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Characteristics of wine produced by mushroom fermentation.

Saccharomyces cerevisiae is the main microorganism used in wine brewing, because this microbe has potent ability to produce alcohol dehydrogenase. We have recently discovered that some genera of mushroom produced alcohol dehydrogenase, and made wine by using a mushroom in place of S. cerevisiae. The highest alcohol concentration in this wine was achieved with Pleurotus ostreatus (2.6 M, 12.2%). In the case of Agaricus blazei, the same alcohol concentration (1.7 M, 8%) was produced under both aerobic and anaerobic conditions. This wine produced by A. blazei contained about 0.68% beta-D-glucan, which is known to have a preventive effects against cancer. The wine made by using Flammulina velutipes showed thrombosis-preventing activity, giving a prolonged thrombin clotting time 2.2-fold that of the control. Thus, the wine made by using mushroom seems to be a functional food which can be expected to have preventive effects against cancer and thrombosis.

Agaricales↗

Apoptosis induction by lectin isolated from the mushroom Boletopsis leucomelas in U937 cells.

A 15-kDa lectin was isolated from the edible mushroom Kurokawa by affinity chromatography using N,N'-diacetylchitobiose-Sepharose 4B. The results of microsequencing analysis indicated that the lectin has a partial amino acid sequence similar to the mushroom lectin, Agaricus bisporus agglutinin (ABA). We found that the Kurokawa lectin inhibited proliferation of human monoblastic leukemia U937 cells dose-dependently. Several lines of evidence indicated that this inhibition was due to its apoptosis induction. We observed that the lectin induced apoptotic bodies formation, chromatin condensation, and DNA ladder formation, features of apoptosis. The DNA ladder formation was inhibited by a general inhibitor of caspases, which are known to play essential roles in apoptosis. In contrast, ABA did not have cell growth-inhibiting or apoptosis-inducing activities. Thus, the Kurokawa lectin is the first mushroom lectin with apoptosis-inducing activity.

Amino Acid Sequence↗

Stimulation of mycelia growth in several mushroom species by rice husks.

When supplemented to the culture medium of mushroom Coprinus cinereus, rice husks soaked beforehand in methanol stimulated mycelia growth up to a concentration of 80 mg/ml dose-dependently, whereas the non-treated stimulated mycelia growth up to 20 mg/ml. This result suggests the existence of both stimulatory and inhibitory substances in rice husks. Since momilactone A (MLA) is recognized as one of the phytoalexins in rice husks, its biological activity against mycelia growth was tested. Momilactone A inhibited mycelia growth at 5 microg/disc, whereas the methanol extract of husks did so at 1 mg/disc, wherein 0.2 microg of MLA was estimated by LC/MS/MS. Thus the phytoalexins including MLA should inhibit mycelia growth. Rice husks stimulated mycelia growth in some edible mushroom species such as Grifola frondosa (maitake), Lentinus edodes (shiitake), Pleurotus eryngii (eringi), and P. ostreatus (hiratake). Our findings might lead to the development of new profitable cultivation methods for mushrooms using rice husks.

Agaricales↗

Stereochemical correlation between 10-hydroperoxyoctadecadienoic acid and 1-octen-3-ol in Lentinula edodes and Tricholoma matsutake mushrooms.

Linoleic acid (LA) incubated with a homogenate of Lentinula edodes or Tricholoma matsutake mushroom significantly increased the amount of (R)-1-octen-3-ol. The alcohol was identified as (S)-10-HODE with 90-87% and >99% enantiomeric excess (ee), respectively. During the incubation of LA with these homogenates in the presence of glutathione-glutathione peroxidase (GSH-GPx), which can reduce hydroperoxy fatty acids to the corresponding hydroxy acids, the formation of (R)-1-octen-3-ol was significantly inhibited, whereas the amount of 10-hydroxy-(8E,12Z)-8,12-octadecadienoic acid (10-HODE) was significantly increased. The acid was identified as (S)-10-HODE with 92-88% ee and >99% ee, respectively. The decrease in the amount of alcohol was approximately the same as the increase in amount of HODE in both mushrooms. These results indicate a stereochemical correlation between (R)-1-octen-3-ol and (S)-10-hydroperoxy-(8E,12Z)-8,12-octadecadienoic acid [(S)-10-HPODE] in both mushrooms.

Agaricales↗

Subcellular distribution of aluminum, bismuth, cadmium, chromium, copper, iron, manganese, nickel, and lead in cultivated mushrooms (Agaricus bisporus and Pleurotus ostreatus).

In this work, the distribution of nine metals in two types of cultivated mushroom had been investigated. For Agaricus bisporus, the biomass was separated into caps and stalks, and for Pleurotus ostreatus, the entire mushrooms were taken for analysis. Electrothermal atomic absorption spectrometry was used for total element determination in acid digests. For accuracy checking, the certified reference material (NIST 1,571, citrus leaves) was analyzed. The results obtained for the two fungi species were within the ranges of concentration reported previously by other authors. Subcellular fractionation was accomplished by centrifugation of cell homogenates, which had been suspended in Tris-HCl buffer. In the first centrifugation (7,300 g, 4 degrees C, 10 min), cell walls were separated (pellet I), and the second centrifugation (147,000g, 4 degrees C, 60 min) yielded mixed membrane fraction (pellet II) and cytosol (supernatant II). Recoveries of the fractionation procedure were in the range 70--100% (with the exception of Fe). For all elements studied, the highest relative contributions were found in cytosol fractions of the fruiting bodies (63--72%, 49--76%, 44--93%, 26--87 pc, 55--85%, 50--68%, 41--78%, 39--78%, 54--67% respectively for Al, Bi, Cd, Cr, Cu, Fe, Mn, Ni, and Pb. Lower contributions were found in cell walls (respectively 22--32%, 24--44%, 6.1--47%, 12--52%, 7.3-- 37%, 7.9--32%, 19--52%, 20--42%, and 25--38%) and only minute amounts in the mixed membrane fraction (3.0--5.8%, 0.7--7.0%, 0.7--8.3%, 1.0--22%, 7.5--14%, 16--24%, 1.1--19%, and 5.1--7.7%). The results obtained indicate that small water-soluble molecules were the primary forms of nine elements in two mushroom species studied. On the other hand, the evidence has been provided on elements binding to larger, water-insoluble molecules contained in the structures of cell wall and membranes. The relative distribution was both element and fungi dependent. Thus, in P. ostreatus, total element levels were higher than in A. bisporus, with the preference for their accumulation in cytosol. On the contrary, total element content in the latter fungi was lower; however, a clear tendency toward more efficient element incorporation to the water-insoluble structures was observed (no apparent differences between stalks and caps).

Agaricus↗

Evolution of the bipolar mating system of the mushroom Coprinellus disseminatus from its tetrapolar ancestors involves loss of mating-type-specific pheromone receptor function.

Mating incompatibility in mushroom fungi is controlled by the mating-type loci. In tetrapolar species, two unlinked mating-type loci exist (A and B), whereas in bipolar species there is only one locus. The A and B mating-type loci encode homeodomain transcription factors and pheromones and pheromone receptors, respectively. Most mushroom species have a tetrapolar mating system, but numerous transitions to bipolar mating systems have occurred. Here we determined the genes controlling mating type in the bipolar mushroom Coprinellus disseminatus. Through positional cloning and degenerate PCR, we sequenced both the transcription factor and pheromone receptor mating-type gene homologs from C. disseminatus. Only the transcription factor genes segregate with mating type, discounting the hypothesis of genetic linkage between the A and B mating-type loci as the causal origin of bipolar mating behavior. The mating-type locus of C. disseminatus is similar to the A mating-type locus of the model species Coprinopsis cinerea and encodes two tightly linked pairs of homeodomain transcription factor genes. When transformed into C. cinerea, the C. disseminatus A and B homologs elicited sexual reactions like native mating-type genes. Although mating type in C. disseminatus is controlled by only the transcription factor genes, cellular functions appear to be conserved for both groups of genes.

Agaricales↗

From edible to useful mushrooms--an attempt for the new economical assessment of large fungi.

According to the stand of the modern applied mycological research the most commonly used term "edible mushroom" does not express all significant aspects large fungi can be used for. Additionally to bioconversion for food and animal feed production there are at least three other fields where large fungi may also get economical relevance: for establishing of ectomycorrhiza, for medical application and for soil decontamination including environmental engineering. This new situation justifies the introduction of a new, all-embracing designation for large fungi. The term "useful mushroom" will be suggested. The various options of the use of mushrooms will be introduced and briefly discussed in this paper.

Agaricales↗

Study of heavy metals in wild edible mushrooms under different pollution conditions by X-ray fluorescence spectrometry.

In this work we studied and compared the metal uptake in edible mushrooms (Lepiota procera, Boletus badius, Boletus edulis, Tricholoma equestry, Lactarius deliciosus, Cantarelus tubalformis and Cantarelus edulis), relative to sampling sites submitted to different pollution conditions: car traffic, soil pollution due to pesticides and fertilizers used in old vineyards, and incineration of hospital waste. Soil was also collected in some places, and its content was correlated to the corresponding one in some mushrooms species. All samples, without any chemical treatment, were analyzed by an X-ray fluorescence set-up. This technique is based on a monochromatic X-ray beam ionizing the atoms of the sample. Following this ionization, the emitted radiation is characteristic of the element, allowing its identification and quantification. Vineyards are normally submitted to very high amounts of sulfating, containing high copper concentrations. This metal is accumulated on the soil, and can be up-taken by vegetation. Very high levels of Fe and Cu were found in Lepiota procera species in old vineyards. Zinc was found to be always higher than Cu by factors ranging from 1.5 to 8 in clean wood taken as a reference for the whole analyzed species, while in old vineyards the ratio Zn/Cu reach 0.25 for Lepiota procera. This is correlated to the soil content for both elements. In addition, pollution induced by car traffic was checked in some samples, collected in the proximity of highways. Pb was the main contaminant in these areas, and presenting values 10 times higher than the corresponding ones in sites not submitted to pollution, for some species. Mushrooms contamination due to incineration of hospital waste was also studied, but we did not observe any contamination involving heavy metals in the several analyzed species around these areas. This is in agreement with what was expected, taking into account that hospital waste is mostly organic and, in principle, no heavy metals would be observed.

Agaricales↗

Hypersensitivity pneumonitis induced by spores of Penicillium citrinum in a worker cultivating Enoki mushroom.

A 47-year-old Japanese woman was admitted to our hospital with a 2-week history of dry cough and shortness of breath. She had been engaged in Enoki mushroom production for 22 years. Chest X-ray and chest computed tomography (CT) scan showed bilateral fine-nodular shadows and ground glass opacity. Bronchoalveolar lavage fluid demonstrated an increase of total cell counts with predominant lymphocytosis. Pathological specimens obtained by video-assisted thoracoscopic surgery revealed alveolitis and noncaseating granuloma with giant cells. Lymphocyte stimulation test showed positive responses with Enoki mushroom, culture medium, and Penicillium citrinum. On double immunodiffusion test, a precipitation line was observed between patient's serum and Penicillium citrinum antigen. She was found to have hypersensitivity pneumonitis caused by Penicillium citrinum. This is the first report of mushroom worker's lung caused by Penicillium citrinum.

Agaricales↗

Listeria monocytogenes septicemia associated with consumption of salted mushrooms.

Listeria monocytogenes septicemia in an 80-year-old man is described. On the day before clinical symptoms appeared the patient had eaten homemade salted mushrooms, rufous milkcap (Lactarius rufus Fr.). L. monocytogenes serotype 4b was isolated in blood cultures. The mushrooms which had been stored in cold for 5 months before consumption contained the same listeria serotype at a level of 10(6) CFU/g. Salt content (NaCl) of the mushrooms was 7.5%. Fever and diarrhea disappeared with penicillin therapy and the patient was discharged after 4 weeks in the hospital.

Aged↗

Distribution of ascorbic acid analogs and associated glycosides in mushrooms.

Mushrooms contain reducing substances with chemical properties similar to ascorbic acid (AsA). In this study, the four types of reducing substances contained in Flammulina velutipes (Enokitake), Hypsizigus mamoreus (Bunashimeji), Pholiota nameko (Nameko), and Grifola frondosa (Maitake) were respectively purified, and the structure of each was analyzed using nuclear magnetic resonance (NMR) and other methods. The results confirmed that those substances were AsA analogs and associated glycosides (6-deoxy-AsA, 6-deoxy-5-O-(alpha-D-xylopyranosyl)-AsA, 6-deoxy-5-O-(alpha-D-glucopyranosyl)-AsA, and 5-O-alpha-D-glucopyranosyl-erythro-AsA). These substances were characteristic in that saccharide was bonded with the C-5 of the AsA analogs. Osazones were formed from the reducing substances in 19 kinds of edible mushrooms. Using thin-layer chromatography (TLC), they were developed to examine the distribution of the above reducing substances and AsA. The results showed that at least one of the above compounds was certain to be present in any mushroom; that AsA was present in very small quantities if at all; and that several substances similar to the above compounds were present.

Ascorbic Acid↗

Incidence of enterohemorrhagic Escherichia coli, Escherichia coli O157, Salmonella, and Listeria monocytogenes in retail fresh ground beef, sprouts, and mushrooms.

The objective of this study was to determine the prevalence of enterohemorrhagic Escherichia coli (EHEC), E. coli O157, Salmonella, and Listeria monocytogenes in retail food samples from Seattle, Wash. A total of 2,050 samples of ground beef (1,750 samples), mushrooms (100 samples), and sprouts (200 samples) were collected over a 12-month period and analyzed for the presence of these pathogens. PCR assays, followed by culture confirmation were used to determine the presence or absence of each organism. Of the 1,750 ground beef samples analyzed, 61 (3.5%) were positive for EHEC, and 20 (1.1%) of these were positive for E. coli O157. Salmonella was present in 67 (3.8%) of the 1,750 ground beef samples. Of 512 ground beef samples analyzed, 18 (3.5%) were positive for L. monocytogenes. EHEC was found in 12 (6.0%) of the 200 sprout samples, and 3 (1.5%) of these yielded E. coli O157. Of the 200 total sprout samples, 14 (7.0%) were positive for Salmonella and none were positive for L. monocytogenes. Among the 100 mushroom samples, 4 (4.0%) were positive for EHEC but none of these 4 samples were positive for E. coli O157. Salmonella was detected in 5 (5.0%) of the mushroom samples, and L. monocytogenes was found in 1 (1.0%) of the samples.

Agaricales↗

A lectin with mycelia differentiation and antiphytovirus activities from the edible mushroom Agrocybe aegerita.

A lectin named AAL has been purified from the fruiting bodies of the edible mushroom Agrocybe aegerita. AAL consisted of two identical subunits of 15.8 kDa, its pI was about 3.8 determined by isoelectric focusing, and no carbohydrate was discerned. Being treated by pyrogultamate aminopeptidase, the blocked N-terminus of AAL was sequenced as QGVNIYNI. AAL agglutinated human and animal erythrocytes regardless of blood type or animal species. Its hemagglutinating activity was unaffected by acid or alkali treatment and demetalization or addition of divalent metals Mg(2+), Ca(2+) and Zn(2+). AAL was toxic to mice: its LD50 was 15.85 mg per kilogram body weight by intraperitoneal injection. In this study, two novel activities of AAL were proved. It showed inhibition activity to infection of tobacco mosaic virus on Nicotiana glutinosa. The result of IEF suggested that AAL attached to TMV particles. Mycelia differentiation promotion was the other interesting activity. AAL promoted the differentiation of fruit body primordia from the mycelia of Agrocybe aegerita and Auricularia polytricha. AAL antiserum was prepared and immunologically cross-reactived with several proteins from five other kinds of mushrooms. These results suggested that AAL probably was a representative of a large protein family, which plays important physiological roles in mushroom.

Agaricales↗

Validation of microwave digestion method for determination of trace metals in mushrooms.

A microwave digestion method for mushrooms, developed in the study, allows fast preparation of samples and reduces the contamination risk in the process of determining trace metals. Concentrations of six trace elements, Fe, Mn, Cu, Zn, Pb, and Cd were measured in 50 samples of different species of edible mushrooms (fam. Boletaceae) using atomic absorption spectrometry after microwave and dry ashing procedure. The methods were validated through certified Standard Reference Material SRM 1577b (Bovine Liver) which was treated and analysed using the same procedures as for the mushrooms. The samples were either digested with concentrated HNO3 in closed Teflon PFA vessels in a microwave oven, or ashed in quartz crucibles at 450 degrees C. The respective recoveries of Fe, Mn, Zn, Cu, Pb, and Cd obtained by measuring SRM were 112, 107, 104, 115, 111 and 95% after microwave digestion procedure and 86, 101, 109, 111, 98, and 110% after dry ashing procedure. The correlation between concentrations obtained by the two different methods of sample preparation was high for all metals.

Agaricales↗

Induction of apoptosis in human prostatic cancer cells with beta-glucan (Maitake mushroom polysaccharide).

PURPOSE: To explore more effective treatment for hormone-refractory prostate cancer, we investigated the potential antitumor effect of beta-glucan, a polysaccharide of the Maitake mushroom, on prostatic cancer cells in vitro. MATERIALS AND METHODS: Human prostate cancer PC-3 cells were treated with various concentrations of the highly purified beta-glucan preparation Grifron-D(R) (GD), and viability was determined at 24 h. Lipid peroxidation (LPO) assay and in situ hybridization (ISH) were performed to unravel the antitumor mechanism of GD. RESULTS: A dose-response study showed that almost complete (>95%) cell death was attained in 24 h with GD > or = 480 microg/mL. Combinations of GD in a concentration as low as 30 to 60 microg/mL with 200 microM vitamin C were as effective as GD alone at 480 microg/mL, inducing >90% cytotoxic cell death. Simultaneous use with various anticancer drugs showed little potentiation of their efficacy except for the carmustine/GD combination (approximately 90% reduction in cell viability). The significantly (twofold) elevated LPO level and positive ISH staining of GD-treated cells indicated oxidative membrane damage resulting in apoptotic cell death. CONCLUSION: A bioactive beta-glucan from the Maitake mushroom has a cytotoxic effect, presumably through oxidative stress, on prostatic cancer cells in vitro, leading to apoptosis. Potentiation of GD action by vitamin C and the chemosensitizing effect of GD on carmustine may also have clinical implications. Therefore, this unique mushroom polysaccharide may have great a potential as an alternative therapeutic modality for prostate cancer.

Agaricales↗

Parallel organization in honey bee mushroom bodies by peptidergic Kenyon cells.

Antisera against the neuromodulatory peptides, Phe-Met-Arg-Phe-NH(2)-amide (FMRFamide) and gastrin cholecystokinin, demonstrate that the mushroom bodies of honey bees are subdivided longitudinally into strata. Three-dimensional reconstructions demonstrate that these strata project in parallel through the entire pedunculus and through the medial and vertical lobes. Immunostaining reveals clusters of immunoreactive cell bodies within the calyx cups and immunoreactive bundles of axons that line the inside of the calyx cup and lead to strata. Together, these features reveal that immunoreactive strata are composed of Kenyon cell axons rather than extrinsic elements, as suggested previously by some authors. Sorting amongst Kenyon cell axons into their appropriate strata already begins in the calyx before these axons enter the pedunculus. The three main concentric divisions of each calyx (the lip, collar, and basal ring) are divided further into immunoreactive and immunonegative zones. The lip neuropil is divided into two discrete zones, the collar neuropil is divided into five zones, and the basal ring neuropil is divided into four zones. Earlier studies proposed that the lip, collar, and basal ring are represented by three broad bands in the lobes: axons from adjacent Kenyon cell dendrites in the calyces are adjacent in the lobes even after their polar arrangements in the calyces have been transformed to rectilinear arrangements in the lobes. The universality of this arrangement is not supported by the present results. Although immunoreactive zones are found in all three calycal regions, immunoreactive strata in the lobes occur mainly in the two bands that were ascribed previously to the collar and the basal ring. In the lobes, immunoreactive strata are visited by the dendrites of efferent neurons that carry information from the mushroom bodies to other parts of the brain. Morphologically and chemically distinct subdivisions through the pedunculus and lobes of honey bees are comparable to longitudinal subdivisions demonstrated in the mushroom bodies of other insects, such as the cockroach Periplaneta americana. The functional and evolutionary significance of the results is discussed.

Animals↗

[Use of molecular markers for differentiation of cultivated strains of oyster and button mushrooms].

Analysis of commercial strains of two edible mushrooms, Pleurotus ostreatus and Agaricus bisporus, using PCR and isozyme electrophoresis techniques allowed us to differentiate groups of genetically similar and distant strains. Among the commercial strains of P. ostreatus, the level of genetic variation was higher suggesting a broader genetic basis employed in breeding of this mushroom. The cultivars and hybrids of, A. bisporus, showed a higher level of homology. The isozyme markers (nonspecific esterase, leucinaminopeptidase, and phosphoglucoisomerase) are recommended for identification of the commercial strains of edible mushrooms.

Agaricus↗

Mushroom cryopreservation and its effect on survival, yield and genetic stability.

Mycelial stock cultures of Agaricus bisporus, A. Bitorquis, Pleurotus flabellatus, P. Sajor-caju, P. Ostreatus, P. Sapidus, Auricularia polytricha, Lentinula edodes, Morchella esculenta and Volvariella volvacea were maintained by frequent subculturing at an interval of two months and separately as wheat grain spawn in liquid nitrogen with 15 percent glycerol. Preservation of mushroom stock cultures as wheat grain spawn under liquid nitrogen proved to be the better method of maintenance. The percent recoveries of stored samples were unchanged from the first recovery after six months to the last recovery after 42 months in nine out of 11 stock cultures preserved under liquid nitrogen. However, a marginal decline in survival of 10 % was recorded in Auricularia polytricha and Volvariella volvacea. Yields before preservation of mushroom stock cultures and after 30 months of preservation exhibited static biological efficiency and fruitbody weight. The comparison of Random Amplified Polymorphic DNA (RAPD) and Internal Transcribed Spacers (ITS) PCR amplified products did not exhibit DNA fragment variation in banding patterns at the intraspecific level during preservation of stock cultures by either method. The modified cryopreservation protocol and experimental demonstration of genetic stability of stock cultures reported here validate the use of mushroom cryopreservation techniques and supports studies on genetic stability of preserved biological materials.

Agaricales↗