• Journal of Korean Society of Forest Science
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• v.98 no.4
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• pp.357-362
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• 2009

Trials to find a method for overcoming the depletion of Tricholoma matsutake mycorrhiza from the artificially inoculated pine seedling (Pinus densiflora) when it is transplanted in situ were carried out. The culturing jars containing mycorrhizal seedlings were punched at the bottom to make the jar like a pot with keeping the soil structure of the original medium in vitro, and 8 kinds of irrigation solution were applied. Irrigation of distilled water caused disappearance of T. matsutake mycorrhizae, but biweekly irrigation of 2.5% PDMP (24 g potato dextrose broth, 3 g malt extract, 1 g peptone) solution or 0.5% Tween 80 solution until 3 months of acclimation could keep mycorrhizal association stronger. The percentage of mycorrhizal roots out of total length of roots was increased during the acclimation, and the acclimation rate reached up to 70%. Thus, we recommend that the mycorrhizal pine seedlings inoculated with T. matsutake would be acclimated before transplanting into the field.

• The Korean Journal of Mycology
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• v.36 no.2
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• pp.148-152
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• 2008

We inoculated hypal suspension of Tricholoma matsutake and T. magnivelare were examined on Pinus densiflora seedlings grown in a granite soil substrate with 1/2 PDMP (12 g/l potato dextrose broth, 1.5 g/l malt extract, and 0.5 g/l peptone) medium. Four months after inoculation, the pine seedlings were examined for infection rate, matsutake aroma, and Hartig-net formation. The roots of pine seedling formed ectomycorrhizal roots in the 9 isolates from 12 isolates of T. matsutake and T. magnivelare. However, the seedlings showed different ectomycorrhizae forming rates among the 9 isolates. While matsutake aroma was confirmed from the ectomycorrhizal seedlings, the pine seedling contaminated by bacteria or fungi did not form matsutake ectomycorrhizae with sickening smell. Thus, the aroma was chosen as a good way for the verification of mycorrhizal infection. At the early stage, the mycorrhizal roots showed unramified and branched types without root hair. They also showed thin mantle layers, Hartig-nets, and turned into black color at later stage. Among the examined strains, that of Yecheon isolated in 1995 showed the best infection rate, which indicated that we need to pay attention to the selection of isolates for better result.

• Journal of Korean Society of Forest Science
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• v.95 no.3
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• pp.358-364
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• 2006

This study was conducted to develop optimal solid culture medium for Tricholoma matsutake. As the solid matrix, granitic soil, perlite, vermiculate, pine sawdust and peat moss were compared regarding their effected on mycelial growth. Ergosterol content which is a fungal wall component was used as the growth index of the mycelia. Among the various solid matrixes, the granitic soil, perlite and mixture of the two supported the growth most. Barely flour appeared to be very effective on the stimulating of the mycelial growth when added to the solid matrix. An mixture of the matrix contained an even (1:1:1:1, v/v/v/v) mixture of granitic soil, perlite, vermiculate and pine sawdust. T. matsutake started growth 2 weeks after inoculation and reached stationary growth phase after 8th weeks in the solid matrix mixture. The mycelial density in the solid matrix was 7 times higher than that in fairy-ring soil. In addition, 30~70% water content and 10% humus soil in the solid matrix also supported good growth suggesting that T. matsutake needs humus soil for a nutrient sources. The solid matrix developed in the present study could be used to study physiological characteristics of T. matsutake as well.

• Journal of Korean Society of Forest Science
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• v.89 no.3
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• pp.389-396
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• 2000

To determine whether the Tricholoma matsutake (pine mushroom, Songyi) is symbiotic or parasitic to Pinus densiflora, structural change in their natural ectomycorrhizas were examined. The mycorrhizal samples were collected at three progressional points in the natural hypogeous colony(shiro) : colony front edge, near the fruiting point and 20cm back. The fine roots in the colonies were typical ectomycorrhizas with fungal mantle and Hartig net. However, the T. matsutake mycorrhizas had unique characteristics compared to other types of ectomycorrhizas. That is, spatially the fungal mantle and Hartig net of the T. matsutake mycorrhizas continued to develop along the growing tip, while temporally those structures declined to shrink changing to black brown in the older part of the roots behind the actively growing tip portion. However, there was no mark that the fungal hyphae penetrated into either the cortical cells, endodermal cell layers or stele. The apical tips of the blackened roots remained alive to form new mycorrhizas with other fungi later. Therefore, we conclude that the mycorrhiza of T. matsutake+P. densiflora is rather a dynamic symbiosis that changes its position spatiotemporally as the root grows than either a simple parasitism or symbiosis.

• Journal of Korean Society of Forest Science
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• v.95 no.6
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• pp.636-642
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• 2006

To make a new fairy ring of Tricholoma matsutake in situ, the way of production and transplanting of ectomycorrhizal seedlings of T. matsutake using Pinus densiflora was investigated after transplanting from 2000 to 2005 as well as the method to improve their survival rate for the fungus. For the production of ectomycorrhizal pine seedlings, the seedlings planted at the edge of fairy ring of T. matsutake in November showed 97% of the survival rate, while those planted in April showed 80% of the rate. For the complete infection of the T. matsutake, it required more than two years after planting. The infection rate of mycelia for the ectomycorrhizal seedlings was 17.6% when the natural seedlings were used, whereas it was relatively low when the seedlings prepared from the nursery were used. The survival of T. matsutake mycelium reached up to 22% by the transplanting in April, while the mycelium transplanted in October and November showed less than 5% of the survival. The survival of T. matsutake on the transplanted seedlings was the highest in the seedlings having 50% of infection rate before transplanting. Excavation of the ectomycorrhizal seedling to examine the vitality of ectomycorrhizal roots of T. matsutake resulted in the perishing of them. Therefore, the method using a 'rice bag triers' to check living mycelium of T. matsutake without digging of transplanted seedlings was introduced in this study. In addition, it is recommended that the examination has to be conducted at least two years after transplanting.

• Journal of Korean Society of Forest Science
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• v.90 no.6
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• pp.767-773
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• 2001

We investigated the impacts of the fairy-ring of Tricholoma matsutake on the dynamics of soil microflora and soil enzyme activities by grouping the soils around the fairy-ring of T. matsutake into four regions. The regions were grouped as 'zone of decayed mycorrhizae', 'zone of mycorrhizae for fruiting', 'zone of physiologically active mycorrhizae' and 'zone free from mycorrhizal infection'. Soil fungi and actinomycetes were quite little at the soils around the fairy-ring of T. matsutake compared to those of general forest soils, and there were significant differences among the four regions. The soils with the mycelial cluster of T. matsutake showed about one third of microbial population compared to those in the zone free from mycorrhizal infection, which indicated that T. matsutake took a dominant position within the fairy-ring of the fungus. We could manifest that T. matsutake showed a distinctive characteristics of mycorrhizal fungus since the activities of dehydrogenase were significantly different between the zone of physiologically active mycorrhizae and the zone free from mycorrhizal infection. The dehydrogenase activity was the highest at the early season of fruiting around the fairy-ring of T. matsutake, while the acid-phosphatase activity increased from March to June followed by a slight decrease on August and peaked on October. This phenomenon made us infer that the vitality of T. matsutake be sustained after fruiting.

• Journal of Life Science
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• v.18 no.9
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• pp.1290-1298
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• 2008

Tricholoma matsutake, one of edible and medicinal mushroom belonging to Tricholomaceae of Agaricales, has been known to contain some curing effect on gastric cancer and ulcer, and inhibitory effect on sarcoma 180 and Ehrlich sarcoma. Neutral salt soluble (0.9% NaCl), hot water soluble and methanol soluble substances (hereinafter referred to Fr. NaCl, Fr. HW and Fr. MeOH, respectively) were extracted from fruiting body of the mushroom. In vitro cytotoxicity tests, crude polysaccharides were not cytotoxic against cancer cell lines such as Sarcoma 180, HepG2, HT29 and NIH3T3 at the concentration of 2.0 mg/ml. Intraperitoneal injection with crude polysaccharides showed life prolongation effect of 23.4$\sim$37.2% in mice previously inoculated with Sarcoma 180. Fr. MeOH and Fr. HW exhibited the immuno-potentiating activity of B lymphocyte by increasing the alkaline phosphatase activity by 2.2$\sim$11.9 folds compared with control at the concentration of 0.2$\sim$0.5 mg/ml. In case of Fr. NaCl, the numbers of peritoneal exudate cells and circulating leukocytes were increased by 6.0 and 1.5 folds at the concentration of 50 mg/kg, respectively. Therefore, it is concluded that crude polysaccharides extracted from fruiting body of Tricholoma matsutake showed antitumor and immuno-potentiating activity against Sarcoma 180 of mouse.

• The Korean Journal of Mycology
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• v.19 no.2
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• pp.109-119
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• 1991

The vertical distribution of the fungal population for the soil samples from two sites of producing and nonproducing of Tricholoma matsutake, song-yi mushroom, were examined at Yang­yang and Myung-joo, Gangweon province. By the dilution plate method, a total number of propagu­les of fungi per gram of soil was observed to be low at the song-yi producing sites but high at the song-yi nonproducing sites under the communities of Pinus densiflora. The tendency of the number of fungal propagules were decreased with the increasing vertical depth. In the incuhation method at $42^{\circ}C$, six genera and nineteen species of the fungi were isolated from two sites; Aspergillus fumigatus, Acremonium sp., Talaromyces stipitatus, Penicillium lilacinum, P. oxalicum and Westerdykella multispora. The most dominant species by this method was A. fumigatus. From heat treatment method at $70^{\circ}C$, seven genera and nineteen species were isolated; Aspergillus fumigatus, Alternaria alternata, Neurospora sitophila and Mucor sp.. In the ethanol treatment method, one genera and one species was isolated Mortierella sp.. From the three isolation methods, it was found that the total number of the soil fungi and the frequency of species appeared were the highest at the soil of upper layer whereas the lowest at the soils of lower layer in its vertical distribution.

• The Korean Journal of Mycology
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• v.45 no.3
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• pp.188-195
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• 2017

Despite the high commercial value of the pine mushroom (Tricholoma matsutake) around the world, its production still depends upon natural harvesting. In recent years, mushroom researchers in Korea and Japan have been successful in artificially cultivating T. matsutake by producing single mushroom through matsutake-infected pine seedlings. In this study, we determined the minimum seedling height required for the survival of matsutake mycelia in the infected pine seedlings in the field. The survival rate of matsutake mycelia in the matsutake-infected pine seedlings was 50% (14) in two years and 71% (10) in one year after their transplantation. The average height at time of transplantation of the matsutake-infected pine seedlings that bore surviving mycelia after transplantation was 25 cm (minimum 12 cm to maximum 40 cm). In conclusion, the matsutake mycelium of matsutake-infected pine seedlings was able to survive in field conditions if the height of the seedling at the time of T. matsutake infection was at least 12 cm. These results suggest that the height of the host plants used in conventional matsutake-infected pine seedling production should be greatly reduced to improve the matsutake cultivation. Therefore, standardization of the seedling height for artificial cultivation of pine mushrooms by the matsutake-infected pine seedling method is suggested.

• Journal of Korean Society of Forest Science
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• v.72 no.1
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• pp.27-31
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• 1986

To compare the fungal distribution of matsutake-producing (Tricholoma matsutake) stand with that of non-producing stand, mushrooms on forest floor were collected in September of 1984 from 28-year-old non-producing and 55-year-old matsutake-producing stands of Pinus densiflora in Kangwon Province. A total of 18 fungal species in 12 genera were collected from a non-producing stand, and all the species except one were putative ectomycorrhizal. Common genera in non-producing stand included Suillus, Russula, Lactarius, Hydnum, and Amanita. From a matsutake-producing stand a total of 30 fungal species in 22 genera were collected, of which 22 species were ectomycorrhizal. Common genera in the producing stand included Hydnum, Tricholoma, Laccaria, and Cantharellus. Common species in both stands were six species only. It is concluded that matsutake-producing stand had more diversified fungal population than non-producing stand, probably because of higher contents of organic matters in soil and better development of understory vegetation through the ageing of the stand.