• Mycobiology
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• v.51 no.3
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• pp.122-138
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• 2023

Elsinoë are plant pathogenic fungi that cause scabs, spotted anthracnose, and some morphological distortions on various plants, including woody plants, economically important crops, and ornamental plants. Taxonomical reexamination of Elsinoë species in Japan has not yet been conducted based on the modern species criteria. In this study, several Japanese isolates were reexamine based on the morphological and molecular-phylogenetic analysis of the internal transcribed spacer region (ITS), large subunit gene (LSU)m and protein-coding gene such as RNA polymerase II subunit (rpb2) and Translation elongation factor 1-alpha (tef). Japanese isolates were divided into four clades and three new species, Elsinoë hydrangeae, E. sumire, and E. tanashiensis were proposed. One species, Sphaceloma akebiae, was transferred to the genus Elsinoë.

• Microbiology and Biotechnology Letters
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• v.49 no.3
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• pp.403-412
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• 2021

This study aimed to identify plant growth-promoting activity, phytopathogenic fungi growth inhibitory activity, mineral solubilization ability, and extracellular enzyme activity of the genus Bacillus in soil and the rhizosphere. With regards to antifungal activity against phytopathogenic fungi, DDP257 showed antifungal activity against all 10 pathogenic fungi tested. ANG20 showed the highest ability to produce indole-3-acetic acid, a plant growth-promoting factor (70.97 ㎍/ml). In addition, 10 species were identified to have 1-aminocyclopropane-1-carboxylate deaminase production ability, and most isolates showed nitrogen fixation and siderophore production abilities. Thereafter, the isolated strains' ability to solubilize minerals such as phosphate, calcite, and zinc was identified. With extracellular enzyme activity, the activity appeared in most enzymes. In particular, all the strains showed similar abilities for alkaline phosphatase, esterase (C4), acid phosphatase, and naphtol-AS-BI-phosphohydrolase production. This result was observed because the genus Bacillus secreted various organic substances, antibiotics, and extracellular enzymes. Therefore, through the results of this study, we suggest the possibility of using strains contributing to the improvement of the soil environment as microbial agents.

• The Plant Pathology Journal
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• v.20 no.3
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• pp.184-191
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• 2004

Methanol extracts of fresh materials of 57 plants were screened for in vivo antifungal activity against Magna-porthe grisea, Corticium sasaki, Botrytis cinerea, Phyto-phthora infestans, Puccinia recondita, and Blumeria graminis f. sp. hordei. Among them, seven plant extracts showed disease-control efficacy of more than 90％ against at least one of six plant diseases. None of the plant extracts was highly active against tomato gray mold. The methanol extracts of Chloranthus japonicus (roots) (CjR) and Paulownia coreana (stems) (PcS) displayed the highest antifungal activity; the CjR extract controlled the development of rice blast, rice sheath blight, and wheat leaf rust more than 90％, and tomato gray mold and tomato late blight more than 80％. The PcS extract displayed control values of more than 90 ％ against rice blast, wheat leaf rust, and barley powdery mildew and more than 80％ against tomato gray mold. The extract of PcS also had a curative activity against rice sheath blight and that of CjR had a little curative activity against rice blast. On the other hand, the extract of Rumex acetocella roots reduced specifically the development of barley powdery mildew. Further studies on the characterization of antifungal substances in antifungal plant extracts are underway and their disease-control efficacy should be examined under greenhouse and field conditions.

• The Plant Pathology Journal
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• v.27 no.4
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• pp.342-348
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• 2011

Bacillus subtilis EBS05, an endophytic bacteria strain isolated from a medicinal plant Cinnamomum camphor, can produce antagonistic compounds that effectively inhibit plant pathogenic fungi. The greenhouse experiments showed that wheat sharp eyespot disease (WSED) was reduced by 91.2%, 88.2% and 43.0% after the treatment with fermentation broth, bacteria-free filter and a fungicide fludioxonil, respectively. The culture broth of strain EBS05 can more effectively control WSED than can fludioxonil. The fermentation broth and bacteria-free filter ability to suppress WSED was not significantly different, suggesting that an active secreted substance played a major role in controlling WSED. Separation and purification of the active compounds was carried out by serial processes, including hydrochloric acid (pH 2.0) treatment, methanol extraction and Sephadex LH-20 column chromatography, silica gel column chromatography and reverse-phase high-pressure liquid chromatography (HPLC), respectively. The purified compounds, one of active peaks in the HPLC spectrum, were obtained from the collection. Analysis of the chemical structures by time-of-flight mass spectrometry (TOF-MS) and electrospray ionization mass spectrometry/mass spectrometry (ESI-MS/MS) showed that the active substances produced by the endophytic bacteria EBS05 are mixture of the ${\beta}$-hydroxy-C12~C15-$Leu^7$ surfactin A isomers with 1035.65 Da, 1021.64 Da, 1007.63 Da and 993.65 Da molecular weights, respectively.

• The Plant Pathology Journal
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• v.31 no.2
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• pp.108-114
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• 2015

Curvularia lunata is an important maize foliar fungal pathogen that distributes widely in maize growing area in China, and several key pathogenic factors have been isolated. An yeast two-hybrid (Y2H) library is a very useful platform to further unravel novel pathogenic factors in C. lunata. To construct a high-quality full length-expression cDNA library from the C. lunata for application to pathogenesis-related protein-protein interaction screening, total RNA was extracted. The SMART (Switching Mechanism At 5' end of the RNA Transcript) technique was used for cDNA synthesis. Double-stranded cDNA was ligated into the pGADT7-Rec vector with Herring Testes Carrier DNA using homologous recombination method. The ligation mixture was transformed into competent yeast AH109 cells to construct the primary cDNA library. Eventually, a high qualitative library was successfully established according to an evaluation on quality. The transformation efficiency was about $6.39{\times}10^5$ transformants/$3{\mu}g$ pGADT7-Rec. The titer of the primary cDNA library was $2.5{\times}10^8cfu/mL$. The numbers for the cDNA library was $2.46{\times}10^5$. Randomly picked clones show that the recombination rate was 88.24%. Gel electrophoresis results indicated that the fragments ranged from 0.4 kb to 3.0 kb. Melanin synthesis protein Brn1 (1,3,8-hydroxynaphthalene reductase) was used as a "bait" to test the sufficiency of the Y2H library. As a result, a cDNA clone encoding VelB protein that was known to be involved in the regulation of diverse cellular processes, including control of secondary metabolism containing melanin and toxin production in many filamentous fungi was identified. Further study on the exact role of the VelB gene is underway.

• Applied Biological Chemistry
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• v.42 no.4
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• pp.271-276
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• 1999

Several Streptomyces strains were tested for potent antifungal agents active against phytopathogenic fungi. Among the tested, S. hygroscopicus MJM1004 showed a potent antifungal activity when assayed using Candida albicans as indicator organism. With the strain of MJM1004, fermentation medium for the production of an antifungal agent was developed with varying carbon sources, nitrogen sources, and mineral elements, which resulted in the highest productivity in the medium containing 2% soybean meal, 1% glucose, 2% starch, 0.3% $CaCO_3$, 0.05% $MgSO_4{\cdot}7H_2O$, 0.05% $K_2HPO_4$. The active compound showed a broad spectrum of antifungal activity against several plant pathogenic fungi. The antifungal compound was purified and showed the physicochemical characteristics similar to azalomycin F complex in NMR and MS analysis.

• Journal of Microbiology and Biotechnology
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• v.32 no.4
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• pp.504-513
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• 2022

Chitin deacetylase (CDA) inhibitors were developed as novel antifungal agents because CDA participates in critical fungal physiological and metabolic processes and increases virulence in soil-borne fungal pathogens. However, few CDA inhibitors have been reported. In this study, 150 candidate CDA inhibitors were selected from the commercial Chemdiv compound library through structure-based virtual screening. The top-ranked 25 compounds were further evaluated for biological activity. The compound J075-4187 had an IC50 of 4.24 ± 0.16 µM for AnCDA. Molecular docking calculations predicted that compound J075-4187 binds to the amino acid residues, including active sites (H101, D48). Furthermore, compound J075-4187 inhibited food spoilage fungi and plant pathogenic fungi, with minimum inhibitory concentration (MIC) at 260 ㎍/ml and minimum fungicidal concentration (MFC) at 520 ㎍/ml. Therefore, compound J075-4187 is a good candidate for use in developing antifungal agents for fungi control.

• Journal of Microbiology and Biotechnology
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• v.26 no.10
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• pp.1696-1700
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• 2016

In order to discover plant-derived signaling pathway inhibitors with antifungal properties, a two-component screening system utilizing the calcineurin and Hog1 mitogen-activated protein kinase pathways responsible for the virulence networks of Cryptococcus neoformans was employed, owing to the counter-regulatory actions of these pathways. Of the 1,000 plant extracts tested, two bioactive compounds from Miliusa sinensis were found to act specifically on the calcineurin pathway of C. neoformans. These compounds, identified as pashanone and 5-hydroxy-6,7-dimethoxyflavanone, exhibited potent antifungal activities against various human pathogenic fungi with minimum inhibitory concentration values ranging from 4.0 to >128 μg/ml.

• Korean Journal of Medicinal Crop Science
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• v.14 no.4
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• pp.202-205
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• 2006

Ginseng is major medicinal plant in Korea. Because of its long cultivation period the yield losses of 5 years of ginseng is 50% due to various diseases. The objective of this study is to select potential biocontrol agents. As the result of research so far achieved to contribute to rational prevention of ginseag plant disease for the stable cultivation of ginseng, three bacterial strains, Streptomyces lauretii strain B8180, Bacillus subtilis strain 8856, and Burkholderia cepacia strain 7944 were isolated from oak leaf compost. The strains showed antagonistic activities against five ginseng pathogenic fungi (Cylindrocarpon destructans, Rhizoctonia solani, Phytophthora cactorum, Botrytis cinerea, Fusarium solani f. sp. panacis) and control effects on Phytophthora blight.

• Mycobiology
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• v.45 no.4
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• pp.370-378
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• 2017

Cylindrocarpon destructans is an ascomycete soil-borne pathogen that causes ginseng root rot. To identify effective biocontrol agents, we isolated several bacteria from ginseng cultivation soil and evaluated their antifungal activity. Among the isolated bacteria, one isolate (named JH7) was selected for its high antibiotic activity and was further examined for antagonism against fungal pathogens. Strain JH7 was identified as a Chromobacterium sp. using phylogenetic analysis based on 16S rRNA gene sequences. This strain was shown to produce antimicrobial molecules, including chitinases and proteases, but not cellulases. Additionally, the ability of JH7 to produce siderophore and solubilize insoluble phosphate supports its antagonistic and beneficial traits for plant growth. The JH7 strain suppressed the conidiation, conidial germination, and chlamydospore formation of C. destructans. Furthermore, the JH7 strain inhibited other plant pathogenic fungi. Thus, it provides a basis for developing a biocontrol agent for ginseng cultivation.