• Journal of Microbiology and Biotechnology
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• v.20 no.8
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• pp.1179-1184
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• 2010

We report the generation of the first monoclonal antibody that specifically binds to the polysaccharide chitosan. Mice were immunized with a mixture of chitosans, and hybridoma clones were screened for specific binders, resulting in the isolation of a single clone secreting a chitosan-specific IgM, mAbG7. In ELISAs, the antibody could bind to chitosans of varying composition, but demonstrated the highest affinity for chitosans with lower degrees of acetylation (DA) and very poor binding to chitin. We tested the ability of the antibody to bind to chitosan in situ, using preparations of fungal cell walls. Immunofluorescence microscopy confirmed that the antibody bound strongly to the cell walls of fungi with high levels of chitosan, whereas poor staining was observed in those species with cell walls of predominantly chitin or cellulose. The potential use of this antibody for the detection of fungal contamination and the protection of plants against fungal pathogens is discussed.

• Korean Journal Plant Pathology
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• v.10 no.2
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• pp.83-91
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• 1994

Stem tissues of tomato plants (cv. Kwanyang) inoculated with Phytophthora capsici were examined by light and electron microscopy to compare early cytological differences between comaptible and incompatible interactions of tomatoes with the fungus. Twenty four hours after inoculation, the compatible isolate S 197 colonized severely the epidermis, cortex, and xylem vessels of stem tissue, whereas only few fungal cells colonized the stem tissues inoculated with the incompatible isolate CBS 178.26. Fragmented plasma membrane, distorted chloroplast, degraded cell wall, remnants of host cytoplasm were early ultrastructural features of the damaged host cell observed both in the compatible and incompatible interaction, a number of vesicles were distributed in the space between fungal cell walls and plasma membrane. The degradation of host cell walls by P. capsici was more pronounced in the compatible than the incompatible interactions. The incompatible interactions of tomato cells with P. capsici were characterized by formation of host cell wall apposition in the cortical parenchyma cells, indicating that the apposition of electron-dense material from the host cell walls may function as a plant defense reaction to the fungus. The fungal cells encased by wall appositions had abnormal cytoplasm and separated plasma membranes. The haustorium which formed from the fungal hyphae did not further penetrate through the host wall apposition and cytoplasmic aggregation, especially in the incompatible reactions. In contrast, the haustorium of the compatible isolate S 197 was not encased by wall appositions.

• Bulletin of the Korean Chemical Society
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• v.34 no.3
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• pp.943-945
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• 2013

• The Plant Pathology Journal
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• v.33 no.1
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• pp.95-100
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• 2017

Fungicide-resistant Alternaria alternata impede the practical control of the Alternaria diseases in crop fields. This study aimed to investigate cytological fungicide resistance mechanisms of A. alternata against dicarboximide fungicide iprodione. A. alternata isolated from cactus brown spot was cultured on potato-dextrose agar (PDA) with or without iprodione, and the fungal cultures with different growth characteristics from no, initial and full growth were observed by light and electron microscopy. Mycelia began to grow from one day after incubation (DAI) and continued to be in full growth (control-growth, Con-G) on PDA without fungicide, while on PDA with iprodione, no fungal growth (iprodione-no growth, Ipr-N) occurred for the first 3 DAI, but once the initial growth (iprodione-initial growth, Ipr-I) began at 4-5 DAI, the colonies grew and expanded continuously to be in full growth (iprodione-growth, Ipr-G), suggesting Ipr-I may be a turning moment of the morphogenetic changes resisting fungicidal toxicity. Con-G formed multicellular conidia with cell walls and septa and intact dense cytoplasm. In Ipr-N, fungal sporulation was inhibited by forming mostly undeveloped unicellular conidia with degraded and necrotic cytoplasm. However, in Ipr-I, conspicuous cellular changes occurred during sporulation by forming multicellular conidia with double layered (thickened) cell walls and accumulation of proliferated lipid bodies in the conidial cytoplasm, which may inhibit the penetration of the fungicide into conidial cells, reducing fungicide-associated toxicity, and may be utilized as energy and nutritional sources, respectively, for the further fungal growth to form mature colonies as in Ipr-G that formed multicellular conidia with cell walls and intact cytoplasm with lipid bodies as in Con-G.

• Applied Microscopy
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• v.5 no.1
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• pp.31-43
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• 1975

These studies were carried out to detect the presence of infected virus- like particles and also were observed the ultrastructures of Aspergillus ochraseus isolated from kokja and Korean ginseng. The results of ultrastructures of Aspergillus ochraseus are summarized as follows: 1. In fungal cells, nuclei were enclosed by a irregular double membrane and nucleoli in the nucleus. 2. In cytoplasm, mitochondria, rough endoplasmic reticulum with ribosomes and glycogen were scattering distributed and many lomasomes also observed. 3. The osmiophilic bodies of fungal cells existed in the vesicles. 4. The cell walls were composed of a low electron dense materials. 5, Conidia cell walls were extremely thick and possessed the high electron density of outer coat. The virus-like particles were observed in the hyphae of Penicillium chrysogenum Q-176. These virus-like particles measured $350{\AA}$ in diameter. But strains of Aspergillus ochraseus, showing some vesicle particles were also observed about $800{\AA}$ in diameter in the central region of young fungal hyphae. Based on the results of these experiments, it can not be determined virus particles or not. The further studies to determination of virus particles will be proceeded by the chemical, physical and biological assay methods.

• Journal of the Korean Wood Science and Technology
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• v.24 no.2
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• pp.55-60
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• 1996

Tripitaka Koreana were made during Coryo Dynasty from 1236 to 1251 A.D. Buddhist scriptures were engraved on 81.340 wooden plates. Some plates were varnished with Rhus lacquer, but most of them were uncoated. Macroscopically, most of the plates appeared intact due to the storage in a well-ventilated wooden house. Because, they were irregularly used for printings with ink, it can be assumed that they were repeatedly exposed to ink-water and drying processes. The present were made to examine the changes of wood cell structures occurred during long-term aging deterioration processes in these dry archaeological wooden plates. Light, scanning and transmission electron microscopes were employed for this study. Wedge-shaped cracks and delamilations were found from the lumen side toward the compound middle lamellae and they progressed toward primary or secondary walls. A large amount of hypae in vessels and the degradation of vessel-ray pit walls by the fungal hyphae were observed. When compared to the recent wood, the birefringence of wood fibers was considerably lower or completly disappeared, suggesting the degradation of crystalline cellulose in these wood samples. The degradation of the cell wall could be also revealed the calculation of crystallinity with X-ray diffraction and the size of crystalline region was estimated.

• The Plant Pathology Journal
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• v.24 no.4
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• pp.392-399
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• 2008

Detached chili pepper fruits were inoculated with the conidial suspension of Colletotrichum acutatum JC-24 simultaneously (simultaneous inoculation, SI) and at delayed time (delayed inoculation, DI) after wounding with (delayed wound inoculation, DWI) or without additional wounding (delayed non-wound inoculation, DNI) at the inoculation time. Disease severity was significantly lowered by DNI, compared to SI. By DNI, the disease reduction rates were proportional with the length of delayed time, and greater at the high temperature range (18, 23 and $28^{\circ}$) than at the low temperature ($13^{\circ}$) tested. DWI was also effective in reducing the disease severity especially at 18oC; however, its effectiveness was lower than for DNI. In light microscopy, parenchyma cells at the wounding sites were modified structurally, initially forming new cell walls crossing cytoplasm, enlarged with multiple periclinal cell divisions, and finally layered like wound periderms. In DWI, the above structural modifications occurred, showing the restriction of the fungal invasion by the cell walls in enlarged modified cells, while no definite cellular modifications were found with proliferation of fungal hyphae in SI. Sclerenchyma-like cells with thickened cell walls were proliferated around the wounding sites, which were partially dissolved by DWI, probably leading to some disease development. All of these results suggest that the decline of the anthracnose disease in pepper fruit by the delayed inoculations may be derived from the structural modifications related to the healing processes of the previous wound inflicted on the tissues.

• Mycobiology
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• v.42 no.4
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• pp.311-316
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• 2014

Lichen studies, including biodiversity, phylogenetic relationships, and conservation concerns require definitive species identification, however many lichens can be challenging to identify at the species level. Molecular techniques have shown efficacy in discriminating among lichen taxa, however, obtaining genomic DNA from herbarium and fresh lichen thalli by conventional methods has been difficult, because lichens contain high proteins, polysaccharides, and other complex compounds in their cell walls. Here we report a rapid, easy, and inexpensive protocol for extracting PCR-quality DNA from various lichen species. This method involves the following two steps: first, cell breakage using a beadbeater; and second, extraction, isolation, and precipitation of genomic DNA. The procedure requires approximately 10 mg of lichen thalli and can be completed within 20 min. The obtained DNAs were of sufficient quality and quantity to amplify the internal transcribed spacer region from the fungal and algal lichen components, as well as to sequence the amplified products. In addition, 26 different lichen taxa were tested, resulting in successful PCR products. The results of this study validated the experimental protocols, and clearly demonstrated the efficacy and value of our KCl extraction method applied in the fungal and algal samples.

• Journal of the Korean Wood Science and Technology
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• v.34 no.2
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• pp.68-77
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• 2006

The heartwood of cengal (Neobalanocarpus heimii) is known to have a high degree of decay resistance by virtue of its high extractive content. After 30 years in ground contact an utility pole of this tropical hardwood was found to be degraded only in the surface layers by cavity-forming soft rot fungi. The present work was undertaken 1) to characterize the degradation of cengal heartwood from the aspect of ultrastructure and chemistry and 2) to investigate the correlation between soft rot decay and its extractive microdistribution in wood tissues. The chemical analysis of cengal heartwood revealed the presence of a high amount of extractives as well as lignin. The wood contained a relatively high amount of condensed lignin and the guaiacyl units. Microscopic observations revealed that vessels, fibers and parenchyma cells (both ray and axial parenchyma) all contained extractives in their lumina, but in variable amounts. The lumina of fibers and most axial parenchyma were completely or almost completely filled with the extractives. TEM micrographs showed that cell walls were also impregnated with extractives and that pit membranes connecting parenchyma cells were well coated and impregnated with extractives. However, fungal hyphae were present in the extractive masses localized in cell lumina, and indications were that the extractives did not completely inhibit fungal growth. The extent of cell wall degradation varied with tissue types. The fibers appeared to be more susceptible to decay than vessels and parenchyma. Middle lamella was the only cell wall region which remained intact in all cell types which were severely degraded. The microscopic observations suggested a close correlation between extractive microdistribution and the pattern and extent of cell wall degradation. In addition to the toxicity to fungi, the physical constraint of the extractive material present in cengal heartwood cells is likely to have a profound effect on the growth and path of invasion of colonizing fungi, thus conferring protection to wood by restricting fungal entry into cell walls. The presence of relatively high amount of condensed lignin is also likely to be a factor in the resistance of cengal heartwood to soft rot decay.

• IMMUNE NETWORK
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• v.13 no.5
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• pp.205-212
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• 2013

Dectin-1, which specifically recognizes ${\beta}$-glucan of fungal cell walls, is a non-Toll-like receptor (TLR) pattern recognition receptor and a representative of C-type lectin receptors (CLRs). The importance of Dectin-1 in innate immune cells, such as dendritic cells and macrophages, has previously been well studied. However, the function of Dectin-1 in B cells is very poorly understood. To determine the role of Dectin-1 in B cell activation, we first investigated whether mouse B cells express Dectin-1 and then assessed the effect of Dectin-1 stimulation on B cell proliferation and antibody production. Mouse B cells express mRNAs encoding CLRs, including Dectin-1, and surface Dectin-1 was expressed in B cells of C57BL/6 rather than BALB/c strain. Dectin-1 agonists, heat-killed Candida albicans (HKCA) and heat-killed Saccharomyces cerevisiae (HKSC), alone induced B cell proliferation but not antibody production. Interestingly, HKSC, HKCA, and depleted zymosan (a selective Dectin-1 agonist) selectively enhanced LPS-driven IgG1 production. Taken together, these results suggest that, during fungal infection, ${\beta}$-glucan-stimulated Dectin-1 may cooperate with TLR4 to specifically enhance IgG1 production by mouse B cells.