• The Korean Journal of Mycology
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• v.48 no.4
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• pp.369-379
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• 2020

Black mold was frequently observed on tomatoes grown in a greenhouse in Suwon, Korea in July 2018. The incidence of the disease was 5.0-25.0% (average 12.3%) and 2.0-7.0% (average 3.7%) in the context of tomato and cherry tomato fruits, respectively. Sixteen single-spore isolates of Alternaria sp. were obtained from the diseased fruits and investigated for their morphological characteristics. Among the isolates, eight were used for sequencing analysis. All of the isolates were identified as Alternaria alternata based on their morphological and molecular characteristics. The pathogenicity of four isolates of A. alternata was investigated using three varieties each of tomato and cherry tomato via artificial inoculation. All of the isolates induced black mold symptoms on the inoculated tomato fruits. Notably, the symptoms were similar to those observed in the greenhouse. However, the pathogenicity tests revealed that different tomato varieties presented distinct patterns of susceptibility to the isolates. This is the first report of A. alternata causing black mold on tomato fruits in Korea.

• Journal of Microbiology and Biotechnology
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• v.32 no.2
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• pp.160-169
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• 2022

In the present study we evaluated the efficacy of a bioformulation of Streptomyces cameroonensis for control of black pod disease in cocoa and enhancement of seedling growth. The formulation developed using talc powder and cassava starch as carriers showed high shelf-life of 1.07 × 10⁶ CFU/g after six months storage at 4℃. The formulation was tested for inhibition of spore germination in Phytophthora megakarya and showed 100% inhibition at 10% (w/v) of formulation. To determine the efficacy of the formulation, we performed an in planta assay in the greenhouse on two hybrids of cocoa seedlings, the tolerant SNK413 × (♂) T79/467 and the susceptible UPA 134× (♂) SCA 12. Detached leaf assay showed a significant reduction in the disease severity index of about 67% for the tolerant hybrid and 55% for the susceptible hybrid compared to non-treated plants. A significant enhancement in stem length, leaf surface area and root weight was observed. Analysis of biochemical markers of defense showed a significant increase in total polyphenol, flavonoid, and total protein contents. There was also significant upregulation of PR-proteins such as chitinases, peroxidases and β-1, 3-glucanases following treatment of both tolerant and susceptible hybrids, though with a higher level of synthesis in the tolerant hybrids. A significant increase was also observed in polyphenol oxidase activities in plants treated with the formulation. This work demonstrated the stability and effectiveness of the S. cameroonensis powder formulation in suppressing black pod disease in cocoa and subsequently enhancing the growth of seedlings.

• Journal of Microbiology and Biotechnology
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• v.33 no.5
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• pp.607-620
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• 2023

The biocontrol approach using beneficial microorganisms to control crop diseases is becoming an essential alternative to chemical fungicides. Therefore, new and efficient biocontrol agents (BCA) are needed. In this study, a rhizospheric actinomycete isolate showed unique and promising antagonistic activity against three of the most common phytopathogenic fungi, Fusarium oxysporum MH105, Rhizoctonia solani To18, and Alternaria brassicicola CBS107. Identification of the antagonistic strain, which was performed according to spore morphology and cell wall chemotype, suggested that it belongs to the Nocardiopsaceae. Furthermore, cultural, physiological, and biochemical characteristics, together with phylogenetic analysis of the 16S rRNA gene (OP869859.1), indicated the identity of this strain to Nocardiopsis alba. The cell-free filtrate (CFF) of the strain was evaluated for its antifungal potency, and the resultant inhibition zone diameters ranged from 17.0 ± 0.92 to 19.5 ± 0.28 mm for the tested fungal species. Additionally, the CFF was evaluated in vitro to control Fusarium wilt disease in Vicia faba using the spraying method under greenhouse conditions, and the results showed marked differences in virulence between the control and treatment plants, indicating the biocontrol efficacy of this actinomycete. A promising plant-growth promoting (PGP) ability in seed germination and seedling growth of V. faba was also recorded in vitro for the CFF, which displayed PGP traits of phosphate solubilization (48 mg/100 ml) as well as production of indole acetic acid (34 ㎍/ml) and ammonia (20 ㎍/ml). This study provided scientific validation that the new rhizobacterium Nocardiopsis alba strain BH35 could be further utilized in bioformulation and possesses biocontrol and plant growth-promoting capabilities.

• Journal of Food Hygiene and Safety
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• v.23 no.4
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• pp.343-347
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• 2008

Recently, Sunshik has been issued because of easy-cook and well-being food. Sunshik basically was made of the heated cereals. Amount of spore-forming Bacillus cereus was detected and it has been caused some problem of food safety. B. cereus was isolate from 57 out of 161 Sunshik samples resulting in the isolation rate of 35.4%. Quantitative analysis of 57 samples showed that 21 samples were less than 100 CFU/g, 33 samples were between 100 and 1,000 CFU/g and distinctively even 3 (1.9%) samples had over 1,000 CFU/g. Typical morphology of B. cereus isolated from Sunshik was observed on MYP agar and then further characteristics was identified by using VITEK 2 (Biomeriux, France). 53 strains out of 57 strains isolated from Sunshik (about 93.0%) produced diarrheal enterotoxin in brain heart infusion broth which was detected by the Bacillus cereus enterotoxin reversed passive latex agglutination test kit (Oxoid England). The D-values of the B. cereus spores were $75^{\circ}C$ (37.1mim), $80^{\circ}C$ (22.5mim), $85^{\circ}C$ (4.9mim), and $90^{\circ}C$ (3.1mim) respectively. The Z-value was calculated $12.8^{\circ}C$ in Sunshik sample inoculated with B. cereus. Therefore, the management of B. cereus in Sunshik is required for the food-safety.

• 한국균학회소식:학술대회논문집
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• 2014.10a
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• pp.11-11
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• 2014

Fungal pathogens have huge impact on health and economic wellbeing of human by causing life-threatening mycoses in immune-compromised patients or by destroying crop plants. A key determinant of fungal pathogenesis is their ability to undergo developmental change in response to host or environmental factors. Genetic pathways that regulate such morphological transitions and adaptation are therefore extensively studied during the last few decades. Given that epigenetic as well as genetic components play pivotal roles in development of plants and mammals, contribution of microbial epigenetic counterparts to this morphogenetic process is intriguing yet nearly unappreciated question to date. To bridge this gap in our knowledge, we set out to investigate histone modifications among epigenetic mechanisms that possibly regulate fungal adaptation and processes involved in pathogenesis of a model plant pathogenic fungus, Magnaporthe oryzae. M. oryzae is a causal agent of rice blast disease, which destroys 10 to 30% of the rice crop annually. Since the rice is the staple food for more than half of human population, the disease is a major threat to global food security. In addition to the socioeconomic impact of the disease it causes, the fungus is genetically tractable and can undergo well-defined morphological transitions including asexual spore production and appressorium (a specialized infection structure) formation in vitro, making it a model to study fungal development and pathogenicity. For functional and comparative analysis of histone modifications, a web-based database (dbHiMo) was constructed to archive and analyze histone modifying enzymes from eukaryotic species whose genome sequences are available. Histone modifying enzymes were identified applying a search pipeline built upon profile hidden Markov model (HMM) to proteomes. The database incorporates 22,169 histone-modifying enzymes identified from 342 species including 214 fungal, 33 plants, and 77 metazoan species. The dbHiMo provides users with web-based personalized data browsing and analysis tools, supporting comparative and evolutionary genomics. Based on the database entries, functional analysis of genes encoding histone acetyltransferases and histone demethylases is under way. Here I provide examples of such analyses that show how histone acetylation and methylation is implicated in regulating important aspects of fungal pathogenesis. Current analysis of histone modifying enzymes will be followed by ChIP-Seq and RNA-seq experiments to pinpoint the genes that are controlled by particular histone modifications. We anticipate that our work will provide not only the significant advances in our understanding of epigenetic mechanisms operating in microbial eukaryotes but also basis to expand our perspective on regulation of development in fungal pathogens.

• Korean Journal of Microbiology
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• v.30 no.5
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• pp.339-346
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• 1992

These studies were carried out to identify Bacillus cereus group 1..5-] strain isolated from 5uyeong Bay. This strain was differentiated from B. cereus group using conventional, API system and fatty acid composition analysis. Colony characteristics were opague. mucoid, entire margin. convex. circular and non hemolysis on sheep blood agar plates, and were observed with central spore forming positive bacilli in a Gram stained preparation. and had no motility. The carbohydrates tested; glucose.maltose, and sucrose were assimilated but neither trehalose nor salicin were assimilated. This strain ultilized gelatin and was also inhibited by 6.5% NaCI. The results of biochemical examination were differented from B. cereus group LS-1 compared with others B. cereus group. The fatty acid composition contained major amounts of branched chain acids. iso $C_{15}$ and iso $C_{13}$ and the range of chain length was $C_{12}$ to C"$C_{17}$ and n$C_{15}$, acid was not detected. Automated fatty acid computer profile indicated "B. mycoides GC subgroup B of 0.312 similarity index." The results agreed with other research cases. On the other hand. A TB computer prolile index of API system (API 50 CHB & API 20E) identified" Doubtful profile of 99.7% B. firmus" . These results were presented with considerable discrepancies between API system and fatty acid analysis. With 67 biochemical characters. the similarity matrix of B. mycaides (KCTC 1033). B. thuringiensis (KCTC 1033). B. cereus (5-3) and B. mycoides (S-12) showed 42%. 42%. 59%, and 52%. respectively. Through the key tests and fatty acid analyses. we could notice the appearance of B. mycoides of the B. cereus group and this leads us to suspect the existence of a new biotype B. mycoides.

• Korean Journal of Plant Taxonomy
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• v.38 no.4
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• pp.583-599
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• 2008

Dokdo, small island located in the eastern end of the Korean Peninsula, has been an uninhabited island for long time due to long distance from the land. Moreover, the steep slope of volcanic tuff at Dokdo is well drained for high permeability and Dokdo has few plain areas, swamps and very thin soil layers. In this study, pollen analysis at Dokdo was attempted on the profile of organic sandy soil for the first time in Korea owing to the marine climate environment with high humidity and precipitation including snowfalls in spite of unprofitable condition geologically or geomorphologically. While many historical, political researches have been accumulated for territorial problem of Dokdo with Japan, natural scientific researches with field work are not sufficient, and few paleoecological researches have been done. As a result of pollen analysis, the ratio of the NAP(Non-Arboreal Pollen) and spore was higher than AP(Arboreal Pollen), and the vegetation change existed by showing dramatic decrease of AP at the upper layer. AP was composed of almost Pinus, and a little Alnus, Quercus, Betula, Carpinus, Picea and Ulmus. NAP was composed of Gramineae, Compositae, Chenopodiaceae Cyperaceae, Caryophyllaceae, Liliaceae, Umbelliferae, Artemisia, Lobelia, Rumex, Polygonum. Increase of the NAP such as Gramineae, Artemisia and Chenopodiaceae, they would mostly have been transported from the other areas because agricultural activity is impossible at Dokdo. While one reason of Pinus thunbergii decrease could be regarded as environmental change, the other reason would be lumbering of pine trees by human activity which is adapted to Dokdo environment.

• Research in Plant Disease
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• v.22 no.2
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• pp.72-80
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• 2016

Gummy stem blight, caused by the fungus Didymella bryoniae, is major disease of watermelons worldwide. The objective of the present study was to establish an efficient screening system to identify watermelon resistant to D. bryoniae. An GSB3 isolate was prepared from a watermelon plant showing typical symptoms of gummy stem blight in Haman-gun and identified as D. bryoniae based on molecular analysis of internal transcribed spacer sequence. A simple mass-production technique of inoculum was developed based on spore production of D. bryoniae GSB3 under several incubation conditions and their virulence on watermelon plants. Resistance degrees of 22 commercial watermelon cultivars to the GSB3 isolate were evaluated. Among them, four watermelon cultivars showing different degree of resistance response were selected for further study. Development of disease on the cultivars according to various conditions including inoculum concentrations, incubation periods in dew chamber, and incubation temperatures was investigated. From the results, we suggest an efficient screening method for resistant watermelon cultivars to gummy stem blight. Seeds of watermelon cultivar are sown and grown in a greenhouse until plant stage of 2-fully expanded leaves. Seedlings are inoculated with D. bryoniae by spraying spore suspension of the fungus at a concentration of $5.0{\times}10^5spores/ml$. The infected plants are incubated in humidity chamber at $25^{\circ}C$ for 48 hours and then transferred to a growth chamber at $25^{\circ}C$ and 80% relative humidity with 12-hour light a day. Three to four days after inoculation, disease severity of the plant are measured using percentage of infected leaf area.

• Research in Plant Disease
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• v.18 no.3
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• pp.201-209
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• 2012

The multi-function of 18 Bacillus subtilis isolates collected from agricultural extension centers of local government and National Academy of Agricultural Science was investigated by measuring their antifungal activities against five plant pathogens, such as Rhizoctonia solani, Colletotrichum acutatum, Fusarium oxysporum, Magnaporthe oryzae and Phytophthora capsici, phosphorus solubilization ability, production of indole acetic acid (IAA) and siderophore, and nitrogen fixation. The B. subtilis isolates showed antifungal activity against several plant pathogens and nitrogen fixation activity, and produced siderophore and IAA. They could control pepper powdery mildew (Leveillula taurica), but there was no difference in control efficacy among the B. subtilis isolates. In fields, the control efficacy of B. subtilis R2-1 ($10^8$ cells/ml) was compared with two microbial fungicides, Q-pect and Topsid. In 2009, the control efficacy of B. subtilis R2-1 (37.7%) was lower than that of Topsid (47.6%), but higher than that of Q-pect (25.7%). In 2010, the control efficacy of B. subtilis R2-1 (83.3%) was higher than that of Topsid (67.9%). In order to elucidate mode of action of B. subtilis R2-1 for controlling pepper powdery mildew, spore germination rates of pepper powdery mildew pathogen collected on treated leaves was investigated when suspensions of B. subtilis R2-1 and two microbial fungicides (Q-pect and Topsid) were foliar-sprayed. They highly suppressed spore germination of the pathogen with inhibition values of 84.2% for B. subtilis R2-1, 97.9% for Q-pect and 94.7% for Topsid. Further study on the mass-culturing method and formulation is needed for development of a microbial fungicide.

• Korean Journal of Plant Taxonomy
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• v.39 no.1
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• pp.12-23
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• 2009

To verify hybridity of Asplenium castaneo-viride, external morphology, spore morphology, anatomy and chromosomes of the species and of the two presumed parental species, A. incisum and A. ruprechtii, were examined. A. castaneo-viride usually had 1-pinnately divided frond. However, some individuals had almost simple fronds with pinnatisect basal parts similar to A. ruprechtii, while others had fronds similar to A. incisum in having oblanceolate blades and basal pinnae with triangular, 2-3 lobed apices. On the surface of the spores, sculpturing consisted of folds that were usually prominent; forming long wings, and irregular or incomplete reticulation. However, reticulation patterns varied among species. A. castaneo-viride showed a wide range of variation from sparse to dense patterns, whereas A. incisum showed only from sparse to intermediate patterns. A. ruprechtii showed from intermediate to dense patterns. The spore size of A. castaneo-viride was $54.63{\mu}m$, larger than other two species ($47.81{\mu}m$ in A. incisum and $44.22{\mu}m$ in A. ruprechtii). The level of undulation of epidermal cell wall was also different. A. incisum had the most shallowly undulated wall, and A. castaneo-viride had a pattern intermediate between the two presumed parental species. This same patterns was recognized in the density of stomata. The density of $45.91/mm^2$ in A. castaneo-viride was intermediate between the two presumed parental species ($67.00/mm^2$ in A. incisum, and $37.86/mm^2$ in A. ruprechtii). Chromosome number was constant (2x =2n = 72) as in A. incisum and A. ruprechtii. However, A. castaneo-viride showed a different ploidy level. The populations of Mt. Mai (Jeonbuk province) and Mt. Duryun (Jeonnam province) were diploid (2n = 72) which is a new record for this taxon, whereas the population of Mt. Buram (Seoul) was tetraploid (2n = 144). Conclusively, A. castaneo-viride was revealed to be a hybrid of A. ruprechtii and A. incisum based on evidence involving leaves, spores, epidermal cells, stomata and chromosome number.