Journal of Fungi

28 pages, 60735 KB

Open AccessArticle

Morphological and Phylogenetic Analyses Reveal Four New Species of Hydnellum from China

by Yonglan Tuo, Yiming Li, Libo Wang, Hang Chu, Zhengxiang Qi, Jiajun Hu, Xiao Li, Bo Zhang and Yu Li

J. Fungi 2026, 12(4), 267; https://doi.org/10.3390/jof12040267 - 8 Apr 2026

Abstract

Hydnellum is an ectomycorrhizal fungus with important ecological and medicinal value. However, the species diversity of Hydnellum in China remains poorly understood. To deepen the understanding of the diversity of Hydnellum species in China, this study, based on a combination of morphological observations [...] Read more.

Hydnellum is an ectomycorrhizal fungus with important ecological and medicinal value. However, the species diversity of Hydnellum in China remains poorly understood. To deepen the understanding of the diversity of Hydnellum species in China, this study, based on a combination of morphological observations and molecular phylogenetic analysis of the internal transcribed spacer (ITS) and nuclear ribosomal large subunit (nrLSU) regions, identified and described four new species: H. aureoluteum sp. nov., H. aureotomentosum sp. nov., H. fuscozonatum sp. nov., and H. pileospinosum sp. nov. For each new species, we provided detailed morphological descriptions, hand-drawn illustrations, and comparisons with closely related taxa. In addition, this study systematically compiled key morphological characteristics and ecological distribution data for all known Hydnellum species in China and constructed a dichotomous identification key. This work provides an important basis for taxonomic research on the genus Hydnellum and enhances our understanding of its ecological distribution patterns in China. Full article

(This article belongs to the Special Issue Taxonomy, Phylogeny, and Ecology of Fungi from Different Ecological Niches)

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15 pages, 1455 KB

Open AccessArticle

Kog1 Represses Lipid Accumulation in Mucor circinelloides: A Transcriptomic Analysis Across Nitrogen Conditions

by Zhen Wang, Ying Gao, Wenrui Dang, Lanlan Zhu and Huaiyuan Zhang

J. Fungi 2026, 12(4), 266; https://doi.org/10.3390/jof12040266 - 7 Apr 2026

Abstract

Oleaginous microorganisms usually accumulate large amounts of lipids under nitrogen limitation and in a carbon-abundant environment. However, how cells sense changes in nitrogen and carbon levels in the culture medium remains a research hotspot. Previous studies have found that the target of rapamycin [...] Read more.

Oleaginous microorganisms usually accumulate large amounts of lipids under nitrogen limitation and in a carbon-abundant environment. However, how cells sense changes in nitrogen and carbon levels in the culture medium remains a research hotspot. Previous studies have found that the target of rapamycin complex 1 (TORC1) plays a core role in lipid accumulation in oleaginous microorganisms. The results of the Kog1 (the member proteins of TORC1) knockout strain constructed earlier by our group showed that the Kog1 negatively regulated lipid accumulation in the oleaginous fungus Mucor circinelloides. In this study, transcriptomic analysis of the knockout and control strains under nitrogen-limited and nitrogen-sufficient culture was carried out to investigate significant differences in lipid accumulation. Kog1 knockout led to a significant decrease in cell dry weight and an increase in lipid content in M. circinelloides. The transcriptomic results showed that genes encoding the glyoxylic acid cycle and genes encoding acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), and Δ9 desaturase in lipid synthesis were upregulated to varying degrees under both conditions, indicating enhanced lipid metabolism that ultimately led to increased lipid accumulation. The knockout of the Kog1 gene also activated the pyruvate–acetaldehyde–acetate metabolic axis and significantly modified the branched-chain amino acid metabolic network, suggesting that Kog1 knockout reprograms the pathway of branched-chain amino acid synthesis and degradation, shifting the carbon flux from amino acid metabolism to acetyl-CoA accumulation. In addition, the gene encoding the SSK1p transcription factor, which participates in the nutrient stress response, was upregulated 41.9- and 51.9-fold in the Kog1 knockout strain compared with the control strain under nitrogen-limited and nitrogen-sufficient conditions, respectively. Full article

(This article belongs to the Section Fungal Cell Biology, Metabolism and Physiology)

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3 pages, 160 KB

Open AccessCorrection

Correction: Kurzbaum et al. Exploring Psilocybe cubensis Strains: Cultivation Techniques, Psychoactive Compounds, Genetics and Research Gaps. J. Fungi 2025, 11, 99

by Eyal Kurzbaum, Tomáš Páleníček, Amiel Sharchaton, Sara Azerrad and Yaron Dekel

J. Fungi 2026, 12(4), 265; https://doi.org/10.3390/jof12040265 - 7 Apr 2026

Abstract

In the original publication [...] Full article

(This article belongs to the Special Issue Current and Future Research Trends on Medicinal Mushrooms, 2nd Edition)

30 pages, 1810 KB

Open AccessArticle

Pathogenicity and Pre-Characterised Putative Effectors of Fusarium oxysporum and F. proliferatum in Garlic (Allium sativum) and Other Allium spp.

by Jessie Rose Harper, Saidi Achari, Tonga Li, Cherie Gambley, Stephen Harper and Victor Galea

J. Fungi 2026, 12(4), 264; https://doi.org/10.3390/jof12040264 - 6 Apr 2026

Abstract

Allium spp. (alliums) are susceptible to rot-diseases caused by pathogenic Fusarium spp., including F. proliferatum (FP) and F. oxysporum (FO), which can cause severe crop losses. A series of pathogenicity tests of four FP isolates from garlic (Allium sativum), four FO [...] Read more.

Allium spp. (alliums) are susceptible to rot-diseases caused by pathogenic Fusarium spp., including F. proliferatum (FP) and F. oxysporum (FO), which can cause severe crop losses. A series of pathogenicity tests of four FP isolates from garlic (Allium sativum), four FO isolates from garlic and three FO isolates from onion (Allium cepa var. cepa) were conducted on garlic seedlings and cloves, onion seedlings and bulbs, and shallot (Allium cepa var. aggregatum) bulbs to determine the virulence of the isolates. A combination of PCRs and whole-genome sequencing (WGS), using ONT long-read technology, was used to identify genes encoding putative effectors. The FP isolates caused moderate to severe symptoms in garlic and contained homologues of SIX2, CRX1 and CRX2, and either SIX9 or SIX13. The FOC ex onion isolates caused severe disease symptoms in all allium species tested, while FO from garlic caused moderate to severe disease in garlic but only mild symptoms in onion and shallot. Fusarium oxysporum f. sp. cepae ex onion potentially contained homologues of SIX3, SIX5, SIX7, SIX9, SIX10, SIX12, SIX14, C5, CRX1 and CRX2. The most pathogenic FO isolate to garlic was Fo_VPRI44630 ex garlic, which contained SIX9, SIX13, C5, CRX1 and CRX2. The difference in virulence and putative effector profiles suggests evidence of host-associated differentiation, and as such, the f. sp. or race designation between FO ex garlic and FO ex onion should be investigated further. This is an important finding for future research into best management practices and breeding for disease resistance to FO and FP in garlic. Full article

(This article belongs to the Section Fungal Pathogenesis and Disease Control)

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41 pages, 4735 KB

Open AccessReview

Natural Products of Freshwater Fungi from a Chemical and Bioactive Perspective

by Xiao-Jie Chen, Guo-Jun Zhou, Qian-Hong Yan, Xi Tan, Feng Xu, Fu-Ji Qian, Xu-Hang Fan, Bei Jiang, Cai-Juan Zheng, Hai-Feng Li and Kai-Ling Wang

J. Fungi 2026, 12(4), 263; https://doi.org/10.3390/jof12040263 - 3 Apr 2026

Abstract

Freshwater fungi have attracted considerable attention as a potential source of lead compounds with diverse and novel chemical structures and biological activities in drug discovery. This review summarizes 307 natural products of freshwater fungi from 1988 to the end of October 2025. These [...] Read more.

Freshwater fungi have attracted considerable attention as a potential source of lead compounds with diverse and novel chemical structures and biological activities in drug discovery. This review summarizes 307 natural products of freshwater fungi from 1988 to the end of October 2025. These compounds are categorized into fourteen structural types, including fatty acids and their lactones (compounds 1–18), furans and furanones (compounds 19–31), pyrans and pyranones (compounds 32–109), benzoquinones, phenols and phenolic acids (compounds 110–141), naphthalenes and naphthalenones (compounds 142–192), authraquinones and xanthones (compounds 193–211, depsidones (compounds 212–217), macrolides (compounds 218–234), polyesters (compounds 235–237), alkaloids (compounds 238–251), peptides (compounds 252–280), terpenoids (compounds 281–300), steroids (compounds 301 and 302), and other compounds (compounds 303–307). Some of them displayed promising biological activity, mainly comprising antibacterial, cytotoxic, and nematicidal activities. The preliminary analysis of the Structure––Activity Relationship (SAR) of important compounds is also discussed. In the last section, current challenges and prospective research perspectives are briefly proposed based on opinions from previous reviews. This review would contribute to the understanding of the utilization and development of natural products derived from freshwater fungi as potent medical resources in the future. Full article

(This article belongs to the Section Fungi in Agriculture and Biotechnology)

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14 pages, 1242 KB

Open AccessArticle

Antibacterial Activity of Guttation Droplets from Penicillium pimiteouiense and Penicillium menonorum Against Clinically Relevant Bacterial Pathogens

by Carlos Eduardo Barajas-Saucedo, Mariana Torres-Cruz, Juan Carlos Sánchez-Rangel, Abraham Vidal-Limon and Juana María Jiménez-Vargas

J. Fungi 2026, 12(4), 262; https://doi.org/10.3390/jof12040262 - 3 Apr 2026

Abstract

Antimicrobial resistance (AMR) represents a major global health challenge, driving the search for novel antimicrobial compounds from natural sources. Filamentous fungi are prolific producers of bioactive metabolites, yet the biological potential of fungal guttation droplets remains relatively underexplored. In this study, guttation droplets [...] Read more.

Antimicrobial resistance (AMR) represents a major global health challenge, driving the search for novel antimicrobial compounds from natural sources. Filamentous fungi are prolific producers of bioactive metabolites, yet the biological potential of fungal guttation droplets remains relatively underexplored. In this study, guttation droplets produced by Penicillium pimiteouiense and Penicillium menonorum, isolated from rhizospheric soils of Opuntia spp. in Colima, Mexico, were evaluated for antibacterial activity against clinically relevant pathogens. Fungal isolates were identified through macromorphological characterization and sequence analysis of ITS and β-tubulin genes. Antibacterial activity of the guttation droplets was evaluated using agar well diffusion and microbroth dilution assays to determine inhibition zones, minimum inhibitory volume (MIV), and minimum bactericidal volume (MBV). The exudates exhibited measurable activity against several Gram-negative and Gram-positive bacteria, including Escherichia coli, Salmonella enterica, Klebsiella pneumoniae, Serratia marcescens, and Staphylococcus aureus. Guttation droplets from P. pimiteouiense showed the highest inhibition, with zones up to 24.4 mm against S. enterica, and activity comparable to gentamicin. MBV/MIV ratios indicated bactericidal activity against selected pathogens, including E. coli, K. pneumoniae, and S. aureus. These findings demonstrate that fungal guttation droplets represent a promising and underexplored source of antibacterial compounds and support their potential for antimicrobial discovery. Full article

(This article belongs to the Special Issue Ascomycete Fungi: From Biosystematics and Biodiversity to Biotechnology)

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19 pages, 32171 KB

Open AccessArticle

Effects of Cultivation Substrate Differences on Quality Formation and Polysaccharide Composition Characteristics of Tremella fuciformis

by Jianqiu Chen, Yating Deng, Yujie Chen, Keming Zhu, Xun Yao, Shenqiao Yang, Liding Chen and Shujing Sun

J. Fungi 2026, 12(4), 261; https://doi.org/10.3390/jof12040261 - 3 Apr 2026

Abstract

Cultivation substrate critically affects the quality of Tremella fuciformis. Five substrates, including cottonseed hulls (MZKs), Machilus pauhoi Kanehira sawdust (BNM), lotus seed hulls (LZKs), Corethrodendron scoparium sawdust (HB), and palm fiber (ZL), were evaluated for their effects on agronomic traits, nutritional composition, [...] Read more.

Cultivation substrate critically affects the quality of Tremella fuciformis. Five substrates, including cottonseed hulls (MZKs), Machilus pauhoi Kanehira sawdust (BNM), lotus seed hulls (LZKs), Corethrodendron scoparium sawdust (HB), and palm fiber (ZL), were evaluated for their effects on agronomic traits, nutritional composition, texture, and taste characteristics. Untargeted metabolomics was applied to elucidate substrate-associated metabolic variations, and polysaccharide monosaccharide composition was quantitatively analyzed. The results showed that the BNM group exhibited the highest fresh weight, whereas the LZK group presented the highest dry weight and crude polysaccharide content. The ZL group displayed the greatest ear piece thickness and fruiting body elevation. Higher protein contents were observed in the ZL and LZK groups, with no differences in crude fiber content. Texture analysis indicated that hardness was highest in the LZK group, whereas the MZK group showed better springiness, cohesiveness, and chewiness. Regarding taste characteristics, the MZK group exhibited the strongest sweetness, the LZK group showed a markedly higher bitterness, and umami levels were comparable across all groups. Metabolomic analysis revealed that substrate-induced variations in amino acids, saccharides, and taste-related metabolites were significantly associated with nutritional quality and taste attributes of T. fuciformis. Polysaccharides of fruiting bodies cultivated on the five substrates consisted of six monosaccharides, with composition ratios similar to those of spore extracellular polysaccharides; among them, differences in glucuronic acid (GlcA) proportion represented a key indicator distinguishing fruiting body polysaccharides from spore polysaccharides. This study revealed the metabolic basis and polysaccharide composition underlying substrate-dependent quality of T. fuciformis, supporting substrate optimization for high-quality production. Full article

(This article belongs to the Special Issue Research Progress on Edible Fungi)

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29 pages, 3513 KB

Open AccessArticle

Spatiotemporal Dynamics and Co-Occurrence Patterns of Marine Fungal Communities Along Nutrient Gradients in the Leizhou Peninsula, China

by Yingyi Fan, Menghan Gao, Bihong Liu, Junyu Wei, Jianming Li and Zhangxi Hu

J. Fungi 2026, 12(4), 260; https://doi.org/10.3390/jof12040260 - 3 Apr 2026

Abstract

Marine fungi are pivotal components of coastal ecosystems, facilitating essential biogeochemical cycling and trophic dynamics. However, the complex mechanisms governing their spatiotemporal community patterns in tropical–subtropical coasts remain largely unexplored. In this study, we characterized marine fungal diversity across a comprehensive seasonal cycle [...] Read more.

Marine fungi are pivotal components of coastal ecosystems, facilitating essential biogeochemical cycling and trophic dynamics. However, the complex mechanisms governing their spatiotemporal community patterns in tropical–subtropical coasts remain largely unexplored. In this study, we characterized marine fungal diversity across a comprehensive seasonal cycle (spring (March), summer (June), autumn (August), and winter (December)) at 21 representative sites along the Leizhou Peninsula, China. These sites were strategically selected to encompass a broad range of dissolved inorganic nitrogen (DIN) gradients. Fungal community composition was characterized via high-throughput sequencing of the internal transcribed spacer 2 (ITS2) region, followed by functional guild profiling using the FUNGuild database. A total of 8777 amplicon sequence variants (ASVs) were identified, encompassing a broad taxonomic breadth of 10 phyla and 358 genera. Ascomycota, Basidiomycota, and Chytridiomycota emerged as the predominant phyla across all samples. Our results revealed significant spatiotemporal heterogeneities: seasonal succession fundamentally reshaped community composition, with DIN exerting its most pronounced influence during the winter. Furthermore, fungal functional structures exhibited distinctive clustering across regions defined by DIN enrichment levels. Co-occurrence network analysis revealed a highly modular and robust architecture, characterized by predominantly positive interactions and dense inter-taxon connectivity. These findings underscore the synergistic influence of temporal dynamics and DIN enrichment in shaping marine fungal community assembly and functional compositions. Our study provides critical baseline insights into the ecological resilience of coastal mycobiota in the South China Sea. Full article

(This article belongs to the Special Issue Emerging Investigators in Marine Fungi)

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21 pages, 1116 KB

Open AccessReview

Biomarkers in the Diagnosis of Aspergillosis

by Laura Rivera-Agudelo, Diego H. Cáceres, Julián C. Arango-Rincón, Julio C. Jaramillo-Alzate, Alejandra Zuluaga-Rodriguez and Álvaro L. Rúa-Giraldo

J. Fungi 2026, 12(4), 259; https://doi.org/10.3390/jof12040259 - 3 Apr 2026

Abstract

Aspergillosis is a group of different invasive and non-invasive diseases affecting the lung and other organs, caused by species of the genus Aspergillus. Additionally, complications associated with treatment and the increasing emergence of antifungal-resistant strains of Aspergillus spp. are high-impact problems. For [...] Read more.

Aspergillosis is a group of different invasive and non-invasive diseases affecting the lung and other organs, caused by species of the genus Aspergillus. Additionally, complications associated with treatment and the increasing emergence of antifungal-resistant strains of Aspergillus spp. are high-impact problems. For these reasons, a combined approach of diagnostic tests is necessary to reach an accurate and timely diagnosis. This review aims to describe some biomarkers and their usefulness for the diagnosis of aspergillosis. Among the findings obtained in different studies, the nature, analytical performance and usefulness for diagnosis in different clinical forms of this disease are described. Currently, the main biomarkers used in the diagnosis of Aspergillus disease fall into three categories: membrane components, specific DNA sequences and metabolic products. The detection of biomarkers is one of the most important innovations of recent decades in the field of medical mycology, as it is a diagnostic aid that allows the timely detection of infections and decreases the time of administration of antifungal therapy. Full article

(This article belongs to the Special Issue Invasive Fungal Disease: Advances and Challenges in Diagnosis and Management from a Global Perspective)

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19 pages, 6684 KB

Open AccessArticle

Metabolic Engineering of Rhodotorula toruloides for Biosynthesis of Retinal

by Huihui Qiu, Linyue Tian, Lin Hu, Lianwu Wu, Yu Huang, Ran Ge, Yufan Xing, Alexander A. Kamnev, Ning He and Mingfeng Cao

J. Fungi 2026, 12(4), 258; https://doi.org/10.3390/jof12040258 - 2 Apr 2026

Abstract

Rapid advancements in biotechnology have enabled biomanufacturing to emerge as a feasible approach for industrial chemical production. By harnessing synthetic biology and metabolic engineering, engineered microbial cell factories can convert renewable resources into valuable chemicals, providing a sustainable alternative to traditional chemical methods. [...] Read more.

Rapid advancements in biotechnology have enabled biomanufacturing to emerge as a feasible approach for industrial chemical production. By harnessing synthetic biology and metabolic engineering, engineered microbial cell factories can convert renewable resources into valuable chemicals, providing a sustainable alternative to traditional chemical methods. This study focuses on the microbial production of retinal, an important retinoid used in pharmaceuticals, food, and cosmetics. The oleaginous yeast Rhodotorula toruloides NP11 was genetically modified to synthesize retinal by incorporating and optimizing three β-carotene 15,15′-dioxygenase genes from various sources. Several genetic modifications were made to enhance retinal yield, including the overexpression of isopentenyl-diphosphate isomerase (IDI1), geranylgeranyl diphosphate synthase (BTS1), phytoene synthase (CARRP), and phytoene dehydrogenase (CARB), which led to increased β-carotene levels and boosted retinal production. Furthermore, fermentation conditions such as temperature, antioxidants, and extractants were fine-tuned. The engineered strain Rt13 ultimately achieved a maximum retinal concentration of 20.38 mg/L through fed-batch fermentation. This study highlights the potential of R. toruloides as a cell factory for terpenoid biosynthesis, providing valuable insights for future metabolic engineering endeavors. Full article

(This article belongs to the Special Issue Synthetic Biology and Metabolic Engineering of Yeast)

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16 pages, 19588 KB

Open AccessArticle

Biocontrol Potential and Functional Characteristics of Bacillus sonorensis A-5 Against Watermelon Fusarium Wilt

by Jian-Wei Jiang, Yue Qiu, Liu-Tong Ye, Jing-Xue Luo, Qianwen Nie and Yi Zhou

J. Fungi 2026, 12(4), 257; https://doi.org/10.3390/jof12040257 - 2 Apr 2026

Abstract

Fusarium wilt, caused by Fusarium oxysporum f. sp. niveum (Fon), severely restricts the sustainable development of the global watermelon industry. While conventional chemical fungicides of this disease have triggered prominent ecological issues, Bacillus-based microbial biocontrol, which combines inherent environmental compatibility [...] Read more.

Fusarium wilt, caused by Fusarium oxysporum f. sp. niveum (Fon), severely restricts the sustainable development of the global watermelon industry. While conventional chemical fungicides of this disease have triggered prominent ecological issues, Bacillus-based microbial biocontrol, which combines inherent environmental compatibility with stable control efficacy, has emerged as a key green alternative to chemical management. However, the biocontrol potential of Bacillus sonorensis against this disease has not yet been fully investigated. In this study, we isolated 56 bacterial strains from healthy watermelon rhizosphere soil, and obtained a Fon-antagonistic strain A-5 with the strongest activity (70.15% mycelial inhibition rate), which was identified as B. sonorensis via polyphasic taxonomic analysis. In vitro assays showed that the sterile fermentation filtrate of strain A-5 had a maximum 81.05% inhibition rate against Fon, and its volatile organic compounds also significantly suppressed Fon growth, with broad-spectrum antifungal activity against four common phytopathogenic fungi. Functional tests confirmed that strain A-5 could secrete cell wall-degrading enzymes, produce siderophores and synthesize indole-3-acetic acid, and 17 antimicrobial secondary metabolite biosynthetic gene clusters were identified in its genome. Pot experiments verified that strain A-5 had a 78.04% relative control efficacy against watermelon Fusarium wilt, which significantly reduced seedling disease incidence and upregulated defense-related antioxidant enzyme activities in watermelon leaves. In general, B. sonorensis A-5 is a promising novel biocontrol agent for green management of watermelon Fusarium wilt. Full article

(This article belongs to the Section Fungal Pathogenesis and Disease Control)

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18 pages, 1994 KB

Open AccessArticle

MgO and Zn/MgO Nanoparticles as Direct Antifungal Compounds and Elicitors Against Sclerotinia sclerotiorum in Pea

by Hanaa Omar, Ahmed Mohamed, Nehad El-Gammal, Warda Hussain, Saleh Alhewairini, Mahmoud Abdelfatah, Abdelhamed Elshaer, Reda Omara, Ayman Omar and Aly Derbalah

J. Fungi 2026, 12(4), 256; https://doi.org/10.3390/jof12040256 - 1 Apr 2026

Abstract

The purpose of this study was to assess the effectiveness of two nanostructures (MgO and Zn/MgO) against Sclerotinia sclerotiorum, which causes white mold disease in peas, as direct antifungal agents or resistance inducers in pea plants. The direct antifungal activity of these [...] Read more.

The purpose of this study was to assess the effectiveness of two nanostructures (MgO and Zn/MgO) against Sclerotinia sclerotiorum, which causes white mold disease in peas, as direct antifungal agents or resistance inducers in pea plants. The direct antifungal activity of these nanostructures was evaluated by assessing their ability to inhibit S. sclerotiorum growth in vitro and reduce white mold severity in the greenhouse. The induction of resistance in pea plants was examined by assessing the expression of three defense-related genes using quantitative real-time PCR and measuring the phenolic compounds content in treated pea plants relative to untreated controls. The effect of the tested control agents on the growth and yield of pea plants was investigated. In comparison to the untreated control, S. sclerotiorum growth was markedly suppressed following treatment with the investigated compounds. The complete suppression (100%) of S. sclerotiorum growth was achieved with concentration levels of 100 mg/L for both MgO and Zn/MgO nanostructures. In greenhouse conditions, pea plants treated with the investigated chemicals showed a considerable reduction in the severity of white mold disease when compared to the untreated control plants. The transcript levels of 12-oxophytodienoate reductase 11 (OPR1), antioxidant peroxide (PsOXII), and chlorophyll a-b binding protein genes increased significantly in treated plants with MgO (3.1, 2.7, and 3.5-fold), fungicide (3.2, 2.8, and 2.8-fold), and Zn/MgO (3.5, 3, and 5-fold) compared to control, respectively. Pea plants treated with the tested nanoparticles generated more phenolic content than untreated controls. The application of fungicide and tested nanoparticles to peas greatly enhanced their growth properties. In light of our results, the application of these nanoparticles may represent a novel approach for controlling this pathogen. Full article

(This article belongs to the Special Issue Plant Pathogenic Fungal Infections, Biocontrol and Novel Fungicides)

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10 pages, 2333 KB

Open AccessCommunication

Agrobacterium-Mediated Genetic Transformation of the Edible and Medicinal Cauliflower Mushroom Sparassis latifolia

by Wen Cao, Xinyu Zhou, Ruiheng Yang, Yingying Wu, Yan Li, Chenli Zhou, Jianing Wan, Rongping Li, Xiangying Luo, Zhenhui Shen, Dapeng Bao, Lihua Tang and Junjun Shang

J. Fungi 2026, 12(4), 255; https://doi.org/10.3390/jof12040255 - 1 Apr 2026

Abstract

Sparassis latifolia is an edible and medicinal mushroom with significant economic value, now commercially cultivated on a large scale in China. However, current cultivars face challenges, including an extended mycelial growth period and unstable fruiting body yields. Advances in molecular breeding and functional [...] Read more.

Sparassis latifolia is an edible and medicinal mushroom with significant economic value, now commercially cultivated on a large scale in China. However, current cultivars face challenges, including an extended mycelial growth period and unstable fruiting body yields. Advances in molecular breeding and functional genomics for this species are hindered by the absence of a reliable genetic transformation system. In this study, we first determined that S. latifolia is highly sensitive to carboxin and hygromycin, two selective agents commonly used in fungal genetics. We subsequently constructed a novel binary vector, pCbxHyg, harboring a carboxin resistance cassette driven by its native Pleurotus eryngii promoter and a hygromycin resistance cassette under the control of the P. eryngii Glycerol 3-phosphate dehydrogenase (GPD) gene promoter. Initial transformation attempts using Agrobacterium-mediated transformation of liquid-cultured mycelial pellets were unsuccessful. During microscopic examination, we discovered that S. latifolia mycelia produce abundant asexual chlamydospores. Using these chlamydospores as recipient material, we efficiently and reproducibly obtained transformants with the pCbxHyg vector under both carboxin and hygromycin selection. This method highlights the advantage of using asexual spores of Basidiomycetes as recipients for genetic transformation. PCR analysis confirmed the stable integration of the exogenous resistance genes into the fungal genome. The functionality of the system was further validated by transforming chlamydospores with a vector carrying a β-glucuronidase (GUS) reporter gene, whose expression was confirmed via histochemical staining of the resulting transformant mycelia. This work establishes the first successful Agrobacterium-mediated genetic transformation system for S. latifolia, providing a foundational platform for future gene function studies and molecular breeding efforts. Full article

(This article belongs to the Section Fungal Genomics, Genetics and Molecular Biology)

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16 pages, 554 KB

Open AccessArticle

Targeted Screening to Predict Magnusiomyces Infections in Hematopoietic Cell Transplant Recipients: Evidence from an Outbreak Setting

by Claudia Bartalucci, Chiara Russo, Anna Maria Raiola, Massimiliano Gambella, Vincenzo Di Pilato, Paola Morici, Elena De Carolis, Bram Spruijtenburg, Eelco F. J. Meijer, Monica Melchio, Elisa Balletto, Anna Marchese, Emanuele Angelucci, Matteo Bassetti and Malgorzata Mikulska

J. Fungi 2026, 12(4), 254; https://doi.org/10.3390/jof12040254 - 1 Apr 2026

Abstract

Invasive infections caused by Magnusiomyces spp. are rare, but are associated with severe complications in hematopoietic cell transplantation (HCT) recipients and hospital outbreaks. Following a Magnusiomyces clavatus outbreak in our HCT unit, a prospective targeted screening protocol was implemented, which included pharyngeal and [...] Read more.

Invasive infections caused by Magnusiomyces spp. are rare, but are associated with severe complications in hematopoietic cell transplantation (HCT) recipients and hospital outbreaks. Following a Magnusiomyces clavatus outbreak in our HCT unit, a prospective targeted screening protocol was implemented, which included pharyngeal and rectal swabs cultured on yeast-selective media with prolonged incubation. Clinical and microbiological data were analyzed, and whole-genome sequencing (WGS) was performed on the available isolates. During the study period (September 2022–July 2023), five colonizations and five invasive breakthrough Magnusiomyces infections were identified. Despite prompt initiation of antifungal treatment, 4/5 patients (80%) died. WGS demonstrated clonal relatedness among four M. clavatus isolates, supporting clonal transmission, although no environmental sources were identified. An enhanced two-phase screening strategy involving 71 patients showed limited benefit, identifying only one additional colonization case compared to routine surveillance cultures. A retrospective review (2007–2021) identified 58 Magnusiomyces spp. episodes, with only 10% occurring in patients with hematological malignancies. Our study describes a prolonged clonal outbreak confined to an HCT unit and provides a detailed evaluation of a targeted screening approach in this setting, highlighting the challenges of early identification and prediction of invasive infections. Further studies are needed to define the optimal surveillance and prevention strategies. Full article

(This article belongs to the Section Fungal Pathogenesis and Disease Control)

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15 pages, 2352 KB

Open AccessArticle

Characterization of the Mycoparasite Cladosporium oxysporum and Its Interaction with the Poplar Rust Pathogen Melampsora larici-populina

by Penghua Chen, Kuocheng Shen, Yadan Zhang, Zhongdong Yu and Qiangfeng Li

J. Fungi 2026, 12(4), 253; https://doi.org/10.3390/jof12040253 - 1 Apr 2026

Abstract

Poplar rust, caused by Melampsora larici-populina, remains a primary biotic constraint limiting poplar growth. In this study, we characterized the diversity and antimicrobial potential of fungi associated with the aeciospore and urediniospore pustules of M. larici-populina. We isolated and identified 502 [...] Read more.

Poplar rust, caused by Melampsora larici-populina, remains a primary biotic constraint limiting poplar growth. In this study, we characterized the diversity and antimicrobial potential of fungi associated with the aeciospore and urediniospore pustules of M. larici-populina. We isolated and identified 502 fungal strains, spanning 16 families, 22 genera, and 47 species. Comparative analysis revealed that the urediniospore stage harbored a greater abundance of fungal isolates but exhibited lower overall diversity, whereas the aeciospore stage was characterized by greater species richness despite lower individual strain counts. Notably, Cladosporium oxysporum was identified as the predominant species across both life stages, maintaining a consistent isolation frequency. We demonstrated that C. oxysporum exhibits strong antifungal activity; its metabolites contain bioactive components capable of degrading the cell walls of M. larici-populina urediniospores, resulting in an inhibition rate of 78.59%. Mechanistic observations via electron microscopy revealed that C. oxysporum develops appressoria to penetrate the urediniospore cell wall and subsequently forms haustoria-like structures within the protoplast to facilitate nutrient acquisition. This hyperparasitic interaction ultimately leads to the death of the rust pathogen. Collectively, our results identify C. oxysporum as a mycoparasite of M. larici-populina and highlight its potential as a promising biological control agent for the management of poplar rust. Full article

(This article belongs to the Section Fungal Pathogenesis and Disease Control)

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22 pages, 1463 KB

Open AccessArticle

An Efficient Workflow for Fungal Nucleic Acid Detection in Sputum

by Ying Han, Deding Tang, Yuntong Gao, Lilian Ji, Jie Hu and Yan Gong

J. Fungi 2026, 12(4), 252; https://doi.org/10.3390/jof12040252 - 31 Mar 2026

Abstract

Respiratory fungal infections are challenging to diagnose due to non-specific symptoms and limitations of conventional methods. This study established an integrated, rapid workflow for detecting three common pathogenic fungi, combining an optimized nucleic acid extraction method with a TaqMan-based multiplex real-time qPCR system. [...] Read more.

Respiratory fungal infections are challenging to diagnose due to non-specific symptoms and limitations of conventional methods. This study established an integrated, rapid workflow for detecting three common pathogenic fungi, combining an optimized nucleic acid extraction method with a TaqMan-based multiplex real-time qPCR system. The high-salt lysis method yielded high-purity nucleic acid extracts, with performance comparable to or better than commercial kits. The multiplex qPCR assay was optimized for high specificity, sensitivity, and reproducibility, with detection limits as low as 10 CFU/mL for some fungi. Clinical validation using 14 sputum samples demonstrated high agreement with conventional culture, and the assay detected a low-load infection missed by culture. This integrated approach shows great potential for rapid, sensitive, and specific clinical detection of respiratory fungal infections. Full article

(This article belongs to the Section Fungal Pathogenesis and Disease Control)

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12 pages, 1729 KB

Open AccessArticle

Engineered Saccharomyces cerevisiae for High-Yield and Sustainable Production of α-Bisabolol via Combinatorial Genomic Integration and Pathway Amplification

by Zichen Wu, Baofeng Wan, Chun Li, Shen Zhou, Sishu Huang, Boyang Zhi, Congping Xu, Qin Cheng, Chuansong Zhan and Jie Luo

J. Fungi 2026, 12(4), 251; https://doi.org/10.3390/jof12040251 - 31 Mar 2026

Abstract

α-Bisabolol is naturally occurring in many plants and has great potential in health products and pharmaceuticals. However, the current extraction method from natural plants is unsustainable and cannot fulfil the increasing requirement. This study aimed to develop a sustainable strategy to enhance the [...] Read more.

α-Bisabolol is naturally occurring in many plants and has great potential in health products and pharmaceuticals. However, the current extraction method from natural plants is unsustainable and cannot fulfil the increasing requirement. This study aimed to develop a sustainable strategy to enhance the biosynthesis of α-bisabolol by metabolic engineering. Integration of ERG20 (encoding farnesyl diphosphate synthase) and tHMG1 (encoding truncated 3-hydroxy-3-methylglutaryl-CoA reductase) genes with a constitutive strong promoter into the yeast genome elevated α-bisabolol production from 20.21 mg/L to 98.30 mg/L, representing a 4.86-fold increase. Further optimization of the mevalonate pathway and amplification of ERG20, tHMG1, and OsTPS1 copy numbers enhanced α-bisabolol synthesis to 423.01 mg/L, achieving a 20.93-fold improvement relative to the baseline. This work establishes a reference strategy for high-yield α-bisabolol biosynthesis in engineered yeast. Full article

(This article belongs to the Special Issue Yeast as a Platform for Synthetic Biology)

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21 pages, 2175 KB

Open AccessArticle

Valorization of Spent Brewer’s Yeast by Pulsed Electric Field Treatment Combined with Enzymatic Hydrolysis

by Valentina Ganeva and Boyana Angelova

J. Fungi 2026, 12(4), 250; https://doi.org/10.3390/jof12040250 - 31 Mar 2026

Abstract

Spent brewer’s yeast, a major by-product of the brewing industry, is a valuable source of bioactive compounds. The main technological limitation for their recovery is the rigid yeast cell wall, while the high nucleic acid content may restrict the direct use of yeast-derived [...] Read more.

Spent brewer’s yeast, a major by-product of the brewing industry, is a valuable source of bioactive compounds. The main technological limitation for their recovery is the rigid yeast cell wall, while the high nucleic acid content may restrict the direct use of yeast-derived extracts for human nutrition. In this study, pulsed electric field (PEF) treatment, applied alone or in combination with enzymatic hydrolysis, was investigated for the production of yeast-derived extracts with different compositions. PEF treatment performed in continuous-flow mode resulted in more than 98% of cells with irreversibly permeabilized membranes and enabled the rapid and selective release of low-molecular-weight intracellular compounds during subsequent incubation of the cells in water. Within 4 h, approximately 61% of total antioxidant activity, 65% of glutathione, and around 80% of free α-amino nitrogen and B-group vitamins were recovered at different rates, while the aqueous extracts were characterized by low purine nucleotide content. Electropermeabilized cells exhibited high sensitivity to enzymatic hydrolysis. After 6 h of incubation with 0.2% (v/v) Alcalase, the obtained hydrolysates contained 254 ± 17 mg/g DCW of protein, mostly in the form of peptides, 148.2 ± 17.3 mg/g DCW of free α-amino nitrogen, and a total phenolic content of 16.7 ± 1.9 mg GAE/g DCW. The maximal antioxidant activity (62.7 ± 9.3 mg TE/g DCW) was reached after 4 h of incubation, corresponding to a 2.7-fold increase compared with cell lysates. Overall, PEF treatment, applied alone or in combination with enzymatic hydrolysis, provides an efficient and mild approach for the production of yeast-derived extracts with tailored compositions and potential applications in the food, pharmaceutical, and cosmetic industries. Full article

(This article belongs to the Section Fungi in Agriculture and Biotechnology)

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21 pages, 2219 KB

Open AccessArticle

Chemical and Biological Investigation of the Endophytic Aspergillus terreus (SU5) Inhabiting Date Fruits (Phoenix dactylifera)

by Ahmed Abdel-Hadi, Mohammed Alaidarous, Abdulrahman Alatiq, Yahya Madkhali, Saeed Banawas, Mohamed Abouelela and Abdallah Hassane

J. Fungi 2026, 12(4), 249; https://doi.org/10.3390/jof12040249 - 30 Mar 2026

Abstract

Endophytic fungi associated with date fruits (Phoenix dactylifera) are mostly under-explored, despite their potential as reservoirs of natural compounds. The aims of this study were to characterize the endophytic fungus Aspergillus terreus (SU5) isolated from date fruits, and to investigate its [...] Read more.

Endophytic fungi associated with date fruits (Phoenix dactylifera) are mostly under-explored, despite their potential as reservoirs of natural compounds. The aims of this study were to characterize the endophytic fungus Aspergillus terreus (SU5) isolated from date fruits, and to investigate its biological activities and chemical profile for the first time. Morphological and molecular methods were utilized to identify Aspergillus terreus. A liquid chromatography–mass spectrometry analysis (LC/MS/MS) was conducted to determine the chemical profile of the crude extract. Biological properties were investigated through acetylcholine esterase and butyrylcholine esterase inhibition, cytotoxicity assays against MCF-7 and MCF-7/Adr, and antioxidant assays. LC/MS/MS of the fungal extract resulted in the detection of 39 of established secondary metabolites, primarily comprising polyketides, quinones, and phenolic derivatives. The crude extract demonstrated significant antioxidant activity, especially in the ABTS assay (IC₅₀ = 50.18 μg/mL), considerable cytotoxicity against MCF-7 breast cancer cells, diminished efficacy against the drug-resistant MCF-7/Adr cell line, and preferential inhibition of butyrylcholinesterase compared to acetylcholinesterase. While none of the identified compounds are novel, numerous metabolites are documented here for the first time from an endophytic A. terreus associated with date fruits. The findings underscore date fruits as a prospective ecological niche for a chemically varied endophytic fungus with potential pharmaceutical significance. Full article

(This article belongs to the Special Issue Bioactive Secondary Metabolites from Fungi)

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