Morpho-physiological diversity between Lingzhi or Reishi medicinal mushroom Ganoderma lucidium (W. Curt.:Fr.) P. Karst. and G. carnosum pat

Two weakly differentiated taxa, Ganoderma lucidum and G. carnosum, were compared in their sufficient morphological and physiological features. The obtained results showed that dimensions of basidiospores and pileocystidia were insignificantly different, while pore shape and dimensions have shown greater diversity with average diameter of 138.46 μm in G. carnosum and 238.34 μm in G. lucidum. Mycelial growth rate was higher in G. lucidum (8.39 mm day-1) than in G. carnosum (6.02 mm day-1). G. lucidum was also a slightly better producer of biomass and extracellular polysaccharides (28.16 g L-1 and 1.42 mg mL-1, respectively) than G. carnosum (23.68 g L-1 and 0.35 mg mL-1, respectively). However, a higher amount of synthesized intracellular polysaccharides was noted in G. carnosum than in G. lucidum (40.00 mg g-1 and 30.00 mg g-1 of dry biomass, respectively). Higher activity levels of Mn-oxidizing peroxidases were obtained in G. carnosum, while G. lucidum was a better laccase producer. In G. carnosum, corn stem/NH?NO? medium with nitrogen concentration of 20 mM was the optimum for Mn-dependent peroxidase production (88.00 U L-1), while the highest versatile peroxidase activity was detected in the medium with grapevine sawdust and 10 mM of nitrogen (80.80 U L-1). Wheat straw was the best carbon source for laccase synthesis in G. lucidum (55.75 U L-1).     

Statistical optimization of polysaccharide production by submerged cultivation of Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum (W.Curt.: Fr.) P. Karst. MTCC 1039 (Aphyllophoromycetideae)

Response surface methodology was employed to optimize the concentration of four important cultivation media components such as cottonseed oil cake, glucose, NH4Cl, and MgSO4 for maximum medicinal polysaccharide yield by Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum MTCC 1039 in submerged culture. The second-order polynomial model describing the relationship between media components and polysaccharide yield was fitted in coded units of the variables. The higher value of the coefficient of determination (R2 = 0.953) justified an excellent correlation between media components and polysaccharide yield, and the model fitted well with high statistical reliability and significance. The predicted optimum concentration of the media components was 3.0% cottonseed oil cake, 3.0% glucose, 0.15% NH4Cl, and 0.045% MgSO4, with the maximum predicted polysaccharide yield of 819.76 mg/L. The experimental polysaccharide yield at the predicted optimum media components was 854.29 mg/L, which was 4.22% higher than the predicted yield.     

Immunomodulation of bone marrow macrophages by GLIS, a proteoglycan fraction from Lingzhi or Reishi medicinal mushroom Ganoderma lucidium (W.Curt.:Fr.) P. Karst

The immunomodulatory effect of GLIS (Lingzhi or Reishi medicinal mushroom Ganoderma lucidum immunomodulating substance) on macrophages has been investigated as part of ongoing research into the anticancer properties of this mushroom. Proliferation of bone marrow macrophages (BMMs) was enhanced by GLIS in a dose-dependent manner. Microscopic examination revealed that numerous GLIS-treated BMMs were enlarged and formed pseudopodia. Exposure of BMMs to GLIS resulted in significant increases in NO production, induction of cellular respiratory burst activity, and increased levels of IL-1β, IL-6, IL-12p35, IL-12p40, IL-18, and TNF-α gene expression and levels of TNF-α, IL-1β, and IL-12 secretion. Our data indicate that GLIS activates the immune system by modulating cytokine production.     

A comparative assessment of the potential of polysaccharide production and intracellular sugar composition within Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum (W.Curt.:Fr.)P. Karst. (Aphyllophoromycetideae)

Ganoderma lucidum is a well-known medicinal mushroom species in which polysaccharides are one of the major sources of biological activity. The species was considered as a species-complex due to significant variations in morphological, biochemical, and genetic features among populations with a worldwide distribution. This fact was the basis for setting the aim of this research: to study intraspecific diversity in polysaccharide production and intracellular sugar composition among selected G. lucidum strains. The presence ofintraspecific diversity among 10 G. lucidum strains, from different areas worldwide, was noted. Values of produced mycelia biomass and intracellular polysaccharides were found in wide ranges (3.1 - 28.2 g L(-1) and 20.0 - 53.3 mg g(-1), respectively), while differences in extracellular polysaccharide amounts were minor (0.2 - 1.5 mg mL(-1)). The significant quantitative and qualitative differences in intracellular sugar composition were noted. Glucose was the predominant sugar in almost all strains except one (HAI 447), where sucrose was dominant. The potential of polysaccharide production and intracellular sugar composition could be one more taxonomic criterion for strain characterization within G. lucidum. The differences in intracellular sugar composition and proportions could be reflected in features of produced polysaccharides and also in their biological activities.     

Paddy straw as a substrate for the cultivation of Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum (W.Curt.:Fr.) P. Karst. in India

Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum, is generally cultivated on hardwood logs or sawdust/woodchips based formulations. More than 100 million tonnes of paddy straw is being produced in India per year, and almost 50% of the straw is potentially available for growing mushrooms. In the present study an attempt was made to use paddy straw as a substrate to cultivate G. lucidim. Different proportions of paddy straw were mixed with 0, 22.5%, 45%, and 67.5% sawdust and 10% rice bran. Spawn run period, fruiting initiation period, yield, moisture content, dry recovery, and fruiting body characteristics were recorded and compared. Fructification was observed with all the substrate formulations and they did not show any significant difference in yield. The highest biological efficiency (BE) (29.9%) was observed with the combination sawdust:paddy straw:rice bran 22.5:67.5:10, followed by saw dust:paddy straw:rice bran 45:45:10 with BE 27.3%. The current study demonstrated for the first time that the cultivation of G. lucidum is possible with paddy straw as the base substrate and indicated the enormous potential of paddy straw for the cultivation of G. lucidum.     

Mycosynthesis of silver nanoparticles using Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum (W. Curt.:Fr.) P. Karst. and their role as antimicrobials and antibiotic activity enhancers

Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum, has been used over the ages as highly medicinal herb in the Orient. Many useful properties of this fungus are still being studied; we report here a new facet of this "elixir of life" as a mycosource for synthesis of metal nanoparticles. Treating the extracellular suspension filtrate of the mycelia of G. lucidum with silver nitrate reduces the metal ions to nanoparticles. Optical detection followed by confirmation through spectroscopic analysis suggests that this fungus can be used for the purpose of safe and sure synthesis of silver nanoparticles, demand for which is growing day by day in all fields of human life. LM-20 analysis of these G. lucidum-synthesised nanoparticles reveals the polydispersity and distribution of silver nanoparticles in the range of 10-70 nm with an average size of 45 nm and a concentration of 0.37 x 108 particles/mL. FT-IR spectrum confirms the stability of these nanoparticles due to presence of amide linkages and protein capping. These nanoparticles have shown strong bactericidal activity against test pathogens Staphylococcus aureus and Escherichia coli, and also exhibited their efficiency in enhancing the activity of the synthetic antibiotic tetracycline. The method of synthesising silver nanoparticles and its bactericidal effect discussed here can be used for environment-friendly and economically feasible production for different applications where chemically synthesized nanoparticles cause undesirable effects.     

Production of biomass, polysaccharides, and ganoderic acid using non-conventional carbon sources under submerged culture of the Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum (W.Curt.:Fr.)P. Karst. (higher Basidiomycetes)

The effect of different non-conventional carbon sources was studied in the submerged culture of Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum, for simultaneous production of mycelial biomass, bioactive ganoderic acid, and polysaccharides, in less time, using non-conventional carbon sources to minimize the high costs of current culture media. The optimal medium composition was defined as (g/L): 50 of barley flour, 0.2 of KH2PO4, 0.1 of MgSO4?7H2O, and 1 NH4Cl. Cultivated under this complex culture medium, the mycelial biomass production was 23.49 ± 0.37 g/L; the extracellular polysaccharides production was 2.72 ± 0.11 g/L; the intracellular polysaccharides production was 2.22 ± 0.06 g/L; the ganoderic acids production was 299.67 ± 11.63 mg/L. One liter of culture medium developed in this project was priced at USD $ 0.11 if barley flour is used as carbon source or $ 0.13 with oat flour in order to get a good amount of products of interest.     

Determination of the antioxidant activity and polyphenol contents of wild Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum (W.Curt. Fr.) P. Karst. (higher Basidiomycetes) from central Himalayan hills of India

The antioxidant potential of wild strain of Lingzhi or Reishi medicinal mushroom Ganoderma lucidum from Central Himalayan Hills (2000 m MSL) was evaluated, and compared with its in vitro cultured mycelia grown on malt extract broth in the laboratory. Antioxidant activities of both wild and cultivated G. lucidum in terms of IC?? (mg/ mL) were determined against different in vitro radical systems such as DPPH (1, 1-diphenyl-2-picrylhydrazyl), ABTS [2,2'-azinobis (3-ethylenebenzothiazoline-6-sulphonic acid)] and hydroxyl radicals, in addition to ferric reducing antioxidant power assay. Polyphenol contents were also determined, in order to assess their effects on the antioxidant activity of extracts. All the extracts showed significant antioxidant activity, and maximum scavenging was observed in the case of methanolic extracts of wild G. lucidum with minimum IC50 values 0.953 ± 0.040, 0.690 ± 0.014 and 3.295 ± 0.027 mg/mL, respectively, for DPPH, ABTS, and hydroxyl radicals. The efficacy of wild G. lucidum as a rich source of natural antioxidant was established for nutraceutical development.     

Inhibition of quorum sensing in the opportunistic pathogenic bacterium Chromobacterium violaceum by an extract from fruiting bodies of Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum (W.Curt.:Fr.) P. Karst. (higher Basidiomycetes)

Extracts of Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum, inhibited quorum sensing in Chromobacterium violaceum CV026. G. lucidum fruiting bodies were milled and extracted with ethyl acetate. The crude extract was dissolved in an appropriate concentration of methanol, sterilized by filtration through a 0.22-μm membrane filter, and added to Ch. Violaceum CV026 cultures, which were used as an indicator to monitor quorum sensing inhibition. Inhibitory activity was measured by quantifying violacein production using a microplate reader. Methanol-soluble compounds extracted from G. lucidum significantly inhibited quorum sensing-controlled behavior in Ch. Violaceum in a concentration-dependent manner. The results suggest that compounds in G. lucidum might be useful to control and handle detrimental infections caused by human, animal, and plant pathogens. Further studies are in progress in our lab to isolate the specific compounds from G. lucidum extract, evaluate them as quorum sensing inhibitors, and analyze their mechanism of action.     

Preliminary Results of Determination of Chemical Changes on Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (W.Curt.:Fr.)P. Karst. (Higher Basidiomycetes) Carried by Shenzhou I Spaceship with FTIR and 2D-IR Correlation Spectroscopy

Spaceflight represents a complex environmental condition. Space mutagenesis breeding has achieved marked results over the years. The objective of this study is to determine the chemical changes in medicinal mushroom Ganoderma lucidum cultivated after spaceflight in 1999. Fourier transform infrared (FTIR) and two-dimensional infrared (2DIR) correlation spectroscopy were used in analysis. The sample Sx and its control Cx showed the least dissimilarities in one-dimensional FTIR spectra, but absorbance of Sx is twice as high as Cx. Sx presented a clear peak at 1648 cm in 2nd derivative spectra, which could not be detected in the Cx. The 2DIR spectra showed the intensity of Sx in the range 1800-1400 cm-1 for protein is higher than the control. The sample Sx produced some carbohydrate peaks in the area of 889 cm-1 compared with the Cx. The spaceflight set up an extreme condition and caused changes of chemical properties in G. lucidum strain.     

Antioxidant activity and toxicity profile of total triterpenes isolated from Ganoderma lucidum (Fr.) P. Karst occurring in South India

The total triterpene fraction isolated from Ganoderma lucidum, a highly nutritional and popular medicinal mushroom occurring in South India, was evaluated for its antioxidant activity in vitro and in vivo. Total triterpenes successfully scavenged DPPH(+), ABTS(+) and superoxide radicals, showed significant ferric reducing activity and was highly effective in reducing the in vitro lipid peroxidation. Activities of the antioxidant enzymes in blood and tissue were increased by the administration of total triterpenes to Swiss albino mice in vivo. The ability of total triterpenes to scavenge the free radicals and to enhance body's antioxidant defence systems indicates its potential use as an antioxidant. An attempt was also done to gauge the toxicity of total triterpenes using acute and sub acute study models in Swiss albino mice. The results showed that Ganoderma triterpenes did not possess significant toxicity. The findings thus reveal the possible therapeutic use of Ganoderma triterpenes.     

Investigation of antiproliferative effect of ether and ethanol extracts of birch polypore medicinal mushroom, Piptoporus betulinus (Bull.:Fr.) P. Karst. (higher Basidiomycetes) in vitro grown mycelium

This paper describes the study conducted to evaluate the antiproliferative activity of ether and ethanol extracts isolated from Piptoporus betulinus against cancer-derived cells. The fungal material used for extract preparation and further experiments was obtained from in vitro grown strains of P. betulinus. To the best of the authors? knowledge, this is the first study evaluating antiproliferative potential of in vitro cultured birch polypore fungus. The effect of ether and ethanol extracts on cell proliferation, viability, and adhesion was assessed on colorectal adenocarcinoma cancer cell line LS180, whereas the cytotoxicity effect was investigated in normal colon epithelium-derived cell line CCD 841 CoTr. Studied extracts highly decreased the viability of cancer cells, slightly inhibiting proliferation and tumor cell adhesion in a time- and dose-dependent manner. Cytotoxicity of extracts against cells of normal colon epithelium origin was observed only at the highest studied concentration. The obtained results may seem interesting in comparison with previous studies on water extracts from natural grown P. betulinus. Future research on mycelial extract activity, as well as the content analysis, is needed.