Medicinal Mushroom Research Articles

First Report of Pestalotiopsis trachicarpicola Causing Apothecium Wither on Cultivated Morchella sextelata in Sichuan, China.

Morel (Morchella sextelata) is a valuable medicinal and edible mushroom, but the wild yield is seriously insufficient, so several artificial varieties have been developed to alleviate the shortage of wild yield. However, in 2020-2022, apothecium wither symptoms were observed in Nanchong, Sichuan. A total of 30.80% of the morels in the planting base (1.3 km2) showed these symptoms. The initial symptoms were slight white lesions on the surface of apothecium, then the mycelia gradually invaded the interior of the fruiting body, and eventually leading to black and wilt apothecium (Fig. 1a). Fifteen symptomatic morels were collected and ten isolates were obtained using the single spore isolation technique developed by Chomnunti et al. (2014). The morphological characteristics of the ten isolates were similar, which showed dense aerial white mycelia colony texture on PDA, and later forming concentric black mucus (Fig. 1c). The reverse side was yellow (Fig. 1d). The acervulus was floral-shaped and discrete, with smooth walls and measured 120.0 to 400.5 × 15.5 to 40.0 μm (n=10) (Fig. 1e); the conidia were fusiform and hyaline, 21.0 to 28.6 × 6.0 to 7.6 μm in size (n=50), each contained five cells. The apical cell was hyaline, conic and with 2 to 4 tubular apical appendages on the top. The three intermediate cells were brown to olivaceous, doliform to cylindrical, constricted at septa. The basal cell was conic to acute (Fig. 1f). The morphological characteristics were consistent with the published description of Pestalotiopsis trachicarpicola (Maharachchikumbura et al., 2012). PCR was performed with primers ITS1/ITS4 for internal transcribed spacer (ITS) (White et al., 1990), BT2A/BT2B for β-tubulin gene (TUB) (Glass and Donaldson, 1995), and EF1-526F/EF1-1567R for translation elongation factor 1-alpha (TEF-1α) (Roger et al., 1999). The pairwise alignments of ITS, TUB, and TEF-1α sequences was nearly 100% identical to P. trachicarpicola with GenBank accession numbers MT889666.1 (579/585 bp, 99%), MT884145.1 (445/450 bp, 99%), and MW149930.1 (946/958 bp, 99%), respectively. The resulting sequences were deposited in GenBank (Accession no. ITS: OL362082; TUB: OL828342; and TEF-1α: OL905009). Phylogenetic analysis performed with maximum likelihood method used MEGA 7.0 (1000 bootstrap replications) classified WLM5 into the P. trachicarpicola clade (Fig. 2), so we finally confirmed the identity of WLM5 as P. trachicarpicola. To fulfill Koch's postulates, twenty morels were surface disinfected with 2% sodium hypochlorite and then artificially wounded (diameter of 0.5 mm) prior to inoculation with 200 μL conidial suspension (105 conidia/mL), while an equal amount of sterile distilled water was applied to controls. After 4 days, the inoculated fruiting bodies showed symptoms consistent with field infection (Fig. 1b) and P. trachicarpicola was re-isolated using the same protocol, while the control remained asymptomatic. This first report of P. trachicarpicola causing apothecium wither on morel will help develop robust disease management strategies against this emerging fungal pathogen.

Emulsion for stabilizing β-carotene and curcumin prepared directly using a continuous phase of polysaccharide-rich Schizophyllum commune fermentation broth

In this study, we examined the effect of Schizophyllum commune fermentation broth (SCFB) rich in polysaccharides (SCFP) on the stability and bioaccessibility of β-carotene and curcumin. An SCFB-stabilized oil-in-water (o/w) emulsion (SCFBe) was prepared using SCFB as the continuous phase, and then evaluated for storage stability using an SCFP-based emulsion (SCFPe) as the control. The findings revealed that SCFBe is more stable at 60 °C than SCFPe, and stratification or droplet size varied at differing pH levels (3–9) and concentrations of Na+ (0.1–0.5 M) and Ca2+ (0.01–0.05 M). Since the absolute value of the zeta potential of SCFBe is much lower at 60 °C than that at 4 °C and 25 °C, a higher temperature (60 °C) may enhance the reactivity of polysaccharides and proteins in SCFB to improve the stability of SCFBe. Both the protective impact of SCFB on functional food molecules and their capacity to block lipid oxidation increased as polysaccharide content improved. The bioaccessibility of β-carotene after in vitro simulated gastrointestinal digestion is 11.18 %–12.28 %, whereas that of curcumin is 31.64 %–33.00 %. By fermenting edible and medicinal fungi in liquid, we created a unique and environmentally friendly approach for getting food-grade emulsifiers without extraction.

Isolation and Identification of Chemical Compounds from Agaricus blazei Murrill and Their In Vitro Antifungal Activities

This study explores the antifungal properties of Agaricus blazei Murrill, a valuable medicinal and edible fungus. Six compounds (1–6) were first isolated from A. blazei using various isolation techniques and identified using spectroscopic methods. These compounds include linoleic acid, 1,1′-oxybis(2,4-di-tert-butylbenzene), glycerol monolinoleate, volemolide (17R)-17-methylincisterol, (24s)-ergosta-7-en-3-ol, and dibutyl phthalate. This study also assesses the antifungal activities of these compounds against Trichophyton mentagrophology, Trichophyton rubrum, Candida albicans, and Cryptococcus neoformans. The results demonstrate varied sensitivities against these pathogenic fungi, with compound 2 showing significant inhibition against T. mentagrophology, compound 3 showing significant inhibition against T. rubrum, and compound 6 showing significant inhibition against C. albicans. This study underscores the medicinal potential of A. blazei as an antifungal agent and sheds light on its valuable research implications.

Fungi and their medicinal properties: a scientometric study

Fungi are eukaryotic, heterotrophic, unicellular or multicellular organisms that feed by absorption. Some of their attributes stand out in the medicinal and pharmaceutical sector since several genera of fungi have metabolic properties and compounds related to the manufacture of numerous medicines. In this context, the objective of this study was to carry out a scientometric analysis to verify the application and properties of medicinal fungi as well as their contributions to the knowledge of medicinal fungi. A survey of applied literature, which included the word “Medicinal fungi”, was carried out based on the articles indexed in the Thomson Reuters base (www.isiwebofknowledge.com), considering the period from 2003 to 2021. One hundred and thirty-nine scientific articles were used for the scientometric analysis, and these were divided into six time intervals (2003-2006, 2007-2009, 2010-2012, 2013-2015, 2016-2018, 2019-2021). An increase was noted in the scientific production on medicinal fungi from 2007 onwards, mostly presenting publications with an experimental approach. The phylum Basidiomycota, followed by Ascomycota, presented the highest number of records in the study, mainly with applications aimed at antitumor and antibacterial activity. Therefore, the importance of studies that aim to expand the knowledge of the properties of medicinal fungi is highlighted, providing effective results for various diseases, since their cultivation and development can positively generate equitable economic growth, with regional and national impacts.

Extraction, isolation, identification, and bioactivity of polysaccharides from Antrodia cinnamomea

Antrodia (A) cinnamomea is a precious edible and medicinal mushroom and has attracted attention because of its rare resources and unique bioactivities. The bioactive compounds from the fruit body, mycelium, and fermentation broth of A. cinnamomea include triterpenoids, polysaccharides, antroquinonols, benzenoids, and succinic and maleic acid derivatives. Among these, polysaccharides are very important and are high-content bioactive compounds. The A. cinnamomea polysaccharides (APSs) present numerous biological activities, such as antiviral, anticancer, anti-inflammatory, antivascular, immunoregulation, antioxidant, and nerve protection. However, only few studies have focused only on APSs so far. Therefore, the cultivation methods, extraction, isolation, composition, structure, biological activity, and application of APSs are summarized in this review to provide a comprehensive and convenient reference for further research and development on APSs.

Nature's Own Pharmacy: Mushroom-Based Chemical Scaffolds and Their Therapeutic Implications.

Mushrooms are new potential sources of valuable medicines, long neglected because of difficulties experienced in their cultivation. There is a large variety of medicinal mushrooms which possess significant therapeutic properties and are used as medications for various diseases because they contain several novel highly bioactive components. Medicinal mushrooms can be identified based on their morphology, size, mass, and the color of the stalk, cap and spore, and attachment to the stalk. Medicinal mushrooms possess a variety of important biological activities and are used as antioxidants, hepatoprotectors, anticancer, antidiabetic, anti-inflammatory, antiaging, antiviral, antiparasitic, and antimicrobial agents, among others. This review provides a basic overview of the chemical scaffolds present in mushrooms and their therapeutic implications in the human body.

Study on some biological characteristics of the albino strain of medicinal fungus Cordyceps militaris isolated in Vietnam

Cordyceps militaris is one of the important medicinal fungi in traditional medicine. The fruiting body of C. militaris contains various bioactive compounds that have health-enhancing effects and support the treatment of various diseases. Therefore, this fungus is a potential source of medicinal materials for the development of healthcare products. The search for new strains of C. militaris with high yield, quality, and uniqueness is always of great interest. This study has provided new findings on some biological characteristics of an albino strain of C. militaris isolated in Vietnam. The albino strain of C. militaris SHBTQ was isolated and accurately identified based on morphological characteristics and the ITS (internal transcribed spacer) sequence of ribosomal DNA. The mycelial of C. militaris SHBTQ is white and remains stable for five successive generations. The most suitable carbon and nitrogen sources for the growth of the albino SHBTQ strain were sucrose and yeast extract, respectively. Notably, the SHBTQ strain is capable of forming fruiting bodies, with an average fresh fruiting body yield of 30 g/box, a biological efficiency of 12.2%, and a cordycepin content of 10.45 mg/g of dried fruiting bodies. This newly discovered strain offers great potential for further applications in production.

Valorization of Grape Pomace for Trametes versicolor Mycelial Mass and Polysaccharides Production

Polysaccharides and protein–polysaccharide complexes produced from the fungal strain Trametes versicolor have demonstrated bioactive properties that depend on the substrate, the fermentation conditions and also the fungal strain. Likewise, the submerged and controlled fermentation of medicinal mushrooms elicits numerous advantages over traditional processes to produce mycelia and added-value products, along with the exploitation of biodiversity. This study evaluated the growth profile of an indigenous T. versicolor isolate using commercial nutrients that were subsequently replaced with renewable resources, specifically, grape pomace extract (GPE), under static and shaking conditions. The effect of elicitor addition was also assessed using GPE. The process productivity was significantly improved, yielding 21 g/L of biomass. Agitation proved beneficial for all examined cases regarding biomass productivity and substrate consumption rates. The chemical and antioxidant profile of crude intracellular and extracellular polysaccharides was determined, whereby intracellular extracts indicated >50% antioxidant activity. FTIR analysis validated the preliminary chemical characterization of the extracts, whereas the amino acid profile of IPS extracts was also included. Evidently, our study elaborates on the development of a bioconversion concept to valorize wine-making side-streams to formulate added-value products with potential bioactive attributes.

Ganoderma microsporum immunomodulatory protein as an extracellular epidermal growth factor receptor (EGFR) degrader for suppressing EGFR-positive lung cancer cells

Epidermal growth factor receptor (EGFR) abnormalities relevant to tumor progression. A newly developed strategy for cancer therapy is induction of EGFR degradation. GMI, an immunomodulatory protein from the medicinal mushroom Ganoderma microsporum, exhibits anticancer activity. However, its role in the intracellular trafficking and degradation of EGFR remains unclear. In this study, we discovered that GMI inhibits the phosphorylation of multiple tyrosine kinases. Specifically, GMI was discovered to suppress lung cancer cells harboring both wild-type and mutant EGFR by inhibiting EGFR dimerization and eliminating EGFR-mediated signaling. Functional studies revealed that GMI binds to the extracellular segment of EGFR. GMI interacts with EGFR to induce phosphorylation of EGFR at tyrosine1045, which triggers clathrin-dependent endocytosis and degradation of EGFR. Furthermore, in the mouse models, GMI was discovered to suppress tumor growth. Knockdown of EGFR in lung cancer cells abolishes GMI's anticancer activity in vivo and in vitro. Our results reveal the interaction mechanisms through which GMI induces EGFR degradation and abolishes EGFR-mediated intracellular pathway. Our study indicates that GMI is an EGFR degrader for inhibiting EGFR-expressing tumor growth.

The Effect of Mushroom Dietary Fiber on the Gut Microbiota and Related Health Benefits: A Review.

Mushroom dietary fiber is a type of bioactive macromolecule derived from the mycelia, fruiting bodies, or sclerotia of edible or medicinal fungi. The use of mushroom dietary fiber as a prebiotic has recently gained significant attention for providing health benefits to the host by promoting the growth of beneficial microorganisms; therefore, mushroom dietary fiber has promising prospects for application in the functional food industry and in drug development. This review summarizes methods for the preparation and modification of mushroom dietary fiber, its degradation and metabolism in the intestine, its impact on the gut microbiota community, and the generation of short-chain fatty acids (SCFAs); this review also systematically summarizes the beneficial effects of mushroom dietary fiber on host health. Overall, this review aims to provide theoretical guidance and a fresh perspective for the prebiotic application of mushroom dietary fiber in the development of new functional foods and drugs.

Characterisation and identification of chemical constituents in aqueous extract of Fomes officinalis Ames based on ultrahigh-performance liquid chromatography tandem quadrupole-Orbitrap high-resolution mass spectrometry.

Fungal species are an attractive resource for physiologically functional food and drug precursor. Fomes officinalis Ames, a medicinal fungus, is traditionally used as a folk medicine in traditional Chinese medicine prescription for the therapy of cough and asthma. The water-soluble substances in Chinese herbal medicines are likely to play an important physiological function. However, information on probing and identifying chemical components of the aqueous extract of Fomes officinalis Ames (AFO) remains unknown. This study was conducted to screen and characterise the chemical components of AFO. An effective and sensitive ultrahigh-performance liquid chromatography tandem quadrupole-Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap-HRMS) method with the Full MS/PIL/dd-MS2 acquisition approach was applied for the profiling of chemical components in AFO. An HSS T3 column was used for component separation, and a strategy of simultaneous targeted and untargeted multicomponent characterisation was implemented. Multiple identification approaches were used, including accurate molecular mass and elemental composition matching, literature and database searching, and fragmentation rules elucidation. A total of 115 components, including 20 amino acids and derivatives, six nucleobases, nine nucleosides, 75 dipeptides, two tripeptides, and three other components, were tentatively identified. Among them, the targeted exploring method screened six nucleobases and nine nucleosides including modified nucleosides. To our best knowledge, this is the first time a report has been done on the presence of the 115 compounds in AFO. Profiling and characterisation compounds of AFO enriched its material basis, which would lay the foundation for improving potential medicinal and nutritional values and effecting comprehensive quality control of Fomes officinalis Ames.

Profiling of tyrosinase levels, antioxidant and DNA protective properties in different edible and medicinal mushroom species from India

AbstractBackgroundTyrosinase is a polyphenol oxidase, one of the globally attracted enzymes for its multi‐mechanistic approaches in various fields such as pharma, food, environment, medicine, and agriculture. Tyrosinase from natural nutritive edible mushrooms gained more attention for their rich levels and inexpensive nature. Our previous work has revealed that mushroom tyrosinase has strong similarities in structure and function with that of skin tyrosinase, which has significance as a skin tumor marker. Besides, mushrooms are a richer source of antioxidants. However, it is imperative to analyze all the available varieties of mushrooms for bioactive potentials like tyrosinase and antioxidant levels. Thus, the main goal of the study was to identify the variety of mushrooms that can be selected for continuous multiplication and propagation as a viable source for tyrosinase, which can be employed to screen tyrosinase inhibitors as skin tumor inhibitors.ResultsOf the 18 varieties screened for tyrosinase levels, Boletus edulis, Termitomyces hemii, and Termitomyces mummiformis had high levels of tyrosinase and exhibited potent hydrogen peroxide and nitric oxide scavenging activity as well as DNA protection. The potent DNA and antioxidant abilities appear to be due to the enriched levels of hydroxybenzoates.ConclusionThe current study could provide an economic source of tyrosinase (mushrooms) that can be used to monitor tyrosinase inhibitors, a potent component that enables their use against skin cancer.

Structurally diverse indole alkaloids with cytotoxicity from Lonicera Japonica-associated endophytic fungus Penicillium ochrochloron YT2022-65

Natural products, especially fungal secondary metabolites, have been served as valuable sources of drug leads in pharmaceutical industry. Medicinal plants-associated endophytic fungi possess a well-developed secondary metabolism. In this study, chemical investigation on Penicillium ochrochloron YT2022-65, an endophytic fungus associated with Lonicera Japonica, led to the isolation of six structurally diversified indole alkaloids, including a new one, namely peniochroloid A (1), as well as five previously reported alkaloids, flavonoid B (2), brocaeloid C (3), isoroquefortine C (4), roquefortine C (5), and dihydrocarneamide A (6). Their structures, including the absolute configuration of 1, were determined by a combined analysis of HRESIMS, NMR spectroscopic data, and calculation of the optical rotation. Their cytotoxicity against A549, HepG2, MCF-7, and THP-1 cell lines were evaluated in vitro. The new compound 1 was found to possess considerable cytotoxicity against MCF-7 and THP-1 cell lines with IC50 values of 10.2 and 11.0 μM, respectively.

First report of emerging fungal pathogens of Cordyceps militaris in Vietnam

Cultivation of Cordyceps militaris, a valuable medicinal and edible fungus, has dramatically increased in Vietnam since 2010. During industrial production, parasitic white molds were found to infect the mycelia and fruiting bodies of C. militaris causing significant quality and yield losses. Two different fungal strains were obtained from the mycelia and fruiting bodies of C. militaris in Danang mushroom farms and were characterized by morphological and multiple DNA markers analysis. The sequence alignment of ITS, LSU and rpb2 markers revealed that the pathogens are related to the type species Lecanicillium coprophilum and Calcarisporium cordycipiticola with more than 99% sequence identities. The growth characteristics and pathogenic activities of the two isolated species on their host C. militaris were also investigated. The phylogenetic analysis based on the ITS sequences showed that L. coprophilum WF2611 is closer to its host C. militaris than C. cordycipiticola NT1504. To our knowledge, this is the first worldwide report of C. militaris infected by L. coprophilum which would be an useful information on prevention and control of the disease and be helpful for the industrial cultivation of C. militaris.

Assessment of cytotoxicity, acute, subacute toxicities and antioxidant activities (in vitro) of Sanghuangporus vaninii crude polysaccharide

Ethnopharmacology relevanceSanghuangporus vaninii (S. vaninii), as a traditional large medicinal fungus, has a history of more than 2000 years in Chinese history and has been widely used to treat female diseases such as vaginal discharge, amenorrhea, and uterine bleeding, and recent pharmacological studies have also found that it has antioxidant, anti-inflammatory, and anti-tumor physiological activity, which has received more and more attention. Aim of the studyThe objective was to evaluate cytotoxicity and the acute, subacute toxicity, and in vitro antioxidant activity of S. vaninii crude polysaccharide (SVP). Materials and methodsThe monosaccharide composition of SVP was determined by HPLC (high-performance liquid chromatography). The cytotoxicity of different concentrations of SVP on three types of cells (HT-22, Kupffer macrophages, HEK293) was assessed using CCk-8. The acute toxicity in vivo was evaluated for 14 days after the administration of SVP (2500,5000, or 10,000 mg/mL). For the evaluation of subacute toxicity, mice were daily treated for 28 days with SVP (2500,5000, or 10,000 mg/mL). In addition, DPPH, hydroxyl radical, and superoxide anion radical were used to evaluate the in vitro antioxidant activity of SVP. ResultsSVP was not toxic in all three cell lines tested. In vitro antioxidant tests on the extracts showed that SVP possessed a strong antioxidant capacity in vitro. In the acute study, the no-observed-adverse-effect level (NOAEL) in male and female rats was 10，000 mg/kg body weight. There were also no deaths or severe toxicity associated with SVP in subacute studies. However, SVP treatment had a decreasing effect on body weight in mice of both sexes (2500, 5000, and 10000 mg/kg). At doses (5000 and 10,000 mg/kg), SVP had a reduced effect on food intake in both male and female mice. In addition, there were significant effects on organ coefficients of the liver, lung, and kidney. Hematological analysis showed significantly lower LYM (%) values in mice of both sexes, with significantly lower MCH (pg) values obtained in males (5000 mg/kg and 10000 mg/kg) and higher GRAN (%) values in females. In addition, the RDW-SD (fL) values were significantly lower in the male mice given the highest dose. Biochemical tests showed that there were no significant changes in ALT, AST, TP, and Cr levels after SVP treatment. In histopathological analysis, mild liver toxicity was observed in both female mice treated with 10,000 mg/kg SVP. ConclusionThe extract of SVP showed a predominance of polysaccharide compounds, with non-toxic action in vivo. Our approach revealed SVP on the chemical composition and suggests a high margin of safety in the popular use of medicinal fungi. In conclusion, our results suggest that SVP is safe, and can be used as health care products and food.

Determination of Phenolic Acids Using Ultra-High-Performance Liquid Chromatography Coupled with Triple Quadrupole (UHPLC-QqQ) in Fruiting Bodies of Sanghuangporus baumii (Pilát) L.W. Zhou and Y.C. Dai.

Sanghuangporus, a medicinal mushroom, has gained significant attention due to its beneficial properties. Phenolic acids are among the major bioactive compounds in Sanghuangporus, known for their antioxidant and anti-inflammatory activities. To precisely quantify the phenolic acid content, we developed a method utilizing ultra-high-performance liquid chromatography with triple quadrupole (UHPLC-QqQ). This study optimized the UHPLC-QqQ conditions to simultaneously separate and detect eight phenolic acids in Sanghuangporus baumii (Pilát) L.W. Zhou and Y.C. Dai, including chlorogenic acid, p-coumaric acid, caffeic acid, cryptochlorogenic acid, protocatechuic acid, ferulic acid, sinapic acid, and syringic acid. The separation process utilized a ZORBAX Eclipse Plus C18 column using 0.01% formic acid and 2 mmol/L ammonium formate in water as the aqueous phase and methanol containing 0.01% formic acid and 2 mmol/L ammonium formate as the organic phase. Calibration curves were constructed using standard solutions to quantitatively determine the phenolic acid content. The results showed significant variation in phenolic acid content among S. baumii fruiting bodies, with Protocatechuic acid, p-coumaric acid, and caffeic acid being the most abundant. This method is valuable for quantifying phenolic acid compounds under different cultivation conditions. It provides excellent sensitivity, selectivity, and reproducibility for the quantification of phenolic acids in Sanghuangporus, contributing to a better understanding of its chemical composition and potential health benefits. This approach represents a novel technical means for the simultaneous analysis of compound phenolic acids in Sanghuangporus fruiting bodies.

Exogenous Application of Coconut Water to Promote Growth and Increase the Yield, Bioactive Compounds, and Antioxidant Activity for Hericium erinaceus Cultivation

Hericium erinaceus (Bull.:Fr) Pers. is a medicinal mushroom that has various health benefits and is a rich source of bioactive compounds and antioxidant activity. In recent years, H. erinaceus has been considered for its many medicinal properties and is widely consumed in Asian countries. This work aimed to explore the potential improvement of coconut water utilization in order to promote growth and increase the yield, as well as the enhancement of bioactive compounds and antioxidant activity of H. erinaceus. The application of coconut water at a concentration of 20% (v/v) resulted in the optimal development and production of H. erinaceus. In terms of biological efficiency, it was demonstrated that the 22.09% increase over the control treatment resulted in a higher yield. Moreover, H. erinaceus cultivated from coconut water intake at all concentrations resulted in higher protein content. Additionally, bioactive compounds such as total triterpenoid content and total phenolic content of H. erinaceus were 67.87–89.24 mg Urs/g DW and 16.62–17.39 mg GAE/g DW, respectively, while H. erinaceus grown on a control treatment had the lowest at 56.84 mg Urs/g DW and 14.42 mg GAE/g DW, respectively. Applying coconut water at all concentrations resulted in H. erinaceus IC50 values of 0.58–0.69 mg/mL exhibiting higher DPPH activities than those grown on control treatment IC50 value of 0.77 mg/mL. Therefore, this finding of the study indicated that the utilization of coconut water as a therapy resulted in a significant enhancement in the development, production, and bioactive compounds of H. erinaceus, in comparison to the control treatment.

Bioactivity and Metabolomic Profile of Extracts Derived from Mycelial Solid Cultures of Hypsizygus marmoreus.

The beech mushroom (Hypsizygus marmoreus) is a highly nutritious, edible medicinal mushroom native to East Asia. The present research investigated the impact of different substrates on the metabolite compositions of H. marmoreus mycelia cultivated in vitro. The substrates tested included malt extract agar, malt extract agar enriched with barley malt, and malt extract agar enriched with grape pomace. The study also assessed antimicrobial and antiradical activities of the extracts against gram-positive bacteria (Bacillus subtilis and Staphylococcus aureus), gram-negative bacteria (Escherichia coli, Salmonella typhi, and Pseudomonas aeruginosa), yeasts (Candida albicans, C. tropicalis, and C. parapsilosis), and dermatophytes (Trichophyton mentagrophytes, T. tonsurans, T. rubrum, Arthroderma quadrifidum, A. gypseum, A. curreyi, and A. insingulare). The results revealed that the H. marmoreus mycelia extracts demonstrated antibacterial and antifungal activities against the tested microorganisms. Extracts obtained from the cultivation in substrates enriched with either barley malt or grape pomace exhibited the highest antibacterial activity among all the tested bacterial strains except for P. aeruginosa. The same extracts showed the highest inhibitory effect against C. albicans and C. parapsilosis. Noteworthy, the extract from the mushroom cultivated in the substrate enriched with grape pomace also exhibited remarkable efficacy against T. mentagrophytes and T. tonsurans. Terpenoid and carbapenem compounds could be related to the antimicrobial properties of the extracts from mushrooms cultivated in substrates enriched with grape pomace. In comparison, the higher antiradical properties could be related to the content of indole compounds. In conclusion, growth substrate selection affects the nutritional and medicinal properties of H. marmoreus, making it a valuable contribution to the understanding of the cultivation of this mushroom.

Interspecific Hybridization between Ganoderma lingzhi and G. resinaceum by PEG-Induced Double-Inactivated Protoplast Fusion

Ganoderma lingzhi is an important medicinal fungus, and it is particularly important to select strains with high yields and active substance contents. In this study, protoplasts of G. lingzhi were thermally inactivated to destroy intracellular enzyme proteins and preserve DNA. The DNA of G. resinaceum was damaged by ultraviolet (UV) radiation, and other components of the protoplasm except DNA were preserved. Then, the protoplast was induced using polyethylene glycol (PEG) for fusion. The results showed that the optimal thermal inactivation conditions for G. lingzhi were 30 min in a 45 °C water bath, and the optimal UV inactivation conditions for G. resinaceum were 70 s of irradiation using a 20 W UV lamp at a vertical distance of 15 cm. Antagonistic tests, internal transcribed space (ITS) and mitochondrial DNA identification, intersimple sequence repeat (ISSR) molecular markers and morphology were used to distinguish the parents from the fusants. Four true fusants were obtained, and the yield was 2.5%. The fruiting body yield of the fusants was significantly higher than that of G. lingzhi, and the polysaccharide and triterpene contents of the RAD-64 fusant were significantly higher than those of G. lingzhi. The results presented in this paper show that protoplast fusion technology can effectively improve G. lingzhi varieties and support the breeding of new varieties.

Unveiling the distribution of chemical constituents at different body parts and maturity stages of Ganoderma lingzhi by combining metabolomics with desorption electrospray ionization mass spectrometry imaging (DESI)

Ganoderma lingzhi is an important medicinal fungus, which is widely used as dietary supplement and for pharmaceutical industries. However, the spatial distribution and dynamic accumulation pattern of active components such as ganoderic acids (GAs) among different parts of G. lingzhi fruiting body are still unclear. In this study, desorption electrospray ionization mass spectrometry imaging (DESI-MSI) with untargeted metabolomics analysis was applied to investigate the metabolites distribution within G. lingzhi fruiting body at four different maturity stages (squaring, opening, maturation and harvesting stage). A total of 132 metabolites were characterized from G. lingzhi, including 115 triterpenoids, 11 fatty acids and other component. Most of the GAs content in the cap was significantly higher than that in the stipe, with six components such as ganoderic acid B being extremely significant. GAs in the cap was mainly present in the bottom edge of the mediostratum layer, such as ganoderic A-I and ganoderic GS-1, while in the stipe, they were mainly distributed in the shell layer and the context layer, such as ganoderic A-F. Most ganoderic acids content in both the stipe and the cap of G. lingzhi was gradually decreased with the development of G. lingzhi. The GAs in the stipe was gradually transferred from the shell layer to the content layer, while the distribution of GAs among different tissues of the cap was not significantly changed. In addition, linoleic acid, 9-HODE, 9-KODE and other fatty acids were mainly accumulated in the opening and maturing stage of the caps. This study further clarifies the spatial dynamic distribution of GAs in G. lingzhi fruiting body at four different maturity stages (squaring, opening, maturation and harvesting stage), which provides a basis for the rational utilization of the medicinal parts of G. lingzhi. Furthermore, mass spectrometry imaging combined with non-target metabolome analysis provides a powerful tool for the spatial distribution of active substances in the different regions of the medicinal edible fungi.