• Mycobiology
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• v.44 no.1
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• pp.7-13
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• 2016

The genets of Suillus granulatus in a Pinus strobus stand ($13m{\times}60m$) were identified using random amplified polymorphic DNA molecular markers and the DNA of mushrooms that fruited for two years, and variations in genet size and distribution were analyzed. From a total of 116 mushrooms, 73 genets were identified and were grouped into three locations. The genets of mushrooms in close proximity differed from each other. The genet sizes varied at any of the three locations. The lengths of the identified genets in the pine stand ranged from 0.09 to 2.90 m. The average number of mushrooms per genet was 1.2 to 2.3, and the percentage of genets that were represented by a single mushroom was 44% to 94%. This variation in the genets of mushrooms in close proximity suggests that the ectomycorrhizal mycelial bodies of S. granulatus propagated sexually by fusing haploid spores derived from the mushrooms gills with below-ground mycelia. Therefore, it is necessary further to investigate the formation of new genets through spores in ectomycorrhizal fungal colonies.

• Journal of Microbiology and Biotechnology
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• v.18 no.3
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• pp.483-489
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• 2008

Nine newly isolated mushroom strains were tested to assess both their zinc tolerance and potential for zinc removal from an aqueous solution. Four strains of ectomycorrhizal fungi, namely Clavariadelphus truncatus (T 192), Rhizopogon roseolus (T 21), Lepista nuda (T 373), and Tricholoma equestre (T 174), along with five strains of white rot fungi, Lenzites betulina (S 2), Trametes hirsuta (T 587), Ganoderma spp. (T 99), Polyporus arcularius (T 438), and Ganoderma carnosum (M 88), were investigated using zinc-amended solid and liquid media. Their biosorption properties were also determined. The colony diameter and dry weight were used as tolerance indices for fungal growth. C. truncatus and T. equestre were not strongly inhibited at the highest concentrations of (225 mg/l) zinc in solid media. The most tolerant four strains with solid media, C. truncatus, G carnosum, T. hirsuta, and T. equestre, were then chosen for tolerance tests in liquid media. An ectomycorrhizal strain, C. truncatus, was also detected as the most tolerant strain in liquid media. However, the metal-tolerant strains demonstrated weak activity in the biosorption studies. In contrast, the highest biosorption activity was presented by a more sensitive strain, G. carnosum. In addition, seven different biosorbent types from G. carnosum (M 88) were compared for their Zn (II) biosorption in batch experiments.

• The Korean Journal of Mycology
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• v.42 no.2
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• pp.150-158
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• 2014

Four ectomycorrhizal fungi were tested for their ability to utilize seven carbon sources in modified Melin-Norkrans medium. After 60 days of mycelial culture, the highest mycelial growth in Hygrophorus russula (KFRI 1987), Sarcodon aspratus, Leccinum extremiorientale (KFRI 1194), and Tricholoma matsutake (KFRI 1256) was observed with use of dextrin used as a carbon source. H. russula, S. aspratus (KFRI 1676), and L. extremiorientale showed the lowest mycelial growth on nutrient medium with pectin. The utilization of homoglycans (starch, dextrin) in seven strains (except for T. matsutake KFRI 1256) was higher than that of heteroglycan (pectin). The final pH values of all culture media were decreased by pH 1.1~3.0 compared with the initial pH values of culture media. The dominant color of mycelia was white and varied according to the carbon sources (yellow, brown, and purple) in some strains. A single colony was observed in L. extremiorientale cultured in liquid media containing four or five different types of carbon sources, whereas multiple colonies were formed in liquid media containing six different types of carbon sources by six strains.

• Mycobiology
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• v.37 no.1
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• pp.10-16
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• 2009

The purpose of this study is to understand spatio-temporal changes of active fungal biomass and water in Tricholoma matsutake soil colonies during the mushroom fruiting season. The active fungal biomass was estimated by analyzing ergosterol content at four different points within four replicated locations in a single circular T. matsutake colony at Ssanggok valley in the Sogri Mt. National Park in Korea during 2003 to 2005. The four points were the ahead of the colony, the front edge of the colony and 20 cm and 40 cm back from the front edge of the colony. Ergosterol content was 0.0 to 0.7 ${\mu}g$ per gram dried soil at the ahead, 2.5 to 4.8 ${\mu}g$ at the front edge, 0.5 to 1.8 ${\mu}g$ at the 20 cm back and 0.3 to 0.8 ${\mu}g$ at the 40 cm back. The ergosterol content was very high at the front edge where the T. matsutake hyphae were most active. However, ergosterol content did not significantly change during the fruiting season, September to October. Soil water contents were lower at the front edge and 20 cm back from the front edge of the colony than at the ahead and 40 cm back during the fruiting season. Soil water content ranged from 12 to 19% at the ahead, 10 to 11% at the edge, 9 to 11% at the 20 cm back and 11 to 15% at the 40 cm back. Our results suggest that the active front edge of the T. matsutake soil colony could be managed in terms of water relation and T. matsutake ectomycorrhizal root development.

• The Korean Journal of Quaternary Research
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• v.17 no.2
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• pp.129-133
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• 2003

Water is critically important for Tricholoma matsutake(Tm) growth because it is the major component of the mushroom by over 90%. The mushroom absorbs water through the below ground hyphal colony. Therefore, the objectives of our study were to investigate spatio-temporal water changes in Tm colonies. This study was carried out at Tm fruiting sites in Sogni Mt National Park, where the below-ground mushroom colonies have been irrigated. To identify spatial water status within the Tm soil colony soil moisture and ergosterol content were measured at six positions including a mushroom fruiting position on the line of the colony radius. To investigate temporal soil moisture changes in the soil colony, Time Domain Reflectometry(TDR) sensors were established at the non-colony and colony front edge, and water data were recorded with CR10X data logger from late August to late October. Before irrigation, whereas it was 12.8% at non-colony, the soil water content within Tm colony was 8.0% at 0-5cm from the colony front edge, 6.2% at 10-15cm and 6.5-7.5% at 20-40cm. And the content was 12.1% at 80cm distance from the colony edge, which is similar to that at the non-colony. In contrast, ergosterol content which is proportional to the live hyphal biomass was only 0.4${\mu}g$/g fresh soil at the uncolonized soil, while 4.9 $\mu\textrm{g}$/g fresh soil at the front edge where the hyphae actively grow, and 3.8 ${\mu}g$/g fresh soil at the fruiting position, l.1${\mu}g$/g at 20cm distance and 0.4${\mu}g$/g in the 40cm rear area. Generally, in the Tm fungal colony the water content changes were reversed to the ergosterol content changes. While the site was watered during August to October, the soil water contents were 13.5∼23.0% within the fungal colony, whereas it was 14.5∼26.0% at the non-colony. That is, soil water content in the colony was lower by 1.0∼3.0% than that in the non-colonized soil. Our results show that Tm colony consumes more soil water than other parts. Especially the front 30cm within the hyphal colony parts is more critical for soil water absorption.

• 한국균학회소식:학술대회논문집
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• 2015.11a
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• pp.16-16
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• 2015

Ectomycorrhizal (ECM) mushrooms play a major role in plant growth promotion through symbiotic association with roots of forest trees. They also provide an economically important food resource to us and therefore they have been studied for their artificial cultivation for decades in Korea. We have secured bio-resources of ECM mushrooms from Korean forests and performed their physiological studies. To investigate the cultural characteristics, the fungi were cultured under different conditions (medium, temperature, pH of the medium, inorganic nitrogen source). More than 90% of total 160 strains grew on three solid media (potato dextrose agar, PDA; sabouraud dextrose agar, SDA; modified Melin-Norkrans medium, MMN). The rate of mycelial growth on malt extract agar (MEA) was lower than those of three media (PDA, SDA, MMN). None of the Tricholomataceae strains grew on MEA. Many strains of ECM mushrooms were able to grow at the temperature range of $15{\sim}25^{\circ}C$ on PDA, while they showed poor growth at $10^{\circ}C$ or $30^{\circ}C$. In particular, the growth rates of both Gomphaceae and Tricholomataceae were significantly lower at $10^{\circ}C$ than at $30^{\circ}C$. The optimal pH of many strains was pH 5.0 when they cultured in potato dextrose broth (PDB). Fifty-seven percent of tested strains grew well on medium containing ammonium source than nitrate source. Many strains of Tricholomataceae showed a notable growth on ammonium medium than nitrate medium. Twenty-three percent of strains preferred nitrate source than ammonium source for their mycelial growth. The production and activity of two enzymes (cellulase and laccase) by ECM fungi were also assayed on the enzyme screening media containing CMC or ABTS. Each strains exhibited different levels of enzymatic activities as well as enzyme production. The number of laccase-producing strains was less than that of cellulase-producing strains. We found that 77% of tested strains produced both cellulase and laccase, whereas 2% of strains did not produce any enzymes. The morphological characteristics of mycelial colony were also examined on four different solid media. Yellow was a dominant color in mycelial colony and followed by white and brown on all culture media. ECM mushrooms formed mycelial colonies with a single or multiple colors within a culture medium depending on the strains and culture media. The most common shape of mycelial colony was a circular form on all media tested. Other families except for Amanitaceae formed an irregular colony on MMN than PDA. All strains of Tricholomataceae did not form a filamentous colony on all media. The pigmentation of culture media by mycelial colonies was observed in more than 50% of strains tested on both PDA and SDA. The degree of pigmentation on PDA or SDA was higher than MMN and brown color was dominant than yellow color. The production of exudates from mycelial colony was higher on PDA than MMN. Brown exudates were mainly produced by many strains on PDA or SDA, whereas transparent exudates were mainly produced by strains on MMN. We observed the mycelial colonies with a single or multiple textures in just one culture plate. Wrinkled or uneven colony surfaces were remarkably observed in many strains on PDA or SDA, while an even colony surface was observed in many strains on MMN. Sixty percent of Tricholomaceae strains formed wrinkled surface on PDA. However, they did not form any wrinkle on MMN plate. Cottony texture was observed in mycelia colonies of many strains. Velvety texture was often observed in the mycelial colonies on SDA than PDA and accounted for 60% of Suillaceae strains on SDA.

• 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.87 no.3
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• pp.429-444
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• 1998

Objective of this study was to select ectomycorrhizal fungi for black spruce(Picea mariana) inoculation to overcome the growth inhibitory effects of Kalmia angustifolia. Nineteen isolates representing 11 species of ectomycorrhizal fungi were tested for their abilities to grow and form mycorrhizae with black spruce seedlings in the presence of water leachate of leaves of Kalmia. Mycelium growth of 9 isolates were inhibited by the leaf leachate. Colony diameter and biomass of the other 10 isolates were either increased or unaffected under the same conditions. Acidic pH of the culture medium(pH 3 and 4) inhibited some of the fungi, but a combination of acidic pH and the leaf leachate was more inhibitory. Thirteen isolates were able to form ectomycorrhizae with black spruce in presence of 25% leaf leachate in pure culture. Four isolates, Paxillus involutus(NF4), Cenococcum geophilum(GB12), Laccaria laccata(GB23), and E-strain(GB45) formed mycorrhizae more successfully than the others in presence of up to 50% Kalmia leaf leachate. Black spruce seedlings pre-inoculated with these fungi were grown with Kalmia leaf leachate and live Kalmia plants during a four month greenhouse experiment. Abundant mycorrhizae(77-91% of root tips) were developed on seedlings pre-inoculated with P. involutus, L. laccata and E-strain but relatively poor mycorrhization(32% of root tips) resulted with C. geophidum. Over 90% of the short root mycorrhizae were attributed to the inoculated fungi although indigenous mycorrhizae also occurred on most seedlings. Persistence of the mycorrhizae was not affected by living Kalmia plants. Over 80% of the mycorrhizae on seedlings inoculated with P. involutus, L. laccata and E-strain and 53% of the mycorrhizae on seedlings inoculated with C. geophilum were attributable to the inoculant fungi. Control seedlings formed about 45% ectomycorrhizal short roots with indigenous fungi. The L. laccata and C. geophilum inoculated seedlings exhibited enhanced mycorrhizae formation in presence of Kalmia leaf leachate. Mycorrhizae formation with inoculant fungi was 4-15% lower at pH 4 than at pH 5, with the greatest inhibition occurring for L. laccata. Seedlings inoculated with P. involutus had the greatest shoot and root growth followed by L. laccata and E-strain inoculated seedlings. The P. involutus and L. laccata inoculated seedlings were significantly taller with more shoot dry biomass than the uninoculated(control) seedlings. E-strain inoculated seedlings had significantly higher shoot dry biomass and significantly lower number of first order lateral roots compared to the control but other growth parameters such as height, root dry weight and number of short root tips were not significantly different from the control. Seedlings inoculated with C. geophilum were not significantly different from the uninoculated seedlings in any of the growth parameters except for the number of first artier lateral roots which was significantly less than the control seedlings.