Antitumor and Immunostimulating Activities of Elfvingia applanata Hot Water Extract on Sarcoma 180 Tumor-bearing ICR Mice

Elfvingia applanata, a medicinal mushroom belonging to Basidiomycota, has been used in the effort to cure cancers of the esophagus and stomach, and is also known to have inhibitory effects on hepatitis B virus infection. The hot water soluble fraction (as Fr. HW) was extracted from fruiting bodies of the mushroom. In vitro cytotoxicity tests showed that hot water extract was not cytotoxic against cancer cell lines such as Sarcoma 180, HT-29, HepG2, and TR at concentrations of 10~2,000 microg/mL. Intraperitoneal injection with Fr. HW resulted in a life prolongation effect of 45.2% in mice previously inoculated with Sarcoma 180. Treatment of Fr. HW resulted in a 2.53-fold increase in the numbers of murine spleen cells at a concentration of 50 microg/mL, compared with control. Incubation of murine spleen cells with Fr. HW at a concentration of 500 microg/mL resulted in improved immune-potwntiating activity of B lymphocytes through an 8.3-folds increase in alkaline phosphatase activity, compared with control. Fr. HW generated 12.5 microM of nitric oxide (NO) when cultured with RAW 264.7, a mouse macrophage cell line, at the concentration of 50 microg/mL, while lipopolysaccharide, a positive control, produced 15.2 microM of NO. Therefore, the results suggested that antitumor activities of Fr. HW from E. applanata might, in part, be due to host mediated immunostimulating activity.

Hypolipidemic Activities of Dietary Pleurotus ostreatus in Hypercholesterolemic Rats

This work was conducted to investigate dietary supplementation of oyster mushroom fruiting bodies on biochemical and histological changes in hyper and normocholesterolemic rats. Six-week old female Sprague-Dawley albino rats were divided into three groups of 10 rats each. Feeding a diet containing a 5% powder of Pleurotus ostreatus fruiting bodies to hypercholesterolemic rats reduced plasma total cholesterol, triglyceride, low-density lipoprotein (LDL), total lipid, phospholipids, and LDL/high-density lipoprotein ratio by 30.18, 52.75, 59.62, 34.15, 23.89, and 50%, respectively. Feeding oyster mushrooms also significantly reduced body weight in hypercholesterolemic rats. However, it had no adverse effects on plasma albumin, total bilirubin, direct bilirubin, creatinin, blood urea nitrogen, uric acid, glucose, total protein, calcium, sodium, potassium, chloride, inorganic phosphate, magnesium, or enzyme profiles. Feeding mushroom increased total lipid and cholesterol excretion in feces. The plasma lipoprotein fraction, separated by agarose gel electrophoresis, indicated that P. ostreatus significantly reduced plasma beta and pre-beta-lipoprotein but increased alpha-lipoprotein. A histological study of hepatic cells by conventional hematoxylin-eosin and oil red O staining revealed normal findings for mushroom-fed hypercholesterolemic rats. These results suggest that a 5% P. ostreatus diet supplement provided health benefits by acting on the atherogenic lipid profile in hypercholesterolemic rats.

Antioxidant Activities and Tyrosinase Inhibitory Effects of Different Extracts from Pleurotus ostreatus Fruiting Bodies

We evaluated the antioxidant activity and tyrosinase inhibitory effects of Pleurotus ostreatus fruiting bodies extracted with acetone, methanol, and hot water. The antioxidant activities were tested against beta-carotene-linoleic acid, reducing power, 1,1-diphenyl-2-picrylhydrazyl free radical scavenging activity, and ferrous chelating ability. Furthermore, phenolic acid and flavonoid contents were also analyzed. The methanol extract showed the strongest beta-carotene-linoleic acid inhibition as compared to the other exracts. The acetone extract (8 mg/mL) showed a significantly high reducing power of 1.54 than the other extracts. The acetone extract was more effective than other extracts for scavenging on 1,1-diphenyl-2-picrylhydrazyl radicals. The strongest chelating effect (85.66%) was obtained from the acetone extract at 1.0 mg/mL. The antioxidant activities of the extracts from the P. ostreatus fruiting bodies increased with increasing concentration. A high performance liquid chromatography analysis detected seven phenolic compounds, including gallic acid, protocatechuic acid, chlorogenic acid, naringenin, hesperetin, formononetin, and biochanin-A in an acetonitrile and 0.1 N hydrochloric acid (5 : 1) solvent extract. The total phenolic compound concentration was 188 microg/g. Tyrosinase inhibition of the acetone, methanol, and hot water P. ostreatus extracts increased with increasing concentration. The results revealed that the methanol extract had good tyrosinase inhibitory ability, whereas the acetone and hot water extracts showed moderate activity at the concentrations tested. The results suggested that P. ostreatus may have potential as a natural antioxidant.

Cultural Conditions for Mycelial Growth and Molecular Phylogenetic Relationship in Different Wild Strains of Schizophyllum commune

The common split-gilled mushroom, Schizophyllum commune is found throughout the world on woody plants. This study was initiated to evaluate conditions for favorable vegetative growth and to determine molecular phylogenetic relationship in twelve different strains of S. commune. A suitable temperature for mycelial growth was obtained at 30degrees C. This mushroom grew well in acidic conditions and pH 5 was the most favorable. Hamada, glucose peptone, Hennerberg, potato dextrose agar and yeast malt extract were favorable media for growing mycelia, while Lilly and glucose tryptone were unfavorable. Dextrin was the best and lactose was the less effective carbon source. The most suitable nitrogen sources were calcium nitrate, glycine, and potassium nitrate, whereas ammonium phosphate and histidine were the least effective for the mycelial growth of S. commune. The genetic diversity of each strain was investigated in order to identify them. The internal transcribed spacer (ITS) regions of rDNA were amplified using PCR. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 129 to 143 bp and 241 to 243 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, while the 5.8S sequences were identical. A phylogenetic tree of the ITS region sequences indicated that the selected strains were classified into three clusters. The reciprocal homologies of the ITS region sequences ranged from 99 to 100%. The strains were also analyzed by random amplification of polymorphic DNA (RAPD) with 20 arbitrary primers. Twelve primers efficiently amplified the genomic DNA. The number of amplified bands varied depending on the primers used or the strains tested. The average number of polymorphic bands observed per primer was 4.5. The size of polymorphic fragments was obtained in the range of 0.2 to 2.3 kb. These results indicate that the RAPD technique is well suited for detecting the genetic diversity in the S. commune strains tested.

Mycelial Propagation and Molecular Phylogenetic Relationships of Commercially Cultivated Agrocybe cylindracea based on ITS Sequences and RAPD

This study evaluated the optimal vegetative growth conditions and molecular phylogenetic relationships of eleven strains of Agrocybe cylindracea collected from different ecological regions of Korea, China and Taiwan. The optimal temperature and pH for mycelial growth were observed at 25degrees C and 6. Potato dextrose agar and Hennerberg were the favorable media for vegetative growth, whereas glucose tryptone was unfavorable. Dextrin, maltose, and fructose were the most effective carbon sources. The most suitable nitrogen sources were arginine and glycine, whereas methionine, alanine, histidine, and urea were least effective for the mycelial propagation of A. cylindracea. The internal transcribed spacer (ITS) regions of rDNA were amplified using PCR. The sequence of ITS2 was more variable than that of ITS1, while the 5.8S sequences were identical. The reciprocal homologies of the ITS sequences ranged from 98 to 100%. The strains were also analyzed by random amplification of polymorphic DNA (RAPD) using 20 arbitrary primers. Fifteen primers efficiently amplified the genomic DNA. The average number of polymorphic bands observed per primer was 3.8. The numbers of amplified bands varied based on the primers and strains, with polymorphic fragments ranging from 0.1 to 2.9 kb. The results of RAPD analysis were similar to the ITS region sequences. The results revealed that RAPD and ITS techniques were well suited for detecting the genetic diversity of all A. cylindracea strains tested.

Comparative Effects of Oyster Mushrooms on Lipid Profile, Liver and Kidney Function in Hypercholesterolemic Rats

Comparative effects of oyster mushrooms on plasma and fecal lipid profiles and on liver and kidney function were evaluated in hyper and normocholesterolemic rats. Feeding of hypercholesterolemic rats a 5% powder of oyster mushrooms (Pleurotus ostreatus, P. sajor-caju and P. florida) reduced the plasma total cholesterol level by 37%, 21% and 16%, respectively and reduced the triglyceride level by 45%, 24% and 14%, respectively. LDL/HDL ratio decreased by 64%, 45% and 41% for P. sajor-caju, P. ostreatus and P. florida fed rats, respectively. Mushroom feeding also reduced body weight in hypercholesterolemic rats. However, it had no adverse effect on plasma bilirubin, creatinin and urea nitrogen level. Mushroom feeding also increased the total lipid and cholesterol excretion in the feces. The present study reveals that feeding of 5% oyster mushroom powder does not have detrimental effects on the liver and kidneys rather may provide health benefits for the cardiovascular-related complication by decreasing the atherogenic lipid profiles.

Vegetative Growth of Four Strains of Hericium erinaceus Collected from Different Habitats

Vegetative growth of four different strains of Hericium erinaceus was observed. The temperature suitable for optimal mycelial growth was determined to be 25degrees C, with growth observed in the extend temperature range of 20~30degrees C. The different strains of this mushroom showed distinct pH requirements for their optimum vegetative growth, with the most favorable growth observed at pH 6. Considering vegetative mycelial growth, PDA, YM, Hennerberg, Hamada, and Glucose peptone were the most favorable media, and Czapek Dox, Hoppkins, Glucose tryptone, and Lilly were the most unfavorable media for these mushroom strains. With the exception of lactose, most of the carbon sources assayed demonstrated favorable vegetative growth of H. erinaceus. For mycelial growth, the most suitable nitrogen source was alanine and the most unsuitable was histidine. Oak sawdust medium supplemented with 10~20% rice bran was the best for mycelial growth of the mushroom.

Favorable Culture Conditions for Mycelial Growth of Korean Wild Strains in Ganoderma lucidum

Ganoderma lucidum (Fr.) Karst (Polyporaceae), belonging to basidiomycota, is one of the most famous medicinal mushrooms. This study was carried out to investigate favorable mycelial growth conditions, such as pH, temperature, growth media, carbon sources and nitrogen sources of Korean strains in G. lucidum. The most suitable temperature for the mycelial growth was obtained at 30degrees C. In general, optimal temperature range for the mycelial growth was found at 25~30degrees C. This Mushroom has a broad pH range (5~9) for its mycelial growth and mostly favorable growth was found at pH 5. Generally, Hamada, Glucose peptone, YM, Mushroom complete and Lilly media were the most suitable for the mycelial growth of G. lucidum. Among 10 different carbon sources, dextrin, galactose and fructose were best but the rest of other carbon sources also facilitated the growth of mycelia. The most suitable nitrogen sources were ammonium acetate, glycine, arginine and calcium nitrate, but to a certain extent, all of the supplemented nitrogen sources also stimulated the mycelial growth.

Physicochemical Requirement for the Vegetative Growth of Schizophyllum commune Collected from Different Ecological Origins

Schizophyllum commune is an edible and medicinal mushroom widely distributed in the world. The optimal growth conditions for the mycelia of 10 strains of the fungus were investigated. The temperature suitable for the mycelial growth and density was obtained at 30~35degrees C. Among the tested conditions, the minimum mycelial growth was found at 15degrees C. In case of pH, the most favorable growth was found at pH 5. The results indicated that this mushroom well adapted to high temperature and low pH for its mycelial growth. Considering growth phenotype of mycelia, Hamada, Hennerberg, PDA and YM were the most suitable and Lilly, Glucose triptone, Glucose peptone and Hoppkins were the most unfavorable among tested media for the mycelial growth of S. commune. Out of tested carbon sources, dextrin and fructose were the most suitable and lactose, mannose and sorbitol were the unsuitable for the fungus. Compact mycelial density was obtained from most of the carbon sources. Among used nitrogen sources, calcium nitrate, potassium nitrate and alanine were the most appropriate and the most incompatible were ammonium phosphate, histidine, urea and arginine for mycelial growth of S. commune on the culture media. Calcium nitrate, histidine and potassium nitrate showed moderately thin or thin, and rest of nitrogen sources showed compact or moderately compact mycelial density.

The Artificial Cultivation of Oudemansiella mucida on the Oak Sawdust Medium

To produce fruiting bodies of Oudemansiella mucida, porcelain fungus, on the oak sawdust medium, additives suitable for the mycelial growth and fruiting body formation were screened. In general, the mycelial growth of the three strains of O. mucida used in this study have been good on oak sawdust mixed rice bran of 20~30%. The mycelia incubated in potato dextrose broth for 7 days were inoculated on oak sawdust medium supplemented with various ratios of rice bran and incubated for 30 days at 25degrees C in the dark condition until the mycelia of O. mucida fully colonized the media from top to bottom. Then, top surface of the media in the bottles were horizontally scratched with a spatula and filled with tap water for 3 hours. To induce the primordial formation of O. mucida, the bottles were transferred to the mushroom cultivating room under 12 hrs of light (350 lux) and dark condition with relative humidity of 95% at 17degrees C. The primordia of O. mucida were formed on the surface of oak sawdust media after 7 days of incubation. The mature fruiting bodies were observed 5 days after primordial formation. The fruiting bodies O. mucida were formed on oak sawdust medium mixed with 5 to 30% rice bran. However, abundant fruiting-bodies of O. mucida were produced in oak sawdust medium supplemented with 20% rice bran. This is the first report associated with an artificial fruiting body production of O. mucida in Korea.

The Fruiting Body Formation of Oudemansiella radicata in the Sawdust of Oak (Quercus variabilis) Mixed with Rice Bran

To screen additives and their mixed ratio suitable for the mycelial growth and fruiting body formation of Oudemansiella radicata in the oak sawdust, additives such as rice bran, fermented soybean powder and wheat bran were used. Generally, the mycelial growth of O. radicata has been stable on oak sawdust mixed with rice bran of 5~20%. In case that O. radicata was cultured for about 30 days at 22 +/- 1degrees C under the illumination (350 lux) of 12 hours and moisture condition of 90 +/- 5%, the primordia have been formed gradually from red-brown crusts covering the surface of oak sawdust media. Based on the experimental results from 9 strains of O. radicata, fruiting bodies were produced widely on oak sawdust medium mixed with rice bran of 5 to 30%. Even though fruiting bodies of O. radicata have been produced well on oak sawdust media mixed with rice bran, fruiting bodies of O. radicata were produced intensively on oak sawdust media mixed with rice bran of 10%. Therefore, this result will provide a basic information for commercial production of fruiting body of wild O. radicata. This result is the first report associated with an artificial fruiting body formation of O. radicata in Korea.

The Fruiting Body Formation of Armillaria mellea on Oak Sawdust Medium Covered with Ground Raw Carrots

To produce an artificial fruiting body of Armillaria mellea on the oak sawdust medium, seven strains of A. mellea were used. The top surface of oak sawdust medium covered with ground raw carrot was inoculated with each of 7 strains and cultured for 30 days at 25degrees C in the dark condition until the mycelia of A. mellea completely colonized the medium from top to bottom. Then, the mycelia which were fully covered on the top surface of the medium were scratched slightly with a spatula and filled with tap water for 3 hours. To induce the primordial formation, the 7 strains of A. mellea were transferred to the growth chamber under the illumination (350 lux) of 12 hours and relative humidity of 85 +/- 5% in a day and then cultured at 16 +/- 1degrees C. Only A. mellea IUM 949 could form primordia on the sawdust medium, but the other strains did not make primordia at the same condition. The primordia of A. mellea IUM 949 were formed 10 days after complete colonization of the medium and the fruiting bodies were produced 7 days after a primordial formation. The experimental results suggested that IUM 949 strain might be a good candidate for mass production of fruiting bodies of A. mellea.

Genes Expressed During Fruiting Body Formation of Agrocybe cylindracea

Agrocybe cylindracea, an edible mushroom belonging to Bolbitiaceae, Agaricales, is widely used as invaluable medicinal material in the oriental countries. This study was initiated to find the genes expressed during the fruiting body formation of A. cylindracea. The cDNAs expressed differentially during fruiting body morphogenesis of A. cylindracea were isolated through subtractive hybridization between vegetative mycelia and fruiting bodies. The cDNAs expressed in the fruiting body morphogenesis of A. cylindracea were cloned and twenty genes were identified. Eleven were homologous to genes of known functions, three were homologous to genes in other organism without any function known. Six were completely novel genes specific to A. cylindracea so far examined. Some genes with known functions were a pleurotolysin, a self-assembling poreforming cytolysins; Aa-Pri1 and Pir2p, specifically induced genes during fruiting initiation of other mushroom, Agrocybe aegerita; an amino acid permease; a cytochrome P450; a MADS-box gene; a peptidylprolyl isomerase; and a serine proteinase. For other clones, no clear function was annotated so far. We believe the first report of the differentially expressed genes in fruiting process of A. cylindracea will be great helps for further research.

The Optimal Culture Conditions for the Mycelial Growth of Oudemansiella radicata

Oudemansiella radicata, one of edible mushrooms belonging to Tricholomataceae of Basidiomycota, has been known to exhibit outstanding therapeutic effects on the hypertension caused by high blood pressure and inhibitory effects on the sarcoma 180 and Erhrlich carcinoma of mice. As one of preliminary experiments for producing fruiting-body of O. radicata, this study was carried out to obtain the basic information for culture conditions of mycelial growth of the fungus. The optimal temperature and pH for the mycelial growth were 25degrees C and pH 6, respectively. The medium for favorable mycelial growth of O. radicata was shown in the Lilly medium, whereas compact mycelial density was found in Hamada medium. The carbon and nitrogen sources promoting for mycelial growth of O. radicata were xylose and alanine, respectively. The optimum C/N ratio was about 20 : 1 in case that 3% glucose was supplimented to the basal medium as a carbon source.

The Characteristics of Cultural Conditions for the Mycelial Growth of Macrolepiota procera

Macrolepiota procera, one of edible mushrooms belongs to Agaricaceae of Basidiomycota, has a good taste and good medicinal value. As a preliminary study for the development of artificial cultivation method of edible mushroom, cultural characteristics of M. procera was investigated on various culture media under different environmental conditions. Mycelial growth was compared on culture media composed of various carbon and nitrogen sources, and C/N ratios. The optimal conditions for the mycelial growth were 30degrees C and pH 7. M. procera showed the rapid mycelial growth in the PDA media. The optimal carbon and nitrogen sources were maltose and glycine, respectively. The optimum C/N ratio was about 10 : 1 in case that 1% glucose was supplemented to the basal media as carbon source.

Culture Conditions Affecting the Optimal Mycelial Growth of Cystoderma amianthinum

Cystoderma amianthinum, one of edible fungi belongs to Agaricaceae of Basidiomycota, has a good taste and flavor. This study was carried out to obtain the basic informations for the optimum mycelial growth of C. amianthinum. The optimal conditions for the mycelial growth were 25degrees C and pH 5 in potato dextrose agar (PDA). C. amianthinum showed the favorable growth in the PDA and yeast malt extract agar (YMA). The favorable carbon and nitrogen sources promoting mycelial growth were fructose and histidine, respectively. The optimum C/N ratio was about 30 : 1 in case that 1% glucose was supplemented to the basal medium as a carbon source.

The Optimal Culture Conditions Affecting the Mycelial Growth and Fruiting Body Formation of Paecilomyces fumosoroseus

The fruiting body of Paecilomyces fumosoroseus was collected at Mt. Mani, Ganghwa Island, Korea in September, 2001. This study was carried out to obtain the basic informations for the mycelial growth and fruiting body production of P. fumosoroseus in artificial media. The optimal conditions for the mycelial growth were obtained at 25degrees C and in the range of pH 6~9, respectively. P. fumosoroseus showed the favorable growth on Hamada medium. The carbon and nitrogen source favorable for mycelial growth were dextrin and histidine, respectively. Optimum C/N ratio suitable for optimal growth of P. fumosoroseus was observed on the culture media adjusted to the ratio of 40:1. The mycelial growth of P. fumosoroseus was optimal on corn meal agar supplemented with 30% of silkworm pupae. The most favorable fruiting body formation of P. fumosoroseus was obtained in the medium containing unpolished rice supplemented with 20% (w/w) silk worm pupae at 25degrees C under 100 lux.