• Journal of Mushroom
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• v.15 no.4
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• pp.164-167
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• 2017

Biological efficiency (BE), the ratio of fresh mushrooms harvested per dry substrate weight, expressed as the percentage of Lentinula edodes, also known as shiitake, was determined using the 'Sanjo 701' strain stored in the Department of Mushroom at the Korea National College of Agriculture and Fisheries. The mycelia were grown in glass columns with varying levels of moisture content and varying mixing periods of 0.5, 1, 2, and 3 hours. The substrate was sterilized using a steam pressure autoclave sterilizer at normal and high pressure to avoid contamination. The results showed that mycelial growth (126 mm/15 days) was optimized at 55% moisture content. The best mycelial growth of 117 mm/15 days was obtained with 2 hours of mixing time. Normal pressure sterilization yielded better results with mycelial growth of 96 mm/15 days at $100^{\circ}C$ compared to 88 mm /15 days with sterilization at $121^{\circ}C$. Mycelial density was higher, i.e. 3(+++), with normal pressure sterilization compared to 2(++) with high pressure sterilization. Furthermore, sawdust mixed with 5% woodchips increased the substrate porosity and yielded higher mycelial growth. Thus, we demonstrated that the optimum harvest or potential increased yield of shiitake can be obtained by modulating moisture content, mixing time, and substrate porosity.

• Mycobiology
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• v.45 no.2
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• pp.105-109
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• 2017

Sixteen genomic DNA simple sequence repeat (SSR) markers of Lentinula edodes were developed from 205 SSR motifs present in 46.1-Mb long L. edodes genome sequences. The number of alleles ranged from 3-14 and the major allele frequency was distributed from 0.17-0.96. The values of observed and expected heterozygosity ranged from 0.00-0.76 and 0.07-0.90, respectively. The polymorphic information content value ranged from 0.07-0.89. A dendrogram, based on 16 SSR markers clustered by the paired hierarchical clustering' method, showed that 33 shiitake cultivars could be divided into three major groups and successfully identified. These SSR markers will contribute to the efficient breeding of this species by providing diversity in shiitake varieties. Furthermore, the genomic information covered by the markers can provide a valuable resource for genetic linkage map construction, molecular mapping, and marker-assisted selection in the shiitake mushroom.

• BMB Reports
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• v.35 no.5
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• pp.465-471
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• 2002

From fruiting bodies of L. edodes strain L-54, single-spore isolates (SSIs) were collected. Two parental types of L-54 were regenerated via monokaryotization. By means of random-amplified polymorphic DNA (RAPD), DNA samples from L-54, its two parental types, and 32 SSIs were amplified with arbitrary primers. Dedikaryotization was demonstrated, and 91 RAPD-based molecular markers were generated. RAPD markers that were segregated at a 1:1 ratio were used to construct a linkage map of L. edodes. This RAPD-linkage map greatly enhanced the mapping of other inheritable and stable markers [such as those that are linked to a phenotype (the mating type), a known gene (priA) and a sequenced DNA fragment (MAT)] with the aid of mating tests, bulked-segregant analysis, and PCR-single-strand conformational polymorphism. These markers comprised a genetic map of L. edodes with 14 linkage groups and a total length of 622.4 cM.

• Journal of Mushroom
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• v.7 no.3
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• pp.110-114
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• 2009

Sawdust bag cultivation of Shiitake mushroom (Lentinula edodes) is getting increase. The mycelia browning on the substrate surface is important for the stable production. The development of methods for the rapid mycelia browning is quite required. In this study changes in intracellular enzyme during the mycelial browning were investigated to find the rapid mycelia browning. Mycelia of L. edodes was changed into brown color while it grew in agar and liquid media like sawdust substrates. Mycelia of L. edodes was started to change color at 25 days after inoculation and browning was occurred in whole mycelia colony at 30 days and browning was completed at 40 days. The activities of enzymes was evaluated in these periodically color changing mycelia. Laccase activity was highest at 15 days after inoculation on PDB, but it gradually decreased from 15 days. Tyrosinase activity drastically increased in period between 30 days and 40 days while mycelia browning was progressed. The kinds of phosphotase identified by electrophoresis were esterase, acid phosphotase, and alkaline phosphotase. Activities of phosphotase were increased before the initiation of mycelial browning but they were decreased after browning.

• The Korean Journal of Mycology
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• v.41 no.1
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• pp.28-32
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• 2013

New Shiitake (Lentinula edodes) strain "Chunbaegko" was bred by Di-mon method. Bed-log cultivation of "Chunbaegko" was performed. Fruit-body production of "Chunbaegko" was most at spring and autumn. Optimal temperature of fruit-body formation was $14{\sim}22^{\circ}C$ Baegwhako, the best quality fruit-body, is produced during spring. The diameter of pileus is ca. 52 mm. The total amount of fruit-body production during 4 years(one generation) was 140 $kg/m^3$ log.

• The Plant Pathology Journal
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• v.28 no.4
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• pp.341-356
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• 2012

Shiitake (Lentinula edodes) is the most economically important cultivated mushroom, but yields are impacted by its competitor, Trichoderma spp. We previously found two unidentified Trichoderma species growing in bedlogs and sawdust shiitake media in Korea. Here, we identify and re-describe those two species based on molecular sequence data, morphology, and culture characteristics. Well-supported clades based on phylogenetic analyses of internal transcribed spacer, translation elongation factor 1-${\alpha}$, and RNA polymerase subunit II sequences grouped one of the unidentified Trichoderma spp. with Hypocrea pseudogelatinosa and the other with Hypocrea pseudostraminea, and their morphologies matched well with the original descriptions of the two Hypocrea species. This study reports the first phylogenetic analyses of H. pseudogelatinosa and Japanese strains of H. pseudostraminea. Based on the phylogenetic results, we redescribed these two species using modern taxonomic concepts in Hypocrea/Trichoderma.

• Mycobiology
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• v.49 no.6
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• pp.599-603
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• 2021

CRISPR/Cas9 genome editing systems have been established in a broad range of eukaryotic species. Herein, we report the first method for genetic engineering in pyogo (shiitake) mushrooms (Lentinula edodes) using CRISPR/Cas9. For in vivo expression of guide RNAs (gRNAs) targeting the mating-type gene HD1 (LeA1), we identified an endogenous LeU6 promoter in the L. edodes genome. We constructed a plasmid containing the LeU6 and glyceraldehyde-3-phosphate dehydrogenase (LeGPD) promoters to express the Cas9 protein. Among the eight gRNAs we tested, three successfully disrupted the LeA1 locus. Although the CRISPR-Cas9-induced alleles did not affect mating with compatible monokaryotic strains, disruption of the transcription levels of the downstream genes of LeHD1 and LeHD2 was detected. Based on this result, we present the first report of a simple and powerful genetic manipulation tool using the CRISPR/Cas9 toolbox for the scientifically and industrially important edible mushroom, L. edodes.

• Journal of Mushroom
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• v.19 no.3
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• pp.184-190
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• 2021

With the development of sawdust medium cultivation technology, Lentinula edodes (shiitake mushroom) has become the most extensively produced and consumed mushroom globally. In this study, the approximate composition, nutritional components, and bioactive compounds of L. edodes were analyzed and compared for the five sawdust-cultivated shiitake mushrooms cultivars namely Sanjo701ho, Sanjo707ho, Sanjo715ho, Chamaram, and L808. The approximate range of the composition of freeze-dried shiitake mushrooms was 4.06-5.92 g/100 g of ash, 0.75-1.02 g/100 g of crude fat, and 21.24-29.15 g/100 g of crude protein. Sanjo701ho had the highest trehalose content (9.60±0.08 g/100 g), total polyphenol content (3.49±0.04 mg GAE/g), and total flavonoid content (1.33±0.03 mg QE/g) among the other shiitake mushroom cultivars. The total amino acid content was as follows: glutamic acid>aspartic acid>leucine. Glycine, alanine, valine, isoleucine, leucine, and phenylalanine contents were significantly decreased in the following sequence: Sanjo707ho, Sanjo715ho, Sanjo701ho, Chamaram, and L808. Crude protein, trehalose, and six types of amino acids were identified as classification indicators for the five cultivars of sawdust-cultivated shiitake mushrooms.

• The Plant Pathology Journal
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• v.29 no.4
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• pp.460-464
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• 2013

dsRNA was found in malformed cultures of Lentinula edodes strain FMRI0339, one of the three most popular sawdust cultivated commercial strains of shiitake, and was also found in healthy-looking fruiting bodies and actively growing mycelia. Cloning of the partial genome of the dsRNA revealed the presence of the RdRp sequence of a novel L. edodes mycovirus (LeV), and sequence comparison of the cloned amplicon showed identical sequences sequence to known RNA-dependent RNA polymerase genes of LeV found in strain HKA. The meiotic stability of dsRNA was examined by measuring the ratio of the presence of dsRNA among sexual monokaryotic progeny. More than 40% of the monokaryotic progeny still contained the dsRNA, indicating the persistence of dsRNA during sexual reproduction. Comparing the mycelia growth of monokaryotic progeny suggested that there appeared to be a tendency toward a lower frequency of virus incidence in actively growing progeny.

• The Korean Journal of Mycology
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• v.30 no.1
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• pp.31-36
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• 2002

Since lectin condense more easily cancer cell than normal cell, it has been investigated very actively. Recently, a lot of researchers gave attention to lectin in natural products especially because lectin has effects on T-cell activation and anticancer activity, and specificity on polysaccharide. The specificity is useful to confirm kind of polysaccharide of the cell surface and to study the polysaccharide. In this research, we purified lectin from shiitake, Lentinula edodes, and then characterized it. The molecular weight of the lectin was 23 kDa, and it was stable only under the $40^{\circ}C$ and in a alkaline solution. As for the specificity of polysaccharide, the lectin had specificity on galactose, fucose, glucose, lactose and N-acetyl-D-galactosamine. In addition, it was confirmed to be a glycoprotein.