• Mycobiology
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• v.31 no.1
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• pp.1-8
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• 2003

Detailed structures of ectomycorrhizae formed in Pinus roots were observed with various microscopes: light, fluorescence, and scanning electron microscopes. The mantles and Hartig nets commonly found in the structure of ectomycorrhiza were newly observed according to developmental stage by various staining. The mycelia were observed to be composed of coiled types on the surface of epidermal root during early stage and fused to form mantles of smooth fungal layers, loosing mycelia with some viscous liquid secreted. The ectomycorrhizal hyphae in anatomical roots penetrated the cortical layer and formed obviously mantle and Hartig net. The round spots of ectomycorrhizal mycelia were observed morphological distribution from the cortical layer to vascular bundle of stele in the ectomycorrhizal roots of Pinus species and especially scattered at the area of meristem at the root tip as longitudinal sections. Those mycelia penetrated seemed to move into other roots by means of vascular bundle of ectomycorrhizal roots and newly form ectomycorrhizal roots of dichotomous branches.

• Journal of Korean Society of Forest Science
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• v.94 no.3 s.160
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• pp.146-152
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• 2005

For the application of ectomycorrhizal seedlings on damaged slope lands, studies on cultural characteristics of Lyophyllum shimeji and ectomycorrhizal associations of Lespedeza cyrtobotrya seedlings were carried out by artificial inoculation of L. shimeji. Mycelial growth of L. shimeji was best on MP (1% malt extract, 0.1% peptone, 1% glucose and 1.5% agar) medium. An optimum temperature and pH for the mycelial growth were $25^{\circ}C$ and pH6, respectively. Mycorrhizal root of L. cyrtobotrya seedlings inoculated with L. shimeji showed characteristics of ectomycorrhizas with Hartig net. Growth rate of the mycorrhizal seedlings's roots was higher than that of non-mycorrhizal seedlings. When the mycorrhizal seedlings were transplanted in slope land, survival rate and dry weight were 62% and 850 mg/seedling, respectively. On the other hand, survival rate and dry weight of non-mycorrhizal seedlings were 11% and 430 mg/seedling, respectively.

• Journal of Korean Society of Forest Science
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• v.82 no.4
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• pp.319-327
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• 1993

The objectives of this study were to isolate ectomycorrhizal fungi associated with roots of Pinus rigida ${\times}$ P. taeda trees planted in Chonbuk and Daejon locations and to identify them, and to observe the morphology of mycorrhizae formed by the isolated fungi. Pinus rigida ${\times}$ P. taeda seedlings formed ectomycorrhizae with Rhizopogon rubescens, Rhizopogon sp. and Suillus pictus isolated from the roots of mature Pinus rigida ${\times}$ P. taeda trees, Pinus rigida ${\times}$ P. taeda seedlings formed ectomycorrhizae with Pisolithus tinetorius too. The root surface of inoculated seedlings was covered with mycelia, and Hartig net were well developed in those roots. The size of vacuole was small, and the protoplasm was planty in ectomycorrhizal cell, compared to nonmycorrhizal cell. But the size of nuclei are not changed even in ectomycorrhizal cell.

• 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.94 no.1 s.158
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• pp.16-20
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• 2005

Tricholoma matsutake ectomycorrhizas are unique in their morphology: not bifurcated broom shaped roots with not easily wettable brilliant and profuse white hyphae. To understand these characteristics the ectomycorrhizas were investigated with electron microscopy. T. matsutake ectomycorrhiza have thin mantle and typical Hartig net development in the epidermis and cortex, but no fungal mantle on the root apex. There were no penetrating hyphae inside of the cells of either epidermis, cortex or stele. Inside of the walls of epidermis and cortex cells are lined with ca. $2{\mu}m$ hemispherical amyloplasts. The brilliant hyphal surface was covered with various fine amorphous granules. The hyphal cell wall was thin membrane less than $0.3{\mu}m$ thick. There is no clamp connection on the hyphae. This thin membraneous cell wall with high elasticity can be related to survival strategy of the species without plasmolysis under frequent soil water stress environment. And the coarse hyphal surface with some water repellency can control sudden inrush of water of the hyphae with an extremely low osmotic potential. It is concluded that no mantle on the tip can induce mycorrhizas not bifurcated and that finely coarse surface of T. matsutake hyphae can make the hyphae brilliantly white but less wetted.

• Journal of Forest and Environmental Science
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• v.15 no.1
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• pp.77-88
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• 1999

This experiment was carried out to compare the cultural characteristics of mycorrizal fungi associated with Pinus species, and to form mycorrhizal association with Pinus thunbergii by artificial inoculation of these fungi. Mycorrhizal fungi tested showed great variations in cultural characteristics. Most fungal isolates was best grown on MP medium, except PDA for Lepista sp.(Ln73/92). Hagem for Rhizopogon rubescens(FRI91017), and FDA for Paxillus sp.(Pa60/92). Optimum temperature for these fungi was $25^{\circ}C$, except $30^{\circ}C$ for Pisolithus tinctorius(FRI91004 and Pt1). The range of pH conditions favorable for these fungal isolates were also variable from weak acidic(pH5) to weak alkalic(pH8). Utilization of the carbon sources for these mycorrhizal fungi was different. Fructose, glucose, and maltose were all utilized well, while xylose was not utilized generally. Mycelial growth on the media supplemented with potassium nitrate was better than those on other media with urea, asparagine, or peptone as a nitrogen source, and the poor growth was observed on the media with urea. Pisolithus tinctorius(Pt1) among 7 mycorrhizal fungi artificially inoculated for the mycorrhizal synthesis on pinus thunbergii seedlings in the test tube containing a mixture of peat moss-vermiculite(2:1, v/v) formed mycorrhizae successfully after 3 months. P. tinctorius formed branched and unbranched roots covered with thick fungal mantle and radiating extemal hyphae. Mycorrhizal root cross-sectioned by hand, stained, and observed by Nomarski interference microscope showed typical characteristics of ectomycorrhizae: fungal mantle on epidermal cells and thick Hartig net hyphae around cortex cells.

• 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.82 no.4
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• pp.366-380
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• 1993

Quercus acutissima ectomycorrhizae were classified as apical type, linear type, clavate type, diffuse type, pyramidal type, coralloid type, and nodular type. The surface texture of the fungal mantle at the initial stage of mycorrhizal formation was velvety. The surface texture of Pisolithus tinctorius(Pt) mycorrhizae at 30 September was well-developed felty mantle, Yellowish white. Except Pt all mycorrhizae formed by ectomycorrhizal fungi used to experiment were white with felty mantle. Mycorrhizae at the initial stage of mycorrhizae formation were creamy or creamy brown and swelled with thin mantle. Transverse and longitudinal sections showed radially-elongated cortical cell layers and epidermal cell with Hartig net. The transversal wideth of radially-elongated cortical and epidermal cells in the mycorrhizae with thick mantle on 30 September did not different with the mycorrhizae with thin mantle on the initial stage. Pt #250 formed coralloid mycorrizae but Pt KJ-1 did not although they are same species. On the mean length of linear type ectomycorrhizae of Pt KJ-1(2.21mm) was 1.5 times longer than that of Pt #250(1.32mm). The total dry weight of seedlings inoculated with Pt KJ-1, Pt #250, Lycoperdon pedicellatum, Scleroderma verrucosum were significantly heavier than those of suillus granulatus, Laccaria laccata.