Effect of Cytokinin-Containing Extracts from Some Medicinal Mushroom Mycelia on HepG2 Cells In Vitro.

Endogenous cytokinins in mycelia of medicinal mushrooms Hericium coralloides and Fomitopsis officinalis grown in vitro were identified using high-performance liquid chromatography coupled with mass spectrometry. High amounts of zeatin-type cytokinins and isopentenyladenine were found. The qualitative composition and quantitative content of cytokinins were species-specific traits of mushrooms. Optical microscopy was used to perform a comparison analysis of the influence of crude extracts and purified cytokinin fractions from both species' mycelial biomass on HepG2 tumor cell growth in vitro and morphology. The results showed that purified cytokinin fractions from H. coralloides and F. officinalis mycelia demonstrated a cytotoxic effect on HepG2 cells, unlike crude extracts. Under the influence of all mushroom extracts, similar patterns of changes in HepG2 cell morphology were observed, but they were more pronounced for H. coralloides compared with F. officinalis. Purified fractions of both mushroom species caused an increased level of apoptosis compared to crude extracts. Some increase in glucose uptake by cultured cells was found in all investigated samples wherein the influence of H. coralloides extracts was approximately twice the effect of the corresponding F. officinalis extracts. The data obtained confirm the assumption that cytokinins are involved in the expression of therapeutic effects of medicinal mushrooms and indicate the need to take into consideration the methods of cytokinin extraction when preparing pharmacologically active drugs based on fungal raw materials. Thus, extracts from H. coralloides and F. officinalis mycelial biomass are promising in the search for anticancer agents.

Biology and Nutritional Contents in the Culinary-Medicinal Milky White Mushroom, Calocybe indica (Agaricomycetes), during Cultivation Involving Casing and Scratching Treatments.

The milky white mushroom, Calocybe indica Purkay & A. Chandra, is a tropical species not commercially cultivated in Europe. The influence of casing and scratching treatments on productivity, morphological characteristics, and nutritional contents were investigated during the cultivation of C. indica on locally available substrates. The highest total mushroom yield (43.6 ± 1.7/100 g substrate fresh weight [FW]) and biological efficiency (BE; 128.1 ± 5.0%) were obtained from treatment T6 that received casing (30-mm height) and the scratching treatment before casing. The lowest yield (27.6 ± 2.3/100 g FW) and BE (81.1 ± 6.9%) were in the treatment that had no casing and no scratching treatment (T7). The yield in the first and second flush combined accounted for 82-90% of the total yield in all treatments except in T7, where it was 100%. The longest (61.1 ± 1.7 days) and shortest (39 ± 0 days) cropping cycles were recorded in T6 and T7, respectively. The highest dry matter content in fruiting bodies (12.6%) was in the treatments with casing (10 mm height) and scratching (T4), and the lowest (8.2%) was in T7. The scratching treatment significantly decreased (P < 0.05) the lipid and ash content in fruiting bodies. To our knowledge, this is the first report on tests conducted on tropical mushroom C. indica under simulated industrial cultivation conditions in Europe. Although it requires a high temperature for cultivation (28-32°C), C. indica could become an alternative mushroom for cultivation in temperature climates and during the summer, when high temperatures significantly increase the costs of commercial production and marketing of white button, portabella, and most other exotic mushrooms.

Comparative Analysis of Cytokinins in Mycelial Biomass of Medicinal Mushrooms.

Mushrooms are known to produce phytohormones, in particular cytokinins. Here we studied in vitro production of cytokinins in medicinal mushrooms. Cytokinins were identified and quantified in mycelial biomass of 13 species by using high-performance liquid chromatography-mass spectrometry. Trans-zeatin, zeatin riboside, zeatin-O-glucoside, isopentenyladenosine, and isopentenyladenine were found but only 1 species (Ganoderma lucidum) contained all these forms. Zeatin-type cytokinins predominated. Composition of the cytokinin pool was unique in each species. The largest total amount of cytokinins was detected in Morchella esculenta strain 1755 and the smallest amount, in Flammulina velutipes strain 1878. The productivity of cytokinin biosynthesis in mycelial biomass of mushrooms was the lowest in mycelial biomass of Sparassis crispa strain 314 and highest in Pleurotus ostreatus strain 551. F. velutipes strain 1878 and Cyclocybe aegerita strain 960 mycelial biomass showed the most productive zeatin riboside biosynthesis. We emphasize the need to take into account the biological activity of cytokinins, on the basis of the mycelial biomass of medicinal mushrooms, in the development of drugs or dietary supplements. Macromycetes with high rates of cytokinin biosynthesis are considered to be prospective producers of pharmacologically active compounds.

Influence of Ganoderma lucidum (Curt.: Fr.) P. Karst. on T-cell-mediated immunity in normal and immunosuppressed mice line CBA/Ca.

The article presents the results of the investigation of the effect of biomass powder of the fungus Ganoderma lucidum on T-cell-mediated immunity in normal and immunosuppressed mice CBA/Ca. Delayed-type hypersensitivity assay was used. Experimental immunodeficiency was established with intraperitoneal injection of the immunosuppressant cyclophosphamide at a single dose of 150 mg/kg on the first day of the experiment. Results of the study show that the administration of biomass powder of Ganoderma lucidum in a dose of 0.5 mg/kg orally for 10 days increases the delayed-type hypersensitivity response in normal mice CBA/Ca. Administration of 0.5 mg/kg of biomass powder of the fungus Ganoderma lucidum for 10 days blocked the development of the T-cell-mediated immunosuppression, induced by administration of cyclophosphamide and restored the delayed-type hypersensitivity response in immunosuppressed mice. Key words: fungus Ganoderma lucidum cyclophosphamide immunodeficiency T-cell-mediated immunity delayed-type hypersensitivity.

Some macronutrient content in mycelia and culture broth of medicinal mushrooms cultivated on amaranth flour.

The content of proteins, lipids, and amino and fatty acids was investigated in mycelium and culture broth of medicinal mushrooms Cordyceps sinensis, Pleurotus ostreatus, and Schizophyllum commune. Seven essential amino acids were present in the proteins of all mushroom samples, with aspartic (6.34%-14.29%) and glutamic (15.12%-17.51%) acids predominating in culture mycelium and glutamic acid (16.3%-19.1%) in culture broth. Lipids in the mycelium of species C. sinensis, P. ostreatus, and Sch. Commune consisted of 10 fatty acids and 12 fatty acids in culture broth in our experiments. Major acids in culture mycelium and culture broth of fungi were linoleic (42.43%-67.41%), oleic (10.47%-32.54%), and palmitic (16.43%-20.33%). The proteins and lipids in culture broth of studied species contained a higher level of total non-essential amino acids and unsaturated fatty acids as compared to those in culture mycelium.