• The Plant Pathology Journal
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• v.35 no.6
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• pp.564-574
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• 2019

Like many fungal pathogens, the conidium and appressorium play key roles during polycyclic dissemination and infection of Magnaporthe oryzae. Ran-binding protein microtubule-organizing center (RanBPM) is a highly conserved nucleocytoplasmic protein. In animalia, RanBPM has been implicated in apoptosis, cell morphology, and transcription. However, the functional roles of RanBPM, encoded by MGG_00753 (named MoRBP9) in M. oryzae, have not been elucidated. Here, the deletion mutant ΔMorbp9 for MoRBP9 was generated via homologous recombination to investigate the functions of this gene. The ΔMorbp9 exhibited normal conidial germination and vegetative growth but dramatically reduced conidiation compared with the wild type, suggesting that MoRBP9 is involved in conidial production. ΔMorbp9 conidia failed to produce appressoria on hydrophobic surfaces, whereas ΔMorbp9 still developed aberrantly shaped appressorium-like structures at hyphal tips on the same surface, suggesting that MoRBP9 is involved in the morphology of appressorium-like structures from hyphal tips and is critical for development of appressorium from germ tubes. Taken together, our results indicated that MoRBP9 played a pleiotropic role in polycyclic dissemination and infection-related morphogenesis of M. oryzae.

• Korean Journal Plant Pathology
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• v.3 no.2
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• pp.100-107
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• 1987

Pre-pentration activity of Pyricularia oryzae Cav. from the stage of conidia germination to appressorium formation was examined on rice leaf epidermis under light and scanning electron microscopes to determine the causes· for differences in blast susceptibility between plants grown under three different soil moisture conditions in the greenhouse. No significant differences were found in the external shape of leaf epidermal cells including bulliform cells between plants grown under different soil moisture conditions. Growth and orientation of germ tube and morphology and size of appressorium of P. oryzae did not vary with soil moisture treatment. Site of appressorium formation was consistent over soil moisture treatment with the highest frequency of bulliform cell $(35\~48\%)$, followed by short cell $(19\~27\%)$, and long and guard cells $(13\~20\%)$. No appressorium was formed on trichome. This result suggests that the observed differences in blast susceptibility between plants grown under different soil moisture conditions were not due to the differences in the pre-pentration activity of P. oryzae on those plants.

• 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.38 no.4
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• pp.345-354
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• 2022

NADPH oxidase (Nox) complexes are known to play essential roles in differentiation and proliferation of many filamentous fungi. However, the functions of Noxs have not been elucidated in Colletotrichum species. Therefore, we set out to characterize the roles of Nox enzymes and their regulators in Colletotrichum scovillei, which causes serious anthracnose disease on pepper fruits in temperate and subtropical and temperate region. In this study, we generated targeted deletion mutants for CsNox1, CsNox2, CsNoxR, and CsNoxD via homologous recombination. All deletion mutants were normal in mycelial growth, conidiation, conidial germination, and appressorium formation, suggesting that CsNox1, CsNox2, CsNoxR, and CsNoxD are not involved in those developmental processes. Notably, conidia of 𝜟Csnox2 and 𝜟Csnoxr, other than 𝜟Csnox1 and 𝜟Csnoxd, failed to cause anthracnose on intact pepper fruits. However, they still caused normal disease on wounded pepper fruits, suggesting that Csnox2 and CsnoxR are essential for penetration-related morphogenesis in C. scovillei. Further observation proved that 𝜟Csnox2 and 𝜟Csnoxr were unable to form penetration peg, while they fully developed appressoria, revealing that defect of anthracnose development by 𝜟Csnox2 and 𝜟Csnoxr resulted from failure in penetration peg formation. Our results suggest that CsNox2 and CsNoxR are critical for appressorium-mediated penetration in C. scovillei-pepper fruit pathosystem, which provides insight into understanding roles of Nox genes in anthracnose disease development.

• The Korean Journal of Pesticide Science
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• v.18 no.2
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• pp.115-121
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• 2014

This study was performed to investigate the effect of conidial density, wetness period and temperature on conidial germination, appressoria formation and disease incidence. While there was not significantly correlated between conidial density and temperature, and conidial germination and appressoria formation, there was a significant correlation between those factors and disease incidence. The longer wetness period was, the higher the ratio of conidial germination, appressoria formation and the disease incidence was. The optimum conidial density, temperature and wetness period was $1{\times}10^6$ conidia $mL^{-1}$, $30^{\circ}C$ and 5 days, respectively. In case the wetness period was more than 5 days, the typical symptom was not found on pepper fruits because of the overgrowth of mycelia. Using this fruit assay method, which the pepper anthracnose pathogens were inoculated by spraying spore suspension on non-wounded or wounded pepper fruits, control effect of three fungicides were evaluated against pepper anthracnose by the protective and/or the curative application. Propineb showed high protective control activity, while it showed curative control activity on unwounded fruits, but did not showed curative control activity on wounded fruits. Tebuconazole, one of curative fungicide, showed higher control activity in non-wound inoculation than wound inoculation. Trifloxystrobin, one of strobilurin group, showed high both protective and control activity against anthracnose. In conclusion, we supposed that the newly developed in vitro pepper fruit assay can be used to evaluate antifungal activity of control agents against pepper anthracnose.

• Korean Journal of Agricultural Science
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• v.15 no.1
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• pp.9-14
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• 1988

This experiment was conducted to study the effect of some environmental factors on mycerial growth, sporulation and sclerotial formation of Botrytis elliptica cultured on artificial media. Mycerial growth of B. elliptica was the most favorable on V-8 juice agar among the seven different media tested and sporulation of the fungus was favorable on the medium under NUV light irradiation. Abundant conidia could be obtained from V-8 juice agar medium by NUV light irradiation after 3 days of incubation at $23^{\circ}C$ under darkness. The optimum temperature for mycerial growth and conidial germination was $23^{\circ}C$ and the mycerial growth was favorable at relatively lower temperature ranged $19^{\circ}C$ to $23^{\circ}C$. The optimum pH of the medium for mycerial growth of this fungus ranged 4.5 to 5.0 and that was inhibited at higher pH of the media. Mycerial growth of the fungus was not highly influenced by irradiation of fluorescent light, however sporulation was stimulated under NUV light irradiation. Sclerotia of B. elliptica were formed when it was cultured at lower temperature below $19^{\circ}C$.

• The Plant Pathology Journal
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• v.23 no.3
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• pp.174-179
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• 2007

Seedlot disinfection techniques to control mung bean sprout rot caused by Colletoricum acutatum and C. gloeosporioides were evaluated for commercial production scheme. Soaking seedlots in propolis (100 X) and ethanol (20% for 30 min) appeared promising with control values of 85.5 and 80.8 respectively, but still resulted in up to 20% rot incidence. None of the C. acutatum conidia survived through hot water immersion treatment (HWT) for 10 min at temperatures of 55, 60 and $65^{\circ}C$, whereas the effective range of the dry heat treatment (DHT) was $60-65^{\circ}C$. Tolerance of mung bean seedlot, as estimated by hypocotyl elongation and root growth, was lower for HWT than for DHT. Germination and growth of sprouts were excellent over the range of $55-65^{\circ}C\;at\;5^{\circ}C$ intervals, except for HWT at $65^{\circ}C$ for 5 min. At this marginal condition, heat damage appeared so that approximately 2% of seeds failed to sprout to normal germling and retarded sprouts were less than 5% with coarse wrinkled hypocotyls. These results suggested that DHT would be more feasible to disinfect mung bean seedlots for commercial sprout production. Heat treatment at above ranges was highly effective in eliminating the epiphytic bacterial strains associated with marketed sprout rot samples. HWT of seedlot at 55 and $60^{\circ}C$ for 5 min resulted in successful control of mung bean sprout rot incidence with marketable sprout quality. DHT at 60 and $65^{\circ}C$ for 30 min also gave good results through the small-scale sprouting system. Therefore, we optimized DHT scheme at 60 and $65^{\circ}C$ for 30 min, considering the practical value of seedlot disinfection with high precision and accuracy. This was further proved to be a feasible and reliable method against anthracnose incidence and those bacterial strains associated with marketed sprout rot samples as well, through factory scale mung bean sprout production system.

• Microbiology and Biotechnology Letters
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• v.35 no.2
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• pp.128-134
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• 2007

Previously, we isolated plant growth promoting rhizobacterium (PGPR) Bacillus licheniformis K11 which could produce auxin, cellulase and siderophore. The siderophore of B. licheniformis K11 $(siderophore_{K11})$ was determined to be a catechol type siderophore which is produced generally by Bacillus spp. B. licheniformis K11 could produce the siderophore most highly after 96 h of incubation under nutrient broth at $20^{\circ}C$ with initial pH 9.0. For the production of the $siderophore_{K11}$, trehalose and $NH_4Cl$ were the best carbon and nitrogen sources in Davis minimal medium, respectively. The $siderophore_{K11}$ was Produced in M9 medium (pH 9.0) after 4 days at $20^{\circ}C$, and purified from culture broth of B. licheniformis K11 by using Amberlite XAD-2, Sephadex LH-20 column chromatography, and reversed-phase HPLC. The $siderophore_{K11}$ had the biocontrol activity against spore germination of P. capsici and F. oxysporum on potato dextrose agar (PDA). The results indicate that the $siderophore_{K11}$ is an antifungal mechanism of B. licheniformis K11 against phytopathogenic fungi.

• Korean Journal of Agricultural Science
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• v.19 no.2
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• pp.170-178
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• 1992

Isolates of Botrytis cinerea resistant to dicarboximide fungicides were collected from strawberry fields in greenhouses in spring and early summer of 1990. Five out of 9 isolates of B. cinera were resistant, which showed mycerial growth on PDA containing dicarboximide fungicides(procymidone and vinclozolin) with concentrations of 100, 400 and $1,600{\mu}g/ml$. The minimal inhibitory concentration(MIC) values of the dicaboximide-resistant isolates was more than $6,400{\mu}g/ml$, while that of the sensitive isolates was less than $6.25{\mu}g/ml$. The germination ratio of conidia of the resistant isolates on PDA containing procymidone and vincolozolin was more than 95%, while that of the sensitive was less than 15%. The procymidone-resistant isolates were also resistant to vinclozolin, showing cross-resistant between the fungicides, but cross-resistant was not observed between the dicarboximides and dichlofluanid. Resistance to benomyl was also found in all the dicarboximide resistant isolates. Occurrence frequency of dicarboximide-resistant isolates out of 223 isolates was about 40%. The resistant isolates were widely distributed throughout Korea.

• 한국균학회소식:학술대회논문집
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• 2016.05a
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• pp.13-13
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• 2016

Insects constitute the largest and most diverse group of animals in the world. They also serve as the hosts or nutrient sources for an immense assemblage of pathogens, parasites, and predators. More than 700 fungal species from 100 genera have adopted an entomopathogenic lifestyle. Although entomopathogenic fungi were studied as only biocontrol agents against a variety of pests in various countries, it has been recently focused their additional roles in nature. They are antagonists to/against plant pathogens, endophytes, and possibly even plant growth promoting agents. The potential antimicrobial effect against fungal plant pathogens by an isolate of entomopathogenic fungi including Beauveria bassiana, Lecanicillium spp., and Isaria fumosorosea have been reported since late 1990s, but wasn't reported pathogenicity of the isolate against pests. Later, a Canadian Lecanicillium sp. isolate and L. longisporium isolated from Vertalec$^{(R)}$ showed simultaneous control effect against both aphid and cucumber powder mildew. Therefore, the antimicrobial activities of 342 fungi isolates collected from various regions and conditions in Korea were evaluated against plant pathogenic fungus Botrytis cinerea using dual culture technique on agar plate. As a result, 186 isolates (54.4%) shown the antifungal activity against B. cinerea. The culture filtrates of selected fungi completely suppressed the growth of the microorganisms, indicating that suppression was due to the presence of antimicrobial substances in the culture filtrate. Mode of action of these fungi against insect involves the attachment of conidia to the insect cuticle, followed by germination, cuticle penetration, and internal dissemination throughout the insect. During infection process, secreted enzymes, proteinous toxins, and/or secondary metabolites secreted by entomopathogenic fungi can be used to overcome the host immune system, modify host behavior, and defend host resources. Recently, secondary metabolites isolated from entomopathogenic fungi have been reported as potential bioactive substances. Generally, most of bioactive substances produced by entomopathogenic fungi have reported low molecular weight (lower than 1,000 g/mol) as peptide and, in contrast the high molecular weight fungal bioactive substances are rare. Most substances based on entomopathogenic fungi were shown antimicrobial activity with narrow control ranges. In our study we analyzed the antimicrobial substances having antagonistic effects to B. cinerea. Antimicrobial substances in our fungal culture filtrates showed high thermostability, high stability to proteolytic enzymes, and hydrophilicity and their molecular weights were differed from substance. In conclusion, entomopathogenic fungi showed pathogenicity against insect pests and culture filtrate of the fungi also shown to antimicrobial activity. In the future, we can use the entomopathogenic fungi and its secondary metabolites to control both insect pest control and plant pathogenic fungi simultaneously.