• The Plant Pathology Journal
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• v.38 no.1
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• pp.33-45
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• 2022

To screen antagonistic fungi against plant pathogens, dual culture assay (DCA) and culture filtrate assay (CFA) were performed with unknown soil-born fungi. Among the different fungi isolated and screened from the soil, fungal isolate ANU-301 successfully inhibited growth of different plant pathogenic fungi, Colletotrichum acutatum, Alternaria alternata, and Fusarium oxysporum, in DCA and CFA. Morphological characteristics and rDNA internal transcribed spacer sequence analysis identified ANU-301 as Aspergillus terreus. Inoculation of tomato plants with Fusarium oxysporum f. sp. lycopersici (FOL) induced severe wilting symptom; however, co-inoculation with ANU-301 significantly enhanced resistance of tomato plants against FOL. In addition, culture filtrate (CF) of ANU-301 not only showed bacterial growth inhibition activity against Dickeya chrysanthemi (Dc), but also demonstrated protective effect in potato tuber against soft rot disease. Gas chromatography-tandem mass spectrometry analysis of CF of ANU-301 identified 2,4-bis(1-methyl-1-phenylethyl)-phenol (MPP) as the most abundant compound. MPP inhibited growth of Dc, but not of FOL, in a dose-dependent manner, and protected potato tuber from the soft rot disease induced by Dc. In conclusion, Aspergillus terreus ANU-301 could be used and further tested as a potential biological control agent.

• Microbiology and Biotechnology Letters
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• v.49 no.2
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• pp.217-224
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• 2021

Rice blast caused by Pyricularia oryzae, which is a major threat to food security worldwide, markedly decreases the yield of rice. Some rhizobacteria called 'plant growth-promoting rhizobacteria' inhibit plant pathogens and improve plant growth by secreting iron-chelating siderophores. The decreased availability of iron adversely affects the survival of pathogens, especially fungal pathogens, in the rhizosphere. This study aimed to determine the morphological diversity of siderophore-producing bacteria, analyze the type of siderophores produced by the bacteria, and examine their growth-inhibitory activity against Pyricularia oryzae. The rhizobacteria were isolated from the rhizosphere of Sembada Hitam variety of black rice plants in Pakem, Sleman, Yogyakarta, Indonesia. In total, 12 distinct isolates were screened for the production of siderophores. It was found that 9 out of 12 bacteria produced siderophore and most of them were Gram positive bacteria. The best siderophore-producing isolates with different type of siderophore were used in further studies. The IS3 and IS14 isolates were found to be the best siderophore producer that produced hydroxamate and mixed type of hydroxamate-carboxylate type of siderophore, respectively. In the dual culture assay, IS14 showed a strong antagonistic effect against Pyricularia oryzae by the 81.17% inhibition.

• Food Science and Preservation
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• v.30 no.1
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• pp.42-51
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• 2023

This study aimed to characterize a lytic Salmonella Typhimurium-specific (ST) phage and its biofilm control capability against S. Typhimurium biofilm on polypropylene surface. ST phage was isolated, propagated, and purified from water used in a slaughterhouse. The morphology of ST phage was observed via transmission electron microscopy. Its bactericidal effect was evaluated by determining bacterial concentrations after the phage treatment at various multiplicities of infection (MOIs) of 0.01, 1.0, and 100. Once the biofilm was formed on the polypropylene tube after incubation at 37℃ for 48 h, the phage was treated and its antibiofilm capability was determined using crystal violet staining and plate count method. The phage was isolated and purified at a final concentration of ~11 log PFU/mL. It was identified as a myophage with an icosahedral head (~104 nm) and contractile tail (~90-115 nm). ST phage could significantly decrease S. Typhimurium population by ~2.8 log CFU/mL at an MOI of 100. After incubation for 48 h, biofilm formation on polypropylene surface was confirmed with a bacterial population of ~6.9 log CFU/cm². After 1 h treatment with ST phage, the bacterial population in the biofilm was reduced by 2.8 log CFU/cm². Therefore, these results suggest that lytic ST phage as a promising biofilm control agent for eradicating S. Typhimurium biofilm formed on food contact surfaces.

• Korean Journal of Microbiology
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• v.49 no.4
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• pp.398-402
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• 2013

This study was to investigate beneficial effects of microbial mixture on red pepper which was capable of promoting plant growth by solubilizing insoluble phosphate as well as protecting plants from pathogenic attack. Saccharomyces sp. L13 was isolated for phosphate solubilizing activity on aluminium phosphate, tricalcium phosphate, calcium hydrophosphate, and magnesium hydrophosphate. On the other hand, Bacillus sp. L32 was isolated for antagonistic activity against Phytophthora capsisi and Colletotrichum gloeosporioides, causing Phytophthora blight and Anthracnose disease in pepper, respectively. The strain L32 exhibited antagonistic activities both under dual culture assays and detached leaves assays. The each strain under the condition of mixed cultivation exhibited the same growth rates as one under pure cultivation. In greenhouse study, the mixed culture showed the both effect of plant growth promotion and reduction of disease symptom development against P. capsisi and C. gloeosporioides providing a potential as effective microbial agent for plant husbandry.

• Microbiology and Biotechnology Letters
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• v.44 no.4
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• pp.540-549
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• 2016

Plant residues serve as substrates for the proliferation and overwintering of plant pathogenic fungi in soil. Effects of the biocontrol fungus Trichoderma harzianum on the colonization of wheat straw by the plant pathogenic fungus Fusarium solani were investigated under different soil moisture regimes (-50 vs. -500 kPa) in non-sterile soil. T. harzianum ThzID1-M3 and/or F. solani were added along with wheat straw to non-sterile soils. ThzID1-M3, other Trichoderma species, and F. solani were monitored for a 21-day period using quantitative PCR. ThzID1-M3 reduced the colonization of F. solani on wheat straw (p < 0.05) under both moisture regimes, and F. solani reduced the colonization by ThzID1-M3 and other Trichoderma species (p < 0.05), thus suggesting competitive inhibition between ThzID1-M3 and F. solani. Colonization by ThzID1-M3 and generic Trichoderma was improved in the wet soil (p < 0.05), but colonization by F. solani did not differ between the two moisture conditions. Thus, the inhibitory effect of ThzID1-M3 was greater in the wet soil (p < 0.05). The growth competition between ThzID1-M3 and F. solani to colonize plant debris suggests that the biocontrol fungus T. harzianum may reduce the potential of the plant pathogen, F. solani, to survive and proliferate on crops.

• The Korean Journal of Food And Nutrition
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• v.25 no.1
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• pp.149-155
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• 2012

Although bacterial outbreaks from ready-to-eat foods such as sprouts have increased, the information on microbial biocontrol by means of sanitizers is limited. Twenty sprouts of red cabbage, baby radish, alfalfa, and broccoli each were collected from the wholesale markets in Seoul. Ethanol and organic acids including acetic acid, citric acid, and lactic acid were used to control the amount of mesophilic bacteria and Bacillus cereus. Sanitizer mixtures of ethanol and organic acids showed a remarkable reduction of viable counts and, in particular, the sanitizer mixture comprised of 20% ethanol and 1% lactic acid seemed to be best by a reduction of 7~8 log CFU/g on the sprouts after a 10 minute exposure. At the same time, the sanitizer comprised of 20% ethanol and 1% lactic acid showed the same reduction for Escherichia coli, Salmonella typhimurium, B. cereus, Staphylococcus aureus, and Enterococcus faecalis, with more effect toward the Gram-negative bacteria. Sensory evaluation by texture, browning, off-flavour, and overall acceptability seemed better just after treatment when compared to one day after treatment with the sanitizer. Therefore, the sanitizers of ethanol-organic acid might be an effective means to control the bacterial contamination of sprouts in palce of sodium hypochloric acid.

• Journal of Korean Society of Forest Science
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• v.104 no.3
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• pp.512-518
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• 2015

Pine wilt disease (PWD) caused by pine wood nematode, Bursaphelenchus xylophilus, has become the most serious threat to pine trees in Korea. This study was subjected to investigate effective biological control agent against PWD. To select nematocidal bacteria against PWD, Bacillus licheniformis MH48 was selected among five bacteria due to its high nematocidal potential. B. licheniformis MH48 was tested for cell growth and protease activity to evaluate its nematicidal potential. In the B. licheniformis MH48, cell numbers were highest three days after incubation, while protease activity was highest after seven days. In the effect of different concentrations of B. licheniformis MH48 culture broth against B. xylophilus, 20% concentration of culture broth showed approximately 80% of pine wood nematode mortality compared to the control. Especially, pine wood nematode's cuticle layers were degraded two days after treatment of B. licheniformis MH48 culture broth. The present study suggests that B. licheniformis MH48 can be one of the potential biocontrol candidates against pine wood nematode due to its ability to produce protease.

• Korean Journal of Plant Resources
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• v.29 no.1
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• pp.39-45
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• 2016

Pink Rot on melon and White Stain Symptom on grape are caused by Trichothecium roseum, one of the most important diseases of grape and melon. These diseases have been occurred in national-wide in Korea and causes irreversible damage on the grape and the melon at harvest season. This research presents the evaluation of the capacity of Bacillus subtillis HK2 to protect both melon and grape against T. reseum and establishes its role as a biocontrol agent. In this study, we isolated a Bacillus strain HK2 from rhizosphere soil, identified it as Bacillus subtillis by 16S rRNA analysis and demonstrated its antifungal activity against T. roseum. Under I-plate assay it was observed that the effect of hyphal growth inhibition was not due to production of volatile compounds. The optimum culture condition of HK2 was found at 30℃ and initial pH of 7.0. Application of HK2 culture suspension reduced 90.2% of white stain symptom on grape as compared to control, resulting in greater protection to grape against T. roseum infestation. Butanol extract of HK2 culture purified using flash column chromatography. The antifungal material was a polar substance as it showed antifungal activity in polar elute. Therefore, our results indicated a clear potential of B. subtilis HK2 to be used for biocontrol of Pink rot in melon and white stain symptom on grape caused by T. roseum.

• Korean Journal of Soil Science and Fertilizer
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• v.46 no.2
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• pp.105-111
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• 2013

The experiments were carried out to investigate the biocontrol potential of Lysobacter capsici YS1215 on root-knot nematode and to characterize its lytic enzyme activities. L. capsici YS1215 showed chitinase and gelatinase activities on the medium containing 0.5% chitin or 0.5% gelatin as substrates. Cell growth of L. capsici YS1215 was highest at 6 days, and the highest activities of chitinase (4.0 unit $ml^{-1}$) and gelatinase (7.43 unit $ml^{-1}$) were observed on 3 and 5 days after incubation, respectively. To investigate the effect of L. capsici YS1215 on tomato growth and nematode infection, the plants in pot trial were treated with bacterial culture (BC), half of bacterial culture (HBC), only bacterial medium (BM), tap water (TW) and commercial nematicide (CN). HBC treatd plants showed the higher shoot fresh weight and dry weight on $5^{th}$week after incubation while BM, HBC and BC had consistently higher values than TW at $9^{th}$ week. HBC appeared to be the highest shoot fresh length at $9^{th}$ week. Both CN and BC showed lower number of egg mass, root gall, and population of juveniles in soil compared to BC, HBC, BM and TW. These results suggest that L. capsici YS1215 with its strong ability of lytic enzyme production can be one of the most significant candidates for biocontrol agents against root-knot nematodes.

• Journal of Microbiology and Biotechnology
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• v.31 no.9
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• pp.1231-1240
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• 2021

Members of the genus Bacillus are known to play an important role in promoting plant growth and protecting plants against phytopathogenic microorganisms. In this study, 21 isolates of Bacillus spp. were obtained from the root micro-ecosystem of Suaeda glauca. Analysis of the 16S rRNA genes indicated that the isolates belong to the species Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus subtilis, Bacillus pumilus, Bacillus aryabhattai and Brevibacterium frigoritolerans. One of the interesting findings of this study is that the four strains B1, B5, B16 and B21 are dominant in rhizosphere soil. Based on gyrA, gyrB, and rpoB gene analyses, B1, B5, and B21 were identified as B. amyloliquefaciens and B16 was identified as B. velezensis. Estimation of antifungal activity showed that the isolate B1 had a significant inhibitory effect on Fusarium verticillioides, B5 and B16 on Colletotrichum capsici (syd.) Butl, and B21 on Rhizoctonia cerealis van der Hoeven. The four strains grew well in medium with 1-10% NaCl, a pH value of 5-8, and promoted the growth of Arabidopsis thaliana. Our results indicate that these strains may be promising agents for the biocontrol and promotion of plant growth and further study of the relevant bacteria will provide a useful reference for the development of microbial resources.