• Food Science and Biotechnology
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• 제14권4호
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• pp.543-546
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• 2005

Optimized conditions for protoplast preparation and regeneration from young hyphae of Monascus ruber were established. Heat shock treatment of spores gave rapid and synchronized germination. Spores collected from cultures grown for 7-8 days at $30^{\circ}C$ were germinated until over 70% germ tubes reached to 3-5 spore length. Enzymatic digestion of young hyphae was optimal with 50 mg/mL Glucanex in 0.1 M sodium citrate buffer containing 0.8 M mannitol as an osmotic stabilizer. Regeneration rate was around 10% when 0.8 M sorbitol was used as an osmotic stabilizer in regeneration medium. These conditions will be applied in genetic study of M. ruber that produces citrinin at high level and thus is good model strain for molecular genetic dissection of citrinin biosynthesis.

• The Plant Pathology Journal
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• 제31권3호
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• pp.299-304
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• 2015

Interaction of the the rust fungus Puccinia miscanthi with the biofuel plant Miscanthus sinensis during the teliospore phase was investigated by light and electron microscopy. P. miscanthi telia were oval-shaped and present on both the adaxial and abaxial leaf surfaces. Teliospores were brown, one-septate (two-celled), and had pedicels attached to one end. Transmission electron microscopy revealed numerous electron-translucent lipid globules in the cytoplasm of teliospores. Extensive cell wall dissolution around hyphae was not observed in the host tissues beneath the telia. Hyphae were found between mesophyll cells in the leaf tissues as well as in host cells. Intracellular hyphae, possibly haustoria, possessed electron-dense fungal cell walls encased by an electron-transparent fibrillar extrahaustorial sheath that had an electron-dense extrahaustorial membrane. The infected host cells appeared to maintain their membrane-bound structures such as nuclei and chloroplasts. These results suggest that the rust fungus maintains its biotrophic phase with most mesophyll cells of M. sinensis. Such a nutritional mode would permit the rust fungus to obtain food reserves for transient growth in the course of host alteration.

• Journal of Microbiology and Biotechnology
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• 제10권5호
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• pp.606-611
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• 2000

Dimorphic yeast Candida albicans reversibly switches between the form of yeast and hyphae depending on external conditions. We investigated possible roles of the myosin family in the growth and dimorphic switches of C. albicans with a general myosin ATPase inhibitor, 2,3-butanedione-2-monoxime (BDM). Transition to hyphae as well as proliferation by budding was completely inhibited by BDM at 16 mM. Presence of 16 mM BDM did not affect hyphae-to-bud transition but it blocked budding. The effects of BDM on yeast growth and dimorphic switches were reversible. More than 70% of the BDM-treated cells demonstrated defects in the amount and the polarized localization of F-actin as well as in the shape and migration of the nucleus, suggesting that myosin activities are needed in these cellular processes of C. albicans.

• The Plant Pathology Journal
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• 제17권1호
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• pp.29-35
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• 2001

Early infection process of Phytophthora capsici in pumpkin stems was similar in the compatible and incompatible interactions 24 h after inoculation. Intercellularly growing hyphae penetrated host parenchyma cells by growing hyphae penetrated host parenchyma cells by forming haustoria. An extrahaustorial matrix was found around the haustoria in both compatible and incompatible interactions. No wall appositions were observed at the infection sites in the parenchyma cells. In the compatible interaction, infecting hyphae grew well in the intercellular spaces between xylem vessels in stem tissues. Degraded host cell wall, plasmolysis of plasma membrane, and degenerated chloroplasts were pathological features of pumpkin stem tissues in both compatible and incompatible interactions. A characteristic host response in the resistant pumkin cultivar Danmatmaetdol was rapid cytoplasmic movement of host cells toward the oomycete haustoria.

• 한국산림과학회지
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• 제94권1호통권158호
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• pp.16-20
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• 2005

송이균근은 형태적으로 매우 특이한 외생균근이다. 즉 송이균근은 Y자형이 아닌 빗자루형이며, 균사는 매우 밝은 흰색을 띠고, 메마른 경우 물에 쉽게 젖지 않는다. 송이균근이 왜 이런 특정을 갖는가를 이해하기 위하여 주사전자현미경을 이용하여 형태를 관찰하였다. 송이균근의 정단부에는 균사충이 발달하지 않았고 균근 부위 중에서 균사충이 잘 발달된 부위에서는 Hartig 망이 1-2층 발달하였으나, 세포내부로 침입하여 들어간 균사는 없었다. 그러나 뿌리 외부의 균사충이 덜 발달한 부분에서는 표피세포나 피층세포층에서 Hartig 망이 형성되지 않았다. 이들 세포의 내부에서는 약 $2{\mu}m$ 크기의 반원형 알갱이 형태인 전분체가 세포벽을 전체적으로 덮고 있었다. 송이균사표면은 모양이 일정하지 않은 작은 알갱이로 덮혀있기 때문에 매우 밝은 흰색을 띠며 물의 표면장력으로 인하여 잘 젖지 않는 현상을 부분적으로 설명하는 것으로 생각한다. 송이균사의 세포벽은 매우 얇은 단순한 막질이고, 연결꺽쇄(clamp connection)는 없었다. 송이균근의 정단부에 균사충이 없기 때문에 Y자형으로 갈라지지 않는 것으로 생각된다. 그러나 균근이 가지를 많이치는 것은 송이균근이 소나무뿌리로부터 탄수화물을 받는 표면적을 넓히는 것이라고 생각된다. 송이균사의 세포벽이 매우 얇은 것은 세포벽의 신축성이 커서 주기적인 토양건조에 따라 수분포텐셜을 잘 조절하여 원형질분리가 일어나지 않게 하는 생존전략과 관련된다고 생각한다. 그리고 거친 균사 표면은 균사가 물에 느리게 젖게 함으로써 삼투포텐셜이 매우 낮았던 균사가 수분을 갑자기 많이 흡수하는 것을 막는다고 생각한다. 결론적으로 송이균근은 정단부에서 균사충이 발달하지 않으므로 Y자형으로 갈라지지 않으며, 균사표면의 거친 입자는 흰색을 더욱 밝게하면서 잘 젖지 않도록 한다고 생각한다.

• Journal of Microbiology and Biotechnology
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• 제23권7호
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• pp.1031-1040
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• 2013

Candida albicans is a dimorphic fungus that commensally colonizes human mucosal surfaces. The aim of this study was to assess the role of different C. albicans morphologies in inducing pattern recognition receptors (PRRs) and cytokines in macrophages. Macrophages may respond to pathogen-associated molecular patterns via TLR2 and TLR4 by expressing cytokines. The hyphal transition of C. albicans was induced by 20% serum (S), RPMI-1640 (R), or $39^{\circ}C$ culture (H). Macrophages were then challenged with either yeast (Y) or different hyphae cultures of C. albicans, followed by RT-PCR and FACS analysis of PRRs expression. In addition, macrophages were stimulated with either yeast or different hyphae cultures of C. albicans used by RT-PCR and Bio-Plex analysis of cytokines production. Macrophages expressed high levels of TLR4 and dectin-1 after stimulation with Y cells. In contrast, stimulation with H or R cells strongly increased the expression of TLR2 and dectin-2. Stimulation with Y cells significantly enhanced the expression of IL-$1{\beta}$ and weakly increased the expression of IL-6 and IL-12. Stimulation with hyphal cells (S, R, and H) strongly increased IL-10 expression, but weakly reduced IL-$1{\beta}$ expression. The phagocytosis activity and NO production of macrophages were decreased upon treatment with hyphal cells compared with yeast, and depended on the length of hyphae. In summary, the yeast and hyphae forms of C. albicans resulted in an induction of different PRRs, with accompanying differences in immune cell cytokine profiles.

• 한국균학회지
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• 제22권1호
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• pp.36-45
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• 1994

아버스큘 균근균인, Gigaspora margarita Scutellospora heterogama, S. verrucosa의 포자를 발아시켜서 균사의 생장, 보조세포 발달 그리고 절단된 균사의 회복과정을 조사하였다. 발아된 균사는 배지의 표면과 내부 및 바닥에서 생장하였으며, 배지표면에서는 19-23일째, 배지바닥에서는 40-51일째에 생장을 멈추었다. 보조세포는 포자 발아 후 7-9일째에 2차 균사에서 새로 나온 균사 위에 형성되기 시작하였으며 배지의 표면이나 바닥, 그리고 배지속에서도 발달하였다. G. margarita와 S. heterogama의 원형질 유동속도는 각각 $2.7-3.3\;{\mu}m/s$와 $3.8-4.3\;{\mu}m/s$이었다. 균사가 절단되면 포자쪽 균사의 끝 부분에서 새로운 균사가 뻗어나와 끊어진 부분을 다시 연결하여 계속 생장하였다. 이러한 절단된 균사의 회복 과정은 균근균이 기주식물과 오랜 공생관계를 맺으면서 영양적으로는 절대적으로 식물에 의존함을 나타내는 것이라고 생각한다.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.119.1-119
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• 2003

The glyoxysomal nature of microbodies was determined in Botryosphaeria dothidea hyphae based on morphology and in situ enzyme characteristics by transmission electron microscopy and cytochemistry. Bound by a single membrane, microbodies had a homogeneous matrix and varied in size ranging from 200 to 400 m in diameter. Microbodies had crystalline inclusion(s) which consisted of parallel arrays of fine tubules in their matrices. Microbodies and lipid globules were frequently placed in close association with each other, forming microbody-lipid globule complexes in hyphae. The cytochemical activities of catalase and malate synthase were localized in matrices of microbodies, showing intense electron-density of the organelle. In addition, the immunogold labeling detected the presence of catalase in multivesicular bodies and hyphal cell walls as well as in matrices and crystalline inclusions of microbodies, supporting the enzyme secretion through cell walls. Meanwhile, isocitrate Iyase was localized only in matrices of microbodies. These results suggest that microbodies, particularly complexed with lipid globules, in the fungal hyphae are functionally defined as glyoxysomes, where glyoxysomal enzymes are biochemically active for the glyoxylate cycle to be a metabolic pathway in gluconeogenesis. (Mycology and Fugus Diseases)

• The Plant Pathology Journal
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• 제15권3호
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• pp.144-151
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• 1999

Chitinase activities from Shumard oak tissues were determined to study changes in chitinase activities related to water stress. The enzyme extracted in sodium acetate buffer (0.1M, pH 4.5) was assayed by a colorimetric method. In addition, the fungal hyphae of Hypoxylon atropunctatum in xylem tissues of oak were observed through scanning electron microscopy. The enzyme in oak tissues was mainly endochitinase, and optimum pH for enzyme activity was 5. Specific chitinase activities from both of stems held under high relative humidity (ranges of 0.63-1.11 pKatal/$\mu\textrm{g}$ of protein) and stems held under low relative humidity (ranges of 0.41-0.99 pKatal/$\mu\textrm{g}$ of protein) were significantly increased following fungal inoculation with H. atropunctatum. However, there was no significant difference in chitinase activities between tissues held under high and low humidities, which might be due to fungal chitinase. Scanning electron microscopy showed holes in fungal hyphae in the xylem tissues of stems held under high humidity but not in the stems held under ow humidity, suggesting that hyphae might be hydrolyzed by plant hydolases such as chitinase.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1987년도 식물생명공학 심포지움 논문집 Proceedings of Symposia on Plant Biotechnology
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• pp.157-167
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• 1987

The have been many reports that phenoloxidase are correlated with development in many fungi. C. congregatus, one of nushroom-forming basidiomycetes, which requires light for its development also has phenoloxidases. In C. congragatus, there are two sets of membrane-associated phenoloxidase (PHO I and PHO II) which are differentiated by their isozyme patterns, and each enzyme set consists of two different subtrate specific enzyme protein; o-tolidine reacting enzyme, and DOPA reacting enzyme. PHO I which is localized by a protoplast-concanavalin A technique by using a new solidifying agent, Pluronic Polyol F 127, instead of agar appears in the vegetative hyphae, and PHO II appears at the early primordial stage on agar and at the sclerotial stage of liquid shake cultures. Inhibition of PHO I with the enzyme inhibitors inhibits mushroom formation as well as melanization of the vegetative hyphae at concentrations which do not inhibit the vegetative growth. PHO I deficient mutants do not form mushrooms or melanins, and the mutants show abnormal nuclear migration patterns. PHO II has roles; possibly cementing the adjacent hyphae during the actual three dimensonal structure formation, and melanizing mushrooms and sclerotia. The possible roles of PHO I in the light reception complex and in melanin formation, the function of malanin, and possible roles of postulated post translational modifying enzymes which regulate the phenoloxidases, nuclear migration pattern, and self-nonself recognition mechanism are discussed.