• The Korean Journal of Mycology
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• v.47 no.3
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• pp.181-186
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• 2019

A designated fungal isolate, KNU-US-1802E was isolated from the soil in Uiseong, Korea. To identify characteristics of the isolate, it was cultured on PDA media for 6 days at $35^{\circ}C$. Colonies on PDA are flat, light gray, dense, with entire margins; reverse dark gray to black, with white margins. Aerial mycelia were smooth-walled, hyaline and 40~42 mm diameter after 6 days at $35^{\circ}C$. Conidia were hyaline, one-celled, ellipsoidal to fusiform, forming long chains with average length ${\times}$ width of $5.0{\pm}0.3{\times}2.9{\pm}0.2{\mu}m$. Molecular analysis indicates that the internal transcribed spacer (ITS) region and partial beta-tubulin (tub2) gene sequence showed 100% and 99% similarities, respectively with Acrophialophora ellipsoidea CGMCC 3.15255 collected from China. Phylogenetic analysis by the neighbor-joining (NJ) method shows that the KNU-US-1802E was clustered with A. ellipsoidea CGMCC 3.15255 in a phylogenetic tree constructed using the concatenated sequences of ITS region and tub2 gene sequences with a high bootstrap value. Based on these findings, the isolate KNU-US-1802E was identified as Acrophialophora ellipsoidea, and this is the first report of this isolate in Korea.

• Korean Journal of Plant Resources
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• v.11 no.3
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• pp.296-300
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• 1998

When medicinal mushroom, Poria cocos, is cultured , inoculation method of spawn is cross slice inoculation of which the both sides of pine tree were peeled and spawn of P.cocos was inoculated. However, this method required lots of inoculation amount. This study was carried out to improve the culturing method of P. cocos. A good growth of P.cocos was observed in MCM(mushroom complete medium), showing proper mycelia growth and density. In inoculation amount, conventional method(cross slice inoculation) requires 20 bottles of spawn. In contrast, short log method required 8 bottles of spawn and drilling inoculation method 2~3 bottles, which could save by 60% and 85-90% respectively. In the selectrion of tree species, pine and larch had better condition for spawn culture and sclerotia formation condition.In terms of yield , pine was 33.7kg/3.3$m^2$. In the yield of pine, conventional method was 23.4kg/3.3$m^2$, drilling inoculation 29.4kg/3.3$m^2$, short log inoculation 31.7kg/3.3$m^2$, therefore drilling inoculation could increase by 25% and short log inoculation 35%, In addition, management cost was also saved.

• The Korean Journal of Mycology
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• v.47 no.2
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• pp.125-130
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• 2019

A fungal isolate, designated KNU-JJ-1824, was isolated from a soil sample collected from a field on Jeju, Korea. Colonies of the isolates cultured on PDA, MEA and CMAD for 14 days grew to diameters of 49~51 mm, 60~63 mm, and 47~50 mm, respectively. The colonies of this fungal isolate were dark green in color on both sides, and had irregular margins on the PDA media. Mycelia submerged and exhibited clear lines from the center to the edge on MEA media. On CMAD media, they were dark brown to greenish-brown, with narrow white margins. The shape of the conidia was guttulate, fusiform or clavate, and $1.9{\sim}3.4{\times}6.5{\sim}10.2{\mu}m$ in size. A molecular phylogenetic tree was constructed using dataset from small subunit (SSU) rDNA and the internal transcribed spacer (ITS) regions, and the isolate was found to cluster with Scleroconidioma sphagnicola UAMH 9731. Based on the results of the phylogenetic tree analysis and the cultural and morphological characteristics, the isolate was identified as Scleroconidioma sphagnicola. We report S. sphagnicola for the first time in Korea.

• Journal of the East Asian Society of Dietary Life
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• v.23 no.6
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• pp.833-838
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• 2013

In this study, twelve strains of brewing fungi were individually cultivated on wheat extract broth (WEB), potato dextrose broth (PDB) and malt extract broth (MEB) in order to determine the microorganism with good culture characteristics as well as with amylolytic activity. The strain cultured in PDB exhibited mycelia production from 12.6 g/L (Rhizopus oryzae KACC 45714) to a maximum of 48.0 g/L (Aspergillus oryzae KACC 46959), which was 2.3~9.2 times lower than that of the strain cultured in WEB and 1.7~14.6 times lower than that of the strain cultured in MEB. Accorfing the results, We found that the commercial strains of A. oryzae Suwon, CF1001 and CF1003 had a higher dry cell mass than the wild-type strains KACC 46421, 46423, 46424 and 46959. For Rhizopus sp., the acidity levels in WEB, PDB and MEB were 0.12~0.47%, 0.22~1.0% and 0.16~0.68% (equivalent lactic acid concentration) respectively. For A. oryzae, the acidity levels were 0.06~0.11%, 0.03~0.04% and 0.06~0.08% (equivalent lactic acid concentration), respectively. Amylase enzyme from Rhizopus delemar KACC 46419 exhibited an enzyme activity of 0.013 U and 0.019 U in WEB and MEB cultures, respectively. The enzyme activity of the amylase enzyme from A. oryzae was 0.019~0.037, 0.017~0.033 and 0.028~0.046 U in WEB, PDB and MEB cultures, respectively.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.37 no.4
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• pp.351-356
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• 2011

Citrus unshiu (C. unshiu) Markovich were dried peel of mandarin orange, of which fresh fruit was one of the famous foods in Korea and Eastern Asia. In the oriental medicine, C. unshiu peel was known to have a diuretic effect and to strengthen spleen function. Recently, natural flavonoids of C. unshiu peel have been investigated. In this study, C. unshiu peel extract containing flavonoid-glycosides was cultured with Schizophyllum commune (S. commune) mycelia producing ${\beta}$-glu- cosidase and its biological activities were investigated. ${\beta}$-glucosidase of S. commune mycelia converted the flavonoid-glycosides (rutin and hesperidin) into aglycones (naringenin and hesperetin). Fermented C. unshiu peel extract compounds were analyzed by HPLC system. The photoprotective potential of fermented C. unshiu peel extract was tested in human dermal fibroblasts (HDFs) exposed to UVA. Fermented C. unshiu peel extract extract also showed notable in vitro anti-inflammatory effect on cellular systems generating cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX) metabolites. Also, UVB-induced production of interleukin-$1{\alpha}$ in human HaCaT cells was reduced in a dose-dependent manner by treatment with fermented C. unshiu peel extract. These results suggest that fermented C. unshiu peel extract may mitigate the effects of photoaging in skin by reducing UV-induced adverse skin reaction.

• Journal of Mushroom
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• v.17 no.4
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• pp.191-196
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• 2019

The edible ectomycorrhizal mushroom Amanita javanica is a valuable species protected by forest law in Korea. However, basic characterization data on its use as an important forest resource has been limited. This study was performed to determine mycelia growth characteristics of the domestically isolated Amanita javanica strain NIFoS 1267 on potato dextrose agar media under diverse culture conditions. Physical factors temperature, pH, and light, as well as chemical factors salts, heavy metals, and pesticides were examined for their effects on the growth of the mushroom strain. The mycelia of A. javanica strain exhibited optimal growth when cultured in dark at 30℃ in media with a pH of 5-6. Normal levels of growth were observed in media containing up to 2% saline. At a heavy metal ion content of 50 ppm, mycelial growth was not affected by arsenic ion but was affected by cadmium and lead ions. In the tests performed with two pesticides used in Korean forests, the growth of the mushroom strain was not affected by the presence of abamectin, but was inhibited in media containing acetamiprid, emamectin benzoate, or thiacloprid. These results are expected to facilitate artificial cultivation of A. javanica as a new commercial product.

• Journal of Ginseng Research
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• v.35 no.2
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• pp.235-242
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• 2011

To obtain microorganisms for the microbial conversion of ginsenosides in red ginseng extract (RGE), mushroom mycelia were used for the fermentation of RGE. After fermentation, total sugar contents and polyohenol contents of the RGEs fermented with various mushrooms were not a significant increase between RGE and the ferments. But uronic acid content was relatively higher in the fermented RGEs cultured with Lentus edodes (2155.6 ${\mu}g/mL$), Phelllinus linteus (1690.9 ${\mu}g/mL$) and Inonotus obliquus 26137 and 26147 (1549.5 and 1670.7 ${\mu}g/mL$) compared to the RGE (1307.1 ${\mu}g/mL$). The RGEs fermented by Ph. linteus, Cordyceps militaris, and Grifola frondosa showed particularly high levels of total ginsenosides (20018.1, 17501.6, and 16267.0 ${\mu}g/mL$, respectively). The ferments with C. militaris (6974.2 ${\mu}g/mL$), Ph. linteus (9109.2 ${\mu}g/mL$), and G. frondosa (7023.0 ${\mu}g/mL$) also showed high levels of metabolites (sum of compound K, $Rh_1$, $Rg_5$, $Rk_1$, $Rg_3$, and $Rg_2$) compared to RGE (3615.9 ${\mu}g/mL$). Among four different RGE concentrations examined, a 20 brix concentration of RGE was favorable for the fermentation of Ph. linteus. Maximum biotransformation of ginsneoside metabolites (9395.5 ${\mu}g/mL$) was obtained after 5 days fermentation with Ph. linteus. Maximum mycelial growth of 2.6 mg/mL was achieved at 9 days, in which growth was not significantly different during 5 to 9 days fermentation. During fermentation of RGE by Ph. linteus in a 7 L fermenter, $Rg_3$, $Rg_5$, and $Rk_1$ contents showed maximum concentrations after 5 days similar to flask fermentation. These results confirm that fermentation with Ph. linteus is very useful for preparing minor ginsenoside metabolites while being safe for foods.

• The Korean Journal of Mycology
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• v.18 no.3
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• pp.115-126
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• 1990

To establish basic techniques for protoplast fusion of Coriolus versicolor several factors affecting protoplast formation and regeneration were investigated. Protoplast isolation was at maximum with 2.5-day cultured mycelia of C. versicolor treated with the combination of two enzymes, Novozym 234 (10 mg/ml) and cellulase Onozuka R-10 (15 mg/ml), for 3-4.5 hours at $30^{\circ}C.$ As an osmotic stabilizer for stabilizing the protoplast, 0.6 M sucrose was the best for formation and regeneration of the protoplast from the mycelia of the fungus and the regeneration frequency was 3.48%. Protoplast fusion was made by a modified method of Peberdy using PEG (M.W. 4,000). The fusion frequency between two mutants of C. versicolor was 1.86% and the fusion products showed differences in growth rate and colony morphology.

• Journal of Mushroom
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• v.2 no.1
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• pp.15-20
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• 2004

Pleurotus ostreatus, the oyster mushroom, is one of the most widely cultivated and important edible mushrooms in the world. In order to study the developmental process of P. ostreatus and its regulatory mechanism, a new culturing method needs to be established for inducing the fruiting body and sporulation in the laboratory. In this study, we have examined whether the fruiting body of P. ostreatus can be formed on the plastic petri dish which are commonly used for cell culture in the laboratory. The strain was cultured on $60{\times}15mm$ plastic petri dish with potato dextrose agar media at $28^{\circ}C$ for mycelial growth and then at $18^{\circ}C$ for the formation of primordia and fruiting bodies within plant growth chamber. The development of primordia into fruiting bodies was achieved on cultured dishes under air ventilation. At the primordia stage, the normal formation of fruiting body was blocked by sealing the plastic dish with parafilm. The periods requiring for the formation of primordia and fruiting bodies were examined on the dish culture. About 96% and 76% of cultured samples formed primordia and fruiting bodies under the optimal conditions during ten weeks of culture, respectively. These culturing periods, however, were changed by the mechanical injury treatment to mycelia. As other factors affecting the fruiting body formation, the effects of light and cold shock have been tested. No fruiting formation was observed on the cultured dishes under the dark. The cold shock treatment by storing cultured dishes for one day at $4^{\circ}C$ did not have any significant effects in the fruiting body formation. Spores of fruiting bodies acquired from the petri dishes could be germinated on culture media at $28^{\circ}C$. These results suggest that the fruiting bodies of P. ostreatus can be formed on the experimental petri dish and this dish-culturing method is useful for understanding of the developmental process of P. ostreatus in the laboratory. Furthermore, the dish-culturing method is able to shorten the life cycle of P. ostreatus without requiring large area and expensive device.

• The Plant Pathology Journal
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• v.33 no.1
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• pp.95-100
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• 2017

Fungicide-resistant Alternaria alternata impede the practical control of the Alternaria diseases in crop fields. This study aimed to investigate cytological fungicide resistance mechanisms of A. alternata against dicarboximide fungicide iprodione. A. alternata isolated from cactus brown spot was cultured on potato-dextrose agar (PDA) with or without iprodione, and the fungal cultures with different growth characteristics from no, initial and full growth were observed by light and electron microscopy. Mycelia began to grow from one day after incubation (DAI) and continued to be in full growth (control-growth, Con-G) on PDA without fungicide, while on PDA with iprodione, no fungal growth (iprodione-no growth, Ipr-N) occurred for the first 3 DAI, but once the initial growth (iprodione-initial growth, Ipr-I) began at 4-5 DAI, the colonies grew and expanded continuously to be in full growth (iprodione-growth, Ipr-G), suggesting Ipr-I may be a turning moment of the morphogenetic changes resisting fungicidal toxicity. Con-G formed multicellular conidia with cell walls and septa and intact dense cytoplasm. In Ipr-N, fungal sporulation was inhibited by forming mostly undeveloped unicellular conidia with degraded and necrotic cytoplasm. However, in Ipr-I, conspicuous cellular changes occurred during sporulation by forming multicellular conidia with double layered (thickened) cell walls and accumulation of proliferated lipid bodies in the conidial cytoplasm, which may inhibit the penetration of the fungicide into conidial cells, reducing fungicide-associated toxicity, and may be utilized as energy and nutritional sources, respectively, for the further fungal growth to form mature colonies as in Ipr-G that formed multicellular conidia with cell walls and intact cytoplasm with lipid bodies as in Con-G.