• Research in Plant Disease
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• v.25 no.3
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• pp.136-142
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• 2019

Drought stress is considered as one of major abiotic stresses; it leads to reduce plant growth and crop productivity. In this study, we selected bacterial strains for alleviating drought stress in chili pepper plants. As drought-tolerant bacteria, 28 among 447 strains were pre-selected by in vitro assays including growth in drought condition with polyethylene glycol and plant growth-promoting traits including production of 1-aminocyclopropane-1-carboxylate deaminase, indole-3-acetic acid and exopolysaccharide. Sequentially, 7 among pre-selected 28 strains were screened based on relative water content (RWC); GLC02 and KJ40, among seven strains were finally selected by RWC and malondialdehyde (MDA) in planta trials under an artificial drought condition by polyethylene glycol solution. Two strains GLC02 and KJ40 reduced drought stress in a natural drought condition as well as an artificial condition. Strains GLC02 or KJ40 increased shoot fresh weight, chlorophyll and stomatal conductance while they decreased MDA in chili pepper plants under a natural drought condition. However, two strains did not show biocontrol activity against diseases caused by Phytophthora capsici and Xanthomonas campestris pv. vesicatoria in chili pepper plants. Taken together, strains GLC02 or KJ40 can be used as bio-fertilizer for alleviation of drought stress in chili pepper plants.

• Microbiology and Biotechnology Letters
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• v.49 no.3
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• pp.391-402
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• 2021

Earthworms play an important role in soil fertilization, interacting continually with microorganisms. This study aims to demonstrate the existence of beneficial microorganisms living in the earthworm's immune system, the coelomic fluid. To achieve this goal, a molecular identification technique was performed, using cytochrome c oxidase I (COI) barcoding to identify abundant endogenic earthworms inhabiting the temperate zone of Rabat, Morocco. Then, 16S rDNA and ITS sequencing techniques were adopted for bacteria and fungi, respectively. Biochemical analysis, showed the ability of bacteria to produce characteristic enzymes and utilize substrates. Qualitative screening of plant growth-promoting traits, including nitrogen fixation, phosphate and potassium solubilization, and indole acetic acid (IAA) production, was also performed. The result of mitochondrial COI barcoding allowed the identification of the earthworm species Aporrectodea molleri. Phenotypic and genotypic studies of the sixteen isolated bacteria and the two isolated fungi showed that they belong to the Pseudomonas, Aeromonas, Bacillus, Buttiauxella, Enterobacter, Pantoea, and Raoultella, and the Penicillium genera, respectively. Most of the isolated bacteria in the coelomic fluid showed the ability to produce β-glucosidase, β-glucosaminidase, Glutamyl-β-naphthylamidase, and aminopeptidase enzymes, utilizing substrates like aliphatic thiol, sorbitol, and fatty acid ester. Furthermore, three bacteria were able to fix nitrogen, solubilize phosphate and potassium, and produce IAA. This initial study demonstrated that despite the immune property of earthworms' coelomic fluid, it harbors beneficial microorganisms. Thus, the presence of resistant microorganisms in the earthworm's immune system highlights a possible selection process at the coelomic fluid level.

• Korean Journal of Soil Science and Fertilizer
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• v.45 no.6
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• pp.1037-1042
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• 2012

An auxin-producing bacteria (2SJ8-02) was isolated from waste mushroom bed of Agaricus bisporus in Buyeo-Gun, Chungnam. The strain 2SJ8-02 was classified as a novel strain of Pantoea rwandensis based on a chemotaxanomic and phylogeneticanalyses. The isolate was confirmed to produce indole-3-acetic acid (IAA), one of auxin hormones, by TLC and HPLC analyses. The maximum concentration of IAA, $122mg\;L^{-1}$ was detected from the culture in R2A broth containing 0.1% tryptophan for 24 h at $35^{\circ}C$. The molecular weight of the main peak obtained by LC-mass analysis was 175 corresponding to that of IAA. To investigate the growth-promoting effects to the crops, the culture broth of Pantoea rwandensis 2SJ8-02 was infected to water cultures and seed pots of mung bean. In consequence, the adventitious root induction and root growth of mung bean were two times higher than those of the control.

• The Korean Journal of Mycology
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• v.44 no.1
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• pp.36-47
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• 2016

This study was conducted to evaluate five different strains of rhizobacterial isolates viz. PA1, PA2, PA4, PA5 and PA12 for biological control against Colletotrichum acutatum, C. coccodes, C. gloeosporioides, C. dematium, Botrytis cinerea, Rhizoctonia solani, Sclerotinia minor and Fusarium sp. In vitro inhibition assay was performed on three different growth mediums, potato dextrose agar (PDA), tryptic soy agar (TSA), and PDA-TSA (1:1 v/v) for the selection of potential antagonistic isolates. According to the result, isolate PA2 showed the highest inhibitory effect with 65.5% against C. coccodes on PDA and with 96.5% against S. minor on TSA. However, the same isolate showed the highest inhibition with 58.5% against C. acutatum on PDA-TSA. In addition, an in vivo experiment was performed to evaluate these bacterial isolates for biological control against fungal pathogens. Plants treated with bacteria were analyzed with phytopathogens and plants inoculated with phytopathogens were treated with isolates to determine the biological control effect against fungi. According to the result, all five isolates tested showed inhibitory effects against phytopathogens at various levels. Mode of action of these rhizobacterial isolates was evaluated with siderophore production, protease assay, chitinase assay and phosphate solubilizing assay. Bacterial isolates were identified by 16S rDNA sequencing, which showed that isolates PA1 and PA2 belong to Bacillus subtilis, whereas, PA4, PA5, and PA12 were identified as Bacilus altitudinis, Paenibacillus polymyxa and Bacillus amyloliquefaciens, respectively. Results of the current study suggest that rhizobacterial isolates can be used for the plant growth promoting rhizobacteria (PGPR) effect as well as for biological control of various phytopathogens.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.40-44
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• 2003

Soil-borne infectious disease including Pythium aphanidermatum and Rhizoctonia solani causes severe damage to plants, such as cucumber. This soil-borne infectious disease was not controlled effectively by chemical pesticide. Since these diseases spread through the soil, chemical agents are usually ineffective. Instead, biological control, including antagonistic microbe can be used as a preferred control method. An efficient method was developed to select an antagonistic strain to be used as a biological control agent strain. In this new method, surface tension reduction potential of an isolate was included in the ‘decision factor’ in addition to the other factors, such as growth rate, and pathogen inhibition rate. Considering these 3 decision factors by a statistical method, an isolate from soil was selected and was identified as Bacillus sp. GB16. In the pot test, this strain showed the best performance among the isolated strains. The lowest disease incidence rate and fastest seed growth was observed when Bacillus sp. GB16 was used. Therefore this strain was considered as plant growth promoting rhizobacteria (PGPR). The action of surface tension reducing component was deduced as the enhancement of wetting, spreading, and residing of antagonistic strain in the rhizosphere. This result showed that new selection method was significantly effective in selecting the best antagonistic strain for biological control of soil-borne infectious plant pathogen. The antifungal substances against P. aphanidermatum and R. solani were partially purified from the culture filtrates of Bacillus sp. GB16. In this study, lipopeptide possessing antifungal activity was isolated from Bacillus sp. GB16 cultures by various purification procedures and was identified as a surfactin-like lipopeptide based on the Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR), high performance liquid chromatography mass spectroscopy (HPLC-MS), and quadrupole time-of-flight (Q-TOF) ESI-MS/MS data. The lipopeptide, named GB16-BS, completely inhibited the growth of Pythium aphanidermatum, Rhizoctonia solani, Penicillium sp., and Botrytis cineria at concentrations of 10 and 50 mg/L, respectively. A novel method to prevent the foaming and to provide oxygen was developed. During the production of surface active agent, such as lipopeptide (surfactin), large amount of foam was produced by aeration. This resulted in the carryover of cells to the outside of the fermentor, which leads to the significant loss of cells. Instead of using cell-toxic antifoaming agents, low amount of hydrogen peroxide was added. Catalase produced by cells converted hydrogen peroxide into oxygen and water. Also addition of corn oil as an oxygen vector as well as antifoaming agent was attempted. In addition, Ca-stearate, a metal soap, was added to enhance the antifoam activity of com oil. These methods could prevent the foaming significantly and maintained high dissolved oxygen in spite of lower aeration and agitation. Using these methods, high cell density, could be achieved with increased lipopeptide productivity. In conclusion to produce an effective biological control agent for soil-borne infectious disease, following strategies were attempted i) effective screening of antagonist by including surface tension as an important decision factor ii) identification of antifungal compound produced from the isolated strain iii) novel oxygenation by $H_2O_2-catalase$ with vegetable oil for antifungal lipopeptide production.

• Journal of Ginseng Research
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• v.39 no.3
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• pp.213-220
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• 2015

Background: Korean ginseng (Panax ginseng Meyer) is a perennial herb prone to various root diseases, with Phytophthora cactorum being considered one of the most dreaded pathogens. P. cactorum causes foliar blight and root rot. Although chemical pesticides are available for disease control, attention has been shifted to viable, eco-friendly, and cost-effective biological means such as plant growth-promoting rhizobacteria (PGPR) for control of diseases. Methods: Native Bacillus amyloliquefaciens strain HK34 was isolated from wild ginseng and assessed as a biological control agent for ginseng. Leaves from plants treated with HK34 were analyzed for induced systemic resistance (ISR) against P. cactorum in square plate assay. Treated plants were verified for differential expression of defense-related marker genes using quantitative reverse transcription polymerase chain reaction. Results: A total of 78 native rhizosphere bacilli from wild P. ginseng were isolated. One of the root-associated bacteria identified as B. amyloliquefaciens strain HK34 effectively induced resistance against P. cactorum when applied as soil drench once (99.1% disease control) and as a priming treatment two times in the early stages (83.9% disease control). A similar result was observed in the leaf samples of plants under field conditions, where the percentage of disease control was 85.6%. Significant upregulation of the genes PgPR10, PgPR5, and PgCAT in the leaves of plants treated with HK34 was observed against P. cactorum compared with untreated controls and only pathogen-treated plants. Conclusion: The results of this study indicate HK34 as a potential biocontrol agent eliciting ISR in ginseng against P. cactorum.

• Mycobiology
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• v.34 no.2
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• pp.67-72
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• 2006

Efficacy of resistance induction by the bacterial isolates Pseudomonas putida (TRL2-3), Micrococcus luteus (TRK2-2) and Flexibacteraceae bacterium (MRL412), which were isolated from the rhizosphere of plants growing in Jeju Mountain, were tested in a greenhouse. The disease severity caused by Phytophthora infestans was effectively reduced in the potato plants pre-inoculated with bacterial isolates compared with those of the untreated control plants growing in a greenhouse. In order to estimate the level of protection by the bacterial isolates, Mancozeb WP (Diesen $M^{(R)}$, Kyong nong) and DL-3-amino butyric acid (BABA) were pre-treated, whereas Dimethomorph WP ($Forum^{(R)}$, Kyong nong) and phosphonic acid ($H_{3}PO_{3}$) were post-treated the challenge inoculation with the pathogen. Disease severities of chemical pre-treated as well as post-treated plants were reduced compare to those of the untreated. The disease reduction in the plants pre-treated with Mancozeb WP was the highest, whereas that of post-treated with Dimethomorph WP was the lowest. The yields of plants pre-inoculated with three bacterial isolates were greatly increased than those of control plants. These results suggest that biological control by bacterial isolates might be an alternative strategy against late blight disease in potato plants growing in greenhouse.

• Microbiology and Biotechnology Letters
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• v.40 no.1
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• pp.30-38
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• 2012

We investigated the effects on soil microbial diversity and the growth promotion of red pepper resulting from inoculation with a microbial agent composed of Bacillus subtilis AH18, B. licheniformis K11 and Pseudomonas fluorescens 2112 in a red pepper farming field. Photosynthetic bacteria, Trichoderma spp., Azotobacter spp., Actinomycetes, nitrate oxidizing bacteria, nitrite oxidizing bacteria, nitrogen fixing bacteria, denitrifying bacteria, phosphate solubilizing bacteria, cellulase producing bacteria, and urease producing bacteria are all indicator microbes of healthy soil microbial diversity. The microbial diversity of the consortium microbial agent treated soil was seen to be 1.1 to 14 times greater than soils where other commercial agent treatments were used, the latter being the commercial agent AC-1, and chemical fertilizer. The yield of red pepper in the field with the treated consortium microbial agent was increased by more than 15% when compared to the other treatments. Overall, the microbial diversity of the red pepper farming field soil was improved by the consortium microbial agent, and the promotion of growth and subsequent yield of red pepper was higher than soils where the other treatments were utilized.

• Microbiology and Biotechnology Letters
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• v.31 no.3
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• pp.235-241
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• 2003

For the biological control of Phytophthora blight of red-pepper caused by Phytophthora capsici, an antibiotic-producing plant growth promoting rhizobacteria (PGPR) Bacillus sp. KL 39 was selected from a local soil of Kyongbuk, Korea. The strain KL 39 was identified as Bacillus megaterium by various cultural, biochemical test and API and Microlog system. B. megaterium KL 39 could produce the highest antifungal antibiotic after 40 h of incubation under the optimal medium which was 0.4% fructose, 0.3% yeast extract, and 5 mM KCl at 30 C with initial pH 8.0. The antifungal antibiotic KL 39 was purified by Diaion HP-20 column, silica gel column, Sephadex LH-20 column, and HPLC. Its RF value was confirmed 0.32 by thin-layer chromatography with Ethanol:Ammonia:Water = 8:1:1. The crude antibiotic KL39 was active against a broad range of plant pathogenic fungi, Rhizoctonia solani, Pyricularia oryzae, Monilinia fructicola, Botrytis cinenea, Alteranria kikuchiana, Fusarium oxysporum and Fusarium solani. The purified antifungal antibiotic KL39 had a powerful biocontrol activity against red-pepper phytophthora blight disease with in vivo pot test as well as the strain B. megaterium KL 39.

• Microbiology and Biotechnology Letters
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• v.40 no.3
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• pp.190-196
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• 2012

In this study, a total of more than 1,000 bacteria, including 739 species of aerobic bacteria, 80 species of urease producing bacteria and 303 species of photosynthetic bacteria, were isolated from red-pepper field soils located in the Gyeongsan Province of the Republic of Korea. Amongst these, 158 species of aerobic bacteria, 70 species of urease producing bacteria and 228 species of photosynthetic bacteria were found to be auxin producing soil bacteria through quantification analysis with the Salkowski test. The latter groupings were then tested for antifungal activities to ${\beta}$-Glucanase and siderophore using CMC congo red agar and CAS blue agar media. In addition, the selected strains were examined for antifungal activity against various phytopathogenic fungi on PDN agar media. Six strains; BCB14, BCB17, C10, HA46, HA143, and HJ5, were noted for their ability to both produce auxin and act as antifungal substances. 16S rDNA sequence comparison analyses of these six strains identified them as Bacillus subtilis BCB14, B. methylotrophicus BCB17, B. methylotrophicus C10, B. sonorensis HA46, B. subtilis HA143, and B. safensis HJ5.