• Mycobiology
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• v.49 no.2
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• pp.173-182
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• 2021

Recently, Cryptococcus pseudolongus has been reported as a new pathogen of shiitake (Lentinula edodes). However, its pathological properties are not much known. To further understand its impact on the mushroom, we investigated the pathogen's interactions with the mycelium of shiitake, histopathological properties, host range, and sensitivity to diverse antifungal agents. The strain C. pseudolongus DUCC 4014 inhibited the mycelial growth of L. edodes strain (cultivar Sanjo 701ho) and caused browning in the mycelia confronted with the yeast on PDA. Spray inoculation of the yeast caused an abnormal browning symptom on the cap and/or gills of three shiitake cultivars grown on sawdust media in vinyl bags. Scanning electron microscopic images of the abnormally browned parts of shiitake fruit body illustrated that mushroom tissues were loosed and dispersed in the middle and edge of the cap and the arrangement of basidiospores borne on basidia in the gills was disturbed compared to those of normal shiitake fruit body. Spray inoculation also led to developing abnormal browning on the harvested fruit body, indicating C. pseudolongus could be a problem during mushroom storage. But the yeast was not able to induce abnormal browning on mushrooms of Pleurotus ferulae, Pleurotus fostreatus, and Agaricus bisporus. But it induced browning only on button mushroom (A. bisporus) when they were inoculated after wounding. Tests with 16 kinds of fungicides revealed that the cell growth of C. pseudolongus could be inhibited by benzalkonium chloride at MIC 7 ㎍/ml and benomyl at MIC 3 ㎍/ml.

• Journal of Mushroom
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• v.18 no.3
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• pp.189-200
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• 2020

Pleurotus djamor var. roseus is an edible mushroom isolated from the wild and cultivated on paddy straw substrates. The present study was carried out to compare the nutritional composition and antioxidant properties of P. djamor var. roseus at different growth stages (primordia, basidiomata, and mycelia). The protein content was is in the range of 31.48 to 35.50 g/100g dw. The crude fiber content ranged from 8.0 to 14.60 g, and that of total carbohydrates ranged from 44.75 to 48.90 g. Sodium, magnesium, and calcium reached the maximum levels in basidiomata, and selenium was detected in basidiomata and mycelia (0.47 - 0.22 mg/Kg). The amino acid profile showed that all essential and nonessential amino acids and glycine showed maximum levels in basidiomata and 15.98 ± 0.01 g/100g. The fatty acid profile showed the presence of saturated and unsaturated fatty acids; the unsaturated fatty acid content was maximum in all of the samples, ranging from 76 - 40.41%. The total phenol and flavonoid contents as well as the scavenging (DPPH), ferric thiocyanate (FTC), and thiobarbituric acid (TBA) activities in the context of methanol and water extracts from primordia, basidiomata, and mycelium were determined. Among them, basidiomata and mycelial methanol extracts exhibited significant antioxidant activity. Overall, these findings show that P. djamor var. roseus can be used as a functional food for daily consumption.

• Journal of Mushroom
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• v.13 no.3
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• pp.270-273
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• 2015

Hypsizigus marmoreus is commercially the most important edible mushroom in Japan. This mushroom is usually cultivated for a longer period (about 85~120 days) than other mushroom. In order to develop a new cultivar that has a shortened cultivation period, the genome analysis of this strain has been considered. This study aims to obtain parental monokaryotic strains reproducing 'Haemi' cultivar in Hypsizigus marmoreus for reference genome sequencing. The mycelia were cultured in MCM and MYG media for various incubation periods. Homogenized mycelia were treated with commercial cell wall degrading enzymes to maximize protoplasts production yield from Hypsizigus marmoreus. The greatest number of protoplasts was obtained from mycelia cultured in MCM media for 3 days using Novozyme enzyme. The isolated protoplasts were grown in regeneration agar media after two weeks. Regenerated colonies were picked and moved on separated dishes for microscopic observation. Neohaplonts regenerated from dikayotic strains were identified by the absence of clamp connections. We confirmed that one of monokaryotic strains is a parental strain by crossing with an original compatible strain of 'Haemi' cultivar. This parental strain will be used for reference genome sequence analysis.

• Journal of Mushroom
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• v.17 no.4
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• pp.230-234
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• 2019

This study was conducted to prepare healthy vinegars using the mycelia of various mushrooms (Cordyceps militaris, Phellinus baumi, Pleurotus cornucopiae, and Ganoderma lucidum). Analyses of the bioactive components of these vinegars revealed that those prepared using mushroom mycelial cultures on fermented brown rice had increased bioactive component levels compared to those cultured on unfermented brown rice. The highest beta-glucan contents (78.7 mg/g) were found in P. baumi cultures and 0.34mg/g of cordycepin was detected in C. militaris cultures. Regarding the chemical properties of these mushroom mycelia vinegars (MMVs), those prepared using C. militaris and P. cornucopiae cultures showed higher acidity and reduced sugar content. Sensory assessments of C. militaris and P. cornucopiae MMVs yielded scores of 7.3 and 6.6, respectively. Analyses of bioactive components revealed that C. militaris MMV contained high levels of polyphenols and 786 mg/L of cordycepin. P. cornucopiae MMV contained the highest amount of beta-glucan (20.9 mg/g).

• Journal of Mushroom
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• v.10 no.3
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• pp.120-128
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• 2012

Recently sawdust cultivation of Shiitake mushroom (Lentinula edodes) is increasing. It is important to make mycelia to be brown on the substrate surface. This browned surface in sawdust cultivation plays an important role like as artificial bark of the oak log, which protects the other pests and suppresses water evaporation in the substrate. In order to isolate genes which related to brown color formation, differential display method was used. Two cDNA fragments obtained by DD-PCR were 1.2 and 1.6kb and these were expressed in white colored mycelia from L. edodes, but not brown colored mycelia. Partial sequencing of these cDNA fragments showed that the 1.6kb cDNA had 100% identity with the microsatellites gene from Dugenia polichroa. However, the other 1.2kb cDNA fragment had poly T tail on 3' region of partial open reading frame on 5' region. The new primer designed based on the sequence of 1.2kb cDNA was constructed. RT-PCR analysis using the newly designed 0.12kb cDNA specific primer showed that the gene was only expressed in white color mycelia, but not in brown color mycelia. Sequence analysis of 5' region of this 1.2kb cDNA revealed that this gene contained partial open reading frame consisted of 110 amino acid. Homology search using DNASIS database showed that this gene had high sequence homology of 66.7% in DNA level and 69.2 % in amino acid level with dTDP-glucose 4,6-dehydratases gene from Arabidopsis thaliata. The dTDP-glucose 4,6-dehydratases gene was known to be function to have tolerance with oxidation stress. These results strongly suggest that this gene isolated from white mycelia of L. edodes might have a function of repressor against mycelia browning. Therefore I designated this gene as BCR (Brown Color Repressor) gene.

• Journal of Mushroom
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• v.15 no.1
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• pp.21-24
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• 2017

In this study, the effect of different substrates of agricultural by-products on the mycelial growth rate and density of Trametes versicolor (Turkeytail mushroom) was analyzed. We found that pepper stem and rice bran with a mixing ratio of 9:1(v/v) produced the best mycelial growth of 101 mm in 10 days, while a mixing ratio of 8:2 resulted in mycelial growth of 83 mm in 10 days. The control group treated with a 9:1 mixing ratio of oak sawdust and rice bran (v/v) produced mycelial growth of 74 mm in 10 days. The following results are in the order of beanstalk, sesame stem, and perilla stem. After the harvest of the mushrooms, the mycelial growth rate and the density of T. versicolor in each substrate were as follows the group with waste substrate of Pleurotus eryngii and rice bran with a mixing ratio of 9:1(v/v) produced the best result of 76 mm in days, while a mixing ratio of 8:2 produced of 61 mm in 10 days. The control group with a 9:1 ratio of oak sawdust and rice bran produced mycelia of 74 mm in 10 days, while a mixing ratio of 8:2 resulted in mycelia of 59 mm in10 days.

• Mycobiology
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• v.39 no.4
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• pp.272-277
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• 2011

Chemical mutagenesis of basidiospores of Hypsizygus marmoreus generated new mushroom strains. The basidospores were treated with methanesulfonate methylester, an alkylating agent, to yield 400 mutant monokaryotic mycelia. Twenty fast-growing mycelia were selected and mated each other by hyphal fusion. Fifty out of the 190 matings were successful (mating rate of 26.3%), judged by the formation of clamp connections. The mutant dikaryons were cultivated to investigate their morphological and cultivation characteristics. Mutant strains No. 3 and No. 5 showed 10% and 6% increase in fruiting body production, respectively. Eight mutant strains showed delayed and reduced primordia formation, resulting in the reduced production yield with prolonged cultivation period. The number of the fruiting bodies of mutant No. 31, which displayed reduced primordial formation, was only 15, compared to the parental number of 65. Another interesting phenotype was a fruiting body with a flattened stipe and pileus. Dikaryons generated by mating with the mutant spore No. 14 produced flat fruiting bodies. Further molecular biological studies will provide details of the mechanism. This work shows that the chemical mutagenesis approach is highly utilizable in the development of mushroom strains as well as in the generation of resources for molecular genetic studies.

• Journal of Mushroom
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• v.18 no.1
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• pp.100-106
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• 2020

Eight morel mushroom species were collected from Korea and other countries. The culture characteristics, genetic relationships, and beta-glucan content of the strains were analyzed. The mycelia of Morchella species exhibited optimal growth when cultured in dark at 25 ℃ in media with pH 7. The mycelia had a distinctive mycelial scent and characteristically changed color, being white initially, and then turning dark yellow to dark brown as it grew. The mycelia were classified into five types based on morphology. The isolates were identified as Morchella conica, two M. sextelata, M. importuna, M. esculenta, and three M. crassipes, based on ITS-rDNA sequences. PCR polymorphisms were variably produced within Morchella spp. using Universal Fungal Fingerprinting Primers (UFPF) and classified into four groups at the intra and inter species level. The strains, KMCC04971 and KMCC04407, showed the same banding pattern as M. conica and M. sextelata, respectively; however, these results were different from those of ITS analysis. Glucan content analysis by strain showed that the KMCC 04973 strain of M. importuna had the highest alpha- and beta-glucan content, at 16.4 g and 33.1 g per 100 g, respectively.

• Journal of Mushroom
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• v.16 no.4
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• pp.347-351
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• 2018

The aim of this study is to investigate the effect of Eleutherococcus senticosus (ES) on the laccase and cellulase activity of Ganoderma lucidum mycelia. Following the addition of ES, the laccase activity of Ganoderma mushroom mycelia was found to be 0.84-2.18 times, 0.61-2.37 times, and 0.78-2.17 times the activity of mycelia treated with sawdust in Yeongji-1 (Y1; ASI-7004), Yeongji-2 (Y2; ASI-7071), and nokgak (GN; ASI-7013), respectively, with the laccase activity of Y2 being the highest at 0.947 U/min. Using the Congo-red assay, a wider clear zone was formed because of the CMCase activity of mycelia treated with ES than that of mycelia treated with sawdust. Cellulase activity was found to be 1.84-2.24 times, 1.77-1.87 times, and 2.74-2.81 times that of mycelia treated with sawdust in Y1, Y2, and GN, respectively, with the cellulase activity of GN being the highest at 0.172 U/min. However, the addition of ES did not affect the growth of G. lucidum mycelia.

• Journal of Mushroom
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• v.21 no.3
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• pp.190-193
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• 2023

To cultivate pine mushroom (Tricholoma matsutake) artificially, co-cultivation with microorganisms has been introduced. Here, experiments were performed to assess the growth-promoting effect of bacteria on T. matsutake mycelia. Bacteria were isolated from soil samples collected in Yangyang County, Korea. Four of the bacterial isolates (Y22_B06, Y22_B11, Y22_B18, and Y22_B22) exhibited a growth-promoting effect on T. matsutake mycelia (154.67%, 125.91%, 134.06%, and 158.28%, respectively). To analyze the characteristics of the bacteria, especially the antifungal activity, 𝛼-amylase and cellulase activity assays were performed. In comparison with the controls, the isolated bacteria exhibited low 𝛼-amylase and cellulase activity. 16S rRNA gene sequencing was performed to identify the four bacterial isolates. The isolates belonged to the Terrabacteria group and were identified as Microbacterium paraoxydans, Paenibacillus castaneae, Peribacillus frigoritolerans, and P. butanolivorans. These bacterial isolates are expected to have contributed to the growth promotion of T. matsutake mycelia and the artificial cultivation of T. matsutake.