Search Results (21)

Lentinula edodes (Berk.) Pegler, also known as Shiitake, is one of the most popular edible mushroom species containing high contents of polysaccharides, proteins and unique aroma, widely cultivated in China, Japan and Korea. A series of studies has been carried out on the extraction and active effect of the L. edodes polysaccharides, but the molecular mechanisms involved in the protein expression profiles during the whole life cycle are relatively unclear. This study employed an iTRAQ-MS/MS proteomic approach, combined with real-time quantitative PCR (qRT-PCR) and enzyme activity assays, to systematically analyze the protein expression profiles and their relationship with lignocellulose degradation in L. edodes across four key developmental stages: mycelia (SF), brown film formation (BF), primordia (YF), and fruiting bodies (MF). A total of 2043 proteins were identified, with 1188 being differentially expressed proteins (DEPs). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses revealed that metabolic processes, carbohydrate metabolism, and related pathways were significantly active during development. The study specifically focused on carbohydrate-active enzymes (CAZymes), identifying 197 CAZyme proteins classified into 78 families. Key families such as glycoside hydrolases (GHs) and carbohydrate esterases (CEs) played crucial roles in lignocellulose degradation. The enzymatic activities of major lignin-degrading enzymes (laccase, manganese peroxidase, and lignin peroxidase) were dynamically regulated across the developmental stages. qRT-PCR results largely corroborated the proteomic data, confirming the reliability of the protein expression profiles. This study provides a comprehensive, stage-resolved proteomic landscape of lignocellulose degradation during L. edodes development, revealing species-specific temporal dynamics, offering a valuable basis for understanding its growth and development, with implications for edible fungus cultivation and biomass conversion applications. Full article

Lentinula edodes (shiitake) is a major edible and medicinal mushroom and a key component of the horticultural mushroom industry in East Asia. During April–June 2024 cropping season, a widespread blue mold outbreak was observed on bag-cultivated shiitake in Xixia County, Henan Province, China. Affected cultivation rooms showed extensive blue-green sporulation on the exposed surfaces of substrate blocks and on developing and mature fruiting bodies, leading to rapid loss of marketability. To clarify the etiology of this disease, we coupled field surveys with morphological, molecular, and pathogenicity analyses. Fifty-five Penicillium isolates were obtained from symptomatic cultivation bags. Three representative isolates (LE06, LE15, and LE26) were characterized in detail. Colonies on PDA produced velutinous to floccose mycelia with blue-green conidial masses and terverticillate penicilli bearing smooth-walled, globose conidia. Sequencing of four loci—the internal transcribed spacer (ITS1-5.8S-ITS2), β-tubulin (benA), calmodulin gene (CaM), and RNA polymerase II second largest subunit (rpb2)—followed by multilocus phylogenetic analysis placed all three isolates in a well-supported clade with the ex-type CBS 227.28 of Penicillium bialowiezense. Inoculation of healthy shiitake cultivation bags with conidial suspensions (1 × 10⁶ conidia mL⁻¹) reproduced typical blue mold symptoms on substrate surfaces and fruiting bodies within 40 days post inoculation, whereas mock-inoculated controls remained symptomless. The pathogen was consistently reisolated from diseased tissues and showed identical ITS and benA sequences to the inoculated strains, thereby fulfilling Koch’s postulates. This is the first confirmed report of P. bialowiezense causing blue mold on shiitake, and it expands the known host range of this species. Our findings highlight the vulnerability of bag cultivation systems to airborne Penicillium contaminants and underscore the need for improved hygiene, environmental management, and targeted diagnostics in commercial shiitake production. Full article

Although the aroma profile of Lentinula edodes has been extensively studied in fruiting bodies, the mycelial stage provides a distinct context for elucidating the fundamental metabolic pathways, free from the complexities of organismal development. To elucidate the mechanism underlying aroma differences between L. edodes strain 808 (the control strain) and its mutant strain ww808 (with almost no shiitake aroma), this study employed GC-IMS combined with PCA and OPLS-DA to identify key aroma biomarkers during the mycelial stage. All analyses were performed with three biological replicates. Furthermore, fatty acids composition, key enzyme activities of the LOX pathway, and their gene expression levels were systematically compared. The results indicated significant differences in the content of volatile aroma compounds in the mycelia of the two strains, primarily stemming from fundamental restructuring of gene expression and enzyme activity in the LOX pathway. The LOX gene expression and LOX activity of 808 mycelium were relatively high, facilitating the accumulation of key aroma compounds such as phenylethanal, benzaldehyde, and ethyl acetate, which constitute its distinctive aromatic profile. However, although the mycelium of ww808 possessed richer fatty acid precursor (C18:2), its lower LOX gene expression restricted the flux of this pathway. The significantly increased expression of ADH2, ADH3, and ADH5 genes and higher ADH activity enhanced the conversion capacity of aldehydes to alcohols and ketones. Given the generally higher odor thresholds of alcohols and ketones compared to aldehydes, distinct aroma profiles emerged between the two strains. Pearson correlation analysis further confirmed the significant correlations between the aroma biomarkers, fatty acids, key genes, and enzyme activities. This study revealed the formation mechanism of aroma differences in the mycelia of the two strains from the perspective of metabolic pathways, providing a theoretical foundation and candidate targets for the directed genetic improvement of L. edodes aroma quality. Full article

With the increases in the global population, demand for meat, and income, plant-based meat alternatives using mushrooms, soybeans, and other ingredients are attracting increasing attention to address food security. Mushroom mycelia have a high moisture content and are prone to quality deterioration; therefore, interventions, such as freeze-drying and optimized packaging, are necessary to improve shelf life. Furthermore, controlling storage temperature is a key factor in preserving product quality. Therefore, in this study, Lentinula edodes (shiitake mushroom) mycelium sausage were stored under various packaging methods (aerobic packaging, vacuum packaging, and modified atmosphere packaging) and storage temperatures (0 and 4 °C). Quality and storability were evaluated at 0, 7, 15, 30, and 50 days using tests for proximate components, pH, storage loss, color, water activity (Aw), aerobic bacterial count, thiobarbituric acid reactive substances (TBARS), and volatile basic nitrogen (VBN) using an electronic nose and an electronic tongue. The vacuum-treated groups showed superior pH, CIE L*, CIE a*, Aw, TBARS, and VBN levels. At 0 °C, each treatment group exhibited significantly lower VBN levels on days 30 and 50 (p < 0.05). Overall, vacuum packaging and low storage temperatures are expected to be beneficial for maintaining the quality and storage characteristics of shiitake mushroom mycelium sausages. Full article

Flammulina filiformis is one of the widely produced edible fungi worldwide. It is rich in γ-aminobutyric acid (GABA), a non-protein amino acid with important physiological functions in humans. To investigate the functions of key genes in the GABA metabolic pathway of F. filiformis, we isolated the monokaryon Fv-HL23-1 from the factory-cultivated F. filiformis strain Fv-HL23 and then sequenced and assembled the genome using the PacBio Sequel and Illumina NovaSeq sequencing platforms. The results showed that the genome comprised 140 scaffolds with a total length of 40.96 Mb, a GC content of 49.62%, an N50 of 917,125 bp, and 14,256 protein-coding genes. Phylogenetic analysis based on the whole genome revealed a close evolutionary relationship of Fv-HL23-1 with Armillaria mellea, Lentinula edodes, and Schizophyllum commune. A total of 589 carbohydrate-active enzymes were identified in the genome of Fv-HL23-1, suggesting its strong lignocellulose degradation ability, and 108 CYP450 gene family members were identified, suggesting important functions such as resistance to stress, secondary metabolite synthesis, and growth and development. The F. filiformis proteins glutamate decarboxylase 1 (Ff-GAD1) and glutamate decarboxylase 2 (Ff-GAD2), which may be responsible for GABA synthesis, were identified by protein alignment. Molecular docking analysis showed that Ff-GAD2 may have better catalytic activity than Ff-GAD1. To verify the function of Ff-gad2, its heterologous expression in the mycelia of the mononuclear Hypsizigus marmoreus was analyzed. Compared with wild type, the GABA content of mycelia was increased by 85.40–283.90%, the growth rate was increased by 9.39 ± 2.35%, and the fresh weight was increased by 18.44 ± 7.57%. Ff-GAD2 may play a catalytic role in GABA synthesis. In addition, the expression of the full-length Ff-gad2 gene was increased by 7.96 ± 1.39 times compared with the exon expression level in H. marmoreus mycelia, suggesting that the intron may contribute to the heterologous expression of Ff-GAD2. Based on whole-genome sequencing, we analyzed the enzyme system related to the important life activities of F. filiformis, focusing on the function of Ff-GAD, a key enzyme in the GABA synthesis pathway. The results lay a foundation for elucidating the GABA metabolism pathway of edible fungi and developing targeted breeding strategies for GABA-producing edible fungi. Full article

In the industrial production of Lentinula edodes and Pleurotus eryngii, slow growth of the mother seed and insufficient hyphal vitality can significantly affect the cultivation process. To shorten the growth period on traditional PDA medium, two strains of L. edodes and P. eryngii were cultured with different proportions of P. tuber-regium and Wolfiporia hoelen sclerotium powders added into the medium to investigate the effect on the mycelial growth. Compared to the PDA, the addition of sclerotia powder significantly enhanced the growth of mycelia, with an optimal addition ratio of 2%. Transcriptome sequencing was performed after culturing L. edodes and P. eryngii on PDA, PDA with 2% P. tuber-regium sclerotium powder, and PDA with 2% W. hoelen sclerotium powder. GO enrichment analysis of the differentially expressed genes (DEGs) of L. edodes and P. eryngii strains cultured in the sclerotia powder media showed significant changes in oxidoreductase and glucosidase activities. Changes were observed in KEGG annotation for carbohydrate metabolism, glycolysis, pyruvate metabolism, and other energy metabolic pathways. Moreover, carbohydrate-active enzyme (CAZyme) family genes were predominantly upregulated. The increase in the activity of CAZyme and oxidoreductases promotes the degradation of nutrients in the sclerotia into small-molecule substances, which explains why the sclerotia powder culture medium promotes mycelial growth. Full article

Spent mushroom substrates (SMSs) and mushroom basal bodies (MBBs) are significant by-products because of their nutrient content even after harvesting. This study aimed to evaluate the effect of these two by-products, derived from Agaricus bisporus (Ab) and Cyclocybe cylindracea (Cc) cultivation, as potential growth and biochemical composition add-value enhancers of edible mushroom mycelia such as Pleurotus ostreatus, C. cylindracea, and Lentinula edodes. Fungal growth substrates enriched with SMS and MBB extracts significantly affected the growth of mushroom mycelia. In particular, on P. ostreatus, the MBBs Ab and Cc extracts determined an increase in mycelial weight by 89.5%. Also, by-products influenced mushrooms’ mycelial texture, which appeared more floccose and abundant in growth. FT-IR analysis showed that L. edodes mycelium, grown on MBB substrates, showed the highest increase in bands associated with proteins and chitin. Results demonstrated that mushroom by-products enhance mycelial growth and confer an enrichment of compounds that could increase mycelial resistance to pathogens and make a nutraceutical improvement. Full article

A standardized extract of cultured Lentinula edodes mycelia (ECLM, AHCC^®) has been shown to have beneficial effects on organ metabolism. ECLM has been indicated to have liver protective properties by suppressing inflammatory responses. The pathogenesis of hepatic ischemia-reperfusion injury is thought to involve the induction of inflammatory mediators. However, whether ECLM affects inflammatory mediators caused by warm hepatic ischemia-reperfusion injury and partial hepatectomy (HIRI+PH) has not been clarified. In this study, we evaluated the protective effects of ECLM against liver damage caused by HIRI+PH. Rats were fed a normal diet (HIRI+PH) or a normal diet with 2% ECLM (HIRI+PH and ECLM) for ten days, then the liver and duodenal ligament were clamped and subjected to 15 min of hepatic ischemia. After 70% hepatectomy, the inflow occlusion was released, and liver and blood samples were collected at 3, 6, and 24 h. The effect of ECLM on mortality induced by 30 min of ischemia and hepatectomy was evaluated. The results showed that ECLM attenuated pathological liver damage, including apoptosis, in the rats treated with HIRI+PH, and decreased serum aminotransferase activity; ECLM decreased mRNA levels of the inflammation-related genes inducible nitric oxide synthase and C-X-C motif chemokine ligand 1, and increased mRNA levels of interleukin 10, an anti-inflammatory cytokine; ECLM increased hepatocyte growth factor mRNA levels and Ki-67 labeled nuclei in the liver at 24 h; ECLM significantly reduced HIRI+PH-induced mortality. In conclusion, ECLM may prevent HIRI+PH-induced liver injury in part by suppressing various inflammatory responses and promoting liver regeneration. Full article

Puberty is a critical developmental period of life characterized by marked physiological changes, including changes in the immune system and gut microbiota development. Exposure to inflammation induced by immune stressors during puberty has been found to stimulate central inflammation and lead to immune disturbance at distant sites from the gut; however, its enduring effects on gut immunity are not well explored. Therefore, in this study, we used a pubertal lipopolysaccharides (LPS)-induced inflammation mouse model to mimic pubertal exposure to inflammation and dysbiosis. We hypothesized that pubertal LPS-induced inflammation may cause long-term dysfunction in gut immunity by enduring dysregulation of inflammatory signaling and epigenetic changes, while prebiotic/probiotic intake may mitigate the gut immune system deregulation later in life. To this end, four-week-old female Balb/c mice were fed prebiotics/probiotics and exposed to LPS in the pubertal window. To better decipher the acute and enduring immunoprotective effects of biotic intake, we addressed the effect of treatment on interleukin (IL)-17 signaling related-cytokines and pathways. In addition, the effect of treatment on gut microbiota and epigenetic alterations, including changes in microRNA (miRNA) expression and DNA methylation, were studied. Our results revealed a significant dysregulation in selected cytokines, proteins, and miRNAs involved in key signaling pathways related to IL-17 production and function, including IL-17A and F, IL-6, IL-1β, transforming growth factor-β (TGF-β), signal transducer and activator of transcription-3 (STAT3), p-STAT3, forkhead box O1 (FOXO1), and miR-145 in the small intestine of adult mice challenged with LPS during puberty. In contrast, dietary interventions mitigated the lasting adverse effects of LPS on gut immune function, partly through epigenetic mechanisms. A DNA methylation analysis demonstrated that enduring changes in gut immunity in adult mice might be linked to differentially methylated genes, including Lpb, Rorc, Runx1, Il17ra, Rac1, Ccl5, and Il10, involved in Th17 cell differentiation and IL-17 production and signaling. In addition, prebiotic administration prevented LPS-induced changes in the gut microbiota in pubertal mice. Together, these results indicate that following a healthy diet rich in prebiotics and probiotics is an optimal strategy for programming immune system function in the critical developmental windows of life and controlling inflammation later in life. Full article

Mycelial cultures of Lentinula edodes, an edible and medicinal mushroom, have been used in our previous research to obtain selenium-containing immunomodulatory preparations. Our current attempts to obtain a new preparation containing both selenium and zinc, two micronutrients necessary for the functioning of the immune system, extended our interest in the simultaneous accumulation of these elements by mycelia growing in media enriched with selenite and zinc(II) ions. Subsequently, we have studied the effects of new L. edodes mycelium water extracts with different concentrations of selenium and zinc on the activation of T cell fraction in human peripheral blood mononuclear cells (PBMCs). Flow cytometry analysis was used to measure the expression of activation markers on human CD4+ and CD8+ T cells stimulated by anti-CD3 and anti-CD3/CD28 antibodies (Abs). It was demonstrated that statistically significant changes were observed for PD-1 and CD25 antigens on CD8+ T cells. The selenium and zinc content in the examined preparations modified the immunomodulatory activity of mycelial polysaccharides; however, the mechanisms of action of various active ingredients in the mycelial extracts seem to be different. Full article

The coronavirus disease 2019 (COVID-19) pandemic has significantly impacted global public health safety and the economy. Multiple antiviral drugs have been developed, and some have received regulatory approval and/or authorization. The use of nutraceuticals can be beneficial for preventing and treating COVID-19 complications. AHCC is a standardized, cultured extract of an edible mushroom Lentinula edodes of the Basidiomycete family of fungi that is enriched in acylated α-1,4-glucans. Here, we evaluated the effects of the oral administration of AHCC on the host response to SARS-CoV-2 infection in two murine models, K18-hACE2 transgenic mice and immunocompetent BALB/c mice. Oral administration of AHCC every other day for one week before and one day post SARS-CoV-2 infection in both strains of mice decreased the viral load and attenuated inflammation in the lungs. AHCC treatment also significantly reduced SARS-CoV-2-induced lethality in the K18-hACE2 mice. AHCC administration enhanced the expansion of γδ T cells in the spleen and lungs before and after viral infection and promoted T helper 1-prone mucosal and systemic T cell responses in both models. In AHCC-fed BALB/c mice, SARS-CoV-2 specific IgG responses were also enhanced. In summary, AHCC supplementation enhances host resistance against mild and severe COVID-19 infection primarily via the promotion of innate and adaptive T cell immune responses in mice. Full article

Mycelial ageing is associated with ROS and autophagy in Lentinula edodes. However, the underlying cellular and molecular mechanisms between ROS and autophagy remain obscure. This study induced autophagy in L. edodes mycelia through exogenous H₂O₂ treatment. Results showed that 100 μM H₂O₂ treatment for 24 h significantly inhibited mycelial growth. H₂O₂ caused the depolarisation of MMP and accumulation of TUNEL-positive nuclei, which was similar to the ageing phenotype of L. edodes mycelia. Transcriptome analysis showed that differentially expressed genes were enriched in the mitophagic, autophagic, and MAPK pathways. LeAtg8 and LeHog1 were selected as hub genes. RNA and protein levels of LeATG8 increased in the H₂O₂-treated mycelia. Using fluorescent labelling, we observed for the first time the classic ring structure of autophagosomes in a mushroom, while 3D imaging suggested that these autophagosomes surrounded the nuclei to degrade them at specific growth stages. Phospho-LeHOG1 protein can translocate from the cytoplasm to the nucleus to regulate mycelial cells, resisting ROS-induced oxidative stress. Furthermore, LeATG8 expression was suppressed when LeHOG1 phosphorylation was inhibited. These results suggest that the LeATG8-dependent autophagy in L. edodes mycelial is closely associated with the activity or even phosphorylation of LeHOG1. Full article

The spawn of Lentinula edodes and other basidiomycete fungi tend to age with long-term culture. This causes heavy yield losses if aging spawn is used for propagation. In this study, we cultivated dikaryotic L. edodes mycelia in plates for 60 days to produce intrinsic aging phenotypes. We found that intracellular reactive oxygen species levels increased in contrast to mitochondrial depolarization and also observed greater DNA fragmentation with longer culture time. Transcriptome analysis of mycelia at different growth stages revealed pronounced expression differences between short- and long-term cultures. In particular, “phenylalanine, tyrosine, and tryptophan biosynthesis”, “mitophagy and autophagy”, “MAPK signaling pathway”, and “ABC transporter” were among the enriched terms in the mycelial aging process. Weighted correlation network analysis identified LeAtg8, LeHog1, LePbs2, and LemTOR as key genes during aging. Western blotting confirmed that LeATG8 and phosphorylated LeHOG1 protein levels were significantly upregulated in aging mycelia. Our combined analytical approach provides insights into the mechanisms that regulate mycelial aging, indicating that autophagy/mitophagy plays a major role in counteracting the effects of age on mycelial growth development. Full article

It has been demonstrated that lignin was efficiently degraded by Lentinula edodes (L. edodes). However, the process of lignin degradation and utilization by L. edodes has not been discussed in detail. Therefore, the effects of lignin on L. edodes mycelium growth, chemical compositions, and phenolic profiles were investigated herein. It has been revealed that 0.10% lignin acted as the most effective concentration to accelerate mycelia growth, which yielded the highest biomass of 5.32 ± 0.07 g/L. Furthermore, a 0.10% concentration of lignin promoted the accumulation of phenolic compounds, especially protocatechuic acid, with peak value of 48.5 ± 1.2 μg/g. In contrast, the higher concentration of lignin (0.20%) exerted an inhibitory effect on the growth of L. edodes. Overall, the application of lignin at the optimal concentration of 0.10% could not only enhance the mycelial growth but also accumulate the phenolic acids and raise the nutritional and medical values of L. edodes. Full article

We reported the development of an effective cancer treatment using a multidisciplinary treatment, including photodynamic therapy (PDT) with indocyanine green (ICG) liposomes and a combination of Lentinula edodes mycelia (LEM) and hydrogen gas inhalation therapy. ICG liposomes were prepared by adding 5 mg of ICG to 50 mL liposomes. Later, 25 mL of ICG liposomes were diluted with 250 mL of 5% glucose solution and administered intravenously to the patient. We selected the multi-laser delivery system (MLDS), a laser irradiator for performing PDT. Further, the patients received a combination of LEM and hydrogen gas inhalation therapy throughout the treatment. We reported two cases of PDT therapy, one with middle intrathoracic esophagus carcinoma and the other with hypopharyngeal cancer. In the first case, the MLDS laser was directly attached to the endoscope and directed to the cancer area with wavelengths of 810 nm. After the treatment, a biopsy demonstrated no tumor recurrence. In the second case, the patient was treated with endovascular PDT using ICG liposomes and MLDS fiber optics. Later, tumor shrinkage was demonstrated after the first round and disappeared after six months. In conclusion, the present findings suggest that the effect of PDT using ICG liposomes with LEM and hydrogen gas may eradicate cancer without burdening patients by enhancing tumor immunity. Full article