• Mycobiology
• /
• v.46 no.2
• /
• pp.159-167
• /
• 2018

This study evaluated the in vitro and in vivo hypolipidemic effects of the medicinal mushroom Phellinus pini. The methanol extract (ME) of the fruiting body of Ph. pini was active against pancreatic lipase and cholesterol esterase with 99.14% and 67.23% inhibited activity at 1.0 mg/mL, respectively. It also inhibited 81.81% and 55.33% of ${\alpha}$-glucosidase and ${\alpha}$-amylase activities, respectively, at 2.0 mg/mL. Hyperlipidemia as induced by feeding rats with a high fat and cholesterol diet (HFC). HFC supplemented with a 5% fruiting body powder of Ph. pini (HFC + PhP) significantly reduced plasma total cholesterol, low-density lipoprotein cholesterol, and triglycerides in rats compared with HFC. The reduced levels were comparable to rats fed the normal control diet (NC). The atherogenic index of HFC + PhP rats was significantly lower than that of the HFC rats. The excretion of fecal total lipid and cholesterol in the HFC + PhP rats was significantly higher than those in the NC and HFC rats. Histopathological examinations demonstrated scant deposition of lipids in the liver of rats fed HFC + PhP. The dietary supplementation with the fruiting body powder provided natural plasma lipid and glucose lowering effects in experimental rats without adverse effects on the plasma biochemical parameters and liver function related enzyme activities. Therefore, the hypolipidemic effects of Ph. pini may be due to the inhibitory effects on pancreatic lipase, cholesterol esterase, ${\alpha}$-glucosidase, and ${\alpha}$-amylase, and excretion of excess lipids and cholesterol in the feces.

• Mycobiology
• /
• v.48 no.3
• /
• pp.195-203
• /
• 2020

Marine yeasts have tremendous potential in industrial applications but have received less attention than terrestrial yeasts and marine filamentous fungi. In this study, we have screened marine yeasts for amylolytic activity and identified an amylase-producing strain PH-Gra1 isolated from sea algae. PH-Gra1 formed as a coral-red colony on yeast-peptone-dextrose (YPD) agar; the maximum radial growth was observed at 22 ℃, pH 6.5 without addition of NaCl to the media. Based on the morphology and phylogenetic analyses derived from sequences of internal transcribed spacer (ITS) and a D1/D2 domain of large subunit of ribosomal DNA, PH-Gra1 was designated Sporidiobolus pararoseus. S. pararoseus is frequently isolated from marine environments and known to produce lipids, carotenoids, and several enzymes. However, its amylolytic activity, particularly the optimum conditions for enzyme activity and stability, has not been previously characterized in detail. The extracellular crude enzyme of PH-Gra1 displayed its maximum amylolytic activity at 55 ℃, pH 6.5, and 0%-3.0% (w/v) NaCl under the tested conditions, and the activity increased with time over the 180-min incubation period. In addition, the crude enzyme hydrolyzed potato starch more actively than corn and wheat starch, and was stable at temperatures ranging from 15 ℃ to 45 ℃ for 2 h. This report provides a basis for additional studies of marine yeasts that will facilitate industrial applications.

• Mycobiology
• /
• v.42 no.2
• /
• pp.167-173
• /
• 2014

A ${\beta}$-glucan synthase gene was isolated from the genomic DNA of polypore mushroom Sparassis crispa, which reportedly produces unusually high amount of soluble ${\beta}$-1,3-glucan (${\beta}$-glucan). Sequencing and subsequent open reading frame analysis of the isolated gene revealed that the gene (5,502 bp) consisted of 10 exons separated by nine introns. The predicted mRNA encoded a ${\beta}$-glucan synthase protein, consisting of 1,576 amino acid residues. Comparison of the predicted protein sequence with multiple fungal ${\beta}$-glucan synthases estimated that the isolated gene contained a complete N-terminus but was lacking approximately 70 amino acid residues in the C-terminus. Fungal ${\beta}$-glucan synthases are integral membrane proteins, containing the two catalytic and two transmembrane domains. The lacking C-terminal part of S. crispa ${\beta}$-glucan synthase was estimated to include catalytically insignificant transmembrane ${\alpha}$-helices and loops. Sequence analysis of 101 fungal ${\beta}$-glucan synthases, obtained from public databases, revealed that the ${\beta}$-glucan synthases with various fungal origins were categorized into corresponding fungal groups in the classification system. Interestingly, mushrooms belonging to the class Agaricomycetes were found to contain two distinct types (Type I and II) of ${\beta}$-glucan synthases with the type-specific sequence signatures in the loop regions. S. crispa ${\beta}$-glucan synthase in this study belonged to Type II family, meaning Type I ${\beta}$-glucan synthase is expected to be discovered in S. crispa. The high productivity of soluble ${\beta}$-glucan was not explained but detailed biochemical studies on the catalytic loop domain in the S. crispa ${\beta}$-glucan synthase will provide better explanations.

• Mycobiology
• /
• v.29 no.4
• /
• pp.218-223
• /
• 2001

A rapid radicle assay for prescreening antagonistic bacteria to Phytophthora capsic4 causal agent of Phytophthora blight of pepper was developed. Sixty-four bacterial strains with in vitro antifungal activity selected out of 1,400 strains isolated from soils of Ansung, Chunan, Koyang, and Paju, Korea in 1998 were used for development of the bioassay. Uniformly germinated pepper seeds dipped in bacterial cells for 3 hours were placed near the edges of growing mycelia of P. capsici on water agar containing 0.02% glucose. Five-week-old pepper plants(cv. Nockwang) were inoculated to compare with results of the radicle assay developed in this study. For plant inoculation, pepper seeds were sown in potting mixtures incorporated with the bacterial strains, then transplanted into steam-sterilized soils 3 weeks later. Plants were hole-inoculated with zoospores of P. capsici 2 weeks after transplanting. Disease incidence and severity were determined in radicle and plant assessments, respectively. In radicle assay, six strains, GK-B15, GK-B25, OA-B26, OA-B36, PK-B09, and VK-B14 consistently showed the significant(P=0.05) disease reduction against radicle infection by the fungus, four of which also did in plant assessments. Strains OA-B36 and GK-B15 consistently reduced the fungal infection in both the radicle assay and the plant assessment. Therefore, prescreening strains using the radicle assay developed in this study followed by plant assay could reduce time and labor, and improved the possibility of selecting antagonistic bacteria for control of Phytophthora blight of peppers.

• Mycobiology
• /
• v.40 no.3
• /
• pp.151-158
• /
• 2012

Very few studies have addressed the phylogenetic diversity of fungi from Northeast India under the Eastern Himalayan range. In the present study, an attempt has been made to study the phylogenetic diversity of culturable soil fungi along the altitudinal gradients of eastern Himalayas. Soil samples from 24 m above sea level to 2,000 m above sea level altitudes of North-East India were collected to investigate soil micro-fungal community structure and diversity. Molecular characterization of the isolates was done by PCR amplification of 18S rDNA using universal primers. Phylogenetic analysis using BLAST revealed variation in the distribution and richness of different fungal biodiversity over a wide range of altitudes. A total of 107 isolates were characterized belonging to the phyla Ascomycota and Zygomycota, corresponding to seven orders (Eurotiales, Hypocreales, Calosphaeriales, Capnodiales, Pleosporales, Mucorales, and Mortierellales) and Incertae sedis. The characterized isolates were analysed for richness, evenness and diversity indices. Fungal diversity had significant correlation with soil physico-chemical parameters and the altitude. Eurotiales and Hypocreales were most diverse and abundant group of fungi along the entire altitudinal stretch. Species of Penicillium (D=1.44) and Aspergillus (D=1.288) were found to have highest diversity index followed by Talaromyces (D=1.26) and Fusarium (D=1.26). Fungal distribution showed negative correlation with altitude and soil moisture content. Soil temperature, pH, humidity and ambient temperature showed positive correlation with fungal distribution.

• Mycobiology
• /
• v.36 no.4
• /
• pp.249-254
• /
• 2008

Minimal growth inhibitory concentrations (MICs) of chitosan acetate (M.W. 60 kDa) on heterotrophic bacteria (strains MK1, S, and R) isolated from the soft-rotten tissues of Neungee mushroom (Sarcodon aspratus) were measured. The slimy substance produced by the MK1 strain was responsible for the diseased mushroom’s appearance. The S and R strains were members of the Burkholderia cepacia complex. These strains showed different levels of susceptibility toward chitosan acetate. The MIC of chitosan acetate against the MK1 and S strains was 0.06%. The MIC against the R strain was greater than 0.10%. Survival fractions of the MK1 and S strains at the MIC were $3\;{\times}\;10^{-4}$ and $1.4\;{\times}\;10^{-3}$ after 24 h, and $2\;{\times}\;10^{-4}$ and $7\;{\times}\;10^{-4}$ after 48 h, respectively. Survival fractions of the R strain after 24 and 48 hr at 0.1% chitosan acetate were $1\;{\times}\;10^{-2}$ and $6.9\;{\times}\;10^{-3}$, respectively. Compared to the MK1 and S strains, the low susceptibility of the R stain towards chitosan acetate could be due to the ability of the R strain to utilize chitosan as a carbon source. Thirty-eight percent of Neungee pieces treated in a 0.06% chitosan acetate solution for $2{\sim}3$ second did not show any bacterial growth at 4 days, whereas bacterial growth around untreated mushroom pieces occurred within 2 days. These data suggest that chitosan acetate is highly effective in controlling growth of indigenous microorganisms on Neungee. The scanning electron micrographs of the MK1 strain treated with chitosan revealed a higher degree of disintegrated and distorted cellular structures.

• Mycobiology
• /
• v.31 no.3
• /
• pp.179-182
• /
• 2003

A total of 187 endophytic fungi were isolated from 11 plant species, which were collected from 11 locations in Korea. Their antifungal activities were screened in vivo by antifungal bioassays after they were cultured in potato dextrose broth and rice solid media. Antifungal activity against plant pathogenic fungi such as Magnaporthe grisea(rice blast), Corticium sasaki(rice sheath blight), Botrytis cinerea(tomato gray mold), Phytophthora infestans(tomato late blight), Puccinia recondita(wheat leaf rust), and Blumeria graminis f. sp. hordei(barley powdery mildew) was determined in vivo by observing the inhibition of plant disease development. Twenty(11.7%) endophytic fungi fermentation broths were able to control, by more than 90%, at least one of the six plant diseases tested. Among 187 liquid broths, the F0010 strain isolated from Abies holophylla had the most potent disease control activity; it showed control values of more than 90% against five plant diseases, except for tomato late blight. On the other hand, fourteen(7.5%) solid culture extracts exhibited potent disease control values of more than 90% against one of six plant diseases. The screening results of this study strongly suggested that metabolites of plant endophytic fungi could be good potential sources for screening programs of bioactive natural products.

• 한국균학회소식:학술대회논문집
• /
• 2018.05a
• /
• pp.29-29
• /
• 2018

Sparassis latifolia is called "Cauliflower Mushroom" and is known as an edible mushroom that has high content of ${\beta}$-glucan. Recently, artificial cultivation of S. latifolia has been done by bottle, plastic bag and wood cultivation in Korea. However it is not widely used because there are low incubation ratio and yield. For the high efficiency of production, we aim to find the superior strains and media for better mycelial and fruit body growth. First, we analyzed the genetic relationship among 31 strains and divided five groups with three kinds of URP primers. And then ten strains were selected from five groups based on the experiment of mycelial growth. The suitability of media for mycelial growth was different according to media type. The suitable solid and liquid media for mycelial growth of S. latifolia isolates were PDA and M2, respectively. In addition, with regard to C/N ratio, the mycelial growth increased even until C/N 160. Second, we investigated the production of fruitbody of the strains by plastic bag cultivation. The substrate was mixed with larch sawdust, corn flour, and wheat flour (8:1:1, v/v). Moisture content of substrate was controlled by about 60% with 10% molasses solution. Out of 31 strains, 19 strains formed primordia. The eight strains produced more than 140g/1kg in fresh weight. Third, molasses culture media was selected for the mycelial growth. And molasses suitable sugar content and input aeration were around 8Brix% and 0.3~0.6vvm, respectively. The longer the incubation period is, the more dried weight of mycelia increased, but medium volume decreased. Therefore, the best incubation period was 9 to 11 days depending on strains. In the future, research project entitled development of culture system and new variety for stable production of S. latifolia will be considered as a new item.

• Mycobiology
• /
• v.43 no.4
• /
• pp.458-466
• /
• 2015

Oak tree death caused by symbiosis of an ambrosia beetle, Platypus koryoensis, and an ophiostomatoid filamentous fungus, Raffaelea quercus-mongolicae, has been a nationwide problem in Korea since 2004. In this study, we surveyed the yeast species associated with P. koryoensis to better understand the diversity of fungal associates of the beetle pest. In 2009, a total of 195 yeast isolates were sampled from larvae and adult beetles (female and male) of P. koryoensis in Cheonan, Goyang, and Paju; 8 species were identified by based on their morphological, biochemical and molecular analyses. Meyerozyma guilliermondii and Candida kashinagacola were found to be the two dominant species. Among the 8 species, Candida homilentoma was a newly recorded yeast species in Korea, and thus, its mycological characteristics were described. The P. koryoensis symbiont R. quercusmongolicae did not show extracelluar CM-cellulase, xylanase and avicelase activity that are responsible for degradation of wood structure; however, C. kashinagacola and M. guilliermondii did show the three extracellular enzymatic activities. Extracelluar CM-cellulase activity was also found in Ambrosiozyma sp., C. homilentoma, C. kashinagacola, and Candida sp. Extracelluar pectinase activity was detected in Ambrosiozyma sp., C. homilentoma, Candida sp., and M. guilliermondii. All the 8 yeast species displayed compatible relationships with R. quercus-mongolicae when they were co-cultivated on yeast extract-malt extract plates. Overall, our results demonstrated that P. koryoensis carries the yeast species as a symbiotic fungal associate. This is first report of yeast diversity associated with P. koryoensis.

• Mycobiology
• /
• v.41 no.2
• /
• pp.86-93
• /
• 2013

Sequence characterized amplified region (SCAR) markers are one of the most effective and accurate tools for microbial identification. In this study, we applied SCAR markers for the rapid and accurate detection of Phytophthora katsurae, the casual agent of chestnut ink disease in Korea. In this study, we developed seven SCAR markers specific to P. katsurae using random amplified polymorphic DNA (RAPD), and assessed the potential of the SCAR markers to serve as tools for identifying P. katsurae. Seven primer pairs (SOPC 1F/SOPC 1R, SOPC 1-1F/SOPC 1-1R, SOPC 3F/SOPC 3R, SOPC 4F/SOPC 4R, SOPC 4F/SOPC 4-1R, SOPD 9F/SOPD 9R, and SOPD 10F/SOPD 10R) from a sequence derived from RAPD fragments were designed for the analysis of the SCAR markers. To evaluate the specificity and sensitivity of the SCAR markers, the genomic DNA of P. katsurae was serially diluted 10-fold to final concentrations from 1 mg/mL to 1 pg/mL. The limit of detection using the SCAR markers ranged from $100{\mu}g/mL$ to 100 ng/mL. To identify the limit for detecting P. katsurae zoospores, each suspension of zoospores was serially diluted 10-fold to final concentrations from $10{\times}10^5$ to $10{\times}10^1$ zoospores/mL, and then extracted. The limit of detection by SCAR markers was approximately $10{\times}10^1$ zoospores/mL. PCR detection with SCAR markers was specific for P. katsurae, and did not produce any P. katsurae-specific PCR amplicons from 16 other Phytophthora species used as controls. This study shows that SCAR markers are a useful tool for the rapid and effective detection of P. katsurae.