• Journal of Life Science
• /
• v.20 no.5
• /
• pp.782-788
• /
• 2010

The protective effects of a powder mixed with solid-cultured and liquid-cultured Lentinus edodes mycelia (2 : 1, w/w) (designate LED) with different doses of carbon tetrachloride ($CCl_4$) on induced hepatotoxicity in male Sprague-Dawley (SD) rats was investigated. The rats were divided into seven groups (6 rats/group) and the following substances were administered orally to each group: Vehicle (0.2 ml distilled water), Control (0.2 ml distilled water), LED (LED 100, 200, 300 and 400 mg/kg BW in 0.2 ml distilled water), and Silymarin (200 mg/Kg BW in 0.2 ml distilled water). After two weeks of daily administration, all groups except for the Vehiclegroup were subjected to abdominal injection with $CCl_4$ ($CCl_4$ : corn oil, 1 : 1 v/v; 0.5 ml/kg BW). One day later, blood and liver samples were collected to analyze biomarkers. All LED treatments elevated hepatic superoxide dismutase (SOD), catalase and glutathione peroxidase (GSH peroxidase) activities, and reduced thiobarbituric reactive substances (TBARS), tumor necrosis factor-$\alpha$ (TNF-$\alpha$), interleukin-$1{\beta}$ (IL-$1{\beta}$) and interleukin-6 (IL-6), resulting in the reduction of glutamate-oxalate transaminase (GOT), glutamate-pyruvate transaminase (GPT) and lactic acid dehydrogenase (LDH) activities in plasma. These results indicate that LED effectively protected SD rat hepatotoxicity induced by $CCl_4$ through its antioxidative activity and reduction of some cytokines. The highest efficacy was found in LED 200 mg/kg BW, showing potential as a useful material for protection from hepatotoxicity in humans.

• The Korean Journal of Mycology
• /
• v.35 no.2
• /
• pp.81-84
• /
• 2007

In some cases, the problem with the mycelium of Tricholoma giganteum is delayed mycelial growth or non-regeneration. Therefore, this study was conducted to understand the mycelial storage condition of T. giganteum and to investigate the regeneration ratio of mycelial growth. T. giganteum obtained from KACC in RDA was evaluated for its simple preservation at $10^{\circ}C$ and subcultured on different media. Mycelium of T. giganteum was cultured on the PBA (potato bamboo extract medium) for seven days at $30^{\circ}C$. Using simple preservation method, the mycelium of T. giganteum (MKACC 50852) and Pleurotus ostreatus (Chunchu No. 2) were stored on six different media in two kinds of storage vessels (tube and vial) for a period of 1-12 months at $4^{\circ}C,\;15^{\circ}C$, and $25^{\circ}C$ storage temperatures. At $4^{\circ}C$ storage condition, mycelial regeneration was failed in all agar media, but was successful in the sawdust medium for 3 months. At $15^{\circ}C$ storage, mycelial activity was maintained up to six months. On the other hand, P. ostreatus produced well-regenerated mycelia both at $4^{\circ}C$ and $15^{\circ}C$ storage up to 12 months. In conclusion, it is estimated that the mycelia storage condition of T. giganteum must be done at $15^{\circ}C$ and subcultured within six months.

• Journal of Mushroom
• /
• v.7 no.3
• /
• pp.115-121
• /
• 2009

Recently sawdust cultivation of Shiitake mushroom (Lentinula edodes ) is getting increased because log cultivation is getting difficult to get oak logs. It is important to make mycelia browning on the substrate surface in sawdust cultivation. This browned surface plays an important role like as artificial bark of the oak log, which protects the other pests and suppresses water evaporation in the substrate. The period for mycelia browning is so long that the sawdust cultivation of Shiitake mushroom can not spread well into the mushroom farms. In this article we would like to discuss about the effect of environmental condition to the mycelial browning during sawdust bag cultivation for the To reduce the period required for browning of substrates, sawdust substrates was illuminated light with difference intensity. One hundred Lux light illumination was needed for producing normal yield of fruit body but fruit body yield was low and abnormally shaped fruit body was produced when cultured under the dark condition of incubation. Illumination over 200lux is necessary for the successful browning of substrates during incubation. Optimum incubation temperature for browning of substrates and fruiting was $25^{\circ}C$. The treatment of cotton plug with different size to identify the effect of aeration on the browning of substrates and fruiting showed rapid mycelial growth and reduced the periods for browning as the size of cotton plug was bigger. However, yield of fruit body was the highest at 16mm diameter cotton plug as compared to 20mm of that. $CO_2$ content in vessel of substrates was low as the size of cotton plug was bigger during incubation. $CO_2$ content during incubation of substrate was highest in periods between 8 week and 14 week after inoculation of shiitake when substrate was changed color into brown. $C_2H_4$ content in vessel with substrates was highest at 8mm diameter cotton plug and it was increased by order of 12, 16, 20, 0, 4 mm diameter cotton plug during substrate incubation. Sawdust substrate was soaked in cold water for different time to identify soaking effect of sawdust substrate on fruit body yield and activities of enzymes in these substrates were investigated. The fruit body yield was increased up to 40% by soaking substrates in comparison with unsoaked substrates. The soaked substrates showed 165, 175g/1,000ml at treatment of 4 and 15 hours, respectively. Cellulose activities in soaked substrates were not changed with soaking time, but activities of laccase, lignin degradation enzyme, were drastically increased up to 4 times in comparison with unsoaked substrates.

• KSBB Journal
• /
• v.21 no.5
• /
• pp.384-388
• /
• 2006

The production stability of high-yielding mutants of Streptomyces lincolnensis immobilized on celite beads was examined in repeated batch cultures. We also explored the feasibility of immobilization of vegetative mycelial cells on pre-wetted celite beads, which is practical method for cell immobilization. Repeated transfer of immobilized cells into fresh medium every 10 days increased productivity of immobilized cells and maximum concentration of lincomycin, 1007 $({\pm}256)$ mg/L, was obtained at the end of the ninth cycle. A 1.4-fold higher productivity was obtained in immobilized-cell culture than that obtained by suspended-cell culture. When pre-wetted beads were inoculated with vegetative mycelia and cultured a slightly higher amount of immobilized cells and lincomycin was obtained more than those obtained by culture of spores immobilized on dry beads. This result indicates that immobilization of mycelial cells on pre-wetted beads was readily available. This technique is simple and no additional facilities are required for cell immobilization.

• Mycobiology
• /
• v.32 no.1
• /
• pp.1-5
• /
• 2004

The present study was carried out to investigate morphological characteristics of pseudosclerotia of Grifola umbellata formed by artificial cultures. Isolate G. umbellata DUM GUS-01 was obtained from sclerotium cultivated in field. The fungal isolate was cultured on PDYM broth, PDYMA(potato dextrose yeast malt agar) and oak sawdust media at $20^{\circ}C$ under the dark condition. G. umbellata DUM GUS-01 showed a volumetric increment of fungal lumps rather than mycelial growth. Particularly, G. umbellata DUM GUS-01 produced a large amount of melanin pigments in all culture treatments. The color of the fungal mass has been changed into grey gradually, and then formed melanized rind-like structure on its superficial part. The fungal structures which were covered with melanized rind-like layer were named as pseudosclerotia of G. umbellata. The pseudosclerotia of G. umbellata DUM GUS-01 formed a new white mycelial mass, which was swollen out of the melanized rind structure for its volumetric increment. When the pseudosclerotia were sectioned, their structure was discriminated from two structures such as a melanized rind-like structure layer formed by aggregation of aged mycelia and a white mycelial mass with high density. As results of scanning electron microscopic examination, the pseudosclerotia of G. umbellata DUM GUS-01 which were formed in in vitro conditions were similar to the sclerotia of G. umbellata cultivated in natural conditions except for the crystals formed in medula layer of natural sclerotia. Although size, solidity of rind structure and mycelial compactness of pseudosclerotia were more poor than those of natural sclerotia, the morphological structure and growth pattern of pseudosclerotia were very similar to those of natural sclerotia. Therefore, it is probable to induce pseudosclerotia to sclerotia of G. umbellata in in vitro conditions. Consequently, it seems that the induced pseudosclerotia can be used as inoculum sources to substitute natural sclerotia in field cultivation.

• The Korean Journal of Mycology
• /
• v.14 no.2
• /
• pp.165-174
• /
• 1986

Conditions for production, fusion and reversion of protoplasts of Aspergillus niger were investigated, and an attempt was made to enhance fusion frequency. Auxotrophic mutants and morphological mutants were induced by U.V. irradiation $(9.9\;erg/mm^2,\;13min)$ on Aspergillus niger. Maximum yield of protoplasts was obtained from 21 hr cultured mycelia by using 1% driselase in 0.6 M KCl or 0.6 M $NH_4Cl$ as osmotic stabilizer. The optimal temperature for mycelium digestion was $30^{\circ}C$, and the optimal pH was 6.0. Protoplasts produced at different digestion period showed heterogeneity in size and vacuole content. Maximal frequency of protoplasts reversion was obtained on 0.6 M KCl stabilized agar medium at pH 5.0. Reversion frequencies of protoplasts produced for 3 hr and 1 hr mycelial digestion were 8.0% and 15.3%, respectively. The optimal concentration of PEG(m.w. 6000) for protoplast fusion was 30%, and that of $CaCl_2$ was $1{\sim}50\;mM$. The optimal pH and period for the reaction of PEG solution were 8.0 and 10 minutes, respectively. Fusion frequencies between auxotrophic protoplasts produced for 3 hr-mycelial digestion were $0.06{\sim}0.42%$, and those for 1 hr-mycelial digestion were $0.09{\sim}0.54%$.

• The Korean Journal of Mycology
• /
• v.22 no.2
• /
• pp.130-137
• /
• 1994

This experiment was carried out to investigate proper conditions for protoplast isolation and regeneration from mycelia of Lyophyllum decastes. Novozym 234(10 mg/ml) with 0.6 M $MgSO_4$ in phosphate buffer(pH 4.0) was proper for protoplast isolation. The optimal reaction time of the mycelium with the lytic enzyme was four hours in shaking condition at 120 strokes per min. When the mycelium of L. decastes was cultured at $24^{\circ}C$ for 5 days, the formation of protoplasts was effective. The liquid medium was more effective for protoplast isolation than the solid medium. In the liquid medium, high yields of protoplasts were obtained from 0.6 M $MgSO_4$ osmotic stabilizer. Protoplasts of L. decastes were regenerated to normal hyphal growth and the regeneration frequency of the protoplasts in the complete agar medium containing Triton X-100(0.0025%) was $5.94{\sim}8.32%$. The regeneration medium stabilized with 0.6 M sucrose was the best for regeneration of the protoplasts. In contrast to protoplast formation, regeneration was inhibited by the inorganic salts used as osmotic stabilizer.

• The Korean Journal of Mycology
• /
• v.40 no.2
• /
• pp.98-103
• /
• 2012

This study has been conducted to screen the decolorization of 4 aromatic synthetic dyes and production of ligninolytic enzymes by 4 white rot fungi such as Bjerkanderia adusta, Cerrena unicolor, Pleurotus pulmonarius and Abortiporus biennis. It was found that B. adusta, C. unicolor, and P. pulmonarius have the ability to efficiently decolorize congo red and moderately decolorized amaranth and orange G in solid and liquid culture media. However, the decolorization rate of 4 synthetic dyes by A. biennis was relatively low. The decolorization of congo red, amaranth, orange G were related to the growth rate of the fungal mycelia in the solid medium. But, the all fungi tested did not efficiently decolorize methylene blue in the liquid culture media. To investigate the production of ligninolytic enzymes in media containing aromatic compounds, fungi were cultured in 1% naphthalene supplemented potato dextrose broth medium. All fungi tested had the capability to produce laccase, lignin peroxidase and manganese peroxidase, and B. adusta was the best ligninolytic enzymes producing white rot fungus among other fungi tested.

• Journal of Mushroom
• /
• v.11 no.2
• /
• pp.63-68
• /
• 2013

The fruiting body of honey mushroom, Armillaria gallica, was collected from Gastrodia elata cultivated fields. Pure cultures were isolated from fruiting body tissue of the mushrooms, and cultured on MCM (mushroom complete medium) or PDA (potato dextrose agar) medium. Then, 12 different types of mycelial growth characteristics such as growth rate, colony morphology and rhizomorph formation were obtained. The vitality of the mycelial growth and rhizomorph formation of the fruiting body culture isolates were better on MCM than PDA, suggesting that the optimal culture medium for A. gallica mycelia was MCM. To observe the feature of colony morphology, the subculture of isolates were incubated on MCM. Consequently, we could find the segregated or differentiated colony morphology from isolate type 11 that was similar morphology to isolate type 12. For phylogenetic analysis of the 12 isolates, RAPD (Random Amplified Polymorphic DNA) were performed. The isolate type 12 was not only shown different band patterns of RAPD variation in other 11 isolates, but also commercial strain known as Chunmagyun No. 1. Among the tissue culture isolates of fruiting, strains with better mycelial growth characteristics than Chunmagyun No. 1 were selected. We expect that the new strain can be substituted to commercial strain Chunmagyun No. 1.

• The Korean Journal of Mycology
• /
• v.15 no.3
• /
• pp.149-157
• /
• 1987

This study was undertaken to investigate the differentiation, from spore germination to hyphae growth and phialide formation, of Aspergillus niger through the method of synchronous and submerged culture. Through continuous experiments by shake culture with potassium acetate medium, we observed the formation of spores at appropriate concentration and pH. Potassium acetate medium was set pH 5.5, 6.0, 6.5 and 7.0 on each scale, and control, 20 mM, and 40 mM, 80 mM and 160 mM concentrations on the other scale. Aspergillus niger was cultured in the defined media at $28^{\circ}C$, and mycelial dry weight, changes of pH and the onset of sporulation were checked. The mycelial dry weight, increased in potassium acetate medium, and pH increased during mycelial growth and gradually decreased after the spore formation. When pH increased excessively in Potassium acetate medium with pH 7.0, the mycelia could not adapt and mycelial dry weight decreased gradually. At pH 5.5, the onset of sporulation was done within one day at 20 mM it took, at 80 mM three days and at 160 mM concentration. in two days, at 40 mM one to four days were taken, 80 mM concentration respectively. At pH 6.5, the onset of sporulation was done in three days and four days at 80 mM concentrations respectively. Spore formation was not shown at pH 7.0. In controlled medium with all levels of pH, spore formation was not shown.