• Microbiology and Biotechnology Letters
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• v.48 no.1
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• pp.38-47
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• 2020

This study was conducted to investigate both plant growth-promoting and plant disease- controlling activities of bacterial strains isolated from soil. All the isolated strains were able to grow at various temperatures. All the strains, except ANG40, showed antagonistic effects against various phytopathogenic fungi. This antagonism can be ascribed to the production of siderophores and antibiotic substances. In addition, all the strains showed abilities such as nitrogen fixation, phosphate solubilization, and siderophore production. These results suggest that nitrogen, phosphorus, and iron can be converted into forms that can be easily absorbed by the plants for their growth. Analysis of the growth-promoting properties revealed that ANG51 produced 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase and indole-3-acetic acid (IAA) both of which are related to ethylene production. In contrast, the other strains were found to have only IAA-producing ability. Therefore, this study suggests that Pseudomonas geniculata ANG3, Exiguobacterium acetylicum ANG40, and Burkholderia stabilis ANG51, which were selected through analysis of comparative advantages for both plant growth promotion and disease-controlling activity, may be used as biological agents.

• The Korean Journal of Mycology
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• v.47 no.4
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• pp.417-425
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• 2019

Biocontrol agents (BCAs) are widely used to protect plants from diverse biotic and abiotic stresses in agricultural and ecological fields. Among the various microbes, many subspecies of the gram-positive genus, Bacillus, have been successfully industrialized as eco-friendly biological pesticides and fertilizers. In the current study, we demonstrated that Bacillus thuringiensis C25 exhibited antagonistic effects on the mycelial growth of Rosellinia necatrix, a fungal phytopathogen. Scanning electron microscopy analysis revealed that B. thuringiensis C25 degraded the cell wall structures of R. necatrix mycelia. In the functional genomic analysis of B. thuringiensis C25, we annotated 5,683 genes and selected the gene sets that potentially encoded fungal cell wall degrading enzymes (CWDEs). The growth inhibition effects on R. necatrix were highly correlated with the transcriptional activity of the mycelial cell wall degrading genes of B. thuringiensis C25. The transcript levels of CWDEs, including CshiA, B, and Glycos_transf_2 genes in B. thuringiensis C25, were enhanced following co-cultivation with R. necatrix. In conclusion, our study suggested that B. thuringiensis C25 could serve as a suitable candidate for controlling R. necatrix and could facilitate elucidating the mechanisms underlying the antifungal activities of BCAs against phytopathogens.

• Korean Journal of Environmental Biology
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• v.20 no.2
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• pp.165-172
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• 2002

The combined action of two factors on organisms can be either antagonistic, non-effective, additive or synergistic. Although synergism is of biological importance, the common features of synergistic interaction between harmful environmental factors are largely unknown. The purpose of this study is to establish general rules describing the response of various organisms to the combined action of heat with another inactivating agent. Synergistic interaction due to the simultaneous treatment of hyperthermia with ionizing or non-ionizing radiation has been analyzed using the experimental data mainly obtained with yeast cells. In addition, the results reported by others for viruses, bacterial spores, cultured mammalian cells, plants and animals were also analyzed to check the regularities revealed. The common rules of the synergistic interaction obtained in this study can be summarized as follows. For any constant rate of exposure, the synergy can be observed only within a certain temperature range. An increase in exposure rate resulted in an increase of this specific temperature and vice versa. For a constant temperature at which the irradiation occurs, synergy can be observed within a certain dose rate range. As the exposure temperature is reduced, the optimal intensity decreases and vice versa. A new conception taken into consideration those regularities can make a clue for environmental disaster preventive analysis of the synergy of radiation with the other factor.

• The Korean Journal of Pesticide Science
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• v.10 no.1
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• pp.56-65
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• 2006

An antagonistic bacterial isolate, that inhibits the growth of plant pathogens, was selected and identified from 5,000 isolates screened from the rhizosphere of various crop plants. An isolate Bacillus sp. N32, tested against Colletotrichum gloeosporioides causing anthracnose disease in hot pepper, produced both a heat resistant antifungal protein and a heat sensitive antifungal protein. The heat resistant protein was partially purified by Ammonium sulfate fractionation and gel filtration chromatography. The bioautography showed that the proteins possessed high antifungal activity. The biosynthetic gene cluster responsible for the heat resistant antifungal protein was cloned from cosmid library using DNA probe obtained from PCR product with the primers targeting the conserved nucleotide sequence of the synthetic genes reported earlier, Most of the clones obtained showed higher homology to fengycin antibiotic synthetic gene family reported earlier. On the other hand, the heat sensitive protein was isolated from SDS-PAGE and electroblotting to determine the N-terminal amino acid sequences. The heat sensitive antifungal protein gene was cloned from the ${\lambda}-ZAP$ libraries using a DNA probe based on the N-terminal amino acid sequences of the heat sensitive protein. We are contemplating to clone and sequence the whole gene cluster encoding the heat sensitive protein for further analysis.

• Korean Journal of Soil Science and Fertilizer
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• v.43 no.4
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• pp.490-497
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• 2010

A chitinolytic bacterium having a strong antagonistic activity against various pathogens including Phytophtora capsici was isolated from rhizosphere soil, and identified as Lysobacter enzymogenes (named as LE429) based on 16S rRNA gene sequence analysis. This strain produced a number of substances such as chitinase, ${\beta}-1$, 3-glucanase, lipase, protease, gelatinase and an antibiotic compound. This antibiotic compound was purified by diaion HP-20, sephadex LH-20 column chromatography and HPLC. The purified compound was identified as phenylacetic acid by gas chromatography-electron ionization (GC-EI) and gas chromatography-chemical ionization (GC-CI) mass spectrometry. In field experiment, pepper plants were treated by the strain LE429 culture (CB), neem oil solution (NO), combination (CB+NO) or control (CON). Plant height and number of branches, flowers and pods of pepper plant in CB treatment were generally highest, and followed by CB+NO, CON and NO. The fungal pathogens were strongly inhibited, while several insect pests were discovered in CB treatment. Any insect pests were not found, while all fungal pathogens tested were not suppressed in NO treatment. However, in CB+NO treatment, non incidence of fungal pathogens and insect pests were found. The strain LE429 producing secondary metabolites with neem oil should be a potential agent to control fungal diseases and insect pests.

• The Korean Journal of Mycology
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• v.24 no.4 s.79
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• pp.293-304
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• 1996

A bacterial strain, KSl, possessing strong antifungal activity was isolated from soil samples of ginseng fields and identified as Bacillus subtilis. In greenhouse test, the culture filtrate of B. subtilis KS1 showed strong protective effect against several fungal diseases of agricultural plants such as cucumber gray mold and wheat leaf rust. In addition, the crude butanol fraction of the culture filtrate exhibited antagonistic effect against several fungi including plant or human pathogens, such as Botrytis maydis, Chytridium lagenarium and Candida albicans. The antifungal compound, SW1, produced by B. subtilis KS1 was purified through consecutive chromatographic separations on a pep-RPC column and a ${\mu}$ Bondapak $C_{18}$ reverse phase column. Temperature and pH showed little effect on the stability of the compound in the ranges $-20-121^{\circ}C$ and pH 4.0-10.0, respectively. The composition and structural characteristics of SW1 were analysed by HPLC and by $^1H-,\;^1H-^1H-COSY$, NOESY, COSY-NOESY and HOHAHA NMR spectroscopy, respectively, which revealed that the compound belongs to iturin A, a typical cyclic antifungal compound produced by B. subtilis. In contrast to the previously reported iturin A compounds which have one or no $-CH_3$ side chain in the hydrophobic hydrocarbon chain of ${\beta}-amino$ acids, SW1 was shown to have a ${\beta}-amino$ acid containing 12-carbon skeleton with two $-CH_3$ side chains.

• Research in Plant Disease
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• v.15 no.3
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• pp.209-216
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• 2009

During the period of June, 2005 to May, 2008, 44 host plants infected with powdery mildew were collected in the Jeon-ju and Jang-su districts of Jeonbuk province and in the Jang-sung district of Jeonnam province. The hyperparasites, Ampelomyces were confirmed in 12 plant species. Most of the pycnidium shapes of Ampelomyces were circular or oval shaped, and the sizes were different even within the same host plant, and also the color of pycnidium was ranged from light brown to dark brown. Ampelomyces species were isolated from 4 hosts including Impatiens balsamina L., Cucurbita pepo, Rudbeckia laciniata var. elatier and Youngia sonchifolia, and thus the most appropriate 12 Ampelomyces strains for the current experiment were selected. When analyzing the selected 12 strains' incubational and nutritional characteristics, the malt extract agar was the most appropriate media. When investigating the effect of osmotic pressure on the spore germination, 0.15M NaCl concentration was the optimum germination concentration. When the isolated Ampelomyces sp. was tested in-vitro, it was found to be effective to control in other plant pathogens, isolated Ampelomyces showed no pathogenicity to the plant. strains isolated . studied on rDNA ITS sequence analysis. The rDNA ITS sequence data of Ampelomyces sp. isolate BSLAH16 from Impatiens balsamina L. were analyzed and identified.

• Journal of Life Science
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• v.33 no.10
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• pp.842-850
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• 2023

Pyrrolnitrin, pyrrolomycin, and pyoluteorin are functional halogenated phenylpyrrole derivatives (HPDs) derived from microorganisms with diverse antimicrobial activities. Pyrrolnitrin is a secondary metabolite produced from L-tryptophan through four-step reactions in Pseudomonas fluorescens, Burkholderia cepacia, Serratia plymuthica, etc. It is currently used for the treatment of superficial dermatophytic fungal infections, has high antagonistic activities against soil-borne and foliar fungal infections, and has many industrial applications. Since pyrrolnitrin is easily decomposed by light, it is difficult to widely use it outdoors. As an alternative, fludioxonil, a synthetically produced non-systemic surface fungicide that is structurally similar and has excellent light stability, has been commercialized for seed and foliar treatment of plants. However, due to its high toxicity to aquatic organisms and adverse effects in human cell lines, many countries have established maximum residue levels and strictly control its levels. Pyrrolomycin and pyoluteorin, which have antibiotic/antibiofilm activity against Gram-positive bacteria and high anti-oomycete activity against the plant pathogen Pythium ultimum, respectively, were isolated and identified from microorganisms. This review summarizes the biosynthesis and production of natural pyrrolnitrin derived from bacteria and the characteristics of synthetic fludioxonil and other natural phenylpyrrole derivatives among the HPDs. We expect that a plethora of highly effective, novel HPDs that are safe for humans and environments will be developed through the generation of an HPD library by microbial biosynthesis and chemical synthesis.

• Korean Journal of Ecology and Environment
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• v.41 no.spc
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• pp.68-76
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• 2008

This study examined the inhibition effects of a freshwater bivalve (Unio douglasiae) and a submerged plant (Potamogeton crispus) on the cyanobacterial bloom (Oscillatoria sp.). The experiment were conducted in aquarium $(50cm{\times}65cm{\times}120cm)$ with lake sediments in the bottom of the aquarium in 10 cm thick. Before the experiments, artificial cyanobacterial bloom was induced with the addition of lake sediment and CB medium. Total 12 transparent acrylic cylinders (${\Phi}19cm$, height 40 cm) were placed in the aquarium, and within which bivalves and plants were placed in various conditions such as the control (C), plant addition (P:5 stems), mussel addition (U:2 individuals), and both mussel and plant addition (PU: the same quantity as used in each treatment). The experiment was conducted in triplicate during 7 days. pH, dissolved oxygen (DO), electric conductivity (EC), salinity, cyanobacterial cell density, chlorophyll-${\alpha}$ concentration, and mussel filtering rate were monitored daily. At the end of the experiment, total phosphorus (TP), total nitrogen (TN), and plant height and weight were measured. Overall, a large degree of cyanobacterial growth inhibition appeared in both P and U treatments, and the effect was highest in the U treatment, followed by P and PU. The combined treatment of both U and P did not show any synergic effects compared to the effect in separated treatment. In all enclosures of the treatments chlorophyll-${alpha}$ (Chl-${alpha}$) concentration decreased until 36 hours after the additions of the plants and mussels. In contrast, Chl-${alpha}$ concentrations increased in PU enclosures after 36 hours. The same trend was shown in the cell density of Oscillatoria. pH and DO gradually decreased until 120 and 144 hours, respectively, in the P and PU enclosures. TP concentration increased in the mussel enclosures (U and PU), while TN concentration largely decreased in the plant enclosures (P and PU). Our results suggest that applied bivalve (Unio) and submerged plant (Potamogeton) seemed to have a potential effect on the growth inhibition of cyanobacteria, but their combined application may have an antagonistic effect to diminish the degree of the inhibition.

• The Korean Journal of Mycology
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• v.32 no.1
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• pp.31-38
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• 2004

An endophytic bacterial strain EB215 that was isolated from cucumber (Cucumis sativus) roots displayed a potent in vivo antifungal activity against Colletotrichum species. The strain was identified as Burkholderia cepacia based on its physiological and biochemical characteristics, and 16S rDNA gene sequence. Optimal medium and incubation period for the production of antifungal substances by B. cepacia EB215 were nutrient broth (NB) and 3 days, respectively. An antifungal substance was isolated from the NB cultures of B. cepacia EB215 strain by centrifugation, n-hexane partitioning, silica gel column chromatography, preparative TLC, and in vitro bioassay. Its chemical structure was determined to be pyrrolnitrin by mass and NMR spectral analyses. Pyrrolnitrin showed potent disease control efficacy of more than 90% against pepper anthracnose (Colletotrichum coccodes), cucumber anthracnose (Colletotrichum orbiculare), rice blast (Magnaporthe grisea) and rice sheath blight (Corticium sasaki) even at a low concentration of $11.1\;{\mu}g/ml$. In addition, it effectively controlled the development of tomato gray mold (Botrytis cinerea) and wheat leaf rust (Puccinia recondita) at concentrations over $33.3\;{\mu}g/ml$. However, it had no antifungal activity against Phytophthora infestans on tomato plants. Further studies on the development of microbial fungicide using B. cepacia EB215 are in progress.