• Mycobiology
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• v.47 no.4
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• pp.466-472
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• 2019

For the purpose of protecting the rights of Lentinula edodes breeders, we developed a new simple sequence repeat (SSR) marker set consisting only of genetically independent tetranucleotide or longer core motifs. Using available genome sequences for five L. edodes strains, we designed primers for 13 SSR markers that amplified polymorphic sequences in 20 L. edodes cultivars. We evaluated the independence of every possible marker pair based on genotype data. Consequently, eight genetically independent markers were selected. The polymorphic information content values of the markers ranged from 0.269 to 0.764, with an average of 0.409. The markers could distinguish among 20 L. edodes cultivars and produced highly repeatable and reproducible results. The markers developed in this study will enable the precise identification of L. edodes cultivars, and may be useful for protecting breeders' rights.

• Mycobiology
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• v.48 no.2
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• pp.115-121
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• 2020

In this study, the genetic diversity and the population structure of 77 wild strains and 23 cultivars of Lentinula edodes from Korea were analyzed using 20 genomic SSRs, and their genetic relationship was investigated. The tested strains of L. edodes were divided into three sub-groups consisting of only wild strains, mainly wild strains and several cultivars, and mainly cultivars and several wild strains by distance-based analysis. Using model-based analysis, L. edodes strains were divided into two subpopulations; the first one consisting of only wild strains and the second one with mainly cultivars and several wild strains. Moreover, AMOVA analysis revealed that the genetic variation in the cultivars was higher than that in the wild strains. The expected and observed heterozygosity and values indicating the polymorphic information content of L. edodes cultivars from Korea were also higher than that of the wild strains. Based on these results, we presume that the cultivars in Korea have developed by using numerous strains from other countries. In conclusion, the usage of wild strains for the development of new cultivars could improve the adaptability of L. edodes to biotic and abiotic stress.

• The Korean Journal of Mycology
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• v.49 no.1
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• pp.33-44
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• 2021

Lentinula edodes (Berk.) Pegler, the most produced mushroom in the world, is an edible mushroom with very high nutritional and pharmacological value. Currently, interest in the protection of genetic resources is increasing worldwide, and securing the distinction between new cultivars is very important. Therefore, the development of efficient molecular markers that can discriminate between L. edodes cultivars is required. In this study, we developed cleaved amplified polymorphic sequence (CAPS) markers for the identification of L. edodes cultivars (Sanbaekhyang and Sulbaekhyang). These markers were developed from whole genome sequencing data from L. edodes monokaryon strain B17 and resequencing data from 40 cultivars. A nucleotide deletion existed in scaffold 19 POS 214449 in Sanbaekhyang (GT→G), and a single nucleotide polymorphism changed in scaffold 7 POS 215801 in Sulbaekhyang (G→A). The restriction enzymes Hha I and HpyCH4IV distinguished Sanbaekhyang and Sulbaekhyang, respectively, from other cultivars. Thus, we developed two CAPS markers for the identification of the L. edodes cultivars Sanbaekhyang and Sulbaekhyang.

• The Korean Journal of Mycology
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• v.50 no.3
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• pp.225-233
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• 2022

Pyogo (Shiitake, Lentinula edodes) is one of the most important edible mushrooms because of its outstanding nutritive and medicinal value. In the registration and protection procedure for newly developed mushroom cultivars, the application of molecular markers that can supplement the morphological characteristic-based distinction has been strongly requested. Sanjo 701 and Chamaram, newly developed at the Federation Forest Mushroom Research Center of Korea, have been characterized as innovative cultivars suitable for customer demands because of their high yields and cultivation rates. However, no technical tools can protect the rights to these important cultivars. In this study, using comparative genomic information from 23 commercially available pyogo cultivars, we identified single nucleotide polymorphisms (SNPs) that accurately differentiated Sanjo701 and Chamaram from the other cultivars. We also developed high-resolution melting analysis (HRM)-based SNP markers that discriminate among the tested 23 pyogo cultivars. The developed SNP markers can be utilized for rapid, accurate identification of pyogo cultivars with low genetic diversity and to prevent cultivar contamination caused by illegally distributed inocula. In addition, these markers can serve as a crucial scientific basis for securing the right to conserve new cultivars in international markets.

• Mycobiology
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• v.42 no.3
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• pp.301-304
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• 2014

Shiitake mushrooms (Lentinula edodes) containing ${\beta}$-glucans may be beneficial for human health; they have been used in the treatment of cancer, hypertension, and high cholesterol levels. The objective of this study was to determine the ${\beta}$-glucan content in different sections of the fruiting bodies and mycelia of ten shiitake mushroom cultivars. The measured ${\beta}$-glucan content ranged from $20.06{\pm}1.76%$ to $44.21{\pm}0.13%$ in the pileus sections, and from $29.74{\pm}1.40%$ to $56.47{\pm}4.72%$ in the stipe sections. The results of this study indicate that the variance in ${\beta}$-glucan content dependent on the shiitake cultivar, and that the ${\beta}$-glucan content is higher in the stipe than in the pileus.

• Journal of Mushroom
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• v.16 no.1
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• pp.51-56
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• 2018

In Korea, the oak mushroom (Lentinula edodes) is highly preferred by consumers in the food industry and makes up about 97.7% of the total forest mushroom production. This indicates that the oak mushroom is an important non-timber forest product in Korea. Recently, the breeding and development of new cultivars of L. edodes have been actively initiated, and the development of molecular markers that are able to identify and discriminate the new cultivars is crucial for protecting the breeder's rights. This study was carried out to develop a cleaved amplified polymorphic sequence (CAPS) marker for the identification and discrimination of a new cultivar, Sanmaru 2ho from the 37 other oak mushroom cultivars. A single nucleotide polymorphism (SNP) was identified at the $1,803,483^{rd}$ position of scaffold2 in the genome of Sanmaru 2ho. The amplified DNA containing the SNP of Sanmaru 2ho was uniquely not cleaved by the restriction enzyme, Hha I, and thus Sanmaru 2ho was successfully distinguished from the other oak mushroom cultivars.

• Mycobiology
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• v.51 no.1
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• pp.26-35
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• 2023

The diversity of A mating type in wild strains of Lentinula edodes was extensively analyzed to characterize and utilize them for developing new cultivars. One hundred twenty-three A mating type alleles, including 67 newly discovered alleles, were identified from 106 wild strains collected for the past four decades in Korea. Based on previous studies and current findings, a total of 130 A mating type alleles have been found, 124 of which were discovered from wild strains, indicating the hyper-variability of A mating type alleles of L. edodes. About half of the A mating type alleles in wild strains were found in more than two strains, whereas the other half of the alleles were found in only one strain. About 90% of A mating type combinations in dikaryotic wild strains showed a single occurrence. Geographically, diverse A mating type alleles were intensively located in the central region of the Korean peninsula, whereas only allele A17 was observed throughout Korea. We also found the conservation of the TCCCAC motif in addition to the previously reported motifs, including ATTGT, ACAAT, and GCGGAG, in the intergenic regions of A mating loci. Sequence comparison among some alleles indicated that accumulated mutation and recombination would contribute to the diversification of A mating type alleles in L. edodes. Our data support the rapid evolution of A mating locus in L. edodes, and would help to understand the characteristics of A mating loci of wild strains in Korea and help to utilize them for developing new cultivars.

• The Korean Journal of Mycology
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• v.50 no.4
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• pp.301-310
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• 2022

Lentinula edodes is a type of mushroom widely consumed in East Asia, and many of the components contained in them are used as raw materials for pharmaceuticals or as health supplements. Ergothioneine, a sulfur-containing amino acid in L. edodes, is attracting attention due to its antioxidant and anti-inflammatory effects. L. edodes generally contain an ergothioneine content of about 200 to 800 mg per kg; there are some differences in this value depending on the cultivation conditions. In this study, 24 domestic sawdust-cultivated L. edodes cultivars were cultured under the same conditions, and the morphological characteristics and ergothioneine content of the fruiting bodies were investigated. The yield and morphological characteristics of the fruiting bodies were different in each cultivar. Taehyanggo and Sanjo 713ho had the largest and thickest pileus. The pileus of L. edodes had a higher ergothioneine content than the stipe. Sanjo 715ho had the highest ergothioneine content in the pileus and the stipe at 1,225 mg/kg and 753 mg/kg, respectively. An ergothioneine content of more than 1,000 mg/kg in the pileus was observed in Bambithyang, Sansanhyang, Sulbaekhyang, Taehyanggo, Sanjo 705ho, Sanjo 709ho, Sanjo 715ho, and Sanjo 716ho. These cultivars can be used as parental strains to develop new cultivars with enhanced ergothioneine functionalities.

• 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.

• 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.