The study on macrofungi diversity in Motuo county, Xizang Autonomous Region, was conducted from 2021 to 2025, and 2286 vouchered fungal collections were obtained from local forest ecosystems. A total of 534 species, representing two phyla, eight classes, 28 orders, 96 families, and 241 genera were identified according to the morphological characters and molecular evidence. The orders Agaricales (174), Polyporales (91), Xylariales (74) and Russulales (69) were the dominant orders, collectively accounting for 76.40% of the total species listed. Fifteen dominant families with more 10 species each were identified, namely Russulaceae (58), Polyporaceae (37), Xylariaceae (34), Hypoxylaceae (30), Boletaceae (17), Psathyrellaceae (13), Mycenaceae (12), Agaricaceae (11), Hymenochaetaceae (11), Auriculariaceae (11), Stromphariaceae (11), Marasmiaceae (10), Omphalotaceae (10), Steccherinaceae (10), and Pleurotaceae (10), which occupied approximately 53.37% of the total species. Additionally, 21 dominant genera with more than five species each were recorded, including Russula (27), Xylaria (25), Hypoxylon (17), Lactarius (16), Lactifluus (15), and other 16 genera (101), accounting for around 37.64% of all species. We further compared species composition, distribution patterns, dominant taxa and trophic modes of macrofungi between tropical monsoon forest and evergreen broad-leaved forest. A total of 439 species (82.21%) were endemic to a single forest type, while only 95 species (17.79%) were shared by the two forest habitats. The evergreen broad-leaved forest exhibited slightly higher macrofungal species richness than the tropical monsoon forest in Motuo County. Moreover, it harbored a higher proportion of ectomycorrhizal fungi (25.32% vs. 14.06%) and a lower proportion of saprotrophic fungi (71.52% vs. 83.71%). This study fills the research gap in systematic macrofungal diversity investigation in Motuo County, and provides fundamental data for revealing the evolutionary mechanisms of macrofungi on the Qinghai-Xizang Plateau margin and clarifying the biodiversity formation process across the Himalayan region.
Colombia ranks third among South American countries in terms of fungal diversity (including lichens); however, the lichen diversity of tropical dry forest ecosystems, particularly those in the Caribbean region, remains relatively understudied. The typical lichens found in tropical dry forest are crustose/corticolous microlichens (adapted to the xerophytic environments), which produce certain secondary metabolites that act as protective agents against ultraviolet radiation/decomposition/depredators; nonetheless, secondary metabolites of most of these lichens have not yet been described. This study focused on the compositional analysis and the extrolite content of seven lichens: Cryptothecia sp., Cryptothecia scripta, Graphis dendrogramma, Leucodecton occultum, Helminthocarpon leprevostii, Pyrenula ochraceoflava, and Allographa seminuda, using thin-layer chromatography, high-performance liquid chromatography-diode array detector, liquid chromatography-electrospray-mass spectrometry and/or liquid chromatography-tandem mass spectrometry. The main findings were: (i) two Cryptothecia spp. contained 2'-O-methylperlatolic acid (97%) or ovoic/gyrophoric acids (67%/28%); (ii) two specimens of G. dendrogramma had stictic/norstictic acids (33-60%/40%); (iii) three samples of L. occultum were characterized by norstictic acid (42-82%); (iv) schizopeltic/3-O-methylschizopeltic acids (58%/32%) were found in one specimen of H. leprevostii; (v) one sample of P. ochraceoflava was represented by unidentified xanthone-type metabolite/7-chloroemodin/ (26%/24%); and, (vi) the extrolite content in one specimen of A. seminuda was negligible. Finally, 10 of the 11 lichen specimens studied from the Department of Atlántico within Colombia contained certain secondary metabolites that likely reflect the common types of extrolites, namely, di-/tri-depsides, depsidones, dibenzofurans, and chlorinated anthraquinones/xanthones, which could be hypothetically related to the evolutionary patterns of this type of organism.
Polygonatum cyrtonema (P. cyrtonema) Hua is an important economic crop with both edible and medicinal value. However, frequent root rot severely restricts its industrial development, resulting in sharp yield reduction and quality deterioration. To clarify the primary pathogenic fungi causing root rot of P. cyrtonema Hua, 58 fungal strains from naturally diseased P. cyrtonema Hua plants in different habitats were isolated in this study. By combining morphological observation and molecular identification based on 18S rDNA and ITS rDNA sequences, the species of 22 pathogenic fungi were identified, among which 10 strains belonged to the genus Fusarium, accounting for 45.45% of the identified isolates. The pathogenicity of 21 pathogenic fungi was verified according to Koch's postulates, with findings indicating that Fusarium species exhibited significant pathogenic potential. Meanwhile, six previously identified endophytic Paenibacillus strains isolated from P. cyrtonema Hua were employed to perform dual culture assays and antifungal evaluations of their fermentation supernatants against representative strains including F. concentricum F2, Neopestalotiopsis sp. F3 and F. oxysporum F8. The results indicated that the antagonistic activity exhibited by the six strains exceeded 50%, with the inhibition rates of their fermentation supernatants against strains F2, F3 and F8 surpassing 73%. This study confirmed that Fusarium is the dominant pathogenic fungal group causing root rot of P. cyrtonema Hua. Furthermore, highly effective antagonistic endophytes were preliminarily identified, offering candidate strains and a theoretical foundation for the green management of root rot in P. cyrtonema Hua.
Despite increasing interest in fungal remediation systems for the treatment of persistent contaminants, the mechanisms governing fungal transformation of halogenated organic compounds remain poorly resolved. The aim of this study was to determine whether the transformation of halogenated nitrophenols is driven by isolated extracellular enzymes and cofactor-dependent oxidative activity or instead reflects coordinated system-level fungal metabolism. To address this question, we investigated the transformation of 2-chloro-4-nitrophenol (2C4NP) and 5-fluoro-2-nitrophenol (5F2NP) by ascomycete fungi Caldariomyces fumago (C. fumago) and Curvularia sp. under varying nutrient and cofactor conditions. Whole-culture transformation, crude supernatant activity, purified enzyme assays, intracellular detoxification responses, and genome-resolved functional annotation were integrated to evaluate the relative contributions of extracellular and intracellular processes. Transformation was strongly dependent on fungal species, substrate identity, nutrient availability, and cofactor composition. C. fumago achieved complete transformation of 2C4NP and up to 85.3% transformation of 5F2NP, whereas Curvularia sp. exhibited strict Na3VO4-dependent transformation of 5F2NP. Crude supernatants retained partial transformation capacity, achieving ~40-45% substrate depletion under conditions supporting whole-culture activity. Purified chloroperoxidase and laccase showed negligible independent activity and did not reproduce whole-culture transformation behavior. Lignin peroxidase activity was consistently induced during contaminant exposure and peaked during periods of maximum transformation. Cytochrome P450 inhibition did not prevent transformation. Baseline glutathione S-transferase activity was detected in both fungi, and comparative genome analysis identified conserved intracellular detoxification-associated enzyme alongside divergent extracellular oxidative enzyme repertoires. Together, these findings demonstrate that transformation of halogenated nitrophenols by fungi cannot be explained by isolated extracellular enzymes alone but is consistent with coordinated extracellular and intracellular system-level metabolism. These findings highlight an underexplored role for integrated fungal metabolic systems in bioremediation and provide a mechanistic basis for developing a scalable fungal platform for treatment of persistent halogenated contaminants.
Candidozyma auris (formerly Candida auris) is a critical-priority multidrug-resistant pathogen. Comparative clinical data on first-line echinocandins-micafungin and anidulafungin-in C. auris candidemia remain limited. This retrospective cohort study compared clinical outcomes of micafungin and anidulafungin in adult patients with C. auris candidemia treated between January 2024 and December 2025 at three affiliated hospital campuses in Istanbul, Türkiye. Propensity score matching (PSM) using a 1:1 nearest-neighbor algorithm was performed to balance baseline characteristics. Outcomes included 30-day (primary) and 14-day all-cause mortality, microbiological response, end-of-therapy (EOT) response, relapse, and drug-induced liver injury assessed by the Roussel Uclaf Causality Assessment Method (RUCAM). Among 154 included patients (micafungin, n = 94; anidulafungin, n = 60), no echinocandin resistance was detected. After PSM (55 matched pairs), 30-day all-cause mortality was identical between groups (41.8% vs. 41.8%; mOR 1.00, 95% CI 0.43-2.31; p = 1.000). Fourteen-day all-cause mortality (16.4% vs. 18.2%; p = 0.763), microbiological response (94.5% vs. 90.9%; p = 0.480), EOT response (74.5% vs. 67.3%; p = 0.346), and relapse (12.7% vs. 10.9%; p = 0.763) did not differ significantly between groups. RUCAM-based hepatic safety profiles were descriptively comparable. Micafungin and anidulafungin showed comparable observed outcomes in C. auris candidemia in this cohort.
The date palm is an icon of resilience in arid ecosystems, yet its survival is increasingly challenged by climate change and disease. Belowground, a powerful symbiotic solution exists: Arbuscular Mycorrhizal Fungi (AMF). This review synthesizes eight decades of research, from the pioneering observations in 1940 to contemporary studies, to build a comprehensive understanding of this critical partnership. We explore the vast, largely untapped biogeographic diversity of AMF, from the oases of North Africa to the deserts of Arabia, and detail the physiological and molecular mechanisms that underpin AMF-mediated tolerance to salinity, drought, and pathogens like Fusarium oxysporum. Evidence from individual studies indicates that AMF inoculation can increase plant biomass by more than 100% under specific stress conditions and substantially reduce Bayoud disease severity, although the magnitude of these effects varies with fungal identity, host cultivar, and experimental conditions. We then bridge this fundamental knowledge to cutting-edge biotechnological applications, including the design of precision inoculants and scalable in vitro production systems. We conclude by outlining a strategic vision for future research, focusing on the development of cultivar-specific consortia and the integration of multi-omics, to translate this fundamental ecological interaction into robust, sustainable agricultural solutions.
Grape white rot, primarily caused by Coniella diplodiella and Coniella vitis, is a major fungal disease threatening grape production worldwide, with C. vitis more prevalent in China. Temperature is a critical environmental factor influencing pathogen development and disease outbreaks, but how temperature influences the biological characteristics, pathogenicity, and fungicide sensitivity of these two pathogens remains limited. This study aimed to evaluate the effects of temperature on mycelial growth, conidial germination, extracellular enzyme activity, pathogenicity, and fungicide sensitivity of C. diplodiella 2019 and C. vitis GP1. The growth, conidial germination, extracellular enzyme production, pathogenicity, and fungicide control efficacy against the two pathogens were systematically evaluated under different temperature conditions (5, 15, 25, 35, and 45 °C). Results showed temperature significantly affected all investigated traits in both species. Optimal mycelial growth, conidial germination, and pathogenicity occurred at moderate temperatures (25 °C), whereas extreme temperatures (5 °C and 45 °C) significantly markedly inhibited fungal development and disease severity. C. vitis GP1 showed greater adaptability and consistently higher pathogenicity than C. diplodiella 2019 across the tested temperature range. Fungicide sensitivity also differed between the two species, and the inhibitory activity of difenoconazole was significantly reduced under temperature extremes, indicating that fungicide performance is influenced by environmental temperature. These findings demonstrate that temperature regulates both fungal development and fungicide efficacy, providing new insights into the temperature-dependent epidemiology of grape white rot and a scientific basis for optimizing disease management under changing environmental conditions.
To determine the in vitro anti-Aspergillus activity of monoclonal antibodies (mAb) 1D2 and 4E4 alone and in combination with voriconazole, posaconazole, amphotericin B and caspofungin and to assess whether selected antimicrobial agents interfere with 1D2/4E4 binding to Aspergillus antigens. The in vitro antifungal activity of 1D2, 4E4 and all antifungal drugs was determined in a microplate-based assay. The anti-Aspergillus activity of mAb 1D2 and 4E4 in vitro alone and in combination with voriconazole, posaconazole, amphotericin B and caspofungin was determined using a checkerboard assay. Finally, a competitive ELISA was used to assess whether selected antifungal and antibacterial agents interfered with the binding of 1D2 and 4E4 to immobilised Aspergillus antigens. The in vitro interactions of 1D2 with voriconazole, posaconazole, amphotericin B and caspofungin against four different Aspergillus species were all additive, with a fractional inhibitory concentration index (FICI) between 0.5 and 1. Similarly, the in vitro interactions of 4E4 with these same four antifungals against Aspergillus fumigatus (A. fumigatus) were all additive. An additive interaction was also detected for 4E4 and caspofungin against all four Aspergillus species tested. However, no interaction was found for 4E4 combined with voriconazole, posaconazole and amphotericin B against Aspergillus flavus (A. flavus), Aspergillus niger (A. niger) and Aspergillus terreus (A. terreus), with an FICI between 1 and 4. Moreover, none of the tested antifungal or antibacterial agents significantly interfered with 1D2/4E4 binding to Aspergillus antigens in the competitive ELISA. In this study, 1D2 and 4E4 showed in vitro growth-inhibitory activity against the tested Aspergillus isolates. Specifically, 1D2 showed additive interactions with the tested antifungal agents across all tested species, whereas 4E4 showed a more species- and drug-dependent interaction profile. The tested antimicrobial agents did not measurably interfere with 1D2/4E4 binding under the competitive ELISA conditions examined. Overall, these findings provide preliminary in vitro evidence supporting the further evaluation of 1D2 and 4E4.
Endophytic fungi from Malaysian medicinal plants constitute a metabolically prolific yet underexplored reservoir for natural product discovery. This scoping review of 56 studies published between 2015 and 2025 identified a fundamental methodological divergence within the field: while phenotypic bioactivity screening dominates the literature (>87% of studies), it is weakly supported by chemical characterization (<25%) and entirely disconnected from genomic investigation (0% biosynthetic gene cluster studies). This phenotype-first paradigm has largely confined the field to descriptive reporting, limiting mechanistic understanding and translational potential. Collectively, the evidence reveals a substantial disconnect between reported bioactivities and their underlying biosynthetic foundations. To address this limitation, a practical genotype-to-phenotype workflow is proposed that integrates strain prioritisation, multi-omics-guided activation, chemical mapping, and mechanism-oriented validation. By linking genomic potential with metabolite production and biological function, this framework provides a roadmap for advancing fungal natural product discovery beyond conventional phenotype-driven screening. Adoption of such approaches may improve the identification of chemically novel and biologically relevant metabolites while supporting the sustainable development of Malaysia's endophytic fungal resources for biotechnological and pharmaceutical applications.
This study assessed the pathogenicity and toxigenic chemotypes of Fusarium species causing maize stalk rot. In addition, the ability of selected strains of Trichoderma longibrachiatum, T. koningiopsis, and T. gamsii to inhibit mycelial growth and the biosynthesis of Fusarium mycotoxins was evaluated. Pathogenicity assays indicated that F. graminearum, F. verticillioides, F. boothii, and F. subglutinans, previously characterized via morphological and molecular analyses, caused maize stalk rot symptoms with varying degrees of aggressiveness. PCR-based detection of mycotoxin biosynthetic genes revealed distinct toxigenic chemotypes among the Fusarium isolates, including fumonisin (FUM), 3-acetyl-deoxynivalenol (3-AcDON), zearalenone (ZEN), and beauvericin (BEA). After 7 days of co-culture on potato dextrose agar, all Trichoderma strains significantly inhibited the growth of Fusarium mycelia. The Fusarium pathogen inhibition rate ranged between 28.30 and 71.77%. T. longibrachiatum grew over the mycelium of the pathogen and sporulated. In rice-based co-cultured assays, Trichoderma strains inhibit FUM biosynthesis by about 74% to 92%, 3-AcDON by about 32% to 100%, ZEN by about 11% to 90%, and BEA by about 6% to 57%, depending on the pathogen-antagonist combination. The maximum reduction in mycotoxin production (100% for 3-AcDON) exceeded the maximum inhibition of mycelial growth (71.77%), suggesting that Trichoderma may repress mycotoxin biosynthesis via mechanisms independent of simple growth inhibition. These findings provide a basis for developing green biocontrol strategies for maize stalk rot.
The bZIP type transcription factors (bZIPs) are global regulators governing vegetative growth, development, stress defense and secondary metabolism, including mycotoxin production in filamentous fungi. In this work, we constructed and phenotypically characterized gene deletion and complementation mutants of some bZIPs, including Afap1, AflatfA, LziP, AflatfB and bZIP6 in Aspergillus flavus. Environmental and fungicide stress responses, as well as aflatoxin production of the mutants in both surface cultures and infected maize kernels, were studied. Phenotypic characterization of the mutants revealed that Afap1 and AflatfA were involved in the oxidative (H2O2, menadione, tert-butyl hydroperoxide), cell wall integrity (Congo Red) and heavy metal (CdCl2) stress responses of A. flavus. In addition, the Afap1 and AflatfA gene deletions decreased the diamide and prothioconazole tolerances of the fungus, respectively. The ΔLziP strain showed increased growth in the presence of diamide, while reduced colony diameters were observed after exposure to CdCl2 and fludioxonil. The ΔAflatfB gene deletion mutant was also sensitive to tBOOH, while azoxystrobin and prothioconazole fungicides significantly inhibited the growth of ΔbZIP6. When aflatoxin (AFB1) production was measured in surface cultures, decreased AFB1 levels were detected only in the ΔAflatfA gene deletion mutant strain. However, in corn kernel infection assays, the ΔAfap1, ΔAflatfA, and ΔAflatfB mutants were characterized by significantly reduced aflatoxin production, while the deletion of bZIP6 almost completely abolished AFB1 biosynthesis. Our results suggest that Afap1 and AflatfA appear to be promising targets for the development of new antifungal agents, as their inhibition may increase the sensitivity of A. flavus to environmental stress, simultaneously reducing the aflatoxin production of the fungus and the use of azoles (AflatfA) in the antifungal protection of maize. In addition, bZIP6 may also be considered as an attractive target for further studies when aiming to eliminate aflatoxin production and minimize the use of azoxystrobin and prothioconazole in maize crop protection.
With growing global recognition of the nutritional value of blueberries, their cultivation has expanded significantly across various regions. However, gray mold disease, caused by Botrytis cinerea, remains a major challenge in postharvest storage and transportation. In this study, two strains-Trichoderma asperellum BBR-A and B. cinerea BC7-1-were isolated and identified from the blueberry rhizosphere and diseased leaves. Co-culture assays demonstrated that BBR-A exhibits potent antagonistic activity, rapidly suppressing the growth of the pathogen BC7-1 through mycoparasitic behaviors, including hyphal coiling, penetration, and degradation. Transcriptomic analysis revealed that upon interaction with BC7-1, BBR-A undergoes extensive reprogramming of carbon metabolism, with coordinated upregulation of genes encoding degradative enzymes, transporters, and stress-related proteins. Congo red staining and enzymatic activity assays further confirmed the enhanced secretion of extracellular enzymes and the increased cell wall-degrading capacity of BBR-A. On detached blueberry leaves, seedlings, and fruits, BBR-A treatment significantly delayed lesion expansion and reduced disease severity. Metabolomic profiling further demonstrated that BBR-A induces the marked accumulation of phenolic compounds, polyamines, and other antioxidant- and immunity-related metabolites in blueberry leaves, thereby maintaining host redox homeostasis and enhancing stress resilience. Collectively, these findings indicate that T. asperellum BBR-A effectively suppresses B. cinerea through multiple mechanisms, including antagonism, mycoparasitism, metabolic regulation, and the induction of plant defense responses, highlighting its strong potential for development as a biocontrol agent against blueberry gray mold.
Invasive candidiasis, predominantly driven by multidrug-resistant Candida species and intractable biofilms, represents an escalating global health crisis with mortality rates rivaling major infectious diseases. The clinical efficacy of conventional antifungal agents-azoles, polyenes, and echinocandins-is severely compromised by poor tissue penetration, dose-limiting systemic toxicity, and the rapid evolution of complex resistance mechanisms. Here, we review the two-decade structural evolution of nanotechnological interventions designed to overcome these pharmacological and biological barriers. We systematically analyze advanced nanosystems, including lipid-based formulations, natural polymers, and biogenic metallic nanostructures, highlighting their capacity to penetrate the dense extracellular polymeric substance (EPS), combat potential fungal 'nano-resistance', and significantly reduce metabolically dormant persister cell populations. The literature search was performed using the electronic databases PubMed, Scopus, Web of Science, and Google Scholar. Publications indexed between 2015 and 2025 were primarily considered, while seminal studies published before 2015 were included when necessary to provide historical context and foundational knowledge. We place specific emphasis on next-generation biomimetic and exosome-inspired nanocarriers, which significantly reduce systemic host toxicity while maximizing targeted antifungal efficacy. In this context, the synergistic integration of smart nanocarriers to actively disassemble fungal resistance networks, such as the target of rapamycin (TOR) signaling pathway and sphingolipid biosynthesis. Finally, we outline a strategic roadmap to bridge the translational "Valley of Death". By prioritizing manufacturing standardization, comprehensive long-term biosecurity profiling, and rationally designed biomimetic platforms, we propose an alternative way to outpace the evolutionary adaptations of fungal pathogenesis and translate these innovations into the clinic.
Chagas disease, caused by Trypanosoma cruzi, remains a neglected tropical infection with limited therapeutic options and significant drug-related toxicity. Endophytic fungi are emerging as sustainable sources of bioactive metabolites with potential antiparasitic activity. In this study, endophytic fungi isolated from the medicinal plant Lippia alba were screened for trypanocidal activity. Crude extracts from seven isolates were tested against epimastigote and amastigote forms of T. cruzi, together with cytotoxicity assays in J774A.1 murine macrophages. The crude extract of Montagnula sp. exhibited the strongest trypanocidal activity (IC50 = 22.1 ± 0.5 μg/mL for epimastigotes and 29.2 ± 3.0 μg/mL for amastigotes), comparable to benznidazole, with low toxicity (CC50 > 600 μg/mL) and a high selectivity index (>26). Morphological and mitochondrial analyses demonstrated preservation of host-cell integrity and mitochondrial function, in contrast to the reference drug. UHPLC-ESI-HRMS profiling identified phenolic and terpenoid compounds, including caffeic acid, ferulic acid, naringenin, apigenin, and ursolic acid, which may underlie the observed biological activity. To our knowledge, this study provides the first evidence of trypanocidal activity in an endophytic Montagnula species and highlights endophytic fungi as promising platforms for the discovery of novel anti-T. cruzi agents.
Inonotus is a globally distributed genus within the Hymenochaetales, with certain species widely used in traditional herbal medicine. Here, a new species, I. chrysosporus, was identified as a sister clade to I. hispidus based on morphological, molecular, and vegetative incompatibility characteristics. The growth of I. chrysosporus on two substrate formulations-(1) 60% (wt:wt) Dicranopteris dichotoma, mixed with 18% corncob, 20% wheat bran, 1% gypsum, and 1% lime (GS), and (2) 78% sawdust wood chips (SS)-was compared in terms of the accumulation of bioactive components and the corresponding biological activities. On both substrates, I. chrysosporus exhibited high nutritional value, with high protein and mineral contents, low fat content, and a high proportion of medicinal amino acids. However, significant variations in the profiles of active constituents were observed, with cultivation on the GS formulation promoting the accumulation of bioactive polysaccharides, whereas growth on SS resulted in the accumulation of polyphenols. Aqueous extracts of I. chrysosporus mushrooms derived from both growth substrates exhibited potent antioxidant activities. Correlation analyses indicated that polysaccharide content was positively associated with ·OH scavenging activity and that polyphenol content was correlated with strong DPPH radical scavenging activity and high ferric-reducing antioxidant power (FRAP). This study provides a foundation for the comprehensive development and utilization of the newly described I. chrysosporus species.
Sonoplasma treatment (SPT), which combines hydrodynamic cavitation with low-temperature plasma discharge in water, has been proposed as an advanced oxidation process for reducing biological contamination. By generating physical stressors and reactive oxygen species, including hydrogen peroxide (HP), SPT may inactivate microorganisms, but its effects on stress-resistant filamentous fungi remain insufficiently characterized. We examined two phytopathogenic fungi with contrasting pigmentation: melanized Alternaria alternata and hyaline Fusarium solani. Spore suspensions were exposed to direct and indirect SPT at 30 kHz, and viability, biomass accumulation, conidial production, allelopathic activity, and pigmentation-associated spectral responses were assessed immediately after treatment and after storage. Fungus F. solani showed greater susceptibility, with reduced colony-forming capacity and suppressed biomass production, although surviving propagules showed increased sporulation. In contrast, A. alternata maintained viable growth under the tested conditions and showed stimulation of growth-related and reproductive endpoints, together with darker colony pigmentation. These responses are consistent with pigmentation-associated tolerance to SPT-induced physical and oxidative stress, but do not establish melanin as the sole causal mechanism. SPT efficacy against filamentous fungi is therefore species-dependent and may be limited when resistant melanized taxa are present.
Species of Talaromyces affect human societies in many different ways. Infrageneric classifications of the genus at series level had been established in only two sections, Subinflati and Trachyspermi. In this study, phylogenies of Talaromyces were reconstructed section by section based on separate or concatenated multi-locus datasets: beta-tubulin (BenA), calmodulin (CaM) and RNA polymerase II second-largest subunit (RPB2). Fifty series belonging to nine sections were accordingly recognized, i.e., one in sections Brunneospori and Tenues, two in sections Bacillispori and Helici, three in sect. Subinflati, four in sect. Purpurei, five in sections Islandici and Trachyspermi, and 27 in the speciose section Talaromyces. Among them, forty series were newly established. Three new species were determined phylogenetically and morphologically, i.e., T. fujianensis sp. nov. in sect. Islandici, T. heilongjiangensis sp. nov. in sect. Talaromyces and T. tapisciae sp. nov. in sect. Subinflati. Additionally, three new Chinese records were noted: T. angelicae, T. gautengensis and T. rogersiae. The findings of new species and new records reveal the high diversity of the genus in China. The updated taxonomy of Talaromyces at series level will facilitate a more accurate species identification by means of phylogenetic analysis at a smaller scale, and benefit future studies involving this group of fungi.
Ilyonectria is a common soil-inhabiting fungal genus that comprises numerous plant phytopathogenic species capable of infecting a wide array of crops, medicinal herbs, and horticultural plants. However, the lack of a reliable and efficient genetic transformation method has severely hindered the elucidation of the pathogenic mechanisms of Ilyonectria pathogens. In this study, we established an efficient protoplast-mediated genetic transformation method for two dominant Panax root rot pathogens, I. robusta and I. vredehoekensis. Key parameters governing high-quality protoplast preparation, including mycelium culture time, enzyme composition, osmotic stabilizer type, digestion speed, and digestion time, were systematically optimized. Subsequently, orthogonal experiments were conducted to optimize the PEG-CaCl2-mediated transformation conditions and to screen regeneration media for protoplasts. The optimal enzymatic system is composed of 20 mg/mL driselase and 10 mg/mL lysing enzyme, with 0.7 M NaCl as the osmotic stabilizer. Under these conditions, high-viability and high-quality protoplasts were obtained from I. vredehoekensis after 3 h of digestion at 150 rpm, and from I. robusta after 2 h of digestion at 100 rpm, yielding 5.52 × 107 CFU/mL and 5.75 × 107 CFU/mL protoplasts, respectively. Efficient transformation was achieved using a mannitol-prepared STC buffer mediated by 40% PEG4000. PCR and fluorescence microscopy verified positive transformants. Additionally, pathogenicity assays showed no significant differences in virulence between the transformed and wild-type strains, suggesting that the transformation procedure did not alter virulence. To the best of our knowledge, this is the first study to successfully establish genetic transformation methods for I. robusta and I. vredehoekensis, providing an essential technical platform for functional gene analysis, pathogenicity studies, and host-pathogen interaction research. In addition, the optimized transformation strategy may serve as a valuable reference for studies on other Ilyonectria species.
Fusarium fujikuroi is an industrial producer of gibberellic acid (GA3), a phytohormone of agricultural interest. Despite the high concentration of nutrients used for its production, GA3 yields remain low, highlighting the importance of identifying the major metabolites synthesized during GA3 synthesis. Therefore, the principal aim of this work was to evaluate organic acid production during F. fujikuroi batch cultures by determining GA3 production and organic acid profiles using Fourier transform infrared spectroscopy (FTIR) and liquid chromatography (HPLC) analysis. Significant differences in compound quantification were found; five organic acids, namely lactic, malic, citric, succinic and maleic, were detected by HPLC (in g/L: 101.09, 10.66, 2.80, 6.94 and 1.07, respectively). In addition, eight organic acids were determined by FTIR, namely lactic, butyric, pyruvic, fumaric, malic, succinic, maleic and oxalic (in g/L: 62.97, 19.19, 11.92, 7.54, 2.30, 4.36, 1.25 and 1.06 g/L, respectively). GA3 production was also quantified, reaching nearly 5.0 g/L as determined by HPLC and UV-Vis, and FTIR yielded 2.20 g/L. This report found that the low yields obtained in GA3 production are related to the side conversion of raw materials into organic acids as byproducts. In addition, the FTIR technique can be employed as an innovative strategy for the quantification of metabolites to provide relevant information on F. fujikuroi metabolic regulation. This enables the spread of its application as a biotechnological tool in high-value-added processes with potential for industrial-scale GA3 production.
Rust, caused by the fungus Uromyces pisi, is the most harmful disease of peas in temperate regions. It is necessary to search for sources of resistance with different defense mechanisms in the pea gene pool. A set of 38 Pisum sativum accessions of various origin was studied in Western Siberia in 2021-2024. The aim of the research was to assess the accessions in the field and under controlled conditions using seedlings and adult plants, as well as to study the interaction of U. pisi with resistant varieties, and to determine genetic control of rust resistance. All accessions showed partial (incomplete) resistance to rust in the field. A set of 10 resistant varieties was used for studying U. pisi interaction with peas using cytological methods. The protective mechanisms of Russian varieties led to the inhibition of 50-90% spores on leaf surfaces before penetration into the stomata, and a part of the small colonies died without hypersensitive reaction in the tissues. Hydrogen peroxide and phenolic compounds with red and green autofluorescence appeared by the stage of sporogenesis. Five varieties showed adult resistance to rust. A hybridological analysis revealed monogenic dominant control of resistance in two varieties, and digenic control in two others. The information obtained expands the understanding of the partners' interaction in the pathosystem 'U. pisi-P. sativum', and can also be used for breeding pea varieties with different resistance mechanisms.