• Mycobiology
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• v.43 no.1
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• pp.75-80
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• 2015

We identified single nucleotide polymorphism (SNP) markers in the laccase gene to establish a line-diagnostic system for shiitake mushrooms. A total of 89 fungal isolates representing four lines, including Korean registered, Korean wild type, Chinese, and Japanese lines, were analyzed. The results suggest that SNP markers in the laccase gene can be useful for line typing in shiitake mushrooms.

• Journal of Mushroom
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• v.18 no.1
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• pp.91-94
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• 2020

This study was carried out to develop a method for cultivation of Lentinula edodes that would reduce both labor and waste. Cultural characteristics were studied, and mushroom yields were estimated to identify the most suitable method to culture L. edodes, using bottles and boxes. Among these, box cultivation with 7 kg of substrate had 40 days of browning period and the maximum yield of 945 g fruiting body; its biological efficiency was 32.7%. In contrast, box cultivation after pre-culture, and the bottle cultivation were found to be unsuitable for the production of L. edodes, due to long periods of browning, and low yields with poor quality, respectively. Further studies on box cultivation of L. edodes are necessary for commercial application of this method.

• The Korean Journal of Mycology
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• v.45 no.2
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• pp.114-120
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• 2017

Lentinula edodes is an edible mushroom that is mainly cultivated in Asian countries. Recently, new cultivars of this mushroom have been developed in Korea; variety protection is very important, so the development of efficient molecular markers that can distinguish each variety is required. In this study, we developed cleaved amplified polymorphic sequence (CAPS) markers for the identification of L. edodes cultivars (Sanmaru 1ho and Chunjang 3ho). These markers were developed from whole genomic sequencing data from L. edodes monokaryon strain B17 and resequencing data from 10 dikaryon strains. A single nucleotide polymorphism changed in scaffold 9 POS 1630048 in Sanmaru 1ho($G{\rightarrow}T$), and in scaffold 13 POS 920681 in Chunjang 3ho ($G{\rightarrow}A$). The restriction enzymes TspR I and Xho I distinguished Sanmaru 1ho and Chunjang 3ho, respectively, from other strains. Thus, we developed 2 CAPS markers for the identification of the L. edodes cultivars Sanmaru 1ho and Chunjang 3ho.

• The Korean Journal of Mycology
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• v.48 no.2
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• pp.95-102
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• 2020

This study was carried out to develop a new method of cultivation of 'Hwadam' a new variety of Lentinula edodes, to improve fruiting body productivity and reduce labor. To determine the optimum filter number for box cultivation, L. edodes were assessed for their cultural characteristics and mushroom yield. During spawn running of L. edodes in the box, the temperature gradually increased to the highest value of 25.8℃, 23.7℃, and 23.0℃ on day 18 for the four filter, two filter, and no treatment groups, respectively. Oxygen concentration displayed an opposite trend to carbon dioxide concentration, reaching the lowest value on day 21. The oxygen concentration progressively increased as more filters were used (5.4%, 8.2%, and 8.9% treatment with no, two, and four filters, respectively. Growth of L, edodes in the box resulted in the highest yield (2,228 g/7kg substrate) and biological efficiency (70.7%) using four filters. The biological efficiency of the four filter treatment was 29.4% higher compared to bag cultivation (41.3%). Further studies are necessary to verify stable productivity of fruiting bodies through repeated cultivation.

• The Korean Journal of Mycology
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• v.49 no.4
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• pp.507-514
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• 2021

Lentinula edodes have been cultivated extensively not only in South Korea but also in other Asian countries. In terms of taste and nutrition, it is a valuable mushroom that is comparable to other mushrooms. L. edodes contains various physiologically active compounds that promote human health. One compound, ergothioneine, exerts a powerful antioxidant function that inhibits cellular senescence. As it is not biosynthesized by plants or animals, L. edodes is very important for ergothioneine supply. The L. edodes cultivars Bambithyang (NIFoS 3404) and Sansanhyang (NIFoS 3420) of the National Institute of Forest Science (NIFoS) had higher ergothioneine content than the other cultivars, and 11 strains were obtained using mono-mono hybridization between them. The strains were inoculated into sawdust media to obtain fruiting bodies, and the ergothioneine content of each hybrid strain was confirmed using high-performance liquid chromatography (HPLC) analysis. Among the 11 strains, NIFoS 5108 had a higher ergothioneine content than the parental strains, Bambithyang (NIFoS 3404) and Sansanhyang (NIFoS 3420). This study revealed that it is possible to develop cultivars with improved specific functions using mono-mono hybridization.

• The Korean Journal of Mycology
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• v.49 no.4
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• pp.487-495
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• 2021

Lentinula edodes is a tetrapolar basidiomycete and its mating type is determined by two unlinked genetic loci, A and B. Theoretically, one dikaryotic strain could produce basidiospores with four different mating types in a 1:1:1:1 ratio. Previous studies have described the skewed segregation ratio of mating types among basidiospores of L. edodes. However, they were based only on morphological characteristics, such as clamp connection, to determine mating types. To clarify whether the segregation distortion of mating types is a general phenomenon in L. edodes, we analyzed the mating types of basidiospores obtained from three cultivars of L. edodes using recently developed DNA markers. We found that the skewed segregation of mating types was strain-specific, as reported previously. Among the three cultivars, one cultivar showed balanced segregation, while the other two displayed distorted segregation. We also examined the relationship between mating type and mycelial growth rate of monokaryons derived from each basidiospore. It was found that the monokaryotic mycelial growth rate was related to the A mating type but not to the B mating type. Therefore, homeodomain transcription factor genes that reside on the A locus or other genes linked to the A locus affect the growth rate of monokaryotic mycelia. Considering the importance of mating types in mushroom breeding, this study is informative for establishing an efficient breeding strategy as well as for understanding the mechanism of monokaryotic mycelial growth.

• 한국균학회소식:학술대회논문집
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• 2018.05a
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• pp.37-37
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• 2018

Mating type of Lentinula edodes is determined by two unlinked genetic loci, A and B. To better understand mating behavior of L. edodes, we investigated variations in mating type genes in129 dikaryotic strains collected from East Asia. Through sequence analysis of A locus, we discovered that hypervariable region spanning N-term of HD2-intergenic region-N-term of HD1 could represent A mating type. Mating and hypervariable region analyses revealed 70 unique A mating types: 27 from 98 cultivated strains, 53 from 31 wild strains, and 10 commonly found. It was also revealed that only a few A mating type alleles such as A1, A4, A5, and A7 were prevalent in cultivated strains. Contrarily, A mating type in wild strains was highly diverse: 23 unique A alleles were discovered in small mountainous area in Korean peninsula, suggesting rapid evolution of A mating type in nature. The B locus was assessed by allelic variations in pheromone (PHB) and pheromone receptor (RCB) pairs which constituted subloci Ba and Bb. Sequence analyses and mating assay revealed 5 alleles of RCB1 with 9 associated PHBs in Ba sublocus and 3 alleles of RCB2 with 5 associated PHBs in Bb sublocus. Each RCB was primarily associated with two PHBs. Each PHB-RCB pair was always discovered as a distinct unit. This allowed us to propose 15 B mating types via combinations of five Ba and three Bb subloci. Further investigation on 129 strains confirmed that the B locus, unlike the A locus, was indeed restricted to 15 mating types. Thus, the total number of mating types became 1,050 in L. edodes through a combination of 70 A and 15 B. This number will further increase because of rapid diversification of A mating type. Our findings provide a comprehensive and practical knowledge on mating behaviors of L. edodes.

• The Korean Journal of Mycology
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• v.43 no.1
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• pp.26-32
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• 2015

In this study, a new cultivar of Lentinula edodes was bred from dikaryotic KFRI 490 and monokaryotic KFRI 536 by Di-mon method. The new breed was named "Poongnyunko" and physiological characteristics was investigated. Its optimal growing temperature was $25^{\circ}C$, and it had thin density. Its colony was colorless and did not form any epithelium. On the $7^{th}$ day of mycelial growth, "Poongnyunko" was 56.6 mm, and showed higher cellulase activity compared with "Sanlim 4-ho"(52.9 mm). On the $5^{th}$ day of mycelial growth, "Poongnyunko" was 58.0 mm, and showed higher laccase activity compared with "Sanlim 4-ho" (55.6 mm). Its pH at a substrate was 6.0. Before cultivation, the pH of sawdust substrate was 4.7-5.5 went down to 3.7-3.9 after 120-day cultivation. After 60 days of cultivation, the change of $CO_2$ was the greatest. During the cultivation, $CO_2$ concentration in a growing bag was 4.67% to 3.90%.

• 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.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.12
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• pp.5055-5061
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• 2014

In this study, an anti-oxidant and anti-tumor protein Latcripin-3 of Lentinula edodes C91-3 was expressed in Escherichia coli. for the first time. According to the cDNA library, the full-length gene of Latcripin-3 was cloned by the methods of 3'-full rapid amplification of cDNA Ends (RACE) and 5'-full RACE. The structural domain gene of Latcripin-3 was inserted into the pET32 a(+). The functional protein of Latcripin-3 was expressed in Rosetta-gami (DE3) E. coli, evaluated by Western blotting and mass spectrometry. DPPH testing showed that the protein Latcripin-3 can scavenge free radicals remarkably well. The activity of functional protein Latcripin-3 on A549 cells was studied with flow cytometry and the MTT method. The MTT assay results showed that there was a decreases in cell viability in a dose-dependent and time-dependent manner in protein Latcripin-3 treated groups. Flow cytometry demonstrated that Latcripin-3 can induce apoptosis and block S phase dramatically in human A549 lung cancer cells as compared to the control group. At the same time, the cell ultrastructure observed by transmission electron microscopy supported the results of flow cytometry. This research offers new insights and advantages for identifying anti-oxidant and anti-tumor proteins.