• Korean Journal Plant Pathology
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• v.13 no.4
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• pp.255-256
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• 1997

The mass sporulating method for conidia of Mycosphaerella nawac the causal organism of the spotted leaf casting of persimmon was investigated in this experiment. The conidia of M. nawae were sporulated on artificial media after prolonged period of incubation. The maximum amount of conidia of $39.0{\times}10^4/ml$ was harvested from 90-day old culture on PDA at $25^{\circ}C.$

• Korean Journal Plant Pathology
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• v.14 no.5
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• pp.393-396
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• 1998

The characteristic Ramularia type conidia of Mycosphaerella nawae were formed on naturally infected leaves. Artificial inoculation with the conidia induced typical symptom on leaves, which was not distinguishable from those of ascospore infection, which has been considered as a primary inoculum source in nature. Also the morphology of the conidia produced on PDA was not different from those formed on artificially inoculated leaves or on naturally infected leaves at later stage of symptom development. Accordingly, we report the role of the conidia as a secondary inoculum of the circular leaf spot pathogen of persimmon for the first time.

• Korean Journal Plant Pathology
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• v.14 no.5
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• pp.397-401
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• 1998

Asexual spores of Mycosphaerella nawae were profusely produced on PDA after a prolonged incubation at $25^{\circ}C$ for 90 days. When persimmon trees were artificially inoculated by the conidial suspension, typical symptoms of circular leaf spot of persimmon appeared on the leaves two month later. The imperfect stag of the fungus was identified as Ramularia sp. based on following morphological characteristics examined under a light microscope and a scanning electron microscope. Conidia were mostly ellipsoid, but occasionally cylindrical, elongated oval, taro, peanut or gourd shapes and measured as 12.2~32.6$\times$6.1~10.2 ${\mu}{\textrm}{m}$. erect, hyaline, colorless-light brown. Conidia were formed solitarily or in chains on a medium and infected leaves. Conidiophore was erect, hyaline, colorless-light brown. and the size was 20.4~102.0$\times$3.1~10.2 ${\mu}{\textrm}{m}$, respectively. In this paper, we firstly demonstratrated that asexual spores of M. nawae induced persimmon circular leaf spot in nature as well as sexual spores of the fungus. Therefore, it is hypothesized that the imperfect stage of the fungus plays an important role in nature for epidemics as secondary inoculum.

• The Plant Pathology Journal
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• v.30 no.3
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• pp.229-235
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• 2014

Pear skin stains on 'Niitaka' pears, which occur from the growing stage to the cold storage stage, reportedly negatively influence the marketing of pears. These stains on fruit skin are likely due to a pathogenic fungus that resides on the skin and is characterized by dark stains; however, the mycelium of this fungus does not penetrate into the sarcocarp and is only present on the cuticle layer of fruit skin. A pathogenic fungus was isolated from the skin lesions of infected fruits, and its pathogenicity was subsequently tested. According to the pathogenicity test, Mycosphaerella sp. was strongly pathogenic, while Penicillium spp. and Alternaria spp. showed modest pathogenicity. In this present study, we isolated the pathogenic fungus responsible for the symptoms of pears (i.e., dark brown-colored specks) and identified it as Mycosphaerella graminicola based on its morphological characteristics and the nucleotide sequence of the beta-tubulin gene. M. graminicola was pathogenic to the skin of 'Niitaka' pears, which are one of the most widely growing varieties of pears in South Korea.

• Plant Disease and Agriculture
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• v.1 no.2
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• pp.18-21
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• 1995

Mycosphaerella nawae, the causal organism, of spotted leaf casing disease of persimmon, was isolated from infected leaves showing typical symptom. The cultural characteristics of the fungus were compared on artificial media. Among 24 different combinations of culture media and supplements, oatmeal agar+persimmon leaf extract (PLE) and PAD+ PLE+streptomycin showed the highest rate of isolating as 57.1%. The best medium for mycelial growth of the fungus was PDA+persimmon leaf powder (PLP). The colony diameter was reached 47mm for 30 days at 2$0^{\circ}C$. PDA+PLE also supported good mcyelial growth showing 46mm of diameter in same condition. The optimum growth temperature of this fungus in PDA was recognized fairly low. The mycelial growth was higher at 2$0^{\circ}C$ than 15$^{\circ}C$. The variation of pH between pH 4 to pH 8 did not affect to the mycelial growth of the pathogen.

• Korean Journal Plant Pathology
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• v.14 no.5
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• pp.408-412
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• 1998

In order to illustrate the role of conidia of Mycosphaerella nawae as a secondary inoculum in nature, pseudothecial development on persimmon leaves was investigated microscopically. The fungal ascospores have been believed as the primary or only inoculum source in nature, however, pseudothecia were readily formed on persimmon leaves infected naturally and artificially by conidia. The pseudothecia of M. nawae were found to form in the tissues of infected leaves while the leaves were still hanging on the trees. The size of pseudothecia were approximately 51.0~122.4$\times$51.0~112.2 ${\mu}{\textrm}{m}$ (82.8 $\times$72.5 ${\mu}{\textrm}{m}$in average), the shapes were spherical, ovoid or occidental pear type. The sizes of asci were approximately 30.6~61.2$\times$8.2~10.2 ${\mu}{\textrm}{m}$(46.6$\times$9.4 ${\mu}{\textrm}{m}$ in average) and the shapes were cylinder or banana. The ascospores were mostly spindle type, and the sizes were 10.2~12.2$\times$3.1~4.1 ${\mu}{\textrm}{m}$ (11.4$\times$3.2 ${\mu}{\textrm}{m}$ in average)-like. The pseudothecial formation was initiated before defoliation and morphological characteristics of the pseudothecia, ascus and ascospores on the infected leaves were fully illustrated in this study. Results indicated that conidia of M. nawae induce circular leaf spot of persimmon as much as ascospores, and might play an important role of the disease epidemics in nature.

• Mycobiology
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• v.30 no.3
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• pp.170-174
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• 2002

A total of twenty-nine species and one species variety belonging to 12 genera was isolated from 30 samples of molasses on 1% glucose(10 genera, 22 species and 1 variety) and 50% sucrose(7, 21 and 1) Czapek's agar at $25^{\circ}C$ media. Aspergillus, Mucor, Mycosphaerella and Penicillium were the most common genera on the two types of media. From the above genera, the most prevalent species were: Aspergillus flavus, A. fumigatus, A. niger, Mycosphaerella tassiana, Penicillium chrysogenum, P. oxalicum and P. purpurogenum. Also, some species were only isolated on 50% sucrose such as Eurotium amstelodami, E. chevalieri, E. repens, Humicola fuscoatra, Penicillium aurantiogriseum and P. puberulum. About 65 fungal isolates isolated from 50% sucrose agar were tested for their ability to produce invertase enzyme in liquid medium and 93.8% of the isolates could produce this enzyme. From the positive isolates, 32 showed high invertase activity, 21 had moderate activity and the remaining 8 isolates were of weak activity. Sixty isolates of Aspergillus, Emericella, Eurotium, Mycosphaerella and Penicillium from the preceding study were screened for the presence of their respective mycotoxins. Larva of brine shrimp(Artema sauna L.) were used for toxicity test of the fungal crude extracts. Three isolates out of 60 tested were toxic. Using thin-layer chromatographic technique, 5 different known mycotoxin were detected aflatoxins : B1, B2, G1, G2 and citrinin.

• Korean Journal Plant Pathology
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• v.11 no.4
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• pp.344-347
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• 1995

Factors affecting on the ascospore release of Mycosphaerella nawae, the causal organism of persimmon leaf casting disease, were investigated. The ascospore release of the pathogen occurred following the precipitation of rain. The ascospore release started from the beginning of raining, reached maximum at 2 hours after the precipitation of rain, and then decreased abruptly. When the inoculum source (a file of infected leaves) was submerged in water to imitate raining conditions, 92.5% of the total ascospores were released within 1 hour after submerging, 5.8% were after 2 hours, 1.4% were after 4 hours, 0.1% were after 8 hours, and none detected after 10 hours. The inoculum source overwintered in the field released ascospores much more and earlier than the inoculum source kept in the greenhouse. The first ascospore release was about 10 days earlier, and the amount of the total liberated spores was 3∼4 times higher in the field inoculum source than the greenhouse nioculum source. The early defoliated leaves (in early October) in the previous year produced ascospores twice more than the late defoliated leaves (in early November) produced.

• Mycobiology
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• v.44 no.1
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• pp.38-47
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• 2016

The ascomycete fungus Mycosphaerella graminicola (synonym Zymoseptoria tritici) is an important pathogen of wheat causing economically significant losses. The primary nutritional mode of this fungus is thought to be hemibiotrophic. This pathogenic lifestyle is associated with an early biotrophic stage of nutrient uptake followed by a necrotrophic stage aided possibly by production of a toxin or reactive oxygen species (ROS). In many other fungi, the genes CREA and AREA are important during the biotrophic stage of infection, while the NOXa gene product is important during necrotrophic growth. To test the hypothesis that these genes are important for pathogenicity of M. graminicola, we employed an over-expression strategy for the selected target genes CREA, AREA, and NOXa, which might function as regulators of nutrient acquisition or ROS generation. Increased expressions of CREA, AREA, and NOXa in M. graminicola were confirmed via quantitative real-time PCR and strains were subsequently assayed for pathogenicity. Among them, the NOXa over-expression strain, NO2, resulted in significantly increased virulence. Moreover, instead of the usual filamentous growth, we observed a predominance of yeast-like growth of NO2 which was correlated with ROS production. Our data indicate that ROS generation via NOXa is important to pathogenicity as well as development in M. graminicola.

• Plant Disease and Agriculture
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• v.3 no.2
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• pp.12-15
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• 1997