pathogenic potential under experimental conditions. This study shows that selected wild Rosaceae fruits harbor several members of the FTSC and provides preliminary evidence of cross-host susceptibility under experimental conditions. However, further field-based investigations and non-wound inoculation studies are required to clarify their ecological roles, natural host susceptibility, and potential relevance in cultivated systems.

Journal of Fungi doi: 10.3390/jof12050332

Authors: Yafei Qu Kebin Li Zhimin Wang Huanhuan Dong Athanase Hategekimana Xiaomei Wang Jiao Yin

Soybean root rot, primarily caused by Fusarium oxysporum, leads to severe root decay and substantial yield losses in Glycine max. This study screened ten entomopathogenic nematode-associated symbiotic bacteria for antagonistic activity against F. oxysporum. Among them, Xenorhabdus budapestensis XH-4 exhibited the strongest in vitro inhibition, suppressing mycelial growth by more than 73%. Antifungal activity was primarily attributed to extracellular metabolites, as both fermentation broth and cell-free culture supernatant were effective, whereas bacterial cell suspensions showed no significant inhibition. In greenhouse experiments, 40% (v/v) XH-4 reduced the disease index by 75–80%, comparable to the chemical fungicide metalaxyl–hymexazol. Genome mining revealed 20 biosynthetic gene clusters encoding diverse secondary metabolites, including fabclavine, fabclavine pyrrolizixenamide A, and putrebactin/avaroferrin, which likely underpin the strain antifungal efficacy. Additionally, XH-4 enhanced soybean antioxidant capacity and activated the phenylpropanoid pathway, indicating a dual mechanism involving direct antagonism and induced systemic resistance. These findings support the development of XH-4 as an environmentally friendly biofungicide for sustainable management of soybean root rot.

Journal of Fungi doi: 10.3390/jof12050330

Authors: Melannie Mayorga-Jiménez Roger Moya

The present study aims to determine the properties of sandwich mycelium-based composite panels (sandwich-MBC-panel) fabricated with a lightweight core of mycelium-based composites (MBCs) of Ganoderma lucidum and Pleurotus ostreatus and veneers of Gmelina arborea and Vochysia guatemalensis wood. Physical and mechanical properties, acoustic capacity, chemical composition (determined by FT-IR), thermal degradation, and inorganic components were evaluated. The results showed that the sandwich-MBC-panel presented a density of 0.27–0.40 g/cm3, an MC between 14.56 and 24.71%, and a water absorption between 43.64 and 61.32%. Regarding mechanical characteristics, the sandwich-MBC-panel with the highest MOR, MOE, and internal bond was that composed of G. lucidum and G. arborea. The treatment with the best tensile force value was the mixture of G. lucidum with O. pyramidale. The sandwich-MBC-panel constructed with balsawood showed the lowest noise reduction coefficient, while the panel composed of G. lucidum and P. ostreatus showed good substrate properties and appropriate carbon and nitrogen content. FT-IR spectroscopy revealed substrate degradation by fungal mycelium formation, and TGA curves showed that the MBC containing G. lucidum presented higher thermal degradation than the panel without G. lucidum, without fungal attack. The main results of this study showed that sandwich MBC panels, in which the MBC is used as a lightweight core and wood veneers are used on the faces, have the potential for use as acoustic panels and could represent a sustainable alternative to panels that are generally fabricated from synthetic materials and of low densities.

Journal of Fungi doi: 10.3390/jof12050331

Authors: Paloma Sánchez-Torres

Fungi are the main causative agents of plant diseases and are responsible for substantial and recurrent damage to agricultural systems. Their activity causes significant reductions in crop productivity and food quality, ultimately contributing to plant deterioration and economic losses. It is estimated that phytopathogenic fungi can compromise up to 30% of global agricultural production. To mitigate microbial deterioration, a wide range of control strategies have been employed, with chemical fungicides being one of the most widely used interventions. However, current approaches to fungal control are rapidly transforming owing to the growing prevalence of fungicide resistance, increasingly stringent regulatory frameworks governing chemical applications, and evolving market demands. Taken together, these factors impose new constraints and drive the development of more sustainable alternative options for effective food control. This review examines the diverse strategies used to control fungal diseases in plants, emphasizing advances in biocontrol agents and biofungicides, as well as emerging tools in the molecular biology, genomics, and biotechnology fields. The aim is to highlight recent developments and prospects that can be integrated into comprehensive disease-management approaches.

Journal of Fungi doi: 10.3390/jof12050329

Authors: Leslie Waren Silva de Freitas Layanne de Oliveira Ferro Andre Rodrigues Camila Santana de Oliveira Mateus Oliveira da Cruz Jadson Diogo Pereira Bezerra Hyang Burm Lee Cristina Maria de Souza-Motta Maria Alice Barbosa dos Santos Roger Fagner Ribeiro Melo André Luiz Cabral Monteiro de Azevedo Santiago

During a survey on Mucorales fungi from soil in the world’s most biodiverse savanna, the the Brazilian Cerrado, nine specimens belonging to the Cunninghamellace were isolated. Morphological, multiloci analyses (ITS-nLSU-act) and maximum temperature growth data revealed that those specimens represent four new species: two in Absidia and two in Gongronella. Morphological characteristics of the isolates distinguishes them from other species: Absidia rhizoidea sp. nov. forms rhizopodiform rhizoids at the end of stolons, commonly next to the sporangiophores; A. variabilis sp. nov., mostly with slightly dorsiventrally flattened sporangia; Gongronella longapophysata sp. nov., which forms a long apophysis below sporangia; and G. verticilatta sp. nov., with whorled-branched sporangiophores. The maximum temperatures growth (Tmax) of those new species are as follows: A. rhizoidea (33 °C on MEA and 32 °C on PDA), A. variabilis (31 °C on MEA and 32 °C on PDA), G. longapophysata (32 °C on MEA and 33 °C on PDA), and G. verticilatta (31 °C on MEA and PDA). The present study highlights and discusses the micromorphological, physiological (Tmax) and phylogenetic characteristics of the new species.

Journal of Fungi doi: 10.3390/jof12050328

Authors: Bo-Min Seo Che-Hwon Park Sung-Chul Lee Rae-Won Kang Young-Jin Park

This study aimed to elucidate the molecular mechanisms underlying phenotypic divergence between two Lentinula edodes strains, Le_L and Le_S, which exhibit distinct fruiting body morphologies. While phenotypic variation among mushroom strains has been widely observed, the relative contributions of transcriptional regulation and structural genomic variation to these differences remain poorly understood. Comparative transcriptome analysis identified 8541 differentially expressed genes (DEGs), revealing clear functional divergence between the two strains. Genes upregulated in Le_S were predominantly enriched in ribosomal components and translation-related processes, indicating enhanced protein synthesis activity. In contrast, Le_L-upregulated genes were associated with transporters, transcription factors, and diverse metabolic pathways, suggesting broader regulatory and physiological functions. Protein–protein interaction network analysis further highlighted distinct regulatory architectures, with ribosomal proteins forming highly interconnected hub gene modules in Le_S, whereas Le_L hub genes were functionally diverse and included multiple members of the Major Facilitator Superfamily (MFS). Ortholog analysis across 33 L. edodes strains demonstrated that most hub genes were conserved, indicating their roles as core genetic components. Despite widespread genome-wide variation, including 7931 SNPs and 1149 INDELs, sequence variation within hub genes was limited, and allele-specific expression analysis revealed no significant allelic imbalance. In contrast, presence–absence variation (PAV) analysis identified structural differences affecting MFS transporter genes, which were absent in Le_S but present and upregulated in Le_L. Collectively, these findings suggest that structural genomic variation, particularly involving transporter genes, may play a more prominent role than sequence-level variation in driving phenotypic divergence. This study provides new insights into the genetic basis of strain-specific traits in L. edodes and highlights the importance of integrating multi-level genomic analyses.

Journal of Fungi doi: 10.3390/jof12050327

Authors: Jialiang Ying Huawei Zhang

The co-culture between Trametes sp. D and Aspergillus niger L14 resulted in a distinct orange-brown antagonistic band at their interface. Direct hyphal contact was associated with markedly enhanced production of numerous secondary metabolites (SMs), some of which were absent or decreased in monocultures. T. sp. D induced indolic compounds and cyclic dipeptides, such as Indole-3-acetamide and Cyclo-(Pro-Phe), whereas A. niger L14 overproduced polyketide-derived pigments and organic acids, such as Fonsecin and Kojic acid. These SMs did not inhibit their producer but suppressed the opponent’s growth, indicating reciprocal chemical antagonism. Transcriptomic analysis revealed upregulation of stress-related and metabolic genes, consistent with each fungus activating defense pathways. Biochemical assays showed that the confrontation zone had the highest oxidative stress markers, cell wall-degrading enzyme activity, and acidification (notably by A. niger L14), reflecting intense interfungal antagonism. The stress-response mitogen-activated protein kinase (MAPK) pathway was also activated in both fungi. Our findings supported a mechanistic model of fungal competition involving direct contact, chemical exchange, enzymatic attack, and stress signaling, highlighting that physical interactions likely contributed to triggering cryptic secondary metabolism and robust defense responses.

Journal of Fungi doi: 10.3390/jof12050326

Authors: Francisca J. Grill Karen Pedersen Kenta Reilly Thomas E. Grys Douglas F. Lake Janis E. Blair

The diagnosis of coccidioidomycosis is often achieved serologically by the detection of antibodies against fungal antigens. While several serologic tests are available for coccidioidomycosis, all of them are performed in a laboratory setting causing delays in diagnosis and therapeutic intervention. Point-of-care testing offers the ability to provide a shorter time to result by avoiding specimen send-out, minimizing processing steps, and employing expeditious immunochemical techniques. A preliminary trial of a rapid anti-coccidioidal antibody lateral flow assay (LFA) using fingerstick blood was performed on 22 patients with coccidioidomycosis at the point of care during outpatient clinic visits. Patients were tested longitudinally over the course of one year. An LFA reader was implemented to provide an objective result by quantifying the intensity of the test line band. There was close qualitative concordance observed between positive LFA results with send-out immunodiffusion (89.5%) and complement fixation (78.4%) standard of care clinical laboratory assays. Additionally, the relationship between LFA test line density values and traditional complement fixation antibody titers was assessed.

Journal of Fungi doi: 10.3390/jof12050325

Authors: Camila Mendes Soares Larissa Alves da Silva Luanna de Oliveira e Lima Meryellem Bezerra Soares Raimundo Euzebio da Costa Neto José Maria Barbosa Filho Felipe Queiroga Sarmento Guerra Guilherme Maranhão Chaves Walicyranison Plínio da Silva-Rocha

Candida albicans is a yeast found in the oral cavity, gastrointestinal tract, and vaginal mucosa. This species is the most prevalent and virulent in conditions such as oral candidiasis. Myrtenol is a bicyclic monoterpene alcohol recognized for its antioxidant and anti-inflammatory attributes. Its primary source is the essential oil extracted from plants of the Myrtaceae family. This study evaluated the effect of (−)-myrtenol on the virulence factors of Candida albicans. Ten clinical isolates of Candida albicans and one reference strain (ATCC 90028) were used in this study. The virulence factors examined included adhesion, morphogenesis, and lipase production. Assays were conducted in the presence and absence of (−)-myrtenol, using a concentration corresponding to the minimum inhibitory concentration (MIC; 256 µg/mL). Results: The compound reduced the adherence of C. albicans to human oral epithelial cells (92.24 vs. 28.69), and reduced filamentation in liquid (3.17 vs. 2.57) and solid media. Furthermore, (−)-myrtenol inhibited lipase activity (0.68 vs. 1.00). Virulence factors expressed by C. albicans contribute to increased infection rates and, consequently, increased morbidity and mortality. The present findings demonstrate that (−)-myrtenol affects virulence-associated phenotypes of C. albicans in vitro. This compound represents a promising candidate for further investigation, particularly in studies addressing its mechanisms of action, safety, and potential applicability.

Journal of Fungi doi: 10.3390/jof12050322

Authors: Lucía Maribel Becerra Carolina Torres Palazzolo Selva Valeria Chimeno María Cecilia Lerena Santiago Sari Jorge Prieto Laura Analía Mercado Mariana Combina

Criolla grape varieties are native South American cultivars that represent an important reservoir of genetic and microbiological diversity. This study aimed to investigate the oenological potential of Saccharomyces and non-Saccharomyces yeasts isolated from three criolla grape varieties in order to support the future design of wine starters. Yeasts were isolated at different fermentation stages from four vineyards in Mendoza, Argentina. A total of 485 isolates were recovered and molecularly identified, revealing 12 species belonging to eight genera. Saccharomyces cerevisiae, Hanseniaspora guilliermondii and Hanseniaspora uvarum were the dominant species. Isolates were screened for H2S and acetic acid production, followed by physiological and enzymatic characterisation. Selected strains were further evaluated in small-scale fermentations to assess fermentative kinetics and metabolic performance. Significant variability was observed, particularly among non-Saccharomyces isolates, which generally exhibited lower ethanol yields and acetic acid production compared to S. cerevisiae. Several isolates of H. guilliermondii showed balanced fermentative behaviour and favourable metabolic and enzymatic profiles. Through the applied selection strategy, twelve strains emerged as promising wine starter candidates. These isolates combined a low production of undesirable metabolites, relevant enzymatic activities, and favourable fermentative performance. Overall, the results highlight the oenological potential of autochthonous yeasts as a resource for innovative winemaking strategies.

Journal of Fungi doi: 10.3390/jof12050324

Authors: Zipeng Zhao Xiaona Wang Yafei Ding Taian Hou Xingdian Zhang

Drought and nitrogen deposition are major drivers of global change that can influence forest ecosystems and plant–microbe interactions, yet their combined effects on endophytic fungal communities and the roles of dark septate endophytes (DSE) remain unclear. In this study, we examined the diversity of culturable endophytic fungi in leaves and roots of Quercus dentata under different drought and nitrogen deposition treatments and evaluated the functional effects of representative DSE strains on host growth and physiology. A total of 1488 fungal isolates were obtained, revealing distinct organ-specific community patterns. Root-associated communities showed greater compositional stability across treatments, whereas leaf communities were more responsive to environmental variation. Severe drought reduced the dominance of several genera and promoted community restructuring, while nitrogen deposition had contrasting effects on α-diversity in leaves and roots. Beta diversity analyses indicated significant interaction effects between drought and nitrogen addition. Inoculation with four DSE strains produced strain-dependent effects on plant biomass, photosynthesis, water-use efficiency, physiological traits, and nutrient contents. These results indicate that drought and nitrogen deposition jointly influence endophytic fungal communities and that functional differences among DSE strains may affect host responses to combined stress.

Journal of Fungi doi: 10.3390/jof12050323

Authors: Yu-Yu Shen Yan-Jia Chen Zi-Ruo Deng Jun-Yin Deng Chun-Mei Pang Yao-Bin Song Ming Dong

Cudonia and Pleurocordyceps comprise a relatively limited number of species worldwide. In particular, Pleurocordyceps represents an ecologically important group of entomopathogenic fungi with considerable potential for medicinal applications. In this study, two novel species, Cudonia aurantiaca and Pleurocordyceps tianmushanensis, were collected from the National Nature Reserve of Mount Tianmu, Zhejiang Province, China. Detailed morphological descriptions and illustrations are provided, and genus-level phylogenetic relationships are inferred based on a combined multi-locus sequence dataset comprising the internal transcribed spacer (ITS), small subunit ribosomal RNA (SSU), large subunit ribosomal RNA (LSU), translation elongation factor 1-alpha (tef-1α), RNA polymerase II largest subunit (rpb1), and RNA polymerase II second largest subunit (rpb2) gene regions. This study contributes to a better understanding of the diversity and taxonomy of Cudonia and Pleurocordyceps.

Journal of Fungi doi: 10.3390/jof12050321

Authors: Soravit Chaimongkol Wittayothin Yingkulchao Nattawut Rungjindamai Nguyen Tai Toan Borworn Werapan Kwanruthai Malairuang Phongsawat Khamsuntorn Sayanh Somrithipol Somjit Komwijit Sujinda Sommai Umpawa Pinruan Wai Prathumpai

Twenty strains of Schizophyllum commune from the BIOTEC culture collection were selected for this study. S. commune is characterized by white to gray fan-shaped caps with lobed margins and distinctive split gills. Phylogenetic analysis of combined LSU rDNA and ITS rDNA sequences data using maximum parsimony placed the fungi in a strongly supported clade with S. commune. All strains were primarily screened for exopolysaccharide (EPS) and biomass production using potato dextrose broth (PDB) and peptone yeast glucose medium (PYGM) in 250 mL flasks shaken at 200 rpm for 7 days. The results revealed three strains with high EPS production, each exceeding 2.3 g/L, namely MMCR00487, MMCR00474 and MMCR00256. These strains were selected for media optimization using a Plackett–Burman design. Among them, MMCR00256 exhibited the highest EPS yield of 8.34 ± 1.47 g/L, followed by MMCR00487 and MMCR00474. Therefore, the strain MMCR00256 was further optimized by central composite design. The results revealed that the optimized medium for MMCR00256 increased the production of EPS to 10.39 ± 1.69 g/L, with a biomass yield of 26.28 ± 1.63 g/L (395 mg/g). The 5 L bioreactor optimization tested two inoculum types (mycelial and pellet) and two media (CCD and estimated) using strain MMCR00256. The mycelial inoculum grown in the estimated medium produced the highest EPS yield of 8.37 ± 0.26 g/L after 3 days, with 13.56 ± 2.94 g/L biomass. In conclusion, this study demonstrates that S. commune MMCR00256, when cultivated using the estimated medium and mycelial inoculum, can achieve enhanced exopolysaccharide production with improved efficiency, highlighting its significant potential for the development of efficient and scalable schizophyllan production processes at the industrial scale. Furthermore, this study provides essential insights into the cultivation and optimization of schizophyllan in S. commune.

Journal of Fungi doi: 10.3390/jof12050320

Authors: Liyan Mao Shaozhen Yan Lei Tian Cui Jian Yue Wang Ziyong Sun Zhongju Chen

Background: Trichosporon japonicum is a rare but highly lethal pathogen causing fungemia in immunocompromised patients. With the expanding use of chimeric antigen receptor T (CAR-T) cell therapy, the spectrum of opportunistic fungal infections is changing, yet data on T. japonicum infections in this setting remain scarce. Case Presentation: A 69-year-old man with diffuse large B-cell lymphoma developed catheter-associated fungemia after CAR-T cell reinfusion. He initially presented with neck pain and white oral mucosal patches, followed by fever four days later. T. japonicum was isolated from both peripheral blood and central venous catheter tip cultures, identified by microscopic examination, mass spectrometry, and molecular sequencing. Antifungal prophylaxis was initiated before fever onset based on close monitoring of white blood cell count, procalcitonin, interleukin-6, and C-reactive protein; treatment was subsequently adjusted according to species identification and antifungal susceptibility results. Infection was controlled within two weeks after catheter removal and immune recovery. The patient remained well at six-month follow-up. Conclusion: This case adds to the limited literature on T. japonicum fungemia in patients receiving CAR-T therapy. Our experience, together with a review of the literature, underscores that successful management requires prompt catheter removal, immune restoration, and combination therapy with voriconazole and amphotericin B, as echinocandin monotherapy should be avoided. Awareness of this pathogen in immunocompromised patients is critical.

Journal of Fungi doi: 10.3390/jof12050319

Authors: Chao Lin Zhenxin Wang Na Zhang Yuying Liu Lixiao Song Jin Zhang Khassanov Vadim Haiqiang Wang Minglei Li Jianzhao Qi

Plant pathogens pose a severe threat to global agricultural production, and their pathogenicity is closely linked to the biosynthesis of secondary metabolites. Basidiomycete within the family Ustilaginaceae represent significant plant pathogens, among which Ustilago maydis, as a model species, has been extensively studied for its secondary metabolites. However, the biosynthetic potential of other species within this family remains poorly understood. In this study, we conducted whole-genome bioinformatic analyses of 16 Ustilaginaceae species, including U. maydis, to systematically identify the distribution of biosynthetic gene clusters (BGCs), core gene domain compositions, and interspecies similarities. A total of 181 predicted BGCs were identified, averaging approximately 11 per species. BGCs for mannosylerythritol lipids (MELs), siderophores, and itaconic acid, as well as the melanin-associated genes pks1 and pks2, were widely distributed across most species. Conversely, an additional melanin biosynthetic gene cluster was found exclusively in U. maydis strain 521, indicating species-specific occurrence. Furthermore, this study identified a novel class of polyketide synthase (PKS) gene clusters with uncharacterized functions across 15 species, exhibiting high sequence and structural conservation between species. These findings reveal the rich metabolic diversity and species-specific biosynthetic potential of Ustilaginaceae, and by using U. maydis as a reference model, we highlight several BGCs (e.g., for MELs, siderophores, itaconic acid, and melanin) that are known to contribute to virulence or pathogenicity in plant hosts. This provides new insights into their pathogenic mechanisms.

Journal of Fungi doi: 10.3390/jof12050318

Authors: Alyssa D. Friudenberg Saika Anne Yuan Lu Susan T. Weintraub Ryan L. Peterson

The invasive fungal pathogen Pseudogymnoascus destructans is responsible for the collapse of several North American bat species through an infectious fungal skin disease known as White-Nose Syndrome (WNS). Recent transcriptomic studies have suggested that trace copper ion acquisition is essential for P. destructans propagation on its animal hosts. However, little is known about the mechanistic details of P. destructans adaptation occurring at the protein level. In this study, we report the global proteomic adaptation of P. destructans under chronic Cu-stress growth conditions employing chemically defined media. We identify 4340 P. destructans proteins, or approximately 47.8% of the predicted proteome, spanning a dynamic intensity range of six orders of magnitude. Chronic Cu-withholding stress leads to substantial alterations in the proteome, with 1398 differentially abundant proteins (DAPs) exhibiting statistically significant (p < 0.05) changes in protein levels compared to control growth conditions. We find that Cu-withholding stress induces increased levels of proteins associated with high-affinity Cu-acquisition, changes in intracellular superoxide dismutase (SOD) levels, and alterations in mitochondrial proteins related to aerobic respiration. In contrast, chronic Cu-overload stress leads to 390 DAPs (p < 0.05), which are more widely distributed across the proteome, with several DAPs associated with genomic stability and basic metabolism. Additionally, in this report, we present assessment of antisera products against intracellular and cell-surface protein targets of P. destructans that are effective for indicating Cu-withholding stress by western blotting. Together this report, provides insight into P. destructans adaptability to copper stress and identifies fungal proteins that may alleviate copper stress in the WNS infection niche.

Journal of Fungi doi: 10.3390/jof12050317

Authors: Jinling Lan Yingxue Du Mingxuan Xiong Kaixin Zhang Xiaolin Chen Ying Song Yuejia Song Baohui Lu Changqing Chen Ronglin He Jie Gao

Ginseng Alternaria leaf and stem blight, caused by Alternaria panax, imposes substantial yield and economic losses to the ginseng cultivation industry. Current diagnostic methods for ginseng diseases primarily rely on pathogen isolation from infected tissues, a procedure that is laborious, time-consuming, and inherently low in sensitivity. This study has therefore developed a rapid, specific and sensitive SYBR Green-based quantitative real-time PCR (qPCR) assay for detecting A. panax in plants, seeds, and soil. The developed qPCR assay exhibited high sensitivity and repeatability, with a detection limit of 0.074 fg/μL of target amplicon DNA (0.619 ng/μL of genomic DNA) and a coefficient of variation below 2%. In artificially inoculated tissues (leaves, stems and seeds), Ct values decreased progressively with increasing incubation time, reflecting pathogen proliferation. Analysis of field-collected leaves and stems showed a strong overall correlation between Ct values and visual disease grades. Surveying of ginseng-growing areas revealed that A. panax was detected in asymptomatic leaves and stems at rates of 12.12% and 14.29%, respectively, and in 14.46% of soil samples and 23.73% of seed samples. This qPCR assay presented here provides a robust tool for forecasting early disease, tracking the primary inoculum of the pathogen and its transmission chains, and screening of both ginseng seed lots and candidate soils for ginseng Alternaria leaf and stem blight prior to planting.

Journal of Fungi doi: 10.3390/jof12050316

Authors: Vladimir V. Sokolov Kseniya V. Malysheva Elena N. Bilanenko Natalia N. Markelova Oksana A. Kolpakova Marina L. Georgieva Vera S. Sadykova

Emericellopsis species from extreme environmental conditions provide a rich source of unique and biologically active secondary metabolites. The paper exhibits a comprehensive genomic analysis including complete genome sequencing, phylogenetic reconstruction, and functional annotation of two Emericellopsis species from highly saline and alkaline coastal soil ecosystems. Comparative genomics has been applied to reveal the genetic evolution, metabolic diversity, and environmental adaptation of the Emericellopsis genus. The genomes of E. alkalina E101 and E. fimetaria p24 have been found to encode various enzymes, including carbohydrate-active enzymes such as endoxylanases, that are useful for many ecological adaptations. The genomes of E. alkalina E101 and E. fimetaria p24 feature numerous biosynthetic gene clusters (BGCs), capable of synthesizing both known and potentially novel secondary metabolites with antimicrobial activity. Some BGCs show similarity to those producing known secondary metabolites, such as leucostatin A/B, clavaric acid, ascochlorin, (-)-mellein, and apicidin, among others. However, the majority of BGCs do not display any known similarities. Thus, comparative genomics offers new insights into the biology, adaptation, and evolutionary history of Emericellopsis fungi and may serve as a highly useful tool within biotechnological applications.

Journal of Fungi doi: 10.3390/jof12050315

Authors: Alona Yu. Biketova Isaac Garrido-Benavent Vasco Fachada Giampaolo Simonini Matteo Gelardi Boris Assyov Elias Polemis Antoni Conca Roseina Woods Georgios I. Zervakis Jan Borovička Andrea C. Rinaldi

This study revises the genus Cyanoboletus (Boletaceae) in the Mediterranean Basin, integrating single-locus and multi-locus phylogenetic analyses (ITS, LSU, tef1-α, and rpb2), morphological characterisation, ecology, and arsenic accumulation in basidiomes. Morphological descriptions (including a new form, Cyanoboletus mediterraneensis f. pallidus), comprehensive sampling, type studies, biogeography, macro- and microphotographs, an identification key, and a historical overview of the nomenclatural issues surrounding C. pulverulentus, C. poikilochromus, and C. mediterraneensis are given. An epitype collection is designated for C. pulverulentus. A new method to measure spore suprahilar depression has been proposed, which allowed more clear morphological separation between C. mediterraneensis and C. pulverulentus. This method may prove useful for species delimitation in other fungal groups that have asymmetric basidiospores. Additionally, we generated a new ITS sequence of the C. sinopulverulentus holotype and inferred its conspecificity with the later described C. flavocontextus. Furthermore, notes on the taxonomy of Boletus gabretae are presented, and its placement in the genus Neoboletus is suggested. Cyanoboletus is confirmed as a strongly supported generic clade encompassing 21 monophyletic species-level clades, 14 of which represent known species, and seven are undescribed taxa. The synonymy of Cupreoboletus with Cyanoboletus is also verified. This publication provides the tools to delimit Cyanoboletus species that have important conservation value, which can be used by conservationists, ecologists, and citizen scientists. It also highlights species-specific arsenic hyperaccumulation in C. pulverulentus, contributing to a better understanding of fungal metal uptake. Our study indicates that within Cyanoboletus, only C. pulverulentus demonstrates this characteristic and is the only known member of Boletales that possesses a high arsenic accumulation ability.

Journal of Fungi doi: 10.3390/jof12050314

Authors: Ana Aramburu Núria Beltran-Sanz José Raggio Pradeep K. Divakar Ana Pintado Asunción de los Ríos Leopoldo G. Sancho

Antarctic terrestrial photosynthetic biota is dominated by cryptogamic communities, which are largely restricted to scarce ice-free areas. Among these, nunataks constitute habitats of remarkable biogeographical interest, as they may harbor distinctive biotic assemblages worthy of investigation. This work presents a comprehensive assessment of lichen diversity on Antarctic nunataks. The lichen flora of four nunataks on the Hurd Peninsula (Livingston Island, maritime Antarctica) was investigated. Specimens were identified using an integrative approach combining morphological assessment and DNA barcoding. This survey revealed a high and potentially underestimated species richness, with 39 confidently identified and several additional taxa requiring further taxonomic resolution. A review of published records of lichen occurrence in nunatak and non-nunatak environments throughout Antarctica was used to evaluate patterns in taxonomic, biogeographical, and morphotype composition. This synthesis showed that nunataks support lower species richness than other ice-free environments. Most of their taxa occur in non-nunatak areas, consistent with patterns observed locally on the Hurd Peninsula. Floristic overlap seems greater in continental Antarctica, suggesting a stronger influence of nunatak-associated environmental constraints in the maritime region. These results underscore the ecological significance of nunataks as environmentally filtered habitats and highlight their relevance for understanding biodiversity patterns and community assembly in Antarctica’s terrestrial ecosystems.