• The Korean Journal of Mycology
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• v.44 no.3
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• pp.155-165
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• 2016

Domestic and international production of Tricholoma matsutake has decreased owing to matsutake forests being left alone, host plant disease, forest fires, climate change, and so on. In order to identify strains that are suitable for the production of T. matsutake-inoculated seedlings, Pinus densiflora seedlings were inoculated with T. matsutake after in vitro rooting and mycorrhization was examined in the roots of T. matsutake-inoculated seedlings after 6 months. The mycorrhization rate was greater than 80% for 5 strains (NIFoS 421, 434, 1681, 1984, and 2001) out of 19 total strains. Seven strains (NIFoS 434, 441, 561, 562, 1016, 1807, and 1812) showed shoot/root ratios of less than 3.0 and had a seedling shoot biomass of 2.0 to 4.8 times higher than that of the root. Eight strains (NIFoS 441, 561, 562, 1016, 1807, 1812, 1984, and 2001) stimulated increases in shoot volume and three stains (NIFoS 441, 562, and 1812) promoted the growth of root biomass by mycorrhizal formation. In conclusion, 4 strains (NIFoS 434, 561, 1984, and 2001) out of 19 total strains tested showed higher mycorrhization rates and seedling growth than those of the other strains. We expect that the use of these four strains may contribute to T. matsutake-inoculated seedling production.

• Journal of Mushroom
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• v.22 no.1
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• pp.22-26
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• 2024

Tricholoma matsutake is a traditional favorite food in East Asia, cultivated in fairy rings called "shiro," which are found near Pinus densiflora. For effective artificial cultivation of Tri. matsutake, microorganisms from symbiotic fairy rings are co-cultivated. In this study, one bacterial isolate (Y22_B35) and two fungal isolates (Y22_F64 and Y22_F68) displayed growth-promoting effects on Tri. matsutake mycelium (158.47, 125.00, and 122.26% enhanced growth, respectively). For identification, 16S rRNA or ITS regions from the microorganisms¡¯ genomes were sequenced. Other sequences, including BenA, CaM, and RPB2 were sequenced in the fungal isolates. The bacterial isolate Y22_B35 was identified as Bacillus cereus. Y22_F64 and Y22_F68 were identified as Umbelopsis nana and Aspergillus parvulus, respectively. To identify the effects of the dominant microorganisms on Tri. Matsutake cultivation, metagenomic analyses were performed. Discovery of these Tri. matsutake mycelium growth-promoting microorganisms and metagenomics analyses are expected to contribute to our understanding of Tri. matsutake fruiting body growth and construction of biomimicry.

• The Korean Journal of Mycology
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• v.27 no.1 s.88
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• pp.20-26
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• 1999

The specific DNA band appeared in PCR-RAPD analysis using OPO-2 primer was a very important for the researching Korean pine-mushrooms, Tricholoma matsutake. This DNA band, sequenced to be the 770 base pairs, existed as only a single copy in the whole genomic DNA's of Korean pine-mushrooms. However, this band was not presenting from the PCR-RAPD bands of other ectomycorrhyzal fungi reacted with the OPO-2 primer or the dot blots. Also, this DNA sequence was not matched with those of the other genes known by NCBI and had low homology together with sequence of other proteins compared. Those results suggested that the specific DNA band can be used as probe for identification of T. matsutake and might be related to the informations rather than the gene for the proteins with analysis of protein sequence translated from the DNA sequence.

• Journal of Microbiology and Biotechnology
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• v.18 no.1
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• pp.95-103
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• 2008

Mushrooms are regarded as one of the well-known foods and biopharmaceutical materials with a great deal of interest. Polysaccharide ${\beta}$-glucan is the major component of mushrooms that displays various biological activities such as antidiabetic, anticancer, and antihyperlipidemic effects. In this study, we compared the immunostimulatory potency of polysaccharide fractions, prepared from liquid culture of pine-mushroom Tricholoma matsutake, with a potent immunogen lipopolysaccharide (LPS), and their molecular mechanisms on the functional activation of macrophages. We found that fraction II (TMF-II) was able to comparably upregulate or highly enhance the phenotypic functions of macrophages such NO production and cytokine (IL-$1{\beta}$, IL-6, IL-12, and TNF-${\alpha}$) expression, to LPS. TMF-II triggered the phosphorylation of $I{\kappa}B{\alpha}$, a critical step for NF-${\kappa}B$ activation and translocation. Of the upstream signaling enzymes tested, Src and Akt were thought to be the responsible upstream signaling components in induction of NO production, although TMF-II strongly upregulated the phosphorylation of all MAPK pathways. Therefore, our data suggest that T. matsutake-derived ${\beta}$-glucan may exert its immunostimulating activities with similar potency to LPS via activation of multiple signaling pathways linked to NF-${\kappa}B$ activation.

• Journal of the Korean Wood Science and Technology
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• v.34 no.6
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• pp.96-103
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• 2006

A simple method for identifying of ectomycorrhizal fungi was presented, using the polymerase chain reaction (PCR) to amplify the ITS (Internal transcribed spacer) regions of the nuclear ribosormal repeat. The sequences analyzed 6 species, Pisolithus tinctorius, Chroogomphus rutilus, Leucogyrophana pinastri, Suillus granulatus, Lactarius laeticolorus, and Suillus bovinus at hongreung forest, and analysed 10 species, Craterellus lutescens, Thelephoroid mycorrhizal, Lactarius quieticolor, Tricholoma matsutake, Lactarius chrysorrheus, Sarcodon aspratus, Russula versicolor, Suillus luteus, Tricholoma terreum, and Amanita vaginata at hongcheon forest. Finally, the amplification by PCR with ITS1-ITS4 primers offers good results over classical identification for ectomycorrhizal fungi species.

• The Korean Journal of Mycology
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• v.28 no.1
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• pp.32-37
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• 2000

The objectives of this study was to identify genetic variations of Tricholoma matsutake (S. Ito et Imai) Sing. growing in different geographic ranges in South Korea. Mushrooms were collected during fruiting seasons from 1994 to 1997 from 11 major sites which included four sites (Bonghwa, UIjin, Goryoung, and Chungdo) in Kyongbuk Province, three sites (Changnyung, Hadong, and Hamyang) in Kyongnam Province, two sites (Yangyang and Inje) in Kangwon Province, one site (Goisan) in Choongbuk Province, and one site (Namwon) in Chonbuk Province. Two mushrooms each from three to eight shiros in each sites were collected. Genetic characteristics were analyzed by Inter-Simple Sequence Repeat Polymerase Chain Reaction (I-SSR PCR) method using six primers. With a total of 131 DNA bands identified, Nei's genetic distance and UPGMA tree were constructed. It was estimated that genetic variations between sites amounted to 12.9%, while 87.1% of total variation was explained by variations among individuals within sites. The cluster analysis indicated that the eleven major sites were clustered into four groups, group I (Yangyang, Hamyang, Inje, Hadong and UIjin), group II (Changnyung, Namwon and Chungdo), group III (Goryoung), and group IV (Bonghwa and Goisan). It is concluded that matsutake mushrooms in South Korea have a considerable degree of genetic variations between major sites.

• The Korean Journal of Mycology
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• v.33 no.1
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• pp.22-29
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• 2005

The main objectives of this research were to study the cultural and nutritional characteristics of Tricholoma matsutake and to establish its liquid culture system. The optimum growth of T. matsutake was observed in HA and TMM agar media. Similarly highest growth was observed in PDB and TMM liquid media. The optimal temperature for the mycelial growth was $25^{\circ}C$. The most suitable carbon source was dextrin among 12 different carbon sources tested. Yeast extract and peptone were best nitrogen sources among 17 different sources tested. The optimum mineral salts were $Fe_{2}(SO_{4})_{3}{\cdot}H_{2}O$ and KCl among 9 different sources tested. Shaking culture gave higher mycelial growth compared to stationary culture. Similarly, optimum medium amount for shaking culture was 100 ml per 250 ml flask. The highest mycelial growth was obtained when $5{\sim}7$ mycelial discs were inoculated in 100 ml of medium and incubated for $8{\sim}9$ weeks, respectively. The highest proportion of mycelial growth was observed at 40 : 1 ratio of medium to inoculum volume in 8 l air-lift fermenter.

• 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.

• Journal of Mushroom
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• v.22 no.1
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• pp.27-30
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• 2024

As a member of ectomycorrhizal fungi, Tricholoma matsutake has a symbiotic relationship with its host, Pinus densiflora. To cultivate T. matsutake artificially, the co-cultivation of T. matsutake mycelia and bacteria from shiro was introduced. In this study, bacteria were isolated from soil samples in Bonghwa-gun, and seven bacterial isolates (B22_7_B05, B22_7_B06, B22_7_B07, B22_7_B08, B22_7_B10, B22_7_B13, and B22_7_B14) promoted the growth of T. matsutake mycelia (147.48, 232.11, 266.72, 211.43, 175.17, 154.62, and 177.92%, respectively). Sequencing of the 16S rRNA region of the isolated bacteria was performed. B22_7_B05 and B22_7_B10 were identified as Bacillus toyonensis, B22_7_B06 and B22_7_B08 as Paenibacillus taichungensis, B22_7_B07 and B22_7_B14 as P. gorilla, and B22_7_B13 as P. odorifer. These bacterial isolates were associated with the shiro community and are expected to contribute to the cultivation of T. matsutake.

• Mycobiology
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• v.29 no.4
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• pp.179-182
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• 2001

Chitin synthases(UDP-N-acetyl-D-glucosamine: chitin 4-$\beta$-N-acetyl-D-glucosaminyl transferase, EC 2.4.1.16) catalyze the synthesis of chitin from UDP-N-acetyl-D-glucosamine. Two zymogenic type of chitin synthase gene(TmCHS1 and TmCHS2) were amplified and its nucleotide sequences were determined. By the amino acid comparison and UPGMA tree grouping, TmChs1 and TmChs2 were classified as class II and class IV chitin synthases respectively. The class II type TmChs1 was grouped with others of Agaricales ectomycorrhizal mushroom. Additionally the phylogenetic tree was well adapted to Hymenomycete previously classified by morphological and physiological characteristics.