Search Results (31)

Fungal communities in high plateau ecosystems remain understudied despite their crucial roles in soil ecosystems, and yeasts inhabiting extreme regions have potential for industrial and biotechnological applications. We studied the fungal diversity in soils across 14 Chilean Altiplano sites using amplicon-based metagenomics and isolation of yeasts to assess their growth under various conditions and hydrolytic enzyme secretion. Using the metagenomic approach, the Ascomycota and Basidiomycota phyla were found to be the most abundant (85% and 8%, respectively). Unclassified families and genera prevailed at six and ten sites, respectively. At the other sites, the most abundant families included Cladosporiaceae, Teratosphaeriaceae, and Sporormiaceae, and the genera Oleoguttula, Coniochaeta, and Peziza. Biodiversity indices did not correlate with the soil’s geographic origin, organic matter content, humidity, or pH. Most isolated yeasts belong to the Naganishia, Holtermanniella, and Vishniacozyma genera, growing at temperatures ranging from 4 °C to 26 °C. Most isolates could use glucose, sucrose, and maltose as carbon sources and exhibited amylase, esterase, pectinase, and protease activities at 30 °C and below. Our results indicate that the evaluated soil physicochemical parameters do not explain the fungal distribution in the Altiplano and highlight the region as a reservoir of unknown fungi, including yeasts with industrially relevant enzymes. Full article

Protoplast-based transformation is a vital tool for genetic studies in fungi, yet no protoplast method existed for P. sclerotiorum-scaumcx01 before this study. Here, we optimized protoplast isolation, regeneration, and transformation efficiency. The highest protoplast yield (6.72 × 10⁶ cells/mL) was obtained from liquid mycelium after 12 h of enzymatic digestion at 28 °C using Lysing Enzymes, Yatalase, cellulase, and pectinase. Among osmotic stabilizers, 1 M MgSO₄ yielded the most viable protoplasts. Regeneration occurred via direct mycelial outgrowth and new protoplast formation, with a 1.02% regeneration rate. PEG-mediated transformation with a hygromycin resistance gene and GFP tagging resulted in stable GFP expression in fungal spores and mycelium over five generations. LC/MS-based metabolomic analysis revealed significant changes in glycerophospholipid metabolism, indicating lipid-related dynamics influenced by GFP tagging. Microscopy confirmed successful colonization of tomato roots by GFP-tagged scaumcx01, with GFP fluorescence observed in cortical tissues. Enzymatic (cellulase) seed pretreatment enhanced fungal colonization by modifying root surface properties, promoting plant–fungal interaction. This study establishes an efficient protoplast transformation system, reveals the metabolic impacts of genetic modifications, and demonstrates the potential of enzymatic seed treatment for enhancing plant–fungal interactions. Full article

Barley leaf stripe, caused by Pyrenophora graminea, significantly reduces yield. Polygalacturonase, a key fungal pectinase, facilitates cell wall degradation for nutrition acquisition and colonization. To determine whether P. graminea contains polygalacturonase (PgPG)-encoding genes and their role in pathogenicity, four PgPG genes (PgPG1–PgPG4) were identified in the P. graminea genome. Quantitative RT-PCR revealed that PgPG1 had the highest inducible expression during barley infection, suggesting its critical vital role in pathogenesis. PgPG1 was silenced and overexpressed in P. graminea QWC (wild-type) using CaCl₂-PEG4000-mediated protoplast transformation. The PgPG1 RNAi mutants exhibited slower growth, while overexpression mutants grew faster. Relative to the wild-type, the disease incidence of Alexis, a highly susceptible barley variety, decreased by 62.94%, 42.19%, 45.74%, and 40.67% for RNAi mutants, and increased by 12.73%, 12.10%, 12.63%, and 10.31% for overexpression mutants. Pathogenicity analysis showed decreased disease incidence with PgPG1 RNAi mutants and increased severity with overexpression mutants. Trypan blue staining and polygalacturonase activity assays confirmed that overexpression mutants caused more severe damage compared to wild-type and RNAi mutants. These findings indicate that PgPG1 plays a vital role in the pathogenicity of P. graminea in barley and has great potential as a pathogen target gene to develop a durable resistance variety to P. graminea. Full article

Pectinolytic enzymes play a key role in many beverages manufacturing processes, improving their clarification and filtration steps. Fungal pectinases are considered promising green catalysts for industrial applications, and they can be produced using fruit-processing residues as substrate. In this study, we investigated the optimal conditions to produce polygalacturonase from Aspergillus brasiliensis in a solid-phase bioprocess, using cupuaçu (Theobroma grandiflorum) peel as substrate. Then, the pectinolytic extract was applied in the clarification of cupuaçu juice. A central composite design was used to determine the optimal fungal cultivation conditions. Thus, the optimal fungal cultivation (maximum production of 11.81 U/g of polygalacturonase) was obtained using cupuaçu peel with 80% moisture, at 34 °C, for 7 days in a medium containing 4.2% phosphorus and 2.6% nitrogen. The enzymatic extract showed greater activity at 60 °C and stability at a pH range between 5.0 and 7.0. The pectinolytic extract was able to clarify the cupuaçu juice, causing a 53.95% reduction in its turbidity and maintaining its antioxidant activity. Our results demonstrate that the cupuaçu peel can be used as a substrate to produce polygalacturonase, and the enzymatic extract produced can be applied in the cupuaçu juice processing, contributing to the circular economy. Full article

Rehmannia glutinosa is an important medicinal herb; but its long-term cultivation often leads to continuous cropping problems. The underlying cause can be attributed to the accumulation of and alterations in root exudates; which interact with soil-borne pathogens; particularly Fusarium oxysporum; triggering disease outbreaks that severely affect its yield and quality. It is therefore crucial to elucidate the mechanisms by which root exudates induce F. oxysporum CCS043 outbreaks. In this study; the genome of F. oxysporum CCS043 from R. glutinosa’s rhizosphere microbiota was sequenced and assembled de novo; resulting in a 47.67 Mb genome comprising 16,423 protein-coding genes. Evolutionary analysis suggests that different F. oxysporum strains may adapt to the host rhizosphere microecosystem by acquiring varying numbers of specific genes while maintaining a constant number of core genes.The allelopathic effects of ferulic acid; verbascoside; and catalpol on F. oxysporum CCS043 were examined at the physiological and transcriptomic levels. The application of ferulic acid was observed to primarily facilitate the proliferation and growth of F. oxysporum CCS043; whereas verbascoside notably enhanced the biosynthesis of infection-related enzymes such as pectinase and cellulase. Catalpol demonstrated a moderate level of allelopathic effects in comparison to the other two. Furthermore; 10 effectors were identified by combining the genomic data. Meanwhile; it was found that among the effector-protein-coding genes; ChiC; VRDA; csn; and chitinase exhibited upregulated expression across all treatments. The expression patterns of these key genes were validated using qRT-PCR. Transient overexpression of the two effector-encoding genes in detached R. glutinosa leaves provided further confirmation that ChiC (GME8876_g) and csn (GME9251_g) are key effector proteins responsible for the induction of hypersensitive reactions in R. glutinosa leaf cells. This study provides a preliminary indication that the use of allelochemicals by F. oxysporum CCS043 can promote its own growth and proliferation and enhance infection activity. This finding offers a solid theoretical basis and data support for elucidating the fundamental causes of fungal disease outbreaks in continuous cropping of R. glutinosa and for formulating effective mitigation strategies. Full article

The spectrum of enzymes required for complete lignocellulosic waste hydrolysis is too diverse to be secreted by a single organism. An alternative is to employ fungal co-cultures to obtain more diverse and complete enzymatic cocktails without the need to mix enzymes during downstream processing. This study evaluated the co-cultivation of Aspergillus brasiliensis and Trichoderma reesei RUT-C30 in different conditions using sugarcane bagasse as the carbon source. The resulting enzymatic cocktails were characterized according to the impact of strain inoculation time on enzymatic activities and proteome composition. Data revealed that the profile of each enzymatic extract was highly dependent on the order in which the participating fungi were inoculated. Some of the co-cultures exhibited higher enzyme activities compared to their respective monocultures for enzymes such as CMCase, pectinase, β-glucosidase, and β-xylosidase. Analysis of the T. reesei RUT-C30 and A. brasiliensis co-culture secretome resulted in the identification of 167 proteins, with 78 from T. reesei and 89 from A. brasiliensis. In agreement with the enzymatic results, proteome analysis also revealed that the timing of inoculation greatly influences the overall secretome, with a predominance of T. reesei RUT-C30 proteins when first inoculated or in simultaneous inoculation. Full article

Fusarium head blight (FHB) is a devastating fungal disease caused by Fusarium graminearum. Pectin lyase, a pectinase, acts on the α-1,4-glycosidic linkage of galacturonic acid primarily by β-elimination. In this study, three pectin lyase genes (FgPel1, 2, 3) in F. graminearum were selected, and deletion mutants (ΔFgPel1, 2, 3) were constructed by homologous recombination for functional characterization. The gene deletions affected the morphology and growth rate of F. graminearum on pectin medium at various concentrations, with the growth rate of ΔFgPel1 being more significant. The growth of ΔFgPel1 colonies slowed at pH 4, with optimal growth at pH 6.5, whereas ΔFgPel2 and ΔFgPel3 exhibited greater inhibition at pH 8. Colony morphology and diameter of the deletion mutants showed no significant differences compared to the wild-type strain PH-1, and there was no effect on conidial production or germination rate. Pathogenicity assays demonstrated that gene deletion significantly reduced the ability of F. graminearum to infest corn silks and wheat ears, and that ΔFgPel2 showed a more pronounced reduction in pathogenicity on wheat spikes. In summary, the pectin lyase genes (FgPel1, 2, 3) are involved in pectin utilization and are influenced by external pH conditions, which attenuate the pathogenicity of F. graminearum without affecting its vegetative growth or asexual spore formation. These findings elucidate the roles of these genes and provide a basis for controlling FHB. Full article

Coniella vitis is a dominant phytopathogen of grape white rot in China, significantly impacting grape yield and quality. Previous studies showed that the growth and pathogenicity of C. vitis were affected by the environmental pH. Arrestin-like protein PalF plays a key role in mediating the activation of an intracellular-signaling cascade in response to alkaline ambient. However, it remains unclear whether palF affects the growth, development, and virulence of C. vitis during the sensing of environmental pH changes. In this study, we identified a homologous gene of PalF/Rim8 in C. vitis and constructed CvpalF-silenced strains via RNA interference. CvpalF-silenced strains exhibited impaired fungal growth at neutral/alkaline pH, accompanied by reduced pathogenicity compared to the wild-type (WT) and empty vector control (CK) strains. The distance between the hyphal branches was significantly increased in the CvpalF-silenced strains. Additionally, CvpalF-silenced strains showed increased sensitivity to NaCl, H₂O₂, and Congo red, and decreased sensitive to CaSO₄. RT-qPCR analysis demonstrated that the expression level of genes related to pectinase and cellulase were significantly down-regulated in CvpalF-silenced strains compared to WT and CK strains. Moreover, the expression of PacC, PalA/B/C/F/H/I was directly or indirectly affected by silencing CvpalF. Additionally, the expression of genes related to plant cell wall-degrading enzymes, which are key virulence factors for plant pathogenic fungi, was regulated by CvpalF. Our results indicate the important roles of CvpalF in growth, osmotolerance, and pathogenicity in C. vitis. Full article

Miang is a traditional fermented food made from Assam tea leaves and consumed as a snack. This study investigated the underground Miang fermentation process practiced by the Luar ethnic group in Laos, specifically examining the nutritional composition and microbial dynamics. Lactic acid bacteria and yeast were dominant in the fermentation process, reaching 8.43 and 8.50 log CFU/g after one week before gradually declining, while the coliform bacterial count was at 5.31 log CFU/g in the initial week but became undetectable in the later stages of fermentation. Next-generation sequencing identified Firmicutes (75.02%) and Proteobacteria (23.51%) as the primary phyla. Bacterial genera included Lactobacillus (73.36%) and Acetobacter (21.06%), with fungi mainly represented by Pichia (85.52%) and Candida (13.45%). Fundamental microbes such as Lactobacillus and Acetobacter were predominantly present, alongside Pichia and Candida, in the fungal communities. Microbial activities played a crucial role in generating essential enzymes for Miang’s transformation. The nutritional transformation appears to be complete at 5 weeks of fermentation. The moisture content in the final products was approximately 74% and correlated with a change in nitrogen-free extract (NFE) and crude fiber. The fat content showed a slight increase from 1.3% to 2.52%, but protein content slightly declined from 17.21% to 16.05%, whereas ash content did not change significantly. Key polysaccharide-degrading enzymes, particularly pectinase and β-mannanase, were revealed and peaked at 48.32 and 25.32 U/g Miang, respectively. The total polyphenols increased from 103.54 mg/g dry Miang to 144.19–155.52 mg/g during fermentation. The lowered IC₅₀ value indicated an increase in antioxidant activity. A fermentation period of at least 3 weeks proved to be optimal for enhancing antioxidant properties and bioactive compounds, and mitigating the risk of coliform bacteria. Full article

Endophytic fungi that reside internally in healthy, asymptomatic plants often benefit their hosts by promoting plant growth and/or providing plant protection against abiotic and biotic stresses. However, only a small fraction of the estimated 1.5 million fungal endophytes have been identified. In this study, a total of 369 isolates of fungal endophytes in 59 distinct taxa were isolated from stem samples of Cornus florida (flowering dogwood). All isolates belonged to species of phyla Ascomycota and Basidiomycota distributed across five orders and 11 genera. Isolates belonging to the same family clustered together in a phylogenetic tree generated from a cluster analysis using MEGA 7 software. Diversity indices of the fungi revealed a rich and diverse community that included several species associated with leaf spots, blight, cankers, and/or dieback diseases. Pathogenicity tests confirmed 16 fungal endophytes as C. florida pathogens, including some well-known destructive pathogens Botryosphaera dothidea, Colletotrichum acutatum, and C. gleosporoides. Isolates of the fungal endophytes possess the capacity to produce extracellular hydrolytic enzymes (cellulase, amylase, pectinase, laccase, chitinase, and protease) that are known to function in tissue penetration, plant colonization, nutrient acquisition, and disease suppression in both plant pathogens and endophytes These results support the interchangeable pathogenic–endophytic roles for some taxa. Full article

A fruiting body of a basidiomycete fungus was discovered growing on chopped Ficus nitida tree trunks in the student housing on the Assiut University campus during the course of this inquiry and a normal collecting operation in the Assiut Governorate, Egypt. Following the growth of the basidioma’s inner tissue on PDA, fungal mycelial growth was achieved. Internal transcribed spacer region (ITS) sequencing has allowed for the identification of the fungus as Tomophagus colossus. On the dry weight basis, chemical analysis of T. colossus AUMC 14536 basidioma revealed that it contains 28.81% carbohydrates, 25.34% crude fats, 23.44% crude fibers, 20.64% crude proteins, and 3.02% ash, in addition to potassium, phosphorus, calcium, selenium, iron, and zinc (133.59, 114.46, 6.27, 3.08, 1.28, and 0.73 mg/100 g dry weight, respectively). The total phenolic compounds (39.26 mg/g) and total flavonoids (5.62 mg/g) were also evaluated. The basidioma extract’s antioxidant activity was assessed as %DPPH radical scavenging activity with an IC50 of 4.15 µg/mL compared with a 1.89 µg/mL IC50 of ascorbic acid. In solid-state fermentation (SSF), the fungus could ferment broad bean straw, palm leaf hay, rice husk, rice straw, sugarcane bagasse, and wheat bran to produce endoglucanase, exoglucanase, laccase, pectinase, and xylanase in substantial amounts. Specific activity exhibited the highest values for endoglucanase (81.48 U/mg), exoglucanase (114.35 U/mg), pectinase (81.94 U/mg), and xylanase (70.18 U/mg) on the rice husk, while the peak of laccase activity (94.27 U/mg) was gained on bean straw. This is the first assessment of the organism’s nutritional value, amino acid content, antioxidant activity, and enzymatic capabilities in Egypt. Full article

Coriandrum sativum L. is a globally significant economic herb with medicinal and aromatic properties. While coriander leaf blight disease was previously confined to India and the USA, this study presents new evidence of its outbreak in Africa and the Middle East caused by Alternaria dauci. Infected leaves display irregular chlorotic to dark brown necrotic lesions along their edges, resulting in leaf discoloration, collapse, and eventual death. The disease also impacts inflorescences and seeds, significantly reducing seed quality. Koch’s postulates confirmed the pathogenicity of the fungus through the re-isolation of A. dauci from artificially infected leaves, and its morphology aligns with typical A. dauci features. Notably, this study identified strong lytic activity (cellulase: 23.76 U, xylanase: 12.83 U, pectinase: 51.84 U, amylase: 9.12 U, and proteinase: 5.73 U), suggesting a correlation with pathogenicity. Molecular characterization using ITS (ON171224) and the specific Alt-a-1 gene (OR236142) supports the fungal morphology. This research provides the first comprehensive documentation of the pathological, lytic, and molecular evidence of A. dauci leaf blight disease on coriander. Future investigations should prioritize the development of resistant coriander varieties and sustainable disease management strategies, including the use of advanced molecular techniques for swift and accurate disease diagnosis to protect coriander from the devastating impact of A. dauci. Full article

Plant biomass is one of the most abundant renewable carbon sources, which holds great potential for replacing current fossil-based production of fuels and chemicals. In nature, fungi can efficiently degrade plant polysaccharides by secreting a broad range of carbohydrate-active enzymes (CAZymes), such as cellulases, hemicellulases, and pectinases. Due to the crucial role of plant biomass-degrading (PBD) CAZymes in fungal growth and related biotechnology applications, investigation of their genomic diversity and transcriptional dynamics has attracted increasing attention. In this project, we systematically compared the genome content of PBD CAZymes in six taxonomically distant species, Aspergillus niger, Aspergillus nidulans, Penicillium subrubescens, Trichoderma reesei, Phanerochaete chrysosporium, and Dichomitus squalens, as well as their transcriptome profiles during growth on nine monosaccharides. Considerable genomic variation and remarkable transcriptomic diversity of CAZymes were identified, implying the preferred carbon source of these fungi and their different methods of transcription regulation. In addition, the specific carbon utilization ability inferred from genomics and transcriptomics was compared with fungal growth profiles on corresponding sugars, to improve our understanding of the conversion process. This study enhances our understanding of genomic and transcriptomic diversity of fungal plant polysaccharide-degrading enzymes and provides new insights into designing enzyme mixtures and metabolic engineering of fungi for related industrial applications. Full article

Chitin synthases (CHSs) are vital enzymes for the synthesis of chitin and play important and differential roles in fungal development, cell wall integrity, environmental adaptation, virulence, and metabolism in fungi. However, except for ChsC, a class III CHS, little is known about the functions of CHSs in Aspergillus niger, an important fungus that is widely applied in the fermentation industry and food processing, as well as a spoilage fungus of food and a human pathogen. This study showed the important functions of ChsA, a class II CHS, in A. niger using multi-phenotypic and transcriptional analyses under various conditions. The deletion of chsA led to severe defects in conidiation on different media and resulted in the formation of smaller and less compact pellets with less septa in hyphal cells during submerged fermentation. Compared with the WT, the ΔchsA mutants exhibited less chitin content, reduced growth under the stresses of cell wall-disturbing and oxidative agents, more released protoplasts, a thicker conidial wall, decreased production of amylases, pectinases, cellulases, and malic acid, and increased citric acid production. However, ΔchsA mutants displayed insignificant changes in their sensitivity to osmotic agents and infection ability on apple. These findings concurred with the alteration in the transcript levels and enzymatic activities of some phenotype-related genes. Conclusively, ChsA is important for cell wall integrity and mycelial morphology, and acts as a positive regulator of conidiation, cellular responses to oxidative stresses, and the production of malic acid and some enzymes, but negatively regulates the citric acid production in A. niger. Full article

The main hypothesis of the present research is investigating the efficacy of titanium oxide nanoparticles (TiO₂-NPs) to prevent the growth of fungal strains when applied on leather under an experimental study. Therefore, fifteen fungal strains were isolated from a deteriorated historical manuscript (papers and leathers) and identified by traditional methods and ITS sequence analysis, including Aspergillus chevalieri (one isolate), A. nidulans (two strains), A. flavus (four strains), A. cristatus (one strain), A. niger (one strain), Paecilomyces fulvus (two strains), Penicillium expansum (two strains), and P. citrinum (two strains). The enzymes cellulase, amylase, pectinase, and gelatinase, which play a crucial role in biodegradation, were highly active in these fungal strains. TiO₂-NPs were formed using the cell-free filtrate of the probiotic bacterial strain, Lactobacillus plantarum, and characterized. Data showed that the TiO₂-NPs were successfully formed with a spherical shape and anatase phase with sizes of 2–8 nm. Moreover, the EDX analysis revealed that the Ti and O ions occupied the main component with weight percentages of 41.66 and 31.76%, respectively. The in vitro cytotoxicity of TiO₂-NPs toward two normal cell lines, WI38 and HFB4, showed a low toxicity effect against normal cells (IC₅₀ = 114.1 ± 8.1µg mL⁻¹ for Wi38, and 237.5 ± 3.5µg mL⁻¹ for HFB4). Therefore, concentrations of 100 μg mL⁻¹ were used to load on prepared leather samples before inoculation with fungal strain P. expansum AL1. The experimental study revealed that the loaded TiO₂-NPs have the efficacy to inhibit fungal growth with percentages of 73.2 ± 2.5%, 84.2 ± 1.8%, and 88.8 ± 0.6% after 7, 14, and 21 days, respectively. Also, the analyses including SEM, FTIR-ART, color change, and mechanical properties for leather inoculated with fungal strain AL1 in the absence of NPs showed high damage aspects compared to those inoculated with fungal strains in the presence of TiO₂-NPs. Full article