Journal of Fungi

48 pages, 2975 KB

Open AccessReview

Pulcherrimin and Beyond: The Multifaceted Role of Metschnikowia pulcherrima in Postharvest Disease Management—A Scoping Review

by Juliana Pereira Rodrigues Belas, Caroline Corrêa de Souza Coelho, Leda Maria Fortes Gottschalk, Elisa d’Avila Costa Cavalcanti, Denise Maria Guimarães Freire and Otniel Freitas Silva

J. Fungi 2026, 12(4), 298; https://doi.org/10.3390/jof12040298 - 21 Apr 2026

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Abstract

Postharvest losses of fruits and vegetables are a global problem that directly affect food security, the economy, and the environment. These losses are mainly associated with fungal diseases during storage. Due to the limitations of synthetic fungicides, including the development of resistance and [...] Read more.

Postharvest losses of fruits and vegetables are a global problem that directly affect food security, the economy, and the environment. These losses are mainly associated with fungal diseases during storage. Due to the limitations of synthetic fungicides, including the development of resistance and risks to human health, there is growing interest in sustainable disease control strategies. This scoping review analyzes the potential of the yeast Metschnikowia pulcherrima as a biocontrol agent for postharvest phytopathogens, based on the scientific literature published between 2014 and 2026. The reviewed studies identify several antagonistic mechanisms, including competition for nutrients and space, the production of organic volatile compounds, hydrolytic enzyme activity, biofilm formation, and the induction of resistance in fruits. In vitro and in vivo assays show that M. pulcherrima effectively reduces postharvest disease incidence and severity caused by certain fungi. Furthermore, its synergistic effect when combined with emerging technologies is notable. The results highlight its potential as a sustainable alternative to synthetic fungicides, although further studies are needed for large-scale commercial application. Full article

(This article belongs to the Special Issue Postharvest Fungi: Control of Fungal Diseases in Fruit and Vegetables)

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8 pages, 213 KB

Open AccessCommunication

Feasibility of Allogeneic Hematopoietic Stem Cell Transplantation Following Recent Invasive Mold Disease in Pediatric Patients

by Kevin Martínez Pérez, Daniela Torres-Hernández, Nathalia Sanclemente, Oscar Ramirez, Andrés Portilla, Jorge Buitrago and Eduardo López-Medina

J. Fungi 2026, 12(4), 297; https://doi.org/10.3390/jof12040297 - 21 Apr 2026

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Abstract

A history of invasive mold disease (IMD) often delays or contraindicates allogeneic hematopoietic stem cell transplantation (allo-HSCT) in children. Given the limited data on pediatric patients with pre-allo-HSCT IMD, we aimed to describe the management and clinical outcomes of a cohort of children [...] Read more.

A history of invasive mold disease (IMD) often delays or contraindicates allogeneic hematopoietic stem cell transplantation (allo-HSCT) in children. Given the limited data on pediatric patients with pre-allo-HSCT IMD, we aimed to describe the management and clinical outcomes of a cohort of children with IMD prior to allo-HSCT through day +100 post-transplantation. Between 2021 and 2024, ten pediatric patients were identified with proven or probable IMD. Their median age was 8.5 years. The most common pathogens were Aspergillus (n = 5) and Fusarium (n = 4). Infections most frequently involved the lungs followed by paranasal sinuses, bloodstream, liver, and skin. All patients demonstrated clinical improvement before transplantation, and by day +100 post-HSCT, no IMD relapses or infection-related mortality were observed. These findings suggest that complete radiologic or clinical resolution is not a prerequisite for proceeding with transplantation. Recent IMD should not be considered an absolute contraindication to urgent allo-HSCT when clinical improvement is evident, as transplantation facilitates immune reconstitution necessary for definitive infection control. Full article

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

14 pages, 942 KB

Open AccessArticle

Novel Antifungal Pyridine Alkaloids from Endophytic Fungus Penicillium citrinum VDL118

by Mei Yang, Shan Hu, Zhi-Yu Zhang, Fa-Zhong Yang, Xiao-Qin Yang, Si-Da Xie, Ying-Jun Zhang, Ping Zhao and Guo-Lei Zhu

J. Fungi 2026, 12(4), 296; https://doi.org/10.3390/jof12040296 - 20 Apr 2026

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Abstract

Three novel alkaloids, penicitrioids A–C (1–3), and two known compounds (4–5) were isolated from the ethyl acetate (EtOAc) extract of the solid fermentation of Penicillium citrinum VDL118, an endophytic fungus harbored in the leaves of [...] Read more.

Three novel alkaloids, penicitrioids A–C (1–3), and two known compounds (4–5) were isolated from the ethyl acetate (EtOAc) extract of the solid fermentation of Penicillium citrinum VDL118, an endophytic fungus harbored in the leaves of Vaccinium dunalianum Wight (Ericaceae), a perennial evergreen shrub native to the southwestern regions of China, Myanmar, and Vietnam. Compounds 1 and 2 are novel pyridine alkaloids characterized by an unprecedented dihydrofuro[3,4-c]pyridine core, while 3 features a distinct pyrrolo[3,4-c]pyridine framework. Their structures were unambiguously established by comprehensive spectroscopic analysis and electronic circular dichroism (ECD) calculations. In vitro antifungal assays revealed that compounds 1–5 exhibited moderate to potent inhibitory effects against five tested phytopathogenic fungi, with minimum inhibitory concentrations (MICs) ranging from 3.1 to 100 μg/mL. Notably, four of them (1–4) displayed broad-spectrum and potent activity against Gloeophyllum trabeum, Coriolus versicolor, Fusarium solani, and Botrytis cinerea, with MIC values as low as 3.1–12.5 μg/mL. Furthermore, a plausible biosynthetic pathway for compounds 1–3 was proposed. Full article

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

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19 pages, 1968 KB

Open AccessArticle

Bioremediation Potential of Leaf Endophytic Fungi in Allium ampeloprasum and Brassica oleracea var. capitata

by Dayani Pavalakumar, Sagarika Kannangara, Nadeema Dharmasiri, Chamani Amarasekara, Lanka Undugoda, Kasun M. Thambugala, Jayantha Munasinghe, Sukanya Haituk and Ratchadawan Cheewangkoon

J. Fungi 2026, 12(4), 295; https://doi.org/10.3390/jof12040295 - 20 Apr 2026

Viewed by 475

Abstract

Polycyclic aromatic hydrocarbons (PAHs) are toxic air pollutants mainly released through vehicular emissions and can accumulate on edible plants, posing health risks to humans. This study aimed to isolate and identify endophytic fungi from Allium ampeloprasum and Brassica oleracea var. capitata, which [...] Read more.

Polycyclic aromatic hydrocarbons (PAHs) are toxic air pollutants mainly released through vehicular emissions and can accumulate on edible plants, posing health risks to humans. This study aimed to isolate and identify endophytic fungi from Allium ampeloprasum and Brassica oleracea var. capitata, which are widely cultivated along roadside areas in the upcountry region of Sri Lanka. Sampling sites included Nuwara Eliya town, Nanu Oya, St. Clair’s, and Meepilimana (control), where above-ground parts of the selected vegetables were collected in six replicates. Fungal isolates were obtained through surface sterilization, and their ability to degrade PAHs (naphthalene, phenanthrene, anthracene, and pyrene) was evaluated using plate assays, spectrophotometric analysis, and high-performance liquid chromatography (HPLC). Phyllosphere PAH concentrations were also measured using HPLC. It revealed significantly higher concentrations of all four PAHs in the phyllosphere of both vegetables at polluted sites, with the highest levels recorded in A. ampeloprasum from Nuwara Eliya town: naphthalene (145.92 ng/g), phenanthrene (97.67 ng/g), anthracene (88.71 ng/g), and pyrene (63.82 ng/g). Most endophytic fungal strains isolated from both vegetables were able to grow on Bacto Bushnell–Haas (BBH) medium supplemented with PAHs, producing colonies exceeding 20 mm in diameter. Spectrophotometric analysis showed that Fusarium liriodendri SP2 (PV400499.1) and Trichoderma atroviride SP1 (PV400486.1) achieved approximately 75% degradation of selected PAHs. Furthermore, HPLC analysis confirmed that these isolates effectively degraded all tested PAHs, with degradation rates of approximately 70%. F. liriodendri was the most efficient degrader, achieving degradation rates of 68.50 ± 2.34% for naphthalene, 65.26 ± 1.21% for phenanthrene, 69.21 ± 1.45% for pyrene, and 66.89 ± 1.98% for anthracene. The PAH degradation byproducts of the selected fungal isolates were non-toxic to Artemia salina, confirming their environmental safety. These results highlight the bioremediation potential of endophytic fungi isolated from A. ampeloprasum and B. oleracea var. capitata in PAH-contaminated environments. Full article

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

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22 pages, 1639 KB

Open AccessArticle

Ndt80 Orchestrates Copper Stress Responses and Mitochondrial Homeostasis in Candida albicans

by Hsuan-Yu Chen, Hsiu-Jung Lo, Chi-Jan Lin and Chung-Yu Lan

J. Fungi 2026, 12(4), 294; https://doi.org/10.3390/jof12040294 - 20 Apr 2026

Viewed by 487

Abstract

Copper is a crucial cofactor that sustains multiple cellular electron-transfer reactions, making it an essential element for life. However, cytotoxic levels of copper can cause structural damage and cell death through the production of reactive oxygen species (ROS) and nonspecific attacks on proteins. [...] Read more.

Copper is a crucial cofactor that sustains multiple cellular electron-transfer reactions, making it an essential element for life. However, cytotoxic levels of copper can cause structural damage and cell death through the production of reactive oxygen species (ROS) and nonspecific attacks on proteins. Moreover, immune cells, including neutrophils and macrophages, accumulate copper to induce oxidative bursts that kill engulfed pathogens. Therefore, a well-regulated copper homeostasis system is required for the human commensal fungus Candida albicans to thrive in extreme host environments. Remarkably, C. albicans exhibits higher copper tolerance than the nonpathogenic model yeast Saccharomyces cerevisiae, suggesting the presence of a specific copper tolerance mechanism that supports its adaptability to copper stress. Ndt80 is a versatile transcription factor that regulates several biological processes in C. albicans, ranging from morphological control to drug resistance. This study further reveals that Ndt80 may contribute to copper tolerance by regulating copper transporters and copper-dependent superoxide dismutases (Sods). Additionally, RNA sequencing and complementary approaches uncovered the involvement of Ndt80 in plasma membrane integrity and mitochondrial respiration under copper stress, further linking Ndt80 to copper tolerance. Together, these results broaden our understanding of Ndt80 functions and provide new insights into copper tolerance in C. albicans. Full article

(This article belongs to the Special Issue Candida and Candidemia)

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7 pages, 460 KB

Open AccessBrief Report

Duration of Antifungal Therapy in Disseminated Coccidioidomycosis Patients: A Real-World Treatment Utilization Study

by Craig I. Coleman, Belinda Lovelace and Mark Bresnik

J. Fungi 2026, 12(4), 293; https://doi.org/10.3390/jof12040293 - 20 Apr 2026

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Abstract

Background: Disseminated coccidioidomycosis (DCM) often requires prolonged antifungal therapy (AFT). Real-world data on AFT duration in DCM are limited. We evaluated time to AFT discontinuation among patients with DCM in the United States clinical practice. Methods: This retrospective, longitudinal study used STATinMED data [...] Read more.

Background: Disseminated coccidioidomycosis (DCM) often requires prolonged antifungal therapy (AFT). Real-world data on AFT duration in DCM are limited. We evaluated time to AFT discontinuation among patients with DCM in the United States clinical practice. Methods: This retrospective, longitudinal study used STATinMED data (2016–2024). Patients had ≥1 International Classification of Diseases, Tenth Revision (ICD-10) code for DCM (B38.3, B38.4, B38.7, B38.81) during January 2017–December 2023, ≥1 claim for a triazole or amphotericin B within 21 days of the DCM diagnosis (index date), and continuous medical/pharmacy coverage during the 6-month baseline period. Discontinuation was defined as a ≥21-day gap without AFT. Antifungal agent/formulation switches were not considered discontinuations unless accompanied by a qualifying gap. The Kaplan–Meier methods were used to estimate time to discontinuation. Results: We identified 991 patients with DCM. Median age was 52 years (IQR 36, 65); 60.0% were men. Most resided in California (42.8%) or Arizona (33.6%). Initial AFT consisted predominantly of triazoles (96.8%), primarily fluconazole (83.2%). Discontinuation occurred in 27.6%, 40.0%, 54.2%, and 68.0% of patients by 3, 6, 12, and 36 months. Median AFT duration was 9.9 months. Conclusions: In a large US claims cohort, there was substantial variability in AFT duration in routine practice. Many patients had AFT durations under the lower limit of guideline recommendations for DCM, suggesting potential under-treatment, though appropriate clinical justifications may have existed. Full article

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

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23 pages, 7352 KB

Open AccessArticle

Dual Biocontrol and Plant Growth-Promoting Effects of Trichoderma nordicum V1 Against Oomycete Plant Pathogens

by Songrong Li, Xian Wen, Siqiao Chen, Yishen Zhao, Jinhao Chen, Wanrong Li, Yajuan Chen, Mingyue Ding, Siqi Jiang, Wilfred Mabeche Anjago, Dongmei Zhou, Feng M. Cai, Irina S. Druzhinina, Min Jiu, Lihui Wei and Paul Daly

J. Fungi 2026, 12(4), 292; https://doi.org/10.3390/jof12040292 - 20 Apr 2026

Viewed by 431

Abstract

The potential of Trichoderma nordicum (Hypocreales, Ascomycota), a recently described species, for antagonism and use in the biocontrol of oomycete-caused plant diseases is unknown. Trichoderma is a well-known genus for containing microbial antagonists and biocontrol agents. The T. nordicum in [...] Read more.

The potential of Trichoderma nordicum (Hypocreales, Ascomycota), a recently described species, for antagonism and use in the biocontrol of oomycete-caused plant diseases is unknown. Trichoderma is a well-known genus for containing microbial antagonists and biocontrol agents. The T. nordicum in this study was isolated from decomposing wood, and rpb2 and tef1 barcode sequencing demonstrated that the isolates were a match to the reference T. nordicum and T. nigricans strains. Since T. nordicum was described before T. nigricans, the isolates were assigned to T. nordicum, although taxonomic uncertainty between these species requires future clarification. In dual-culture confrontation assays, T. nordicum overgrew five economically important oomycete plant pathogens (Phytophthora capsici, P. sojae, Pythium aphanidermatum, P. myriotylum, and Globisporangium ultimum). The inability to recover viable P. aphanidermatum and P. capsici from the parts of the plate overgrown by T. nordicum, coupled with protease and endo-cellulase activities, correlates with T. nordicum having antagonistic abilities. Inoculation with T. nordicum preventively reduced the levels of cucumber seedling damping-off caused by P. aphanidermatum by up to 70%. The T. nordicum biocontrol effects against pepper blight caused by P. capsici were greater than 80%, compared to an autoclaved T. nordicum spore control. T. nordicum could also significantly promote the growth of pepper, with plant weight increased by up to 40%, compared to an autoclaved-spore control. In contrast, T. nordicum could not be used to control Pythium soft rot of ginger caused by P. myriotylum, even though P. myriotylum was overgrown by T. nordicum, suggesting host- or pathosystem-specific factors influence biocontrol efficacy. In summary, T. nordicum is a promising biocontrol agent for use in the control of pepper blight caused by P. capsici, and also has potential for use in the control of other oomycete-caused plant diseases in vegetable production systems. Full article

(This article belongs to the Special Issue Fungal Biotechnology and Application 3.0)

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11 pages, 1374 KB

Open AccessBrief Report

Endophytic Fusarium commune G3-29-Mediated dsRNA Delivery for Efficient Control of Western Flower Thrips

by Xueyuan Sheng, Yanfei Wang, Chang Chen, Chao Ma, Shuangchao Wang, Endong Wang, Yan Zhao and Lihua Guo

J. Fungi 2026, 12(4), 291; https://doi.org/10.3390/jof12040291 - 18 Apr 2026

Viewed by 251

Abstract

RNA interference (RNAi) provides a sequence-specific strategy for pest management, but efficient and stable double-stranded RNA (dsRNA) delivery remains a key challenge. Here, we established a plant-probiotic-based gene silencing system using the endophytic fungus Fusarium commune G3-29 as a dsRNA delivery vector against [...] Read more.

RNA interference (RNAi) provides a sequence-specific strategy for pest management, but efficient and stable double-stranded RNA (dsRNA) delivery remains a key challenge. Here, we established a plant-probiotic-based gene silencing system using the endophytic fungus Fusarium commune G3-29 as a dsRNA delivery vector against western flower thrips (WFTs, Frankliniella occidentalis). Recombinant G3-29 strains expressing dsRNA targeting the essential WFT genes ACT and SNF were constructed and confirmed to colonize kidney bean leaves without pathogenicity. Bioassays showed that feeding on leaves colonized by dsRNA-expressing G3-29 significantly decreased survival and downregulated target gene expression in both WFT larvae and adults. Within 4 days, survival of both larvae and adults fell below 10%. In larvae, target gene expression decreased by 63% (ACT) and 33% (SNF), while in adults, reductions of 74% (ACT) and 65% (SNF) were observed. In contrast, in vitro-synthesized dsRNA failed to induce significant gene silencing or mortality in larvae, and its control efficacy against adults was also inferior to that of endophytic fungus-mediated dsRNA delivery. Our findings establish endophytic fungus F. commune G3-29 as an effective and sustainable dsRNA delivery vehicle for RNAi-based pest control, offering distinct advantages over existing strategies such as HIGS and SIGS. This approach provides a promising new direction for managing WFTs and other insect pests. Full article

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

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22 pages, 925 KB

Open AccessReview

Genotype–Phenotype Relationships in Azole-Resistant Aspergillus: Two Sides of the Same Coin

by Merlijn H. I. van Haren, Willem J. G. Melchers, Jianhua Zhang, Sarah Dellière, Christine C. Bii, Felicia A. Stanford, Michael Voetz, P. Lewis White, Paul S. Dyer, Suzan D. Pas, Paul E. Verweij and Jochem B. Buil

J. Fungi 2026, 12(4), 290; https://doi.org/10.3390/jof12040290 - 18 Apr 2026

Viewed by 567

Abstract

Aspergillus fumigatus is a leading cause of invasive fungal disease in humans and is classified as a critical priority threat by the World Health Organization. Triazole antifungals remain the cornerstone of therapy, yet their effectiveness is steadily being eroded by the continuous rise [...] Read more.

Aspergillus fumigatus is a leading cause of invasive fungal disease in humans and is classified as a critical priority threat by the World Health Organization. Triazole antifungals remain the cornerstone of therapy, yet their effectiveness is steadily being eroded by the continuous rise in drug resistance. Most resistance mechanisms trace back to mutations in Cyp51A, spawning well-defined genotypes such as TR₃₄/L98H and TR₄₆/Y121F/T289A. However, the Cyp51A genotype–phenotype landscape in A. fumigatus is far from straightforward. Isolates that share an identical TR genotype can display strikingly divergent susceptibility profiles, and mutational hotspots in Cyp51A, such as G54, M220 and G448, are linked to varying resistances, challenging assumptions about predictable resistance behavior. Complicating matters further, an expanding array of resistance mechanisms, independent of Cyp51A, is now being uncovered. This review summarizes the current state of knowledge on azole resistance in A. fumigatus, dissecting the intricate genotype–phenotype relationships, spotlighting emerging non-Cyp51A pathways and outlining future strategies to enhance the detection and clinical management of antifungal resistance. Full article

(This article belongs to the Special Issue Aspergillus Infections, Diagnostics and Antifungal Treatment)

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21 pages, 1879 KB

Open AccessArticle

Two-Step Engineering of Food-Grade Aspergillus oryzae via Endogenous Signal Peptides and Vesicle Trafficking Proteins to Enhance Carrier-Free Protein Secretion

by Sarocha Panchanawaporn, Nakul Rattanaphan, Sukanya Jeennor, Jutamas Anantayanon, Weerapong Woraprayote, Laphaslada Pumpuang, Thipphiya Karirat, Nuttamon Prompakdee, Kobkul Laoteng and Chanikul Chutrakul

J. Fungi 2026, 12(4), 289; https://doi.org/10.3390/jof12040289 - 18 Apr 2026

Viewed by 307

Abstract

Heterologous protein secretion in filamentous fungi is often constrained by limitations in signal peptide recognition and intracellular trafficking. Aspergillus oryzae, a food-grade industrial fungus, has a robust native secretory system. However, its capacity for recombinant protein secretion remains suboptimal. Here, we developed [...] Read more.

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

(This article belongs to the Special Issue Filamentous Fungi as Excellent Industrial Strains: Development and Applications, 2nd Edition)

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13 pages, 6474 KB

Open AccessArticle

Emergence of Postharvest Strawberry Fruit Rot Caused by Penicillium citrinum in China and Its Whole-Genome Sequencing

by Haohao Yan, Lili Jiang, Tianyu Guo, Mikael Motelica-Heino and Chong Wu

J. Fungi 2026, 12(4), 288; https://doi.org/10.3390/jof12040288 - 17 Apr 2026

Viewed by 351

Abstract

China has the largest strawberry cultivation area worldwide and produces substantial quantities of the fruit. However, postharvest diseases of strawberries occur frequently, limiting their safe storage. In November 2025, a localized occurrence of postharvest fruit rot affecting strawberry (cv. Hongyan) was observed in [...] Read more.

China has the largest strawberry cultivation area worldwide and produces substantial quantities of the fruit. However, postharvest diseases of strawberries occur frequently, limiting their safe storage. In November 2025, a localized occurrence of postharvest fruit rot affecting strawberry (cv. Hongyan) was observed in Tai’an, China. A pathogenic fungus, designated CM-RB5, was isolated from diseased fruits and identified as Penicillium citrinum based on morphological characteristics and molecular analyses. This is the first report of P. citrinum causing postharvest fruit rot in strawberry. The genome of the pathogenic fungal strain CM-RB5 was sequenced using the Illumina MiSeq II and PacBio RS III platforms. Genome assembly analysis revealed the total sequence length of P. citrinum CM-RB5 to be 32,053,718 bp, with a GC content of 46.41%. Additionally, P. citrinum CM-RB5 was found to produce ochratoxin and citrinin. These findings provide insights that may facilitate the development of effective control strategies for postharvest fruit rot in strawberry, thereby ensuring the consumption of safe, high-quality fruit and strawberry-derived products. Full article

(This article belongs to the Special Issue Postharvest Fungi: Control of Fungal Diseases in Fruit and Vegetables)

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15 pages, 4030 KB

Open AccessArticle

Mating-Type Imputation (MTI) Provides an Efficient Tool for the Mating-Type Inference of Tetrapolar Fungi

by Zhenyang Yu, Yu Wang, Haixu Liu, Ruiheng Yang, Weijun Li, Huiyang Xiong, Yu Li, Yongping Fu, Shijun Xiao and Dapeng Bao

J. Fungi 2026, 12(4), 287; https://doi.org/10.3390/jof12040287 - 17 Apr 2026

Viewed by 272

Abstract

Mating-type identification is fundamental to studies of genetic diversity and genetic breeding in fungi, especially for tetrapolar basidiomycetes, whose mating types are determined by two multiallelic loci, A and B. Traditional mating-type identification of monokaryons relies on manual inference based on hybridization experiments; [...] Read more.

Mating-type identification is fundamental to studies of genetic diversity and genetic breeding in fungi, especially for tetrapolar basidiomycetes, whose mating types are determined by two multiallelic loci, A and B. Traditional mating-type identification of monokaryons relies on manual inference based on hybridization experiments; however, this process is highly complex, time-consuming, and error-prone when applied to large-scale studies. In this study, we isolated 30 monokaryons from protoplasts derived from 15 dikaryons of Flammulina velutipes and developed a software tool, Mating-Type Imputation (MTI), to automatically, rapidly, and accurately infer monokaryon mating types in tetrapolar fungi using a combinatorial pruning traversal algorithm. Using a compatibility matrix derived from 435 hybridization experiments involving these 30 monokaryons, MTI required only a few minutes to accurately infer the mating types of all monokaryons-a task that typically takes several days for manual inference by experienced investigators. Furthermore, MTI enabled us to investigate how false-positive and false-negative interactions influence mating-type inference results. Using a simulated compatibility matrix, we found that MTI could accurately detect potential false negatives in compatibility and successfully infer the true mating-type combinations even in the presence of limited false negatives; conversely, the tool was easily misled by any false positives, resulting in incorrect mating-type combinations. This indicates that false-positive records in hybridization experiments must be strictly eliminated during mating-type inference. In summary, MTI provides an efficient tool for inferring the mating types of tetrapolar fungi, offering technical support for mating-type studies of edible and medicinal fungi, and holds significant theoretical value and broad application potential in the fields of fungal genetic diversity and breeding research. Full article

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

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39 pages, 1486 KB

Open AccessReview

An Overview of Major Penicillium Species Associated with Plant Diseases

by Latiffah Zakaria

J. Fungi 2026, 12(4), 286; https://doi.org/10.3390/jof12040286 - 17 Apr 2026

Viewed by 771

Abstract

Species of Penicillium are among the most important fungal pathogens responsible for postharvest diseases of agricultural crops worldwide. This review provides an overview of five economically important Penicillium spp., namely P. expansum, P. digitatum, P. italicum, P. citrinum, and [...] Read more.

Species of Penicillium are among the most important fungal pathogens responsible for postharvest diseases of agricultural crops worldwide. This review provides an overview of five economically important Penicillium spp., namely P. expansum, P. digitatum, P. italicum, P. citrinum, and P. oxalicum. Emphasis is placed on P. expansum, P. digitatum, and P. italicum which are the main causal agents of blue mold and green mold rots in pome fruits and citrus, commodities that dominate global fresh produce trade and long-term storage. While studies on plant-pathogenic Penicillium are mainly focused on these hosts, this review highlights reports of infections in other crops across diverse geographic regions, highlighting the broader host range of these species. The main aspects highlighted include host specificity and diversity, production of mycotoxins and other secondary metabolites, current management and control strategies, and the potential influence of climate change on disease incidence and severity. Understanding the biology and epidemiology of plant-pathogenic Penicillium species is essential, as several species are both pathogens and producers of mycotoxins, leading to quality deterioration and nutrient depletion resulting in economic losses. Full article

(This article belongs to the Special Issue Fungal Pathogens in Postharvest Fruit Diseases: Control and Management)

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33 pages, 2028 KB

Open AccessReview

Regional Cultivation Forms of Hericium erinaceus Across China’s Climatic Zones: A Scoping Review and Analytical Lens for Climate-Adaptive Production

by Dongting Yang, Lin Zhu and Qiaoping Zheng

J. Fungi 2026, 12(4), 285; https://doi.org/10.3390/jof12040285 - 17 Apr 2026

Viewed by 523

Abstract

The cultivation of Hericium erinaceus in China accounts for about 85% of the global supply. Its decentralized production systems have developed across diverse climate zones, leading to distinct, location-specific practices recorded in local technical standards. This scoping review synthesizes these empirical protocols from [...] Read more.

The cultivation of Hericium erinaceus in China accounts for about 85% of the global supply. Its decentralized production systems have developed across diverse climate zones, leading to distinct, location-specific practices recorded in local technical standards. This scoping review synthesizes these empirical protocols from five agro-climatic regions. It illustrates how adaptive strategies such as cold-tolerant strains in the northeast and market-driven precision in the subtropics are associated with yield stability. These practices reflect two interconnected forms of diversity. One is the diversity of cultivation systems themselves, from forest-based methods to industrial-scale production systems. The other is the diversity of locally adapted strains developed for specific environments. We use the Intelligent Germplasm–Cultivation–Processing–Market (GCPM) Integration framework to connect local practices with broader questions of systemic resilience. The evidence draws on field-validated standards, not controlled experiments, reflecting the current state of research. This work presents China’s practical knowledge as a reference for designing context-sensitive, climate-resilient cultivation systems elsewhere, suggesting that resilience may depend more on intelligent adaptation to local conditions than on one-size-fits-all solutions. Full article

(This article belongs to the Special Issue Utilizing Fungal Diversity for Sustainable Biotechnology)

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13 pages, 4277 KB

Open AccessArticle

Colletotrichum Species Causing Anthracnose in Ipê Trees

by Elder F. M. Silva, Ana G. G. Amaral, André N. Oliveira, Luis O. Viteri, Cristiano B. Moraes, Eugênio E. Oliveira, Ailton Reis, Lavínia G. A. Freitas, Gil R. Santos and Marcos P. S. Câmara

J. Fungi 2026, 12(4), 284; https://doi.org/10.3390/jof12040284 - 17 Apr 2026

Viewed by 379

Abstract

Ipê trees (Bignoniaceae), mainly belonging to the genus Handroanthus, are widely used in urban landscaping and reforestation programs in Brazil. Anthracnose, typically associated with species of Colletotrichum, represents one of the major diseases affecting ipê seedlings and ornamental trees. However, the [...] Read more.

Ipê trees (Bignoniaceae), mainly belonging to the genus Handroanthus, are widely used in urban landscaping and reforestation programs in Brazil. Anthracnose, typically associated with species of Colletotrichum, represents one of the major diseases affecting ipê seedlings and ornamental trees. However, the etiological agents involved have not yet been fully clarified using modern phylogenetic tools. In this study, we identified Colletotrichum species associated with anthracnose in ipê trees from Pernambuco, Brazil. A total of 22 isolates were obtained from symptomatic leaves of Handroanthus impetiginosus and H. chrysotrichus. Species identification was based on multilocus phylogenetic analyses using CAL, GAPDH, GS, and TUB2 loci. The isolates were assigned to three species: Colletotrichum siamense, C. tropicale, and C. karsti. Colletotrichum siamense was the most prevalent species (50%), followed by C. tropicale (36.3%), while C. karsti represented 13.7% of the isolates. Pathogenicity tests confirmed that all isolates were pathogenic to both ipê species, producing typical anthracnose symptoms. Aggressiveness differed between hosts, with H. impetiginosus showing higher susceptibility, as indicated by larger lesion development, whereas H. chrysotrichus exhibited lower disease aggressiveness. Thus, our findings represent the first multilocus-based identification of Colletotrichum species causing anthracnose in ipê trees, providing new insights into the diversity and epidemiology of this disease in urban environments. Full article

(This article belongs to the Special Issue Monitoring, Management and Ecotoxicological Implications of Plant Fungal Disease Control)

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21 pages, 17546 KB

Open AccessArticle

Characterization of Penicillium halotolerans with Antagonistic Activity Against Fusarium Root Rot in Astragalus membranaceus

by Yuze Yang, Haiping Jiang, Xunjue Yang, Ke Hao, Yujia Zhao, Qingzhi Yao and Min Li

J. Fungi 2026, 12(4), 283; https://doi.org/10.3390/jof12040283 - 17 Apr 2026

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Abstract

Astragalus membranaceus is an important perennial medicinal plant whose roots constitute its primary medicinal organ; however, its cultivation is severely constrained by root rot caused by Fusarium oxysporum. This study aimed to characterize differences in the rhizosphere microbiome between healthy and diseased [...] Read more.

Astragalus membranaceus is an important perennial medicinal plant whose roots constitute its primary medicinal organ; however, its cultivation is severely constrained by root rot caused by Fusarium oxysporum. This study aimed to characterize differences in the rhizosphere microbiome between healthy and diseased plants, identify antagonistic microorganisms from healthy rhizosphere soils, and investigate their suppressive effects on F. oxysporum and the associated host metabolic responses. High-throughput sequencing was used to compare bacterial and fungal communities in the rhizospheres of healthy and diseased plants. Microorganisms were isolated from healthy rhizosphere soils and screened for antagonistic activity against F. oxysporum, followed by validation in pot experiments. Metabolomic analysis was further conducted to assess host metabolic responses to microbial treatment. Root rot disease significantly altered the dominant composition of rhizosphere microbial communities and was associated with reduced fungal diversity and lower bacterial richness in diseased soils. Co-occurrence network analysis revealed increased complexity in bacterial networks and strengthened positive correlations among fungal taxa under diseased conditions. A total of 81 microbial strains were isolated from healthy rhizosphere soils, among which Penicillium halotolerans exhibited the strongest inhibitory activity against the mycelial growth of F. oxysporum. Pot experiments further supported its suppressive effect on Astragalus root rot. Metabolomic analysis indicated that P. halotolerans treatment was associated with changes in host metabolic profiles related to energy metabolism, defense-associated protein synthesis, and nutrient uptake. Overall, this study identified P. halotolerans as a fungal strain with antagonistic activity against F. oxysporum and provided initial evidence for its association with the suppression of Astragalus root rot. These findings offer candidate microbial resources and mechanistic insights for understanding rhizosphere-associated disease suppression in Astragalus membranaceus. Full article

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

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26 pages, 2855 KB

Open AccessArticle

FcLRR1 Regulates Hyphal Growth and Plant Infection in Fusarium circinatum

by Tingting Dai, Chao Chen, Fangyi Ju, Jiahui Zang, Zhongqiang Qi, Haiwen Wang, Xiaorui Zhang and Chun Yang

J. Fungi 2026, 12(4), 282; https://doi.org/10.3390/jof12040282 - 16 Apr 2026

Viewed by 397

Abstract

Pitch canker caused by the fungus Fusarium circinatum is a destructive disease that affects pines in Europe, South Africa, and North America, particularly along the southeastern and western coasts of the United States. This study systematically elucidated the function of the Leucine-rich repeat [...] Read more.

Pitch canker caused by the fungus Fusarium circinatum is a destructive disease that affects pines in Europe, South Africa, and North America, particularly along the southeastern and western coasts of the United States. This study systematically elucidated the function of the Leucine-rich repeat (LRR) protein FcLRR1 in the pine pitch canker pathogen Fusarium circinatum. A total of 13 LRR proteins were identified via bioinformatic analysis. Using a gene knockout system, we demonstrated that deletion of FcLRR1 significantly impaired vegetative growth, conidiation, and conidium germination; led to a complete loss of macroconidia production; and drastically reduced abiotic stress tolerance and virulence. Transcriptome profiling revealed 612 downregulated genes, which were significantly enriched in pathways such as starch and sucrose metabolism, indicating that FcLRR1 modulated energy supply and pathogenicity through carbon source utilization. Through genome-wide protein structure modeling and yeast two-hybrid assays, we identified and validated the interaction between FcLRR1 and ALG-11, among other candidate proteins, further supporting its involvement in carbon metabolism, cell wall integrity, and pathogenesis. This study represents the first functional characterization of an LRR-containing protein in a forest pathogenic fungus and provides a foundational basis for developing targeted disease control strategies. Full article

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

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23 pages, 4113 KB

Open AccessArticle

Assessment of Disease-Suppressive and Plant Growth-Promoting Capabilities of Thelonectria veuillotiana, an Endophytic Fungus Isolated from Impatiens hawkeri

by Huali Li, Xingyao Xiao, Mengting Luo, Jian Liu, Yajiao Sun, Mengyao Wang, Shuwen Liu, Yunqiang Ma, Hongliang Zhang and Junjia Lu

J. Fungi 2026, 12(4), 281; https://doi.org/10.3390/jof12040281 - 15 Apr 2026

Viewed by 367

Abstract

To investigate the key role of endophytic fungi in maintaining host adaptability and overall health, endophytic fungi were isolated from healthy root, stem and leaf tissues of Impatiens hawkeri, and the dominant strain FG8 with growth-promoting and antagonistic functions was screened. Strain [...] Read more.

To investigate the key role of endophytic fungi in maintaining host adaptability and overall health, endophytic fungi were isolated from healthy root, stem and leaf tissues of Impatiens hawkeri, and the dominant strain FG8 with growth-promoting and antagonistic functions was screened. Strain FG8 was identified as Thelonectria veuillotiana by morphological and molecular biological methods. It exhibited an antifungal rate of 58.57% against Stagonosporopsis cucurbitacearum, the pathogen causing leaf spot disease of I. hawkeri. The broad-spectrum antifungal activity was verified by the plate confrontation method, and FG8 showed inhibitory effects on six common pathogenic fungi, with the highest inhibition rate of 64.5% against Apiospora intestini. Furthermore, strain FG8 displayed remarkable growth-promoting and antagonistic characteristics: it produced indole-3-acetic acid at 12.74 μg/mL, and possessed the abilities of phosphate solubilization, potassium mobilization, nitrogen fixation and siderophore synthesis. Its antagonistic activity was mediated by β-glucanase, amylase, cellulase and pectinase. Meanwhile, FG8 significantly induced the activities of four defensive enzymes in I. hawkeri, including superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and polyphenol oxidase (PPO). Seed growth-promotion experiments demonstrated that the root length, plant height, fresh weight and dry weight of seedlings in the FG8-treated group were significantly higher than those in the control group. These results indicate that strain FG8 has both growth-promoting and biological control functions, which can provide a potential resource for the biological control of I. hawkeri leaf spot and the development of fungal fertilizers. Its field application effect and mechanism of action need to be further explored. Full article

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

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14 pages, 3767 KB

Open AccessArticle

Genetic Diversity and Gene Flow of the Ectomycorrhizal Mushroom Lactarius hatsudake in Southern China: Evidence from SSR Markers

by Shatong Yang, Mingwei Mao, Jieyu Huang, Bing Gu and Kuan Zhao

J. Fungi 2026, 12(4), 280; https://doi.org/10.3390/jof12040280 - 15 Apr 2026

Viewed by 342

Abstract

Lactarius hatsudake is an ecologically and economically significant wild edible mushroom in southern China. To elucidate its population genetic diversity, differentiation, and evolutionary history, we analyzed 172 fruiting bodies from eight geographic populations (AQ, BS, DZ, JS, NC, PT, SG, YX) across seven [...] Read more.

Lactarius hatsudake is an ecologically and economically significant wild edible mushroom in southern China. To elucidate its population genetic diversity, differentiation, and evolutionary history, we analyzed 172 fruiting bodies from eight geographic populations (AQ, BS, DZ, JS, NC, PT, SG, YX) across seven provinces in the western and eastern regions of southern China using five highly polymorphic simple sequence repeat (SSR) markers. Combined with STRUCTURE clustering, discriminant analysis of principal components (DAPC), unweighted pair group method with arithmetic mean (UPGMA), and analysis of molecular variance (AMOVA), the results revealed high polymorphism across the studied loci (mean PIC = 0.842). A total of 75 alleles were identified, averaging 15 alleles per locus. At the population level, the mean effective number of alleles (Ne) was 4.023, and the mean unbiased gene diversity (uH) was 0.768. The NC population exhibited the highest genetic diversity (uH = 0.796), whereas the BS population showed relatively lower diversity (uH = 0.647). Clustering analyses (STRUCTURE, DAPC, and UPGMA) consistently identified two distinct genetic clusters (K = 2). Cluster I consisted of populations AQ, PT, BS, and SG, while Cluster II was composed of the remaining four populations. Notably, individuals from AQ and NC displayed significant genetic admixture, suggesting a transitional zone. AMOVA revealed that the majority of genetic variation (83%) resided within populations and 17% among populations. Moderate population differentiation (ENA-corrected global Fst = 0.102) and admixture signals suggest non-negligible connectivity among populations. Full article

(This article belongs to the Special Issue Edible and Medicinal Macrofungi, 4th Edition)

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12 pages, 478 KB

Open AccessArticle

A Retrospective, Comparative Cohort Analysis of the Utilization of Voriconazole-Containing Regimens Versus Itraconazole in the Treatment of Blastomycosis

by Caitlin C. Schanz, Christina G. Rivera, Omar M. Abu Saleh, Josh Clement, Mark J. Enzler, Danielle Firkus, Kristin Cole, Paschalis Vergidis and Ryan W. Stevens

J. Fungi 2026, 12(4), 279; https://doi.org/10.3390/jof12040279 - 14 Apr 2026

Viewed by 406

Abstract

Itraconazole is the recommended first-line antifungal agent in blastomycosis treatment. However, pharmacokinetic limitations and adverse drug events may necessitate the use of alternative antifungal agents. The primary objective of the study was to retrospectively describe a cohort of patients with blastomycosis who received [...] Read more.

Itraconazole is the recommended first-line antifungal agent in blastomycosis treatment. However, pharmacokinetic limitations and adverse drug events may necessitate the use of alternative antifungal agents. The primary objective of the study was to retrospectively describe a cohort of patients with blastomycosis who received voriconazole (as partial or complete therapy), for complete or partial treatment, compared to those who received itraconazole alone. Secondary objectives included rationale for voriconazole selection, treatment response, mortality, and adverse drug events. This retrospective multicenter cohort study included adult patients with proven/probable blastomycosis who received itraconazole or voriconazole. Treatment response was evaluated at one year or end of therapy, whichever came first. Mortality outcomes were assessed within 7 days of the last documented azole dose. Propensity score weighting and subgroup analysis were utilized to control confounding variables between cohorts. A total of 119 patients receiving itraconazole and 25 receiving voriconazole as complete or partial treatment were included. Voriconazole was often selected for CNS involvement or after intolerance to alternative azoles. After propensity score weighting, no significant difference in complete or partial treatment response was observed. Rates of all-cause mortality and blastomycosis-related mortality were numerically higher in the voriconazole cohort. Adverse drug event rates were similar between cohorts; however, discontinuation due to adverse events was more common in the voriconazole cohort. Voriconazole was most utilized for the treatment of blastomycosis in cases with CNS involvement or intolerance to other azoles. A statistically significant difference in response rate was not identified between voriconazole-containing or itraconazole-treated patients; however, given limited sample sizes, further data is needed to assess the equivalency of these agents in the treatment of blastomycosis. Full article

(This article belongs to the Special Issue Invasive Fungal Infections: Recent Advances in Epidemiology, Diagnosis, and Treatment)

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14 pages, 1904 KB

Open AccessArticle

Temporal Trends and Seasonality of Invasive Candidiasis During and After the COVID-19 Pandemic: An Interrupted Time Series Analysis in Colombia

by José M. Oñate-Gutiérrez, Carlos A. Alvarez-Moreno, Claudia Cañadas-Aragón and Hernán Vergara-Samur

J. Fungi 2026, 12(4), 278; https://doi.org/10.3390/jof12040278 - 14 Apr 2026

Viewed by 528

Abstract

Invasive candidiasis is a severe opportunistic infection whose incidence may be influenced by major disruptive events. The COVID-19 pandemic substantially altered hospital dynamics in Colombia. This study aimed to evaluate temporal trends, seasonality, and potential changes in the incidence of invasive candidiasis between [...] Read more.

Invasive candidiasis is a severe opportunistic infection whose incidence may be influenced by major disruptive events. The COVID-19 pandemic substantially altered hospital dynamics in Colombia. This study aimed to evaluate temporal trends, seasonality, and potential changes in the incidence of invasive candidiasis between 2019 and 2024. We conducted an observational time-series study using confirmed cases of invasive candidiasis from medium- and high-complexity hospitals in three major Colombian cities. Cases were aggregated quarterly. An interrupted time-series (ITS) analysis was performed. A total of 1294 cases were analyzed. An increasing trend was observed until mid-2022, followed by a decline during 2023. Seasonal decomposition revealed persistent seasonality with recurrent peaks in the second and fourth quarters. The ITS analysis did not demonstrate statistically significant changes in level or slope after the interruption (p > 0.05), although clinically relevant fluctuations were observed. No significant differences in temporal trends were identified across Candida species. Invasive candidiasis in Colombia exhibited a complex temporal evolution during and after the COVID-19 pandemic characterized by sustained seasonality and an increase followed by a decline. Although the ITS analysis did not identify statistically significant post-pandemic changes, the findings support the use of time-series models as valuable tools for epidemiological surveillance and trend monitoring. 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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12 pages, 5301 KB

Open AccessArticle

The AfldrnA Transcription Factor Is a Pivotal Regulator of the Conidiation–Sclerotial Formation Balance in Aspergillus flavus

by Mohammed A. Abdo-Elgabbar, Bashir Salim, Sang-Cheol Jun, Yu-Kyung Kim, Saeed Alasmari and Kap-Hoon Han

J. Fungi 2026, 12(4), 277; https://doi.org/10.3390/jof12040277 - 14 Apr 2026

Viewed by 326

Abstract

Aspergillus flavus is a globally distributed filamentous fungus of major agricultural and medical importance, capable of producing carcinogenic aflatoxins and forming two specialized developmental structures, conidia and sclerotia. While the molecular framework governing conidiation has been well characterized in Aspergillus nidulans, the [...] Read more.

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

(This article belongs to the Special Issue Mycological Research in South Korea)

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16 pages, 3570 KB

Open AccessArticle

Engineering a Cold-Active Cellulase Complex with a Novel Mushroom Cellobiohydrolase for Efficient Biomass Saccharification and Juice Flavor Optimization

by Jiaqi Yang, Youran Shao, Ying Wang, Ming Gong, Bing Li, Hongyu Chen, Caizhen Wang, Yan Li, Xiang Zhou and Gen Zou

J. Fungi 2026, 12(4), 276; https://doi.org/10.3390/jof12040276 - 10 Apr 2026

Viewed by 532

Abstract

Cold-active cellulases are highly desirable for temperature-sensitive biomass valorization and food processing, yet they remain scarce in conventional industrial fungal platforms. In this study, a novel cold-induced cellobiohydrolase, VvCBHI-II, was mined from the mushroom Volvariella volvacea and successfully engineered into the industrial [...] Read more.

Cold-active cellulases are highly desirable for temperature-sensitive biomass valorization and food processing, yet they remain scarce in conventional industrial fungal platforms. In this study, a novel cold-induced cellobiohydrolase, VvCBHI-II, was mined from the mushroom Volvariella volvacea and successfully engineered into the industrial workhorse Trichoderma reesei via site-specific homologous replacement. Structural homology modeling revealed that the substitution of the flexible B3 loop with a β-sheet creates a more open substrate-binding cleft in VvCBHI-II. Consequently, the purified VvCBHI-II exhibited robust endoglucanase-like characteristics with superior catalytic efficiency on amorphous cellulose. At 10 °C, the engineered cellulase complex demonstrated an 8.1-fold increase in filter paper activity compared to the wild-type strain. Mechanistic structural analyses indicated that the open cleft architecture elongates and weakens the hydrogen-bonding network with the cellobiose product, facilitating rapid product dissociation and alleviating severe cold-induced product inhibition. In practical applications, the engineered cold-active enzyme complex exhibited an exceptional saccharification capacity on natural pear pomace at 10 °C. Furthermore, when applied to simulated fruit juice processing, it significantly maximized the extraction yield, elevated the sweetness response, and substantially mitigated undesirable bitterness and astringency. This study elucidates the structural-functional paradigm of cold-adapted cellobiohydrolases and provides a promising strategy for formulating highly efficient, energy-saving biocatalysts for the food and biorefinery industries. Full article

(This article belongs to the Special Issue Research and Application of Fungal Enzymes)

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16 pages, 4359 KB

Open AccessArticle

Diversity and Pathogenicity of Neopestalotiopsis Species Associated with Strawberry Leaf Spot and Fruit Rot in Nova Scotia

by Sajid Rehman and Shawkat Ali

J. Fungi 2026, 12(4), 275; https://doi.org/10.3390/jof12040275 - 10 Apr 2026

Viewed by 497

Abstract

We reported the first isolation and characterization of Neopestalotiopsis spp. from symptomatic strawberry plants in Nova Scotia, Canada. Morphological and multilocus sequence analyses confirmed that these isolates were closely related to previously identified aggressive Neopestalotiopsis spp. strains from strawberry and blueberry in the [...] Read more.

We reported the first isolation and characterization of Neopestalotiopsis spp. from symptomatic strawberry plants in Nova Scotia, Canada. Morphological and multilocus sequence analyses confirmed that these isolates were closely related to previously identified aggressive Neopestalotiopsis spp. strains from strawberry and blueberry in the southeastern United States and other countries. Five representative isolates were evaluated for pathogenicity on detached leaves, whole plants, and fruits of multiple strawberry cultivars. The results revealed significant variation in virulence, with isolate NS-1 causing the most severe necrosis across all tissue types. Statistical analysis revealed significant effects of isolate, cultivar, and their interaction on disease severity, indicating differential cultivar responses to the tested isolates. Notably, tissue-specific differences were observed, with some isolates being aggressive on leaves but less virulent on fruit or whole plants, reinforcing the importance of multi-organ phenotyping in resistance screening. Phylogenetic analysis clustered the Nova Scotia isolates within the same clade as Neopestalotiopsis isolate 17–43 L from strawberry and isolates from blueberry, suggesting a potential epidemiological link. The shared nursery propagation system of strawberries and blueberries raises the risk of cross-infection, posing a substantial challenge to disease management strategies in both crops. Collectively, these findings underscore the urgent need for continued surveillance, population-level pathogen analysis, and the development of resistant cultivars to mitigate the spread of this emerging and rapidly evolving pathogen. Full article

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

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19 pages, 7754 KB

Open AccessArticle

The Oligosaccharyltransferase Catalytic Subunit PsSTT3B Is Required for Asexual Development and Pathogenicity in Phytophthora sojae

by Quanhe Ma, Borui Zhang, Tongshan Cui, Shanshan Chen, Shan Geng, Fan Zhang, Can Zhang and Xili Liu

J. Fungi 2026, 12(4), 274; https://doi.org/10.3390/jof12040274 - 9 Apr 2026

Viewed by 448

Abstract

N-glycosylation is a fundamental post-translational modification that contributes to protein folding, stability, and secretion in eukaryotes. The catalytic subunit STT3 of the oligosaccharyltransferase complex mediates the transfer of preassembled oligosaccharides to nascent polypeptides in the endoplasmic reticulum. Here, we identified and functionally [...] Read more.

N-glycosylation is a fundamental post-translational modification that contributes to protein folding, stability, and secretion in eukaryotes. The catalytic subunit STT3 of the oligosaccharyltransferase complex mediates the transfer of preassembled oligosaccharides to nascent polypeptides in the endoplasmic reticulum. Here, we identified and functionally characterized PsSTT3B, one of the STT3 paralogs in Phytophthora sojae (P. sojae). PsSTT3B plays an important role in the growth, development, and pathogenicity of P. sojae. CRISPR/Cas9-mediated deletion of PsSTT3B resulted in reduced vegetative growth, sporangia production, and zoospore production in P. sojae. PsSTT3B deletion mutants demonstrated significantly reduced virulence on soybean leaves and etiolated seedlings. Importantly, PsSTT3B deletion mutants also exhibited reduced zoospore germination and diminished chemotaxis toward soybean isoflavones. Moreover, deletion of PsSTT3B increased sensitivity to tunicamycin and dithiothreitol and influenced the ConA-binding glycoprotein profile. Our findings show that PsSTT3B is associated with asexual development, virulence, and sensitivity to ER stress-related conditions of P. sojae. Our study suggests that PsSTT3B represents a potential candidate gene for the prevention and control of P. sojae. Full article

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

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24 pages, 7122 KB

Open AccessArticle

Transposable Element-Driven Genomic Plasticity: Unveiling the Evolutionary Mechanisms of Lifestyle Transition and Ecological Adaptation in Endophytic Fungi

by Yunfeng Lai, Cunzhong Fan, Zhibin Zhang, Riming Yan, Du Zhu and Huilin Yang

J. Fungi 2026, 12(4), 273; https://doi.org/10.3390/jof12040273 - 9 Apr 2026

Viewed by 469

Abstract

The genomic basis underlying the remarkable ecological flexibility of endophytic fungi (EF), particularly their potential to transition between symbiotic, saprophytic, and pathogenic lifestyles, remains poorly understood. Through comparative genomics of 75 Ascomycota and a validation set of 36 Fusarium genomes, we uncovered a [...] Read more.

The genomic basis underlying the remarkable ecological flexibility of endophytic fungi (EF), particularly their potential to transition between symbiotic, saprophytic, and pathogenic lifestyles, remains poorly understood. Through comparative genomics of 75 Ascomycota and a validation set of 36 Fusarium genomes, we uncovered a distinct pattern of genome evolution in EF, contrasting with the well-known “gene loss” model in obligate symbionts. Our analysis reveals that EF genomes are significantly expanded, primarily driven by the accumulation of DNA transposable elements (TEs). Crucially, this TE-mediated genomic plasticity is coupled with the retention and significant expansion of gene families for both saprotrophy and potential pathogenesis. We propose a novel “dual-trophic potential” model: TE-driven genomic expansion and plasticity provide the genetic raw material for EF to maintain a versatile repertoire of ecological tools, facilitating adaptive shifts across the endophytic–saprophytic–pathogenic continuum. This study reframes our understanding of fungal endophytism from a static symbiotic state to a dynamic, genetically enabled ecological strategy. Full article

(This article belongs to the Special Issue Advances in Fungal Genomics and Biotechnology: From Codon Optimization to Protein Expression)

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14 pages, 2591 KB

Open AccessArticle

Species-Discriminating Diagnostic PCR, Ribosomal Intergenic Spacer-Based Single-Marker Taxonomy and Cryptic Descriptions of the Fungal Entomopathogens Metarhizium hybridum and Metarhizium parapingshaense

by Christina Schuster, Haifa Ben Gharsa, Yamilé Baró Robaina, Romina G. Manfrino, Saikal Bobushova, Alejandra C. Gutierrez, Claudia C. López Lastra and Andreas Leclerque

J. Fungi 2026, 12(4), 272; https://doi.org/10.3390/jof12040272 - 9 Apr 2026

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Abstract

(1) Background: Potentially arthropod-pathogenic and plant-associated Metarhizium fungi are of high interest for basic research, biological pest control and plant growth promotion. Unambiguous species delineation enabling the taxonomic assignment of new isolates and the identification of new Metarhizium species is of crucial importance [...] Read more.

(1) Background: Potentially arthropod-pathogenic and plant-associated Metarhizium fungi are of high interest for basic research, biological pest control and plant growth promotion. Unambiguous species delineation enabling the taxonomic assignment of new isolates and the identification of new Metarhizium species is of crucial importance for both research and application. Recently, the new species Metarhizium hybridum and Metarhizium parapingshaense were introduced on the basis of phylogenomic studies. (2) Methods: Neighbor- joining and Bayesian inference-based phylogenetic reconstruction of ribosomal intergenic spacer (rIGS) sequences were used to critically evaluate new species introductions. A species-discriminating diagnostic PCR tool for Metarhizium was adapted to M. hybridum and M. parapingshaense. GenBank database mining was performed to identify cryptic descriptions of the new species. (3) Results: The introduction of M. hybridum and M. parapingshaense was corroborated by rIGS sequence comparison. Data mining revealed cryptic first descriptions of M. hybridum from Canada, China, Colombia, Costa Rica, Cuba, Honduras, Mexico, New Zealand, the USA and the Philippines, and of M. parapingshaense from China, India, Japan, the Philippines and South Korea. (4) Conclusions: Results support the reliability of rIGS as a single taxonomic marker for species-level identification of Metarhizium fungi. Species-discriminating diagnostic PCR was successfully adapted to enable the sequencing-independent identification of the confirmed new species M. hybridum and M. parapingshaense. Full article

(This article belongs to the Topic Diversity of Insect-Associated Microorganisms)

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22 pages, 5900 KB

Open AccessArticle

Straw-Mediated Restructure of Arbuscular Mycorrhizal Fungal Community by Selectively Shifting Edaphic Biogeochemistry in Tea Plantations of South Henan, China

by Xiangchao Cui, Dongmeng Xu, Jiaju Wang, Yu Zhang, Shuping Huang, Wei Wei, Ge Ma, Mengdi Li and Junhui Yan

J. Fungi 2026, 12(4), 271; https://doi.org/10.3390/jof12040271 - 9 Apr 2026

Viewed by 439

Abstract

Background: Straw application (SP) is an important agronomic practice in sustainable agriculture, yet its effects on arbuscular mycorrhizal (AM) fungal communities in tea plantation soils remain poorly understood. Methods: This study investigated the responses of AM fungi to SP in tea plantations in [...] Read more.

Background: Straw application (SP) is an important agronomic practice in sustainable agriculture, yet its effects on arbuscular mycorrhizal (AM) fungal communities in tea plantation soils remain poorly understood. Methods: This study investigated the responses of AM fungi to SP in tea plantations in south Henan, China, by assessing colonization characteristics, community composition, diversity, co-occurrence networks, and soil environmental drivers. Results: SP significantly increased the mycorrhizal colonization rate (MC), by 59.4%. High-throughput sequencing (26,865 sequences and 406 ASVs) revealed that SP reduced the dominance of Claroideoglomus (32.2% to 10.5%) and Glomus (51.01% to 46.7%) while enriching Paraglomus and Acaulospora. Although the α-diversity was unaffected, the β-diversity significantly shifted, indicating community homogenization under SP. Differential taxa analysis confirmed genus-specific responses, and co-occurrence networks showed a simplified topology (nodes: −18.4%; edges: −33.4%) but maintained stability, with increased module specialization (Z_i and P_i). Soil properties explained 80.0% of the variation in AM fungal parameters, with pH and available phosphorus (AP) as key drivers. SP shifted environmental filters from nitrogen/carbon-related factors to metal ions (Al³⁺ and Ca²⁺), altering geochemical conditions. Conclusions: SP selectively reshapes AM fungal communities by altering soil microenvironments and selectively modulating the AM fungal community while maintaining network stability. This study provides new insights into the microbial mechanisms of SP and a basis for sustainable, AMF-based tea plantation management. Full article

(This article belongs to the Section Environmental and Ecological Interactions of Fungi)

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14 pages, 5689 KB

Open AccessArticle

Genome-Scale Phylogenetic Evidence Supports the Synonymy of Lasiodiplodia brasiliensis with Lasiodiplodia theobromae

by Celynne Ocampo-Padilla, Yoshiki Takata, Shunsuke Nozawa, Yui Harada, Katsuhiko Ando and Kyoko Watanabe

J. Fungi 2026, 12(4), 270; https://doi.org/10.3390/jof12040270 - 8 Apr 2026

Viewed by 781

Abstract

The genus Lasiodiplodia includes numerous plant-pathogenic species whose delimitation is complicated by overlapping morphological traits and limited resolution of common genetic markers. Lasiodiplodia brasiliensis was described as a species closely related to L. theobromae; however, its taxonomic status remains controversial. In this [...] Read more.

The genus Lasiodiplodia includes numerous plant-pathogenic species whose delimitation is complicated by overlapping morphological traits and limited resolution of common genetic markers. Lasiodiplodia brasiliensis was described as a species closely related to L. theobromae; however, its taxonomic status remains controversial. In this study, we re-evaluated the species boundaries between L. theobromae and L. brasiliensis using an integrative approach that combined multilocus and genome-scale phylogenetic analyses with morphological comparisons. Multilocus phylogenetic analyses based on ITS, tef1-α, tub2, and rpb2 revealed an unresolved relationship between the two taxa. The L. theobromae clade had low bootstrap support, whereas the ancestral node connecting both species had high support. In contrast, genome-scale phylogenetic analysis using hundreds of single-copy orthologous genes strongly supported a single monophyletic clade encompassing isolates assigned to both L. theobromae and L. brasiliensis. Morphological analyses further revealed that conidial dimensions and other diagnostic characteristics largely overlapped between the two taxa, rendering them unreliable criteria for species separation. Considering the combined molecular and morphological evidence, our results support treating L. brasiliensis as a synonym of L. theobromae. Clarifying species boundaries within this group helps stabilize the taxonomy of Lasiodiplodia and provides a reliable foundation for accurate pathogen identification and disease management. Full article

(This article belongs to the Section Fungal Evolution, Biodiversity and Systematics)

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Open AccessArticle

Unlocking the Potential for Genetic Engineering of the Straw-Degrading Mushroom Stropharia rugosoannulata by Constructing a CRISPR/Cas9 Gene Editing System

by Haibo Hao, Shuzhen Song, Qian Wang, Zongjun Tong, Wen Xu, Jinxiao Yang, Yihong Yue, Tingting Xiao, Yuchen Zhang, Jinjing Zhang and Hui Chen

J. Fungi 2026, 12(4), 269; https://doi.org/10.3390/jof12040269 - 8 Apr 2026

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Abstract

The artificially cultivated edible mushroom Stropharia rugosoannulata is widely promoted and cultivated in China because of its ability to efficiently decompose agricultural and forestry waste. However, methods for CRISPR/Cas9 genome editing have not yet been established for S. rugosoannulata. In this study, [...] Read more.

The artificially cultivated edible mushroom Stropharia rugosoannulata is widely promoted and cultivated in China because of its ability to efficiently decompose agricultural and forestry waste. However, methods for CRISPR/Cas9 genome editing have not yet been established for S. rugosoannulata. In this study, we identified three SrU6 promoters in S. rugosoannulata and constructed the CRISPR/Cas9 expression vector GPiE-SrU6. Moreover, we found that mutant strains were obtained only when the expression of the single guide RNA (sgRNA) was driven by the SrU6-3 promoter. We subsequently employed a tandemly repeated SrU6-tRNA-sgRNA module to knock out two sites within the ura3 gene. The expression vector was introduced into the mycelium via Agrobacterium-mediated transformation (ATMT). Following dual selection with 60 μg/mL hygromycin (Hyg) and 0.2 mg/mL 5-fluoroorotic acid (5-FOA), stable transformants were obtained and subcultured. The mutation efficiency at the targeted ura3 locus was subsequently assessed. The CRISPR/Cas9 system successfully disrupted the target marker gene (ura3), achieving an editing efficiency of 14.9%. In summary, this study reports the first successful establishment of a CRISPR/Cas9 genome editing system in S. rugosoannulata. This study not only meets a future need for genetic manipulation tools for S. rugosoannulata but also provides a robust platform for engineering superior strains for eco-circular agriculture. Full article

(This article belongs to the Special Issue Molecular Biology of Mushroom, 2nd Edition)

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J. Fungi, Volume 12, Issue 4 (April 2026) – 68 articles

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