Search Results (14)

Antimicrobial metabolite production in basidiomycetes varies by strain and growing conditions. This study compared the antimicrobial activity of extracts from nine fungal strains at both their vegetative and reproductive stages. Wild-growing fungal fruiting bodies were collected and identified through both morphological characterization and molecular sequencing. Extracts from fruiting bodies, mycelia, and culture liquids were tested using the agar well diffusion method and by determining the minimum inhibitory concentration (MIC). Analysis revealed that the highest antimicrobial activity was associated with culture liquid extracts. Antimicrobial properties were detected in the submerged mycelium extracts of only two strains: Stereum hirsutum 1 and Flammulina rossica 16. For fruiting bodies, activity was restricted to extracts of strains from the genus Fomitopsis. The strain S. hirsutum 1 was determined to be the most effective producer of antibacterial compounds. The highest activity was exhibited by the S. hirsutum 1 culture liquid extract, with an MIC of 320 µg/mL against clinical Gram-positive strains (Staphylococcus aureus, S. epidermidis, S. haemolyticus, vancomycin-resistant Enterococcus faecium) and Gram-negative Proteus vulgaris. The studied strains demonstrated higher production of antimicrobial metabolites during vegetative growth, with the active compounds being primarily extracellular. Submerged cultivation of basidiomycetes offers an efficient method for obtaining antimicrobial metabolites, permitting their subsequent isolation, physicochemical characterization, and biomedical evaluation. Full article

Background: The JinEr mushroom results from the heterogeneous symbiosis of Naematelia aurantialba and Stereum hirsutum, with low-temperature storage being key for postharvest quality preservation. However, the species-specific low-temperature response patterns remain unclear. Methods: An integrated approach combining metabolomics, transcriptomics (dual-genome alignment), and spatially resolved enzyme assays was used to dissect responses at 0 °C and 4 °C. Results: The two fungi displayed distinct stress response tendencies under the studied conditions. N. aurantialba showed enhanced stress defense (DNA repair, antioxidant pathways) with defense-related enzyme activities concentrated in its apical/middle enrichment regions. S. hirsutum was observed to maintain overall metabolic activity at the pathway level, and its metabolic enzyme activities were predominant in the basal region. The symbiotic system exhibited temperature-dependent plasticity stress responses. Storage at 0 °C induced a survival-oriented response with slower crude polysaccharide degradation. In contrast, storage at 4 °C supported active metabolic defense coordination but more pronounced polysaccharide loss. Conclusions: These observed defense- and metabolism-biased differential responses suggest a cold stress-specific coordination working model within the symbiotic system under postharvest cold stress. A temperature of 0 °C is more suitable for enabling JinEr mushroom postharvest storage to retain polysaccharides. This study advances our understanding of heterogeneous symbiotic fungi’s postharvest biology and provides a temperature-targeted theoretical basis for storage optimization. Full article

The JinEr mushroom (“Golden Ear”), a globally rare edible and medicinal macrofungus, comprises a symbiotic complex formed by the symbiotic association of Naematelia aurantialba (Tremellomycetes) and Stereum hirsutum (Agaricomycetes). However, the interactions between these fungi and their associated microbiome remain poorly understood. This study employed high-throughput amplicon sequencing, in situ microbial isolation and culture, and microbial confrontation assays to analyze microbial diversity, community structure, and potential functional roles of the endomycotic bacterial community within JinEr basidiomata and its cultivation substrate. Molecular analysis confirmed the heterogenous composition of the basidiomata, revealing N. aurantialba constitutes less than 20% of the fungal biomass, while S. hirsutum predominates, accounting for approximately 80%. Endomycotic fungi accounted for 0.33% (relative abundance) of the fungal community. Prokaryotic analysis identified Delftia and Sphingomonas as the dominant endomycotic bacterial genera within basidiomata, comprising 85.42% of prokaryotic sequences. Endomycotic bacterial diversity differed significantly (p < 0.05) between basidiomata and substrate, indicating host-specific selection. Cultivation-based approaches yielded 140 culturable bacterial isolates (spanning four families and seven genera) from basidiomata core tissues. In vitro co-culture experiments demonstrated that eight representative bacterial strains exhibited compatible growth with both hosts, while one Enterobacteriaceae strain displayed antagonism towards them. These findings confirm that the heterogeneous JinEr basidiomata harbor a specific prokaryotic assemblage potentially engaged in putative symbiotic or commensal associations with the host fungi. This research advances the understanding of microbial ecology in this unique fungal complex and establishes a culture repository of associated bacteria. This collection facilitates subsequent screening for beneficial bacterial strains to enhance the JinEr cultivation system through the provision of symbiotic microorganisms. Full article

Alperujo, a solid by-product from the two-phase olive oil extraction process, poses significant environmental challenges due to its high organic load, phytotoxicity, and phenolic content. At the same time, it represents a promising feedstock for recovering value-added compounds such as phenols and volatile fatty acids (VFAs). When used as a substrate for white rot fungi (WRF), it also produces ligninolytic enzymes. This study explores the use of two native WRF, Anthracophyllum discolor and Stereum hirsutum, for the biotransformation of alperujo under solid-state fermentation conditions, with and without supplementation of copper and manganese, two cofactors known to enhance fungal enzymatic activity. S. hirsutum stood out for its ability to release high concentrations of phenolic compounds (up to 6001 ± 236 mg gallic acid eq L⁻¹) and VFAs (up to 1627 ± 325 mg L⁻¹) into the aqueous extract, particularly with metal supplementation. In contrast, A. discolor was more effective in degrading phenolic compounds within the solid matrix, achieving a 41% reduction over a 30-day period. However, its ability to accumulate phenolics and VFAs in the extract was limited. Both WRF exhibited increased enzymatic activities (particularly Laccase and Manganese Peroxidase) with the addition of Cu-Mn, highlighting the potential of the aqueous extract as a natural source of biocatalysts. Phytotoxicity assays using Solanum lycopersicum seeds confirmed a partial detoxification of the treated alperujo. However, none of the fungi could entirely eliminate inhibitory effects on their own, suggesting the need for complementary stabilization steps before agricultural reuse. Overall, the results indicate that S. hirsutum, especially when combined with metal supplementation, is better suited for valorizing alperujo through the recovery of bioactive compounds. Meanwhile, A. discolor may be more suitable for detoxifying the solid phase strategies. These findings support the integration of fungal pretreatment into biorefinery schemes that valorize agroindustrial residues while mitigating environmental issues. Full article

Mushroom Jin Er has attracted widespread attention in Asia over the past two decades due to its medicinal properties and nutritional values. In the present study, Jin Er basidiocarps were often found to be surrounded by Stereum hirsutum fruiting bodies in their natural habitat and occasionally in artificial cultivation. The observation of two different kinds of mycelia within the hymenium and analyses of ITS sequences confirmed that Jin Er basidiocarps were composed of two fungal species, Naematelia aurantialba and S. hirsutum. This heterogeneity of Jin Er fruiting bodies is indeed distinct from the homogeneous hypha of Tremella fuciformis found in Yin Er mushroom, although its development also requires the presence of another fungus Annulohypoxylon stygium. Basidiospores can germinate on the surface of basidiocarps and produce mycelia. However, basidiospores in PDA medium can only bud into yeast-like conidia. The yeast-like conidia of N. aurantialba can transform into pseudohyphae with a change in temperature from 20 °C to 28 °C or switch into filamentous cells on an induction medium (IDM) at 20 °C, 25 °C and 28 °C. This dimorphic was reported for the first time in N. aurantialba. Haustorium-like structures were abundantly observed both within the hymenium and in the aerial mycelia cultured on the IDM. The developmental process was documented firstly in this study, involving the formation of protuberances with basal clamp connections, elongation at the protuberances, branch production, and eventual maturation. However, further observation is required to determine whether the haustorium-like structures can penetrate S. hirsutum hyphae. These findings are expected to provide valuable insights into the relationship and interaction between these two fungi, thereby advancing the cultivation of fruiting bodies. Full article

Wood-decaying fungi are important study subjects for their ecological role as well as for their biotechnological applications. They break down lignin, cellulose, and hemicelluloses using enzymes that modify the chemical structure of these complex macromolecules. Due to their ability to degrade wood, these fungi can create structural damage to wooden structures and to trees, especially those with very low level of fitness. Previous studies on wood decay fungi in the Maltese Islands are limited to records and checklists described by a handful of authors. The aim of this study was to provide a comprehensive description of wood decay fungal diversity in the Maltese Islands including an updated checklist based on DNA barcoding, as well as to establish the first wood-decay fungal culture collection at the Biology Department Seed Bank of the University of Malta. Several surveys were carried out during the rainy season along wooded areas of the Maltese Islands as well as in historical gardens. Isolates were identified using macro- and micro-morphological features, dichotomous keys, as well as molecular data. Basidiomes were recorded growing on 14 different host plant species, 11 axenic cultures have been made and 9 species of wood decay fungi have been conclusively identified by DNA barcoding. The collection of the axenic isolates includes one of Aurificaria cf. euphoria, three of Ganoderma resinaceum sl., two of Laetiporus sulphureus, one of Inonotus sp., one of Inonotus rickii anamorph, one of Inocutis tamaricis, one of Stereum hirsutum, and one of Pleurotus eryngii. However, the mycelium of Coriolopsis gallica, though collected and identified, could not be isolated. Full article

Over the last decade, fungal conservation has become an increasingly important topic, especially for species tied to forest ecosystems. Among these, wood decay fungi are a group of interesting species from ecological and applicative points of view. Culture collections represent an important tool for the conservation of species and research material. The aim of this study was to establish the first research culture collection of wood decay fungal strains, mainly corticioid and polyporoid species, at Salamanca University (Spain). From two areas of the Iberian Peninsula, a total of 120 basidiomata were collected and morphologically identified. From these, 55 strains were successfully isolated in pure culture and their identity was confirmed by DNA molecular analysis. The average growth rate of each strain was recorded and mycelium characteristics, such as colony morphology and microscopic features, were described. Notable strains in the collection included: (1) Botryobasidium asperulum and Phlebia rufa for taxonomical studies; (2) Hericium erinaceus, Grifola frondosa and Pleurotus species for medicinal properties; (3) Irpex lacteus, Phanerochaete sordida and Trametes versicolor for their degradation capabilities; (4) Stereum gausapatum and Stereum hirsutum for their applicative and enzymatic potential. The new fungal strain culture collection represents a valuable tool for the ex situ conservation of Mediterranean wood decay fungi. Full article

Romania has a long history of grapevine culturing and winemaking. However, like any agricultural sector, viticulture faces devastating biological threats. Fungi responsible for grapevine trunk diseases (GTDs) and grape spoilage lead to considerable yield losses and a decline in grapevine quality. In the actual context, many countries, including Romania, have reoriented their approaches to minimize chemical inputs, which have been proven to be toxic and to have negative impacts on the environment, and to replace them with sustainable biocontrol strategies for the wine-growing sector. Within biocontrol strategies, Bacillus spp. is a well-known plant-protective bacteria with antifungal properties. Within this paper, six endophytic bacteria from various plant sources were studied. The bacterial strains were identified as B. pumilus, B. subtilis, and B. velezensis by sequencing their 16S rDNA region. Regardless of the in vitro test methods (using living bacterial cells, bacterial-cell-free supernatant (CFS), and volatile active compounds (VOCs)), B. velezensis strains revealed strong and broad antifungal activity against grape and grapevine fungal pathogens such as Aspergillus spp., Botrytis cinerea, Penicillium expansum, Diplodia seriata, Eutypa lata, Fusarium spp., Clonostachys rosea, Neofusicoccum parvum, and Stereum hirsutum. The functional antifungal genes encoding for difficidin, fengycin, iturins, macrolactin, and mycosubtilin were molecularly detected, which could support the proven antifungal activity of the endophytic strains. Lytic enzymes involved in fungal growth inhibition, such as chitinase, cellulase, and proteases, were also revealed to be produced by some of these bacterial strains. Various other in vitro tests, such as phosphate and phytate solubilization, phytohormone synthesis, the production of enzymes involved in the polyamine biosynthetic pathway, and pH as well as temperature tolerance tests were carried out to reveal the plant-beneficial potential of these bacterial strains. These results revealed that the B. velezensis strains, especially BAHs1, are the most suitable endophytes for grapevine biologic control, which could lead to the future development of sustainable management strategies. Full article

Wood Decay Fungi (WDF) are fungi specialized in degrading wood. An interesting perspective is their use as a source of Novel Foods or food ingredients. Here, for the first time, the metabolite profiling of hydroalcoholic and organic extracts from A. biennis, F. iberica, S. hirsutum mycelia was investigated by NMR methodology. Amino acids (alanine, arginine, asparagine, aspartate, betaine, GABA, glutamate, glutamine, histidine, isoleucine, leucine, lysine, phenylalanine, threonine, tryptophan, tyrosine, valine), sugars (galactose, glucose, maltose, trehalose, mannitol), organic acids (acetate, citrate, formate, fumarate, lactate, malate, succinate), adenosine, choline, uracil and uridine were identified and quantified in the hydroalcoholic extracts, whereas the ¹H spectra of organic extracts showed the presence of saturated, mono-unsaturated and di-unsaturated fatty chains, ergosterol,1,2-diacyl-sn-glycero-3-phosphatidylethanolamine, and 1,2-diacyl-sasglycero-3-phosphatidylcholine. A. biennis extracts showed the highest amino acid concentration. Some compounds were detected only in specific species: betaine and mannitol in S. hirsutum, maltose in A. biennis, galactose in F. iberica, GABA in F. iberica and S. hirsutum, and acetate in A. biennis and S. hirsutum. S. hirsutum showed the highest saturated fatty chain concentration, whereas DUFA reached the highest concentration in A. biennis. A high amount of ergosterol was measured both in A. biennis and F. iberica. The reported results can be useful in the development of WDF-based products with a high nutritional and nutraceutical value. Full article

Nanoparticles, especially silver nanoparticles (Ag NPs), have gained significant attention in recent years as potential alternatives to traditional antibiotics for treating infectious diseases due to their ability to inhibit the growth of microorganisms effectively. Ag NPs can be synthesized using fungi extract, but the method is not practical for large-scale production due to time and biomass limitations. In this study, we explore the use of chitosan to encapsulate the mycelia of the white-rot fungus Stereum hirsutum and form chitosan fungal beads for use in multiple extractions and nanoparticle synthesis. The resulting nanoparticles were characterized using various techniques, including UV-vis spectrophotometry, transmission electron microscopy, dynamic light scattering, and X-ray diffraction analysis. The analysis revealed that the synthesized nanoparticles were composed of chitosan-silver nanoparticles (CS-Ag NPs) with a size of 25 nm. The chitosan fungal beads were reused in three extractions and nanoparticle synthesis before they lost their ability to produce CS-Ag NPs. The CS-Ag NPs showed potent antimicrobial activity against phytopathogenic and human pathogenic microorganisms, including Pseudomonas syringae, Escherichia coli, Staphylococcus aureus, and Candida albicans, with minimum inhibitory concentrations of 1.5, 1.6, 3.1, and 4 µg/mL, respectively. The antimicrobial activity of CS-Ag NPs was from 2- to 40-fold higher than Ag NPs synthesized using an aqueous extract of unencapsulated fungal biomass. The CS-Ag NPs were most effective at a pH of five regarding the antimicrobial activity. These results suggest that the chitosan fungal beads may be a promising alternative for the sustainable and cost-effective synthesis of CS-Ag NPs with improved antimicrobial activity. Full article

Vitis vinifera, known as the common grape vine, represents one of the most important fruit crops in the world. Romania is a wine-producing country with a rich and long tradition in viticulture. In the last decade, increasing reports of damage caused by grapevine trunk diseases (GTDs) have raised concerns in all wine producing countries. Up to now, no study was performed regarding the GTDs situation in Romania, an important grapevine grower in Europe. In this study, we aim, after a comprehensive presentation of the fungal GTDs worldwide, to review the scientific information related to these diseases in Romania in order to open a national platform in an international framework. In order to achieve this, we consulted over 500 references from different scientific databases and cited 309 of them. Our review concludes that, in Romania, there is little amount of available literature on this matter. Three out of six fungal GTDs are reported and well documented in all of the Romanian viticultural zones (except for viticultural zone 4). These are Eutypa dieback, Phomopsis dieback, and Esca disease. Of the fungal pathogens considered responsible Eutypa lata, Phomopsis viticola and Stereum hirsutum are the most studied and well documented in Romania. Management measures are quite limited, and they mostly include preventive measures to stop the GTDs spread and the removal of affected grapevines. Full article

This study evaluated the use of the white-rot fungi (WRF) Anthracophyllum discolor and Stereum hirsutum as a biological pretreatment for olive mill solid mill waste (OMSW). The WRF strains proposed were added directly to OMSW. The assays consisted of determining the need to add supplementary nutrients, an exogenous carbon source or use agitation systems, and evaluating WRF growth, enzyme activity, phenolic compound removal and lignin degradation. The highest ligninolytic enzyme activity was found at day 10, reaching 176.7 U/L of manganese-independent peroxidase (MniP) produced by A. discolor, and the highest phenolic removal (more than 80% with both strains) was reached after 24 days of incubation. The confocal laser scanning microscopy analysis (CLSM) confirmed lignin degradation through the drop in lignin relative fluorescence units (RFU) from 3967 for untreated OMSW to 235 and 221 RFU, showing a lignin relative degradation of 94.1% and 94.4% after 24 days of treatment by A. discolor and S. hirsutum, respectively. The results demonstrate for the first time that A. discolor and S. hirsutum were able to degrade lignin and remove phenolic compounds from OMSW using this as the sole substrate without adding other nutrients or using agitation systems. This work indicates that it could be possible to design an in situ pretreatment of the valorization of OMSW, avoiding complex systems or transportation. In this sense, future research under non-sterile conditions is needed to evaluate the competition of WRF with other microorganisms present in the OMSW. The main drawbacks of this work are associated with both the low reaction time and the water addition. However, OMSW is seasonal waste produced in one season per year, being stored for a long time. In terms of water addition, the necessary optimization will be addressed in future research. Full article

The biological synthesis of silver nanoparticles (AgNPs) for medical, environmental, and industrial applications is considered an alternative to chemical synthesis methods. Additionally, the reducing, capping, and stabilizing molecules produced by the organisms can play a key role in the further activity of AgNPs. In this work, we evaluated the synthesis of AgNPs by four molecular weight fractions (S1: <10 kDa, S2: 10 to 30 kDa, S3: 30 to 50 kDa, and S4: >50 kDa) of mycelia-free aqueous extract produced by the white-rot fungus Stereum hirsutum and their effect on the antimicrobial activity against Pseudomonas syringae and photocatalytic decolorization of nine synthetic dyes exposed to sunlight radiation. All synthesis assay fractions showed the characteristic surface plasmon resonance (SPR) with 403 to 421 nm peaks. TEM analysis of synthesized AgNPs showed different sizes: the whole mycelia-free extracts S0 (13.8 nm), S1 (9.06 nm), S2 (10.47 nm), S3 (22.48 nm), and S4 (16.92 nm) fractions. The results of disk diffusion assays showed an inverse relation between antimicrobial activity and the molecular weight of compounds present in the mycelia-free aqueous extract used to synthesize AgNPs. The AgNPs synthesized by S0 (14.3 mm) and S1(14.2 mm) generated the highest inhibition diameter of P. syringae growth. By contrast, in the photocatalytic assays, the AgNPs synthesized by the S2 fraction showed the highest discoloration in all the dyes tested, reaching 100% of the discoloration of basic dyes after 2 h of sunlight exposure. The maximum discoloration observed in reactive and acid dyes was 53.2% and 65.3%, respectively. This differentiation in the antimicrobial and photocatalytic activity of AgNPs could be attributed to the capping effect of the molecules present in the extract fractions. Therefore, the molecular separation of synthesis extract enables the specific activities of the AgNPs to be enhanced. Full article

Nitrilases participate in the nitrile metabolism in microbes and plants. They are widely used to produce carboxylic acids from nitriles. Nitrilases were described in bacteria, Ascomycota and plants. However, they remain unexplored in Basidiomycota. Yet more than 200 putative nitrilases are found in this division via GenBank. The majority of them occur in the subdivision Agaricomycotina. In this work, we analyzed their sequences and classified them into phylogenetic clades. Members of clade 1 (61 proteins) and 2 (25 proteins) are similar to plant nitrilases and nitrilases from Ascomycota, respectively, with sequence identities of around 50%. The searches also identified five putative cyanide hydratases (CynHs). Representatives of clade 1 and 2 (NitTv1 from Trametes versicolor and NitAg from Armillaria gallica, respectively) and a putative CynH (NitSh from Stereum hirsutum) were overproduced in Escherichia coli. The substrates of NitTv1 were fumaronitrile, 3-phenylpropionitrile, β-cyano-l-alanine and 4-cyanopyridine, and those of NitSh were hydrogen cyanide (HCN), 2-cyanopyridine, fumaronitrile and benzonitrile. NitAg only exhibited activities for HCN and fumaronitrile. The substrate specificities of these nitrilases were largely in accordance with substrate docking in their homology models. The phylogenetic distribution of each type of nitrilase was determined for the first time. Full article