In the original article [...].
In the original article [...].
Laccaria is a diverse and widespread genus of ectomycorrhizal fungi that form symbiotic associations with various trees and shrubs, playing a significant role in forest ecosystems. Approximately 85 Laccaria species are formally recognised, but recent studies indicate this number may be an underestimation, highlighting the need for further taxonomic studies to improve our understanding of species boundaries. This manuscript focuses on Laccaria affinis, originally described by Singer in 1967 as Laccaria laccata var. affinis, and details a comprehensive study of its morphological and molecular characteristics, including the examination of its holotype and recent collections from Italy and the United Kingdom. Our findings reveal significant micromorphological traits that enhance the original description. Phylogenetic analyses indicate that L. affinis occupies a distinct clade within Northern Hemisphere Laccaria species, although minimal genetic differences challenge its independence from L. macrocystidiata. Consequently, we propose that these two taxa be considered synonymous. This study not only contributes to the understanding of Laccaria diversity but also proposes the formal designation of an epitype for L. affinis, thereby providing a foundation for future research on this ecologically significant genus. Furthermore, a new species named Laccaria albifolia belonging to the "/Laccaria bicolor complex clade" is described here on the base of six collections from Italy and Spain.
While the rapid rise in bioinformatics has facilitated the identification of the domains and functions of many proteins, some still have no domain annotation or largely uncharacterized functions. However, the biological roles of unknown proteins were not clear in oomycetes. An analysis of the Phytophthora sojae genome database identified the protein Ps495620, which has no domain annotations and functional predictions in Phytophthora. This study used a CRISPR/Cas9-mediated gene replacement system to knock out Ps495620 to elucidate its function. The Ps495620-knockout mutants exhibited significantly increased oospore production and decreased sporangium formation compared to the wild-type strain P6497. Transcriptomics showed that it is a key regulator of nitrogen, pyruvate, ascorbate, and adorate metabolism in P. sojae. Our findings indicate that Ps495620 is critical in regulating sporangium formation and oospore production in P. sojae.
Secondary metabolism is one of the main mechanisms Trichoderma uses to explore and colonize new niches, and 6-pentyl-α-pyrone (6-PP) is an important secondary metabolite in this process. This work focused on standardizing a method to investigate the production of 6-PP. Ethanol and ethyl acetate were both effective solvents for quantifying 6-PP in solution and had limited solubility in potato-dextrose-broth media. The 6-PP extraction using ethyl acetate provided a rapid and efficient process to recover this metabolite. The 6-PP was readily produced during the development of Trichoderma atroviride growing in the dark, but light suppressed its production. The 6-PP was purified, and its spectrum by nuclear magnetic resonance and mass spectroscopy was identical to that of commercial 6-PP. Light also induced or suppressed other unidentified metabolites in several other species of Trichoderma. The antagonistic activity of T. atroviride was influenced by light, as suppression of plant pathogens was greater in the dark. The secreted metabolite production on potato-dextrose-agar was differentially regulated by light, indicating that Trichoderma produced several metabolites with antagonistic activity against plant pathogens. Light has an important influence on the secondary metabolism and antagonistic activity of Trichoderma, and this trait is of key relevance for selecting antagonistic Trichoderma strains for plant protection.
Fungal biota represents important constituents of phyllosphere microorganisms. It is taxonomically highly diverse and influences plant physiology, metabolism and health. Members of the order Diaporthales are distributed worldwide and include devastating plant pathogens as well as endophytes and saprophytes. However, many phyllosphere Diaporthales species remain uncharacterized, with studies examining their diversity needed. Here, we report on the identification of several diaporthalean taxa samples collected from diseased leaves of Cinnamomum camphora (Lauraceae), Castanopsis fordii (Fagaceae) and Schima superba (Theaceae) in Fujian province, China. Based on morphological features coupled to multigene phylogenetic analyses of the internal transcribed spacer (ITS) region, the large subunit of nuclear ribosomal RNA (LSU), the partial beta-tubulin (tub2), histone H3 (his3), DNA-directed RNA polymerase II subunit (rpb2), translation elongation factor 1-α (tef1) and calmodulin (cal) genes, three new species of Diaporthales are introduced, namely, Diaporthe wuyishanensis, Gnomoniopsis wuyishanensis and Paratubakia schimae. This study contributes to our understanding on the biodiversity of diaporthalean fungi that are inhabitants of the phyllosphere of trees native to Asia.
Enzyme-mediated protein degradation is a major concern in industrial fungal strain improvement, making low-proteolytic strains preferable for enhanced protein production. Here, we improved food-grade Aspergillus oryzae BCC7051 by manipulating the transcriptional regulation of protease-encoding genes. Genome mining of the transcription factor AoprtR and computational analysis confirmed its deduced amino acid sequence sharing evolutionary conservation across Aspergillus and Penicillium spp. The AoPrtR protein, which is classified into the Zn(II)2-Cys6-type transcription factor family, manipulates both intra- and extracellular proteolytic enzymes. Our transcriptional analysis indicated that the regulation of several protease-encoding genes was AoPrtR-dependent, with AoPrtR acting as a potent activator for extracellular acid-protease-encoding genes and a likely repressor for intracellular non-acid-protease-encoding genes. An indirect regulatory mechanism independent of PrtR may enhance proteolysis. Moreover, AoPrtR disruption increased extracellular esterase production by 2.55-fold, emphasizing its role in protein secretion. Our findings highlight the complexity of AoPrtR-mediated regulation by A. oryzae. Manipulation of regulatory processes through AoPrtR prevents secreted protein degradation and enhances the quantity of extracellular proteins, suggesting the low-proteolytic variant as a promising platform for the production of these proteins. This modified strain has biotechnological potential for further refinement and sustainable production of bio-based products in the food, feed, and nutraceutical industries.
Vulvovaginal candidiasis (VVC) can lead to multiple complications when it occurs during pregnancy, so it is necessary to diagnose it promptly for effective treatment. Traditional methods for identifying Candida spp. are often too time-consuming and have limited specificity and sensitivity. In this work, we evaluated the diagnostic utility of an endpoint PCR assay (Cand PCR) in vaginal swab specimens. Using a cotton swab, 108 vaginal swab samples were taken from pregnant women who consented to participate in the study. The samples were inoculated in Sabouraud agar plates (the gold standard) and subsequently used to extract DNA directly from the exudate. The yeasts isolated from the Sabouraud agar were identified in CHROMagar™ Candida. DNA extracted from vaginal swabs was amplified by Cand PCR. Based on the results of the Cand PCR and the gold standard, sensitivity (S), specificity (E), positive predictive values (PPVs), and negative predictive values (NPVs) were determined. Cand PCR presented an S = 65%, E = 100%, PPV = 100% and NPV = 91%. Cand PCR showed low sensitivity for detecting Candida spp. directly from vaginal swabs, but it was useful for identifying the etiologic agent and reducing the time to obtain the result, which is usually at least 48 h.
Periodic mycelial subculture is a method commonly used for the storage of edible mushrooms, but excessive subculturing can lead to the degeneration of strains. In this study, the Volvariella volvacea strain V971(M0) was successively subcultured on PDA medium every 4 days, and one generation of strains was preserved every 4 months. Thus, five generations of subcultured strains (M1-M5) were obtained after 20 months of mycelial subculturing, their production traits were determined, and transcriptomic analysis was performed using RNA-seq; the differentially expressed genes were verified via RT-qPCR. The results showed that as the number of subcultures increased, the diameter of the mycelium and biological efficiency gradually decreased; in addition, the time in which the primordium formed increased and the production cycle was lengthened, while strains M4 and M5 lacked the ability to produce fruiting bodies. There were 245 differentially expressed genes between the M1-M5 and M0 strains, while the highest number of differentially expressed genes was between M3 and M0, at 1439; the smallest number of differentially expressed genes was between M2 and M0, at 959. GO enrichment analysis showed that the differentially expressed genes were mainly enriched in metabolic processes, organelle components, and catalytic activities. KEGG enrichment analysis showed that the differentially expressed genes were mainly enriched in metabolic pathways. The further annotation of differentially expressed genes showed that 39, 24, and 24 differentially expressed genes were related to substrate degradation, amino acid synthesis and metabolism, and reactive oxygen species metabolism, respectively. The downregulation of the related differentially expressed genes would lead to the excessive accumulation of reactive oxygen species, inhibit nutrient absorption and energy acquisition, and lead to the degradation of V. volvacea. These findings could form a theoretical basis for the degeneration mechanism of V. volvacea, and also provide a basis for the molecular function study of the genes related to strain degradation.
Actinobacteria are well known for their production of metabolites of interest. They have been previously studied to identify new antibiotics in medical research and for their ability to stimulate plant growth in agronomic research. Actinobacteria represents a real source of potential biocontrol agents (BCAs) today. With the aim of reducing the use of phytosanitary products by 50% with the different Ecophyto plans, a possible application is the fight against mycotoxin-producing fungi in food matrices and crops using BCAs. To deal with this problem, the use of actinobacteria, notably belonging to the Streptomyces genus, or their specialized metabolites seems to be a solution. In this review, we focused on the impact of actinobacteria or their metabolites on the development of mycotoxigenic fungi and mycotoxin production on the one hand, and on the other hand on their ability to detoxify food matrices contaminated by mycotoxins.
Pleurotus ostreatus (oyster mushroom) holds excellent promise worldwide, bringing several opportunities and augmenting the tool sets used in the biotechnology field, the food industry, and medicine. Our study explores the antimicrobial and probiotic growth stimulation benefits of freeze-dried P. ostreatus powders (OMP-TF, oyster mushroom powder from the total fresh sample; OMP-CSR, oyster mushroom powder from the cooked solid residue; OMP-CL, oyster mushroom powder from the cooked liquid), focusing on their bioactive compounds and associated activities. Our research examined polysaccharide fractions-specifically total glucans and α- and β-glucans-alongside secondary metabolites, including polyphenols and flavonoids, from freeze-dried mushroom powders. Additionally, carbon nanodots (CNDs) were also characterized. The growth inhibition was tested against Escherichia coli and Staphylococcus epidermidis, while the capacity for stimulating probiotic growth was evaluated using Lactobacillus plantarum and Lactobacillus casei. Evidence indicates that OMP-CL and OMP-CSR exhibit significant antimicrobial properties against S. epidermidis Gram-positive bacteria. OMP-CL notably promoted the growth of L. casei. OMP-CL, containing the most significant number of CNDs, has shown to be a valuable source for gut microbiota modulation, with its antimicrobial and probiotic-stimulating efficacy. However, further in vitro and in vivo studies should be performed to explore CNDs and their behavior in different biological systems.