• The Plant Pathology Journal
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• v.15 no.4
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• pp.223-227
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• 1999

Pre-penetration behavior of Boytryosphaeria dothidea on apple fruits was investigated with scanning electron microscopy. Once conidia were deposited on the fruit surface, they germainted and produced germ tubes from one or both ends of the conidia. Germ tubes grew over the fruit surface and entered the fruits through lenticels or surface cracks formed naturally. Germ tubes of the fungus also appeared to penetrated the fruits directly with or without forming appressoria. Globose appressoria were frequently formed at the tip of germ tubes on the fruit surface, where no lenticels or surface cracks were found. The conidia collapsed and became flattened to the fruit surface after appressorial formation. Cuticles of fruit surface underneath appressoria and tips of some germ tubes were evidently altered, indicating possibility of direct penetration of the fungus by enzymatic degradation of the cuticle layers. This the first report on the formation of appressoria by B. dothidea.

• Korean Journal Plant Pathology
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• v.10 no.4
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• pp.254-260
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• 1994

Magnaporthe grisea, a causal agent of blast, forms a specialized infection structure, an appressorium, to infect host. Hydrophobicity of contact surface and cAMP have been suggested as a primary environmental signal and a second messenger to trigger and mediate appressorium formation in this fungus, respectively. To generalize these factors in field isolates of M. girsea, twenty isolates originated from rice and other gramineous hosts were tested. Seventeen including rice and non-rice isolates formed appressoria on hydrophobic surface, but none of isolates formed appressoria on hydrophilic surface. Eighteen isolates formed appressoria on hydrophilic surface in the presence of IBMX, an inhibitor of phosphodiesterase, except two rice isolates. These results strongly support the hypothesis that appressorium formation by M. grisea is induced by hydrophobic hard surface and regulated by the endogenous level of cAMP in the cells. Understanding fungal development is not only of biological interest but provides new targets for novel disease control strategies.

• The Korean Journal of Mycology
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• v.21 no.3
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• pp.224-229
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• 1993

The optimum temperature for germination of conidia and germ tube elongation were between $20\;and\;30^{\circ}C$ in C. dematium and C. gloeosporioides. Appressoria were fairly formed well at $20^{\circ}C$ despite the delay of conidial germination. At $30^{\circ}C$, both the germination and germ tube elongation are favored, but appressoria were poorly detected to be formed. In C. acutatum, the optimum temperature for germination of conidia was from $20\;to\;30^{\circ}C$, but at $25^{\circ}C$, germ tube elongation are accelerated. The conidia become septate and one or both doughter cells become conidiogenous instead of producing germ tubes and a secondary conidia produced, resulting in an arborescent type of connected conidia. Appressoria are infrequently formed by germinating conida. At $20\;to\;25^{\circ}C$ was the optimum for appressorium formation. But conidia that germinated at $30^{\circ}C$ seemed to lose the ability to form appressoria. The relation of temperature to germination of conidia and appressorium formation in Colletotrichum acutatum, C. dematium and C. gloeosporioides are discussed.

• Korean Journal Plant Pathology
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• v.14 no.6
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• pp.578-582
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• 1998

In greenhouse studies, control efficacy of water chestnut (Eleocharis Kuroguwai) by Epicoccosorus nematosporus was affected by temperature and dew condition. The appressoria were formed abundantly in the range of 20~28$^{\circ}C$. When stem segments o(30 cm long) of the water chestnut were inoculated with the conidial suspension of E. nematorporus, the mean conidial number attacted amounted to 2,545 conidia. Out of 2545 conidia attacted to the stem pieces, 1,733 (68%) conidia formed appressoria. When these stem pieces were treated for 24 hr at 28$^{\circ}C$ under dew condition, 183,1 (7.2%) lesions were formed 10 days after incubation. The time necessary for the death of the plants was about 24 days. Appressoria were formed at 15~35$^{\circ}C$, but decreased rapidly in their numbers at the temperature lower than 1$0^{\circ}C$ or at 35$^{\circ}C$. The appressoria formation seemed to be depended on the dew duration, which was effective to the lesion formation and plant mortality. Under dew duration of 16~24 hr with temperature range of $25^{\circ}C$ to 3$0^{\circ}C$, the weed control was increased up to 93.9%. There were no differences between the first and second or third dew treatments. A delay of 2 or 3 days in dew treatment did not increase the mortality of plants. For the use of E. nematosporus as a mycoherbicide of water chestnut, a conidial suspension should be applied when dew conditions are kept for 12 hr after inoculation.

• The Plant Pathology Journal
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• v.34 no.6
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• pp.470-479
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• 2018

The rice blast pathogen Magnaporthe oryzae is a global threat to rice production. Here we characterized RHO2 gene (MGG_02457) that belongs to the Rho GTPase family, using a deletion mutant. This mutant ${\Delta}Morho2$ exhibited no defects in conidiation and germination but developed only 6% of appressoria in response to a hydrophobic surface when compared to the wild-type progenitor. This result indicates that MoRHO2 plays a role in appressorium development. Furthermore, exogenous cAMP treatment on the mutant led to appressoria that exhibited abnormal morphology on both hydrophobic and hydrophilic surfaces. These outcomes suggested the involvement of MoRHO2 in cAMP-mediated appressorium development. ${\Delta}Morho2$ mutation also delayed the development of appressorium-like structures (ALS) at hyphal tips on hydrophobic surface, which were also abnormally shaped. These results suggested that MoRHO2 is involved in morphological development of appressoria and ALS from conidia and hyphae, respectively. As expected, ${\Delta}Morho2$ mutant was defective in plant penetration, but was still able to cause lesions, albeit at a reduced rate on wounded plants. These results implied that MoRHO2 plays a role in M. oryzae virulence as well.

• The Plant Pathology Journal
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• v.37 no.2
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• pp.87-98
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• 2021

To establish an infection, fungal pathogens must recognize diverse signals from host surfaces. The rice blast fungus, Magnaporthe oryzae, is one of the best models studying host-pathogen interactions. This fungus recognizes physical or chemical signals from the host surfaces and initiates the development of an infection structure called appressorium. Here, we found that protein MoAfo1(appressorium formation, MGG_10422) was involved in sensing signal molecules such as cutin monomers and long chain primary alcohols required for appressorium formation. The knockout mutant (ΔMoafo1) formed a few abnormal appressoria on the onion and rice sheath surfaces. However, it produced normal appressoria on the surface of rice leaves. MoAfo1 localized to the membranes of the cytoplasm and vacuole-like organelles in conidia and appressoria. Additionally, the ΔMoafo1 mutant showed defects in appressorium morphology, appressorium penetration, invasive growth, and pathogenicity. These multiple defects might be partially due to failure to respond properly to oxidative stress. These findings broaden our understanding of the fungal mechanisms at play in the recognition of the host surface during rice blast infection.

• 한국균학회소식:학술대회논문집
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• 2014.10a
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• pp.34-34
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• 2014

Filamentous fungi of the genus Colletotrichum (teleomorph, Glomerella) are considered major plant pathogens worldwide. Cereals, legumes, vegetables, and fruit trees may be seriously affected by this pathogen (1). Colletotrichum species cause typical disease symptoms known as anthracnoses, characterized by sunken necrotic tissue, where orange conidial masses are produced. Anthracnose appears in both developing and mature plant tissues (2). We investigated disease occurrence in apple orchards from 2013 to 2014 in northern Gyeongbuk province, Korea. Typical anthracnose with advanced symptoms was observed in all apple orchards studied. Of late, static fruit spot symptoms are being observed in apple orchards. A small lesion, which does not expand further and remains static until the harvesting season, is observed at the beginning of fruit growth period. In our study, static symptoms, together with the typical symptoms, were observed on apples. The isolated fungus was tested for pathogenicity on cv. 'Fuji apple' (fully ripe fruits, unripe fruits, and cross-section of fruits) by inoculating the fruits with a conidial suspension ($10^5$ conidia/ml). In apple inoculated with typical anthracnose fungus, the anthracnose symptoms progressed, and dark lesions with salmon-colored masses of conidia were observed on fruit, which were also soft and sunken. However, in apple inoculated with fungi causing static symptoms, the size of the spots did not increase. Interestingly, the shape and size of the conidia and the shape of the appressoria of both types of fungi were found to be similar. The conidia of the two types of fungi were straight and cylindrical, with an obtuse apex. The culture and morphological characteristics of the conidia were similar to those of C. gloeosporioides (5). The conidia of C. gloeosporioides germinate and form appressoria in response to chemical signals such as host surface wax and the fruitripening hormone ethylene (3). In this study, the spores started to germinate 4 h after incubation with an ethephon suspension. Then, the germ tubes began to swell, and subsequently, differentiation into appressoria with dark thick walls was completed by 8 h. In advanced symptoms, fungal spores of virtually all the appressoria formed primary hyphae within 16 h. However, in the static-symptom fungus spores, no primary hyphae formed by 16 h. The two types of isolates exhibited different growth rates on medium containing apple pectin, Na polypectate, or glucose as the sole carbon. Static-symptom fungi had a >10% reduction in growth (apple pectin, 14.9%; Na polypectate, 27.7%; glucose, 10.4%). The fungal isolates were also genetically characterized by sequencing. ITS regions of rDNA, chitin synthase 1 (CHS1), actin (ACT), and ${\beta}$-tubulin (${\beta}t$) were amplified from isolates using primer pairs ITS 1 and ITS 4 (4), CHS-79F and CHS-354R, ACT-512F and ACT-783R, and T1 and ${\beta}t2$ (5), respectively. The resulting sequences showed 100% identity with sequences of C. gloeosporioides at KC493156, and the sequence of the ${\beta}$t gene showed 100% identity with C. gloeosporioides at JX009557.1. Therefore, sequence data from the four loci studied proves that the isolated pathogen is C. gloeosporioides. We also performed random amplified polymorphic DNA-PCR, which showed clearly differentiated subgroups of C. gloeosporioides genotypes. The clustering of these groups was highly related to the symptom types of the individual strains.

• The Plant Pathology Journal
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• v.24 no.2
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• pp.112-117
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• 2008

Pre-infection behavior of Fusarium circinatum on stems of pine species was investigated with scanning electron microscopy. Two-year-old stems of Pinus densiflora and p. rigida were inoculated with the fungal conidial suspension and subjected to $25^{\circ}C$ for up to 16 hr. Most microconidia germinated 12 hr after inoculation on pine stems. Conidia produced germ tubes from either one or both ends of microconidia. Germ tubes grew over the stem surface and appeared to enter host tissues through natural openings on pine stems. Surface cracks in the cork were entrance sites of germ tubes of F. circinatum. In addition, host cell wall cracks were often found at the tip of germ tubes. The cuticle appeared to be eroded either at the tip of germ tubes or around germlings. Germ tubes also produced appressoria-like structures, exhibiting swollen tips of germ tubes on the stem surface. There seems to be no significant differences in the pre-infection behavior of F. circinatum on stems between the two pine species.

• The Plant Pathology Journal
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• v.16 no.1
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• pp.19-24
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• 2000

The direct effects of acute $\textrm{O}_3$ on the growth, sporulation and infection of Pyricularia grisea, rece blast pathogen, were investigated to understand the interactions between ozone and the pathogen. Acute exposure of 200 nl $\textrm{l}^{-1}$ ozone for 8 h significantly reduced conidia germination on water atar. Ozone exposure of 200 nl $\textrm{l}^{-1}$ for 8h per day for 5 days had no effect on increase in colony diameter, but severely damaged actively growing aerial mycelia. However, the damage to mycalia was recovered during the following 16 h exposure of unpolluted air. Conidial production was also stimulated by the acute ozone exposure for 5 days. The conidia exposed to the acute ozone for 5 days normally germinates but slightly reduce appressoria formation on rice leaf. However, the conidia produced by artificial stimulation under the same ozone concentration for 10 days showed significant reduction in appressorea for mation on a hydrophobic film. This study suggests that the acute ozone could ingibit appressoria formation as well as vegetative growth of the pathogen, resulting in decrease in rece blast development in the field during summer when high ozone episodes could occur occasionally.

• 한국균학회소식:학술대회논문집
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• 2014.10a
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• pp.43-43
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• 2014

The rice blast disease caused by of Magnaporthe oryzae is one of the most destructive diseases of rice. By the microarray analysis, we profiled expression changes of genes during conidiation and found out many putative genes that are up-regulated. Among those, we first selected MGG_06399 encoding a dual-specificity tyrosine-regulated protein kinase (DYRK), homologous to YAK1 in yeast. To investigate functional roles of MoYAK1, We made ${\Delta}Moyak1$ mutants by homology dependent gene replacement. The deletion mutant showed a remarkable reduction in conidiation and produced abnormally shaped conidia smaller than those of wild type. The conidia form ${\Delta}Moyak1$ were able to develop a germ tube, but failed to form apppressoria on a hydrophobic coverslip. The ${\Delta}Moyak1$ formed appressria on a hydrophobic cover slip when exogenous cAMP was induced, but the appressoria shape was abnormal. The ${\Delta}Moyak1$ also formed appressoria abberent in shape on onion epidermis and rice sheaths and failed to penetrate the surface of the plants. These data indicate that MoYAK1 is associated with cAMP/PKA pathway and important for conidiation, appressorial formation and pathogenic development in Magnaporthe oryzae. Detailed characterization of MoYAK1 will be presented.