Journal of Fungi

18 pages, 2399 KB

Open AccessArticle

Intent to Accept a Valley Fever Vaccine for Humans and Dogs and Factors Influencing Intended Uptake: A Cross-Sectional Survey in Two Endemic Regions

by Julia N. Hermann, Sophia E. Kruger, Natalie Wodniak, Jammie Holland, Veronica Janosick, Asley Sanchez, Julio C. Zuniga-Moya, Dana Brucker, Bianca Torres, Keny Mendoza Melo, Emilse Oliveros, Rasha Kuran, Carlos D’Assumpcao, Royce H. Johnson, R. Scott Van Pelt, Abinash Bhattachan, Abram L. Wagner and Jennifer R. Head

J. Fungi 2026, 12(6), 420; https://doi.org/10.3390/jof12060420 (registering DOI) - 10 Jun 2026

Abstract

As Valley fever (coccidioidomycosis) vaccine candidates progress towards human trials, a challenge to their eventual introduction is understanding vaccine demand and addressing vaccine hesitancy. We assessed intent to accept a Valley fever vaccine for humans and dogs in two populations living in highly [...] Read more.

As Valley fever (coccidioidomycosis) vaccine candidates progress towards human trials, a challenge to their eventual introduction is understanding vaccine demand and addressing vaccine hesitancy. We assessed intent to accept a Valley fever vaccine for humans and dogs in two populations living in highly endemic regions: employees at Kern Medical (KM) in Bakersfield, California (N = 103) and members of the public in West Texas (N = 230). We compared the weighted proportions of each population willing to vaccinate themselves and their dogs by demographic and coccidioidomycosis risk factors and assessed the importance of vaccine-related factors on vaccine uptake in each population. We found that 42% (95% confidence interval (CI): 34–49%) of West Texas residents and 76% (95% CI: 63–85%) of KM employees were willing to accept a coccidioidal vaccine, while 49% (95% CI: 41–58%) of West Texas residents and 74% (95% CI: 58–86%) of KM employees were willing to vaccinate their dogs. Among West Texas residents, vaccination willingness was significantly higher among those with prior awareness of Valley fever (adjusted odds ratio (aOR): 3.83, 95% CI: 1.73, 8.49). Across both study populations, absence of side effects was the most important condition that would increase vaccination willingness. Our results indicate substantial interest in a Valley fever vaccine while suggesting that increased Valley fever awareness and minimal vaccine side effects may be important for increased uptake. Full article

(This article belongs to the Special Issue Basic Science and Clinical Research of Coccidioides and Coccidioidomycosis)

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21 pages, 976 KB

Open AccessArticle

Trichoderma asperellum and T. asperelloides: Comparative Genomic Study for Genes Implicated in Biocontrol and Biofertilizer Activities

by Adnan Ismaiel, Jackson Maul and Patricia Millner

J. Fungi 2026, 12(6), 418; https://doi.org/10.3390/jof12060418 (registering DOI) - 9 Jun 2026

Abstract

Trichoderma asperellum and T. asperelloides are two cryptic species that have potential for use as biocontrol and biofertilizer (B&B) agents. Comparison of the reference genomes of the two species revealed that each species had seven chromosomes, but Trichoderma asperellum has about 1000 more [...] Read more.

Trichoderma asperellum and T. asperelloides are two cryptic species that have potential for use as biocontrol and biofertilizer (B&B) agents. Comparison of the reference genomes of the two species revealed that each species had seven chromosomes, but Trichoderma asperellum has about 1000 more genes than T. asperelloides. The number of genes coding for chitinases, cellulases, xylanases, secreted proteases, and genes involved in soil and plant health was slightly greater in T. asperellum than in T. asperelloides. Moreover, T. asperellum had five more genes than T. asperelloides involved in the synthesis of secondary metabolites like peptaibols and siderophores. The B&B genes were distributed on all the chromosomes. No duplicate genes were found for any of the enzymes searched. The investigation also revealed that T. asperellum had 15 copies of the internal transcribed spacer (ITS) region of ribosomal DNA compared to only seven copies in T. asperelloides. Further transcriptomic, proteomic, and efficacy studies are needed to determine the impact of the missing genes in T. asperelloides on its B&B activities compared to those of T. asperellum. The search for B&B genes in T. asperelloides was hindered by the lack of annotation for the genome. Thus, comparison only involves B&B genes searched in T. asperellum and whether homologs to the genes were available or missing in T. asperelloides. A comparison between additional strains of the two species is essential to show whether the data in this study apply to all intraspecies strains of the two species. Full article

(This article belongs to the Special Issue Biotechnological Applications of Fungi)

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7 pages, 684 KB

Open AccessBrief Report

Bioluminescence in the Edible Mushroom Hypsizygus marmoreus by Transformation with a Fungal Luciferase Gene

by Xinyu Zhou, Yan Li, Yingying Wu, Ruisheng Chen, Lihua Tang, Chenli Zhou, Jianing Wan, Dapeng Bao, Ruiheng Yang and Junjun Shang

J. Fungi 2026, 12(6), 417; https://doi.org/10.3390/jof12060417 (registering DOI) - 9 Jun 2026

Abstract

Following the elucidation of the fungal bioluminescence pathway (FBP), it was quickly adopted as a reporter system in plants; however, no such application has been documented in fungi to date. In this study, we established for the first time a luminescent reporter in [...] Read more.

Following the elucidation of the fungal bioluminescence pathway (FBP), it was quickly adopted as a reporter system in plants; however, no such application has been documented in fungi to date. In this study, we established for the first time a luminescent reporter in the commercially important mushroom Hypsizygus marmoreus by expressing the luciferase gene from the luminous fungus Neonothopanus nambi. Using an established Agrobacterium-mediated transformation method, we separately introduced the wild-type luciferase gene nnLuz and the previously reported optimized variant nnLuz-v4 that can enhance bioluminescence expression into H. marmoreus arthroconidia. Both genes were stably integrated into the genome and expressed under the control of the H. marmoreus Glycerol 3-phosphate dehydrogenase (GPD) gene promoter. Upon addition of exogenous luciferin, transformants carrying the wild-type nnLuz produced clear, readily detectable bioluminescence signals, whereas no luminescence was observed in untransformed controls. Unexpectedly, the wild-type luciferase consistently exhibited substantially higher luminescence intensity than the optimized nnLuz-v4 variant. This finding suggests that codon optimization may be unnecessary or even detrimental when the donor and host are phylogenetically close basidiomycetes. The successful deployment of the fungal luciferase gene in H. marmoreus provides a sensitive and non-invasive genetic tool that does not require external excitation. This system opens new avenues for promoter characterization, real-time gene expression monitoring during mushroom development, and molecular breeding efforts aimed at improving agronomically important traits. Full article

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

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17 pages, 3742 KB

Open AccessArticle

Insights into the Species Diversity and Features of Fungi in the Fusarium heterosporum Species Complex

by Olga P. Gavrilova, Aleksandra S. Orina, Nadezhda N. Gogina and Tatiana Yu. Gagkaeva

J. Fungi 2026, 12(6), 416; https://doi.org/10.3390/jof12060416 (registering DOI) - 8 Jun 2026

Abstract

In this study, four Fusarium strains isolated from Poaceae plants infected by Claviceps spp. and one strain isolated from the stem of Cirsium arvense collected from two regions of Russia that are separated by a long distance were analyzed in detail. These fungi [...] Read more.

In this study, four Fusarium strains isolated from Poaceae plants infected by Claviceps spp. and one strain isolated from the stem of Cirsium arvense collected from two regions of Russia that are separated by a long distance were analyzed in detail. These fungi were accurately identified through a phylogenetic analysis of the fragments of translation elongation factor 1-α and RNA polymerase second largest subunit loci. Four of them belong to F. heterosporum species, and one strain, MFG 13060, together with the historical reference strain BBA 62226, forms a distinct lineage within the F. heterosporum species complex (FHSC). The morphological features of the anamorph structures of the fungi within the FHSC are presented. All the analyzed F. heterosporum strains are heterothallic and require a partner to mate. The fertile perithecia of F. heterosporum were obtained in a crossing experiment, and the teleomorph structures were characterized in detail. The screening of 19 mycotoxins typically produced by Fusarium fungi using high-performance liquid chromatography with tandem mass spectrometry revealed the ability of the strains to produce only moniliformin on an autoclaved rice substrate. A reassessment of the species diversity, distribution, and significance of fungi belonging to the FHSC is necessary to elucidate the unclear relationships between F. heterosporum, Claviceps fungi, and cereal plants. Full article

(This article belongs to the Special Issue Morphology, Phylogeny and Pathogenicity of Fusarium—2nd Edition)

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13 pages, 7084 KB

Open AccessArticle

Efficacies of Conventional Antifungals and Complementary and Alternative Medicine as Single or Combination Therapies Against Candida Biofilms in Recurrent Vaginal Candidiasis: An In Vitro Study

by Yihong Pan, Liumei Ye, Lanqian Chen, Lauren Hermann, Panpan Jin, Yingying Cai, Yali Cheng, Weidan Zhang, Cathy J Watson, David McGiffin, Qiong Luo, Xueqiong Zhu and Yue Qu

J. Fungi 2026, 12(6), 415; https://doi.org/10.3390/jof12060415 - 8 Jun 2026

Abstract

Objectives: Recurrent vulvovaginal candidiasis (RVVC) is a difficult-to-treat infection, most likely due to the growth of Candida biofilms on the human vaginal epithelium. We assessed in vitro efficacy of conventional antifungals and complementary and alternative medicine (CAM) used in clinical settings, and sought [...] Read more.

Objectives: Recurrent vulvovaginal candidiasis (RVVC) is a difficult-to-treat infection, most likely due to the growth of Candida biofilms on the human vaginal epithelium. We assessed in vitro efficacy of conventional antifungals and complementary and alternative medicine (CAM) used in clinical settings, and sought for Candida biofilm-effective single or combination therapies. Methods: Standard broth microdilution assay and XTT (2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide) assay were used for antifungal and anti-biofilm efficacies of three conventional antifungals, and selected CAM including boric acid, povidone-iodine, and allicin (garlic extract), against Candida clinical isolates grown at neutral and acidic pHs respectively. Fractional inhibitory concentration (FIC) indices were assessed to evaluate interactions between fluconazole and different CAM. Viable count-based cell enumeration and confocal laser scanning microscopy (CLSM) were performed to confirm the efficacy of single or combination therapies against Candida biofilms. Results: All selected conventional antifungals and CAM showed efficacies against planktonic Candida cells. Acidic vaginal microenvironments provided agent-specific protection to Candida cells against conventional antifungals and the CAM. Synergistic or additive interactions were observed between fluconazole at serum achievable concentrations and povidone-iodide at topically achievable concentrations against all tested Candida strains. Most antifungal agents except caspofungin had very limited activities against Candida biofilms. Combining fluconazole at 8 mg/L with povidone-iodine at 2048 mg/L effectively killed Candida biofilms in an acidic vaginal microenvironment to a level that is comparable to that of caspofungin. Conclusions: We provided robust in vitro evidence supporting the combinational use of oral fluconazole and topical CAM povidone-iodine against Candida biofilms in managing RVVC. Full article

(This article belongs to the Special Issue Candida Infections and Antifungal Treatment)

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23 pages, 1100 KB

Open AccessReview

Heat Shock Proteins in Medically Relevant Fungal Pathogens: From Molecular Chaperones to Virulence Factors and Therapeutic Targets

by Leonardo Padró-Villegas and Héctor M. Mora-Montes

J. Fungi 2026, 12(6), 414; https://doi.org/10.3390/jof12060414 (registering DOI) - 7 Jun 2026

Abstract

Heat shock proteins (HSPs) are highly conserved molecular chaperones that play a key role in maintaining protein homeostasis and cellular survival under stress conditions. Clinically relevant human pathogenic fungi include opportunistic fungi, dimorphic fungi, dermatophytes, Mucorales, and other pathogenic groups. HSPs, including Hsp90, [...] Read more.

Heat shock proteins (HSPs) are highly conserved molecular chaperones that play a key role in maintaining protein homeostasis and cellular survival under stress conditions. Clinically relevant human pathogenic fungi include opportunistic fungi, dimorphic fungi, dermatophytes, Mucorales, and other pathogenic groups. HSPs, including Hsp90, Hsp70, Hsp60, Hsp40, and Hsp110, are essential for the correct nascent protein folding, aggregation prevention, and degradation of misfolded polypeptides. Fungal pathogens frequently encounter environmental and host-imposed stresses, including oxidative stress, temperature fluctuations, and antifungal treatments. This review synthesizes and critically analyzes current evidence on the role of HSP families in essential processes linked to fungal virulence, including morphogenetic transitions, biofilm formation, maintenance of cell wall integrity, and interactions with host immune cells. Beyond their canonical chaperone functions, HSPs act as central mediators in pathogenic processes, such as morphogenesis transitions, biofilm formation, cell wall integrity, and interactions with host immune cells. Hsp90 stabilizes key signaling proteins involved in stress responses, morphogenesis, and antifungal resistance, while Hsp60 and Hsp70 contribute to mitochondrial function, cell wall integrity, and immune modulation. Disruption of these chaperones impairs growth, reduces virulence, and increases susceptibility to antifungal agents. The rise of antifungal resistance underscores the urgent need for new therapeutic strategies. Targeting fungal HSPs has emerged as a promising approach due to their essential roles in stress tolerance and pathogenesis. Hsp90 inhibitors, including geldanamycin derivatives and other small molecules, have demonstrated the ability to impair fungal growth, reduce virulence traits, and sensitize resistant strains to conventional antifungal drugs. Combining HSP inhibitors with existing antifungal drugs represents a potential strategy to overcome resistance and improve treatment outcomes. This review summarizes the current knowledge on HSPs in pathogenic fungi, focusing on their roles in stress adaptation, virulence, host-pathogen interaction, antifungal resistance, and their potential as targets for novel antifungal therapies. Full article

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

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25 pages, 8928 KB

Open AccessArticle

Diversity of Fusarium Species Causing Storage Rot of Table Beet in the Moscow Region of the Russian Federation

by Svetlana Vetrova, Elena Kozar, Irina Engalycheva, Kseniya Mukhina, Vera Chizhik and Viktor Martynov

J. Fungi 2026, 12(6), 413; https://doi.org/10.3390/jof12060413 - 5 Jun 2026

Abstract

Fusarium fungi are known to infect table beet (Beta vulgaris subsp. vulgaris) plants at various stages of development worldwide. Fusarium root rot, which develops post-harvest during long-term storage, is of particular economic significance. In Russia, there is no up-to-date information about [...] Read more.

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

(This article belongs to the Special Issue Fusarium, Alternaria, and Rhizoctonia: A Spotlight on Fungal Pathogens—3rd Edition)

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21 pages, 15820 KB

Open AccessArticle

Biological Control and Growth-Promoting Potential of the Endophytic Fungus Nigrospora sphaerica Against Anthracnose in Begonia benariensis

by Shuwen Liu, Mian Liu, Jian Liu, Huali Li, Yajiao Sun, Mengyao Wang, Hongliang Zhang, Yunqiang Ma and Junjia Lu

J. Fungi 2026, 12(6), 412; https://doi.org/10.3390/jof12060412 - 5 Jun 2026

Abstract

To explore efficient and sustainable biocontrol resources against anthracnose in Begonia benariensis, endophytic fungi were isolated from healthy host tissues and screened for antagonistic activity against Colletotrichum aotearoa SWBG5. Among 31 isolates, four showed strong inhibition, and the most potent strain, QYN6, [...] Read more.

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

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

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22 pages, 13126 KB

Open AccessArticle

The Role of Mitochondrial Protein UPS1 in Regulating Pathogenicity of Candida albicans

by Qianwen Xu, Changlong Xie, Dinghui Wang, Xiaoxiao Zhu, Wenfan Wei, Xiaojia Niu, Tianming Wang, Hongchen Wang and Daqiang Wu

J. Fungi 2026, 12(6), 411; https://doi.org/10.3390/jof12060411 - 4 Jun 2026

Abstract

The mitochondrial membrane protein UPS1, a conserved intermembrane space protein in Saccharomyces cerevisiae, possesses phosphatidic acid transfer activity and plays a positive regulatory role in processes such as cardiolipin metabolism and transport. The role of UPS1 protein in pathogenic fungi such as [...] Read more.

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

(This article belongs to the Special Issue Fungal Pathogenicity and Host Defense: A Molecular Perspective)

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17 pages, 2636 KB

Open AccessArticle

The Role of ATG8 in Promoting Lipid Accumulation in the Oleaginous Fungus Mucor circinelloides During Nitrogen Limitation

by Hequn Li, Hongjuan Yuan, Bushra Iqbal, Tianyu Wang, Zhen Wang and Huaiyuan Zhang

J. Fungi 2026, 12(6), 410; https://doi.org/10.3390/jof12060410 - 4 Jun 2026

Abstract

Autophagy is a central cellular process that recycles intracellular components and supplies precursors for biosynthesis. As a key regulator of autophagosome formation, autophagy-related protein 8 (ATG8) plays an essential role in macromolecular degradation and in the availability of lipid precursors. However, whether enhanced [...] Read more.

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

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

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Graphical abstract

15 pages, 1929 KB

Open AccessArticle

Establishment of a Visual LAMP Technology and Detection of Cronartium ribicola Infecting Chinese White Pine in Southwestern China

by Xinyi Zhang, Zijia Peng, Ruonan Jing, Xinye Liu, Tauseef Ullah, Min Sheng and Zhongdong Yu

J. Fungi 2026, 12(6), 409; https://doi.org/10.3390/jof12060409 - 4 Jun 2026

Abstract

White pine blister rust disease (WPBR), caused by Cronartium ribicola, ranks among the most destructive pathogens of five-needle pines. We developed a hydroxynaphthol blue (HNB)-based Loop-mediated isothermal amplification (LAMP) assay enabling rapid, visual detection of C. ribicola directly following DNA extraction. LAMP [...] Read more.

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

(This article belongs to the Special Issue Rust Fungi: From Systematics to Sustainable Management)

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3 pages, 160 KB

Open AccessEditorial

Fungal Biotechnology and Application 3.0

by Baojun Xu

J. Fungi 2026, 12(6), 408; https://doi.org/10.3390/jof12060408 - 4 Jun 2026

Abstract

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

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

13 pages, 1735 KB

Open AccessArticle

Coccidioidomycosis Exposure Assessed by Skin Testing and Environmental Factors in Baja California, Mexico

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

J. Fungi 2026, 12(6), 407; https://doi.org/10.3390/jof12060407 - 3 Jun 2026

Abstract

Baja California is the second-highest state in Mexico for hospital discharges attributed to coccidioidomycosis (CM), yet epidemiological information on exposure patterns in affected communities remains limited. To estimate exposure to Coccidioides and assess its association with environmental factors, we conducted intradermal coccidioidin skin [...] Read more.

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

(This article belongs to the Special Issue Basic Science and Clinical Research of Coccidioides and Coccidioidomycosis)

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3 pages, 172 KB

Open AccessCorrection

Correction: Thaochan et al. Fungal-Infected Weeds: A Potential Source of Leaf Spot Disease in Rubber Trees from Southern Thailand. J. Fungi 2025, 11, 220

by Narit Thaochan, Chaninun Pornsuriya, Thanunchanok Chairin, Kodeeyah Thoawan, Putarak Chomnunti and Anurag Sunpapao

J. Fungi 2026, 12(6), 406; https://doi.org/10.3390/jof12060406 - 3 Jun 2026

Abstract

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

(This article belongs to the Special Issue Current Trends in Mycological Research in Southeast Asia)

17 pages, 5864 KB

Open AccessArticle

Synergistic Enhancement of Straw Hydrolysis and Lactic Acid Production in Talaromyces pinophilus Through Combined Random Mutagenesis and Plasmid Reconstruction

by Siyuan Yue, Ya Li, Peng Li, Jing Zeng, Junhui Nie, Cheng Zhang, Tong Wang, Jianjun Guo and Lin Yuan

J. Fungi 2026, 12(6), 405; https://doi.org/10.3390/jof12060405 - 3 Jun 2026

Abstract

Lignocellulosic biorefineries are limited by the high cost of cellulolytic enzymes. Consolidated bioprocessing (CBP), which integrates saccharification and fermentation in one step, offers a solution to this challenge. In this study, a cellulase-hyperproducing mutant of Talaromyces pinophilus, Y117, was generated from the [...] Read more.

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

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

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16 pages, 3275 KB

Open AccessArticle

Identification of Circadian Clock Homologs and Their Rhythmic Expression Differences Among Mating-Type Strains in Morchella sextelata

by Meng-Qian Chen, Jun-Xi Liu, Jia Ling and Xi-Hui Du

J. Fungi 2026, 12(6), 404; https://doi.org/10.3390/jof12060404 - 2 Jun 2026

Abstract

The circadian clock is a widespread rhythmic phenomenon across organisms, characterized by distinct gene expression patterns and behaviors at specific times of the day. Extensive genetic studies in the model fungus Neurospora crassa have yielded critical insights into the components and molecular mechanisms [...] Read more.

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

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

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17 pages, 11075 KB

Open AccessArticle

In Vivo Evaluation of a Nanoemulsion-Delivered Chromium(III)–Triazole Complex Against Fluconazole-Resistant Candida albicans

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

J. Fungi 2026, 12(6), 403; https://doi.org/10.3390/jof12060403 - 2 Jun 2026

Abstract

Candida albicans remains one of the leading causes of invasive fungal infections and is recognized as a critical-priority pathogen by the World Health Organization. The increasing emergence of resistance to azole antifungals such as fluconazole highlights the need for alternative therapeutic strategies. In [...] Read more.

Candida albicans remains one of the leading causes of invasive fungal infections and is recognized as a critical-priority pathogen by the World Health Organization. The increasing emergence of resistance to azole antifungals such as fluconazole highlights the need for alternative therapeutic strategies. In this study, we evaluated the antifungal potential of a chromium(III)–triazole coordination complex (CrL1) against C. albicans. In vitro susceptibility testing showed that CrL1 exhibited notable antifungal activity against the fluconazole-resistant strain with low cytotoxicity in murine macrophages. To facilitate aqueous dispersion and enable in vivo administration, CrL1 was incorporated into an oil-in-water nanoemulsion (NE-CrL1). The antifungal activity of NE-CrL1 was evaluated in a murine model of invasive candidiasis. In mice infected with a fluconazole-resistant C. albicans strain, treatment with NE-CrL1 reduced renal fungal burden and was associated with attenuation of histopathological alterations and changes in local inflammatory responses. Although the present study has limitations, including the absence of mechanistic assays and additional physicochemical characterization, these results suggest in vivo antifungal activity of NE-CrL1 and warrant further preclinical evaluation against drug-resistant Candida infections. 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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16 pages, 5813 KB

Open AccessArticle

In Vivo Characterization of the dia Biosynthetic Gene Cluster Reveals Diaporthinic Acid as Its Main Product

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

J. Fungi 2026, 12(6), 402; https://doi.org/10.3390/jof12060402 - 1 Jun 2026

Abstract

The biosynthetic gene cluster (BGC) responsible for producing diaporthinic acid has remained genetically unassigned despite repeated isolation of this metabolite from several fungal species. In this study, we activated the dia BGC in Trichoderma reesei by overexpressing the cluster-associated zinc cluster protein DiaR1 [...] Read more.

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

(This article belongs to the Special Issue Fungal Biosynthesis)

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15 pages, 2749 KB

Open AccessArticle

Heterologous Expression Unexpectedly Activates the Host Cryptic Genes in Aspergillus nidulans and Enables the Discovery of Novel Natural Products

by Cong Liu, Yinan Hao, Siyuan Qi and Jian Bai

J. Fungi 2026, 12(6), 401; https://doi.org/10.3390/jof12060401 - 1 Jun 2026

Abstract

Aspergillus nidulans, a model filamentous fungus endowed with well-established genetic tools and a repertoire of cryptic secondary metabolite biosynthetic gene clusters (BGCs), is extensively exploited as a microbial chassis for heterologous biosynthesis. Mining of its secondary metabolites facilitates the discovery of novel [...] Read more.

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

(This article belongs to the Collection Bioactive Fungal Metabolites)

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