• Journal of Microbiology and Biotechnology
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• v.25 no.9
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• pp.1528-1531
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• 2015

Sclerotia of Wolfiporia cocos are of medicinal and culinary value. The genes and molecular mechanisms involved in W. cocos sclerotial formation are poorly investigated because of the lack of a suitable and reproducible transformation system for W. cocos. In this study, a PEG/CaCl₂-mediated genetic transformation system for W. cocos was developed. The promoter P_gpd from Ganoderma lucidum effectively drove expression of the hygromycin B phosphotransferase gene in W. cocos, and approximately 30 transformants were obtained per 10 μg DNA when the protoplast suspension density was 10⁶ protoplasts/ml. However, no transformants were obtained under the regulation of the PtrpC promoter from Aspergillus nidulans.

• Mycobiology
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• v.50 no.4
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• pp.254-257
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• 2022

Wolfiporia cocos is a wood-decay brown rot fungus belonging to the family Polyporaceae. While the fungus grows, the sclerotium body of the strain, dubbed Bokryeong in Korean, is formed around the roots of conifer trees. The dried sclerotium has been widely used as a key component of many medicinal recipes in East Asia. Wolfiporia cocos strain KMCC03342 is the reference strain registered and maintained by the Korea Seed and Variety Service for commercial uses. Here, we present the first draft genome sequence of W. cocos KMCC03342 using a hybrid assembly technique combining both short- and long-read sequences. The genome has a total length of 55.5 Mb comprised of 343 contigs with N50 of 332 kb and 95.8% BUSCO completeness. The GC ratio was 52.2%. We predicted 14,296 protein-coding gene models based on ab initio gene prediction and evidence-based annotation procedure using RNAseq data. The annotated genome was predicted to have 19 terpene biosynthesis gene clusters, which was the same number as the previously sequenced W. cocos strain MD-104 genome but higher than Chinese W. cocos strains. The genome sequence and the predicted gene clusters allow us to study biosynthetic pathways for the active ingredients of W. cocos.

• Journal of Mushroom
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• v.15 no.1
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• pp.54-56
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• 2017

Wolfiporia cocos is a well-known traditional medicine in China, Japan, Korea, and other Asian countries owing to its numerous therapeutic properties. With the aim to determine the morphology and genetic characteristics of W. cocosten strains of W. cocos were cultivated in vitro, and subsequently, rapid amplification of polymorphic DNA was performed. To the best of our knowledge, this is the first study to examine the morphology of fruit bodies of W. cocos in Korea. W. cocos were cultured on PDA agar at different temperatures (12, 16, 20, 24, and $28^{\circ}C$) under 12-hour light (600 Lux) / 12-hour dark photoperiod condition for 1 month. Appearance of fruit body was the highest at $28^{\circ}C$ condition in all the strains investigated. Honeycomb-like structure on sclerotia was observed in Andong 01, Andong 02, Andong 03, KFRI 1104, KFRI 1105, KFRI 1106, KFRI 1107, KFRI 1108, and ASI 13007 strains of. The KFRI 1103 strain formed cosmos petal-like structure on sclerotia. The average size of basidiospores was recorded as $7.55{\mu}m$ in height and $3.35{\mu}$ in width.

• Journal of Mushroom
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• v.20 no.4
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• pp.187-192
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• 2022

Wolfiporia cocos is an edible fungus commercially cultivated in Asia. To investigate metabolic changes of W. cocos mycelia under both light and dark culture conditions, gas chromatography mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC) analyses were performed. In terms of the total amount of sugars, alcohols, amino acids, organic acids, fatty acids, and purines, there no significant differences between the W. cocos mycelia cultivated under light (L) or dark (D) conditions (p < 0.05). However, there were some differences with respect to the production of particular sugars and proteins. The levels of trehalose (L: 17.2 ± 0.3% vs. D: 13.9 ± 1.6%), maltose (L: 0.9 ± 0.1% vs. D: 0.3 ± 0.1%), turanose (L: 0.7 ± 0.2% vs. D: 0.1 ± 0.1%), glutamine (L: 1.6 ± 0.3% vs. D: 0.7 ± 0.2%), and proline (L: 0.3 ± 0% vs. D: 0.1 ± 0%) were all significantly higher under light condition (p < 0.05). In contrast, the levels of galactose (L: 13.7 ± 1.2% vs. D: 17.6 ± 2.0%), aspartic acid (L: 0.6 ± 0.1 % vs. D: 0.9 ± 0.1%), cystathionine (L: 0.6 ± 0.1% vs. D: 0.8 ± 0 %), and malic acid (L: 0.7 ± 0.1% vs. D: 1.2 ± 0.1%) were higher under the dark condition. It is worth noting that the amount of pachymic acid, a pharmaceutically active compound of W. cocos, was 1.68 times greater under the light condition (p < 0.05).

• The Korean Journal of Mycology
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• v.49 no.3
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• pp.405-412
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• 2021

Wolfiporia cocos is an important medicinal fungus that has been used in regions of Northeast Asia including Korea, Japan, and China. W. cocos is classified in Korea into two types (red bokryeong and white bokryeong) based on the internal colors (yellow orange-pale pink and white) of the sclerotium. Generally, the W. cocos type cultivated on farms produces white sclerotium. In this study, we endeavored to select strains that form sclerotium in sawdust medium using 2-way cross-breeding among two cultivated strains and three wild strains. Monospores were isolated from the fruiting bodies of cultivated and wild strains on potato dextrose agar. Thirty-nine strains of 338 hybrid strains isolated formed sclerotia with white or yellow colors upon culture for 3 months in Pinus densiflora sawdust medium. Selection for sclerotium forming strains using sawdust culture follows a very simple and easy procedure that is presented for the first time in this paper. We plan to test selected strains in the field to aid in developing new varieties for the future.

• Journal of Mushroom
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• v.18 no.3
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• pp.244-253
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• 2020

The aim of this study was to investigate the biological activity of extracts obtained from the mycelium, sclerotium, and fruiting body of Wolfiporia cocos using different extraction solvents (hot water, 70% ethanol, and 70% methanol). Among the three developmental stages, the mycelium extracts showed the highest DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity, ferric reducing antioxidant power (FRAP), nitrite scavenging activity, and total polyphenolic content. Among the extraction solvents in the context of the W. cocos mycelium, the DPPH radical scavenging activity, FRAP, and total polyphenol content in the hot-water extracts were significantly higher than those in the other extracts. In the case of the sclerotium, the reducing power, nitrite scavenging activity, and total polyphenol content were significantly higher in 70% ethanolic extracts. The fruiting body showed the highest DPPH radical scavenging activity, reducing power, nitrite scavenging activity, and total polyphenol content in the context of hot-water extraction. Moreover, the β-glucan content was significantly higher in the sclerotium versus the mycelium or fruiting body. The total amino acid and total essential amino acid contents were remarkably higher in the mycelium and fruiting body than in the sclerotium; of note, and arginine (Arg) and phenylalanine (Phe) were highly detected among the amino acid components.

• The Korean Journal of Mycology
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• v.46 no.3
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• pp.295-306
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• 2018

In the present study, we compared the effects of 50% ethanolic extracts of Chinese and Korean Wolfiporia cocos (CPE and KPE) on in vitro lipid accumulation in 3T3-L1 cells and their anticancer activities in Sarcoma 180 cells. We further compared the anticancer activities and the 50% inhibitory concentrations ($IC_{50}$) of CPE with KPE with cultivated for one and two years in a landfill and a facility (LPE and FPE), respectively. In addition, the single oral dose toxicities of CPE and KPE were evaluated in mice. Lipid accumulation was inhibited after 48 hours, in CPE and KPE treated 3T3-L1 cells; however, no significant difference was observed between CPE and KPE in their lipid accumulation inhibitory activities. The anticancer activity of KPE was higher than that of CPE at $300{\mu}g/mL$ (p<0.05), revealing the possibility of an auxiliary biological means for origin identification. The anticancer activities of LPE and FPE were significantly stronger than that of CPE (p<0.05) but there was no difference between extracts from one- and two-year-old W. cocos, irrespective of the cultivation method. In single oral dose toxicity tests, CPE and KPE did not induce mortality during the 14-day observation. Thus, the 50% of lethal dose ($LD_{50}$) of CPE and KPE were estimated to be higher than 2,000 mg/kg. Taken together, our results indicate that the anticancer assay could be an auxiliary means of identifying the origin of W. cocos. In addition, artificial cultivation could be an alternative way to reduce the import of W. cocos. Lastly, 50% ethanolic W. cocos extracts could be potential candidates for obesity and cancer managements.

• Journal of Korean Society of Forest Science
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• v.113 no.1
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• pp.73-87
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• 2024

Wolfiporia hoelen (Fr.) Y.C.Dai & V. Papp, commonly known as Poria cocos, is a significant traditional herb used for medicinal and culinary purposes Asian and European countries. Many studies have confirmed that the main components of W. hoelen have pharmacological activities and thatits extract has been shown to affect bone metabolism. This study aimed to the potential of a 50% ethanol extract of the sclerotium of W. hoelen for preventing and treating bone diseases. The ethanol extract was systematically fractionated using n-hexane, dichloromethane, and ethyl acetate. The dichloromethane fraction caused an approximately 29% increase in alkaline phosphatase (ALP) differentiation activity in C2C12 cells compared to the control. Four compounds isolated from this active dichloromethane fraction were identified through instrumental analysis and literature references as 3α-dehydrotrametenolic acid, ergosterol, pachymic acid, and dehydrotumulosic acid. All four compounds were evaluated at increasing concentrations (1, 3, 10, 30, and 100 μM) to determine their effects on ALP differentiation activity in C2C12 cells and RANKL-induced inhibition activity in bone marrow macrophages (BMMs), with a concurrent assessment of cytotoxicity at these concentrations. At a concentration of 3 μM, dehydrotumulosic acid caused a 160% increase in ALP activity, 24% higher than in the BMP-2 control. BMMs treated with dehydrotumulosic acid at concentrations between 10 and 100 μM showed a substantial 15-86% decrease in RANKL-induced inhibition activity compared to the control, with distinct patterns of RANKL inhibition and cytotoxicity observed at 10 μM. These findings suggest that the ethanol extract from the sclerotium of W. hoelen has potential to modulate bone-cell differentiation, while highlighting the possible benefits of dehydrotumulosic acid isolated from the dichloromethane fraction of W. hoelen for preventing and treating osteoporosis.