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PloS One 2012Cordyceps militaris, an ascomycete caterpillar fungus, has been used as a traditional Chinese medicine for many years owing to its anticancer and immunomodulatory...
BACKGROUND
Cordyceps militaris, an ascomycete caterpillar fungus, has been used as a traditional Chinese medicine for many years owing to its anticancer and immunomodulatory activities. Currently, artificial culturing of this beneficial fungus has been widely used and can meet the market, but systematic molecular studies on the developmental stages of cultured C. militaris at transcriptional and translational levels have not been determined.
METHODOLOGY/PRINCIPAL FINDINGS
We utilized high-throughput Illumina sequencing to obtain the transcriptomes of C. militaris mycelium and fruiting body. All clean reads were mapped to C. militaris genome and most of the reads showed perfect coverage. Alternative splicing and novel transcripts were predicted to enrich the database. Gene expression analysis revealed that 2,113 genes were up-regulated in mycelium and 599 in fruiting body. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were performed to analyze the genes with expression differences. Moreover, the putative cordycepin metabolism difference between different developmental stages was studied. In addition, the proteome data of mycelium and fruiting body were obtained by one-dimensional gel electrophoresis (1-DGE) coupled with nano-electrospray ionization liquid chromatography tandem mass spectrometry (nESI-LC-MS/MS). 359 and 214 proteins were detected from mycelium and fruiting body respectively. GO, KEGG and Cluster of Orthologous Groups (COG) analysis were further conducted to better understand their difference. We analyzed the amounts of some noteworthy proteins in these two samples including lectin, superoxide dismutase, glycoside hydrolase and proteins involved in cordycepin metabolism, providing important information for further protein studies.
CONCLUSIONS/SIGNIFICANCE
The results reveal the difference in gene expression between the mycelium and fruiting body of artificially cultivated C. militaris by transcriptome and proteome analysis. Our study provides an effective resource for the further developmental and medicinal research of this promising fungus.
Topics: Cordyceps; Fruiting Bodies, Fungal; Gene Expression; Genome, Fungal; Genome-Wide Association Study; Mycelium; Proteome; Sequence Analysis, RNA; Transcriptome
PubMed: 23251642
DOI: 10.1371/journal.pone.0051853 -
Open Biology Apr 2022The biosynthesis of sterols is critical for the majority of eukaryotes; however, some organisms lack this pathway, including most oomycetes. spp. are sterol...
The biosynthesis of sterols is critical for the majority of eukaryotes; however, some organisms lack this pathway, including most oomycetes. spp. are sterol auxotrophic but, remarkably, have retained a few genes encoding enzymes in the sterol biosynthesis pathway. Here, we show that a gene in predicted to encode Δ7-sterol reductase, displays multiple functions. When expressed in , PcDHCR7 showed the Δ7-sterol reductase activity. Knocking out in resulted in loss of the capacity to transform ergosterol into brassicasterol, which means PcDHCR7 has the Δ7-sterol reductase activity in itself This enables to transform sterols recruited from the environment for better use. The biological characteristics of transformants were compared with those of the wild-type strain and a complemented transformant, and the results showed that plays a key role in mycelium development and pathogenicity of zoospores. Further analysis of the transcriptome indicated that the expression of many genes changed in the transformant, which involve in different biological processes. It is possible that compensates for the defects caused by the loss of by remodelling its transcriptome.
Topics: Mycelium; Oxidoreductases; Phytophthora; Plant Diseases; Sterols; Virulence
PubMed: 35382565
DOI: 10.1098/rsob.210282 -
PloS One 2020Oligomycins are macrolide antibiotics, produced by Streptomyces spp. that show antagonistic effects against several microorganisms such as bacteria, fungi, nematodes and...
Oligomycins are macrolide antibiotics, produced by Streptomyces spp. that show antagonistic effects against several microorganisms such as bacteria, fungi, nematodes and the oomycete Plasmopara viticola. Conidiogenesis, germination of conidia and formation of appressoria are determining factors pertaining to pathogenicity and successful diseases cycles of filamentous fungal phytopathogens. The goal of this research was to evaluate the in vitro suppressive effects of two oligomycins, oligomycin B and F along with a commercial fungicide Nativo® 75WG on hyphal growth, conidiogenesis, conidial germination, and appressorial formation of the wheat blast fungus, Magnaporthe oryzae Triticum (MoT) pathotype. We also determined the efficacy of these two oligomycins and the fungicide product in vivo in suppressing wheat blast with a detached leaf assay. Both oligomycins suppressed the growth of MoT mycelium in a dose dependent manner. Between the two natural products, oligomycin F provided higher inhibition of MoT hyphal growth compared to oligomycin B with a minimum inhibitory concentration of 0.005 and 0.05 μg/disk, respectively. The application of the compounds completely halted conidial formation of the MoT mycelium in agar medium. Further bioassays showed that these compounds significantly inhibited MoT conidia germination and induced lysis. The compounds also caused abnormal germ tube formation and suppressed appressorial formation of germinated spores. Interestingly, the application of these macrolides significantly inhibited wheat blast on detached leaves of wheat. This is the first report on the inhibition of mycelial growth, conidiogenesis, germination of conidia, deleterious morphological changes in germinated conidia, and suppression of blast disease of wheat by oligomycins from Streptomyces spp. Further study is needed to unravel the precise mode of action of these natural compounds and consider them as biopesticides for controlling wheat blast.
Topics: Biological Control Agents; Edible Grain; Food Microbiology; Fungicides, Industrial; Hyphae; Magnaporthe; Mycelium; Oligomycins; Plant Diseases; Spores, Fungal; Triticum
PubMed: 32804955
DOI: 10.1371/journal.pone.0233665 -
Molecules (Basel, Switzerland) Jun 2017An efficient synthesis of a series of 4'-oxyalkyl-isocordoin analogues (-) is reported for the first time. Their structures were confirmed by ¹H-NMR, C-NMR, and HRMS....
An efficient synthesis of a series of 4'-oxyalkyl-isocordoin analogues (-) is reported for the first time. Their structures were confirmed by ¹H-NMR, C-NMR, and HRMS. Their anti-oomycete activity was evaluated by mycelium and spores inhibition assay against two selected pathogenic oomycetes strains: and . The entire series of isocordoin derivatives (except compound ) showed high inhibitory activity against these oomycete strains. Among them, compound exhibited strong activity, with minimum inhibitory concentration (MIC) and minimum oomyceticidal concentration (MOC) values of 50 µg/mL and 75 µg/mL, respectively. The results showed that 4'-oxyalkylated analogues of isocordoin could be potential anti-oomycete agents.
Topics: Antifungal Agents; Catechols; Inorganic Chemicals; Microbial Sensitivity Tests; Mycelium; Saprolegnia; Spores, Fungal
PubMed: 28604594
DOI: 10.3390/molecules22060968 -
PloS One 2019Living fungal mycelium with abolished ability to form fruiting bodies is a self-healing substance, which is particularly valuable for further engineering and development...
Living fungal mycelium with abolished ability to form fruiting bodies is a self-healing substance, which is particularly valuable for further engineering and development as materials sensing environmental changes and secreting signals. Suppression of fruiting body formation is also a useful tool for maintaining the stability of a mycelium-based material with ease and lower cost. The objective of this study was to provide a biochemical solution to regulate the fruiting body formation, which may replace heat killing of mycelium in practice. The concentrations of glycogen synthase kinase-3 (GSK-3) inhibitors, such as lithium chloride or CHIR99021 trihydrochloride, were found to directly correlate with the development of fruiting bodies in the mushroom forming fungi such as Coprinopsis cinerea and Pleurotus djamor. Sensitive windows to these inhibitors throughout the fungal life cycle were also identified. We suggest the inclusion of GSK-3 inhibitors in the cultivation recipes for inhibiting fruiting body formation and regulating mycelium growth. This is the first report of using a GSK-3 inhibitor to suppress fruiting body formation in living fungal mycelium-based materials. It provides an innovative strategy for easy, reliable, and low cost maintenance of materials containing living fungal mycelium.
Topics: Agaricales; Biocompatible Materials; Biomarkers; Enzyme Inhibitors; Fruiting Bodies, Fungal; Fungi; Glycogen Synthase Kinase 3; Lithium Chloride; Mycelium
PubMed: 31083677
DOI: 10.1371/journal.pone.0209812 -
Applied and Environmental Microbiology Aug 2018Among the plethora of unusual secondary metabolites isolated from are the tetrapeptidic endolides A and B. Both tetrapeptides contain 3-(3-furyl)-alanine residues,...
Among the plethora of unusual secondary metabolites isolated from are the tetrapeptidic endolides A and B. Both tetrapeptides contain 3-(3-furyl)-alanine residues, previously proposed to originate from bacterial metabolism. Inspired by this observation, we aimed to identify the presence of endosymbiotic bacteria in and to discover the true producer of the endolides. The endobacterium was initially detected by 16S rRNA gene amplicon sequencing from the fungal metagenome and was subsequently isolated. It was confirmed that the tetrapeptides were produced by the axenic only when in latency. Fungal colonies unable to produce conidia and the tetrapeptides were isolated and confirmed to be free of A second endosymbiont identified as related to was also isolated. imaging of the mycelium supported an endosymbiotic relationship between and the two endobacteria. Besides the technical novelty, our analyses revealed that the two endobacteria are compartmentalized in defined fungal cells, prevailing mostly in latency when in symbiosis. Within the emerging field of intracellular bacterial symbioses, fungi are the least studied eukaryotic hosts. Our study further supports the Fungi as a valuable model for understanding endobacterial symbioses in eukaryotes. The discovery of two bacterial endosymbionts harbored in mycelium, and , is described here. Production of tetrapeptides inside the mycelium is ensured by , and fungal sporulation is influenced by the endosymbionts. Here, we illustrate the bacterial endosymbiotic origin of secondary metabolites in an Ascomycota host.
Topics: Ascomycota; Burkholderia; Mycelium; Peptides, Cyclic; Sphingomonas; Spores, Fungal; Symbiosis
PubMed: 29858203
DOI: 10.1128/AEM.00660-18 -
Communications Biology Jun 2020Fungal mycelium is an emerging bio-based material. Here, mycelium films are produced from liquid shaken cultures that have a Young's modulus of 0.47 GPa, an ultimate...
Fungal mycelium is an emerging bio-based material. Here, mycelium films are produced from liquid shaken cultures that have a Young's modulus of 0.47 GPa, an ultimate tensile strength of 5.0 MPa and a strain at failure of 1.5%. Treating the mycelial films with 0-32% glycerol impacts the material properties. The largest effect is observed after treatment with 32% glycerol decreasing the Young's modulus and the ultimate tensile strength to 0.003 GPa and 1.8 MPa, respectively, whereas strain at failure increases to 29.6%. Moreover, glycerol treatment makes the surface of mycelium films hydrophilic and the hyphal matrix absorbing less water. Results show that mycelium films treated with 8% and 16-32% glycerol classify as polymer- and elastomer-like materials, respectively, while non-treated films and films treated with 1-4% glycerol classify as natural material. Thus, mycelium materials can cover a diversity of material families.
Topics: Biocompatible Materials; Biofilms; Biomass; Glycerol; Microscopy; Microscopy, Electron, Scanning; Mycelium; Schizophyllum; Tensile Strength; Water
PubMed: 32591629
DOI: 10.1038/s42003-020-1064-4 -
Scientific Reports Nov 2020Fomitopsis officinalis is a medicinal mushroom used in traditional European eighteenth and nineteenth century folk medicine. Fruiting bodies of F. officinalis were...
Fomitopsis officinalis is a medicinal mushroom used in traditional European eighteenth and nineteenth century folk medicine. Fruiting bodies of F. officinalis were collected from the natural environment of Świętokrzyskie Province with the consent of the General Director for Environmental Protection in Warsaw. Mycelial cultures were obtained from fragments of F. officinalis fruiting bodies. The taxonomic position of the mushroom mycelium was confirmed using the PCR method. The presence of organic compounds was determined by HPLC-DAD analysis. Bioelements were determined by AF-AAS. The biochemical composition of the tested mushroom material was confirmed with the FTIR method. Antioxidant properties were determined using the DPPH method, and the antiproliferative activity was assessed with the use of the MTT test. The presence of indole compounds (L-tryptophan, 6-methyl-D,L-tryptophan, melatonin, 5-hydroxy-L-tryptophan), phenolic compounds (p-hydroxybenzoic acid, gallic acid, catechin, phenylalanine), and sterols (ergosterol, ergosterol peroxide) as well as trace elements was confirmed in the mycelium and fruiting bodies of F. officinalis. Importantly, a high level of 5-hydroxy-L-tryptophan in in vitro mycelium cultures (517.99 mg/100 g d.w) was recorded for the first time. The tested mushroom extracts also showed antioxidant and antiproliferative effects on the A549 lung cancer cell line, the DU145 prostate cancer cell line, and the A375 melanoma cell line.
Topics: Antioxidants; Cell Proliferation; Coriolaceae; Fruiting Bodies, Fungal; Humans; Mycelium; Neoplasms; Phenols; Tumor Cells, Cultured
PubMed: 33208786
DOI: 10.1038/s41598-020-76899-1 -
FEMS Microbiology Reviews Jul 2007Ectomycorrhizal (ECM) fungi form mutualistic symbioses with many tree species and are regarded as key organisms in nutrient and carbon cycles in forest ecosystems. Our... (Review)
Review
Ectomycorrhizal (ECM) fungi form mutualistic symbioses with many tree species and are regarded as key organisms in nutrient and carbon cycles in forest ecosystems. Our appreciation of their roles in these processes is hampered by a lack of understanding of their soil-borne mycelial systems. These mycelia represent the vegetative thalli of ECM fungi that link carbon-yielding tree roots with soil nutrients, yet we remain largely ignorant of their distribution, dynamics and activities in forest soils. In this review we consider information derived from investigations of fruiting bodies, ECM root tips and laboratory-based microcosm studies, and conclude that these provide only limited insights into soil-borne ECM mycelial communities. Recent advances in understanding soil-borne mycelia of ECM fungi have arisen from the combined use of molecular technologies and novel field experimentation. These approaches have the potential to provide unprecedented insights into the functioning of ECM mycelia at the ecosystem level, particularly in the context of land-use changes and global climate change.
Topics: Biomass; Carbon Dioxide; DNA, Fungal; Ecosystem; Fungi; Mycelium; Mycorrhizae; Plant Roots; Soil Microbiology; Trees
PubMed: 17466031
DOI: 10.1111/j.1574-6976.2007.00073.x -
Molecules (Basel, Switzerland) Mar 2021In recent years, mushrooms have drawn the attention of agro-industries and food-industries as they were considered to be valuable natural sources of health promoting...
In recent years, mushrooms have drawn the attention of agro-industries and food-industries as they were considered to be valuable natural sources of health promoting compounds such as β-glucans, ergothioneine, and lovastatin. The detection and quantification of such compounds by implementing reliable analytical approaches is of the utmost importance in order to adjust mushrooms' cultivation conditions and maximize the production in different species. Toward this direction, the current study focuses on the comparison of ultraviolet-visible (UV-Vis) spectrometry and liquid chromatography-mass spectrometry (LC-MS) methods (a) by evaluating the content of ergothioneine and lovastatin in mushrooms and (b) by highlighting any possible substrate-based interferences that hinder the accurate determination of these two compounds in order to propose the technique-of-choice for a standardized bioactive compounds monitoring. For this purpose, mushrooms produced by three species (i.e., , , and ) on various cultivation substrates, namely wheat straw (WS), winery (grape marc (GM)), and olive oil (OL) by-products, were examined. Among the two applied techniques, the developed and validated LC-MS methods, exhibiting relatively short analysis time and higher resolution, emerge as the methods-of-choice for detecting ergothioneine and lovastatin in mushrooms. On the contrary, UV-Vis methods were hindered due to co-absorbance of different constituents, resulting in invalid results. Among the studied mushrooms, contained the highest amount of ergothioneine (822.1 ± 20.6 mg kg dry sample), whereas contained the highest amounts of lovastatin (1.39 ± 0.014 mg kg dry sample). Regarding the effect of different cultivation substrates, mushrooms produced on OL and WS contained the highest amount of ergothioneine, while mushrooms deriving from GM-based substrates contained the highest amount of lovastatin.
Topics: Agaricus; Ergothioneine; Lovastatin; Mycelium; Pleurotus
PubMed: 33805096
DOI: 10.3390/molecules26071832