• Journal of Mushroom
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• v.20 no.4
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• pp.199-207
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• 2022

The demand for novel strains has been rising in the domestic market to increase the production of sclerotia from Wolfiporia hoelen. To improve strain breeding efficiency, we investigated whether single-nucleotide polymorphisms (SNPs) in the RNA polymerase II subunit (RPB2) gene, which may be linked to the mating type locus, are useful for distinguishing monokaryons from dikaryons in Korean W. hoelen strains. We designed a specific primer set to efficiently amplify a region of RPB2 using PCR with the genomic DNA of 12 cultivated strains and 31 wild strains of W. hoelen collected from Korea. Nucleotide sequences of the PCR-amplified RPB2 genes were determined and analyzed for the presence of SNPs among the 43 W. hoelen strains. Previously reported SNP loci were detected in the RPB2 gene of all W. hoelen strains tested. However, these previously reported SNP loci could not be applied to differentiate monokaryons from dikaryons in approximately one-third of Korean wild strains with homozygous genotypes. Three additional SNPs in the RPB2 gene, which may improve the ability to distinguish monokaryons from dikaryons, were identified by searching through the multiple sequence alignments of the 43 W. hoelen strains. The applicability of these three novel SNPs, together with the previously known SNPs, in the RPB2 gene to W. hoelen strain breeding was verified by examining the hybrid strains and their parental strains.

• 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.