• Journal of Microbiology and Biotechnology
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• v.34 no.5
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• pp.1040-1050
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• 2024

To isolate and analyze bacteria with Verticillium wilt-resistant properties from the fermentation residue of kitchen wastes, as well as explore their potential for new applications of the residue. A total of six bacterial strains exhibiting Verticillium wilt-resistant capabilities were isolated from the biogas residue of kitchen waste fermentation. Using a polyphasic approach, strain ZL6, which displayed the highest antagonistic activity against cotton Verticillium wilt, was identified as belonging to the Pseudomonas aeruginosa. Bioassay results demonstrated that this strain possessed robust antagonistic abilities, effectively inhibiting V. dahliae spore germination and mycelial growth. Furthermore, P. aeruginosa ZL6 exhibited high temperature resistance (42℃), nitrogen fixation, and phosphorus removal activities. Pot experiments revealed that P. aeruginosa ZL6 fermentation broth treatment achieved a 47.72% biological control effect compared to the control group. Through activity tracking and protein mass spectrometry identification, a neutral metalloproteinase (Nml) was hypothesized as the main virulence factor. The mutant strain ZL6ߡNml exhibited a significant reduction in its ability to inhibit cotton Verticillium wilt compared to the strain P. aeruginosa ZL6. While the inhibitory activities could be partially restored by a complementation of nml gene in the mutant strain ZL6CMߡNml. This research provides a theoretical foundation for the future development and application of biogas residue as biocontrol agents against Verticillium wilt and as biological preservatives for agricultural products. Additionally, this study presents a novel approach for mitigating the substantial amount of biogas residue generated from kitchen waste fermentation.

• The Korean Journal of Pesticide Science
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• v.13 no.3
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• pp.185-189
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• 2009

A monoterpenoid compound, benzylideneacetone (BZA), is a metabolite of an entomopathogenic bacterium, Xenorhabdus nematophila. Its primary biological activity is an inhibitor of phospholipase $A_2$, which catalyzes the committed step of biosynthesis of various eicosanoids that are critically important to mediate insect immune responses. When BZA was applied to two-spotted spider mite, Tetranychus urticae, it exhibited a dose-dependent mortality in leaf-disc assay. Subsequently BZA was tested against T. urticae infesting apples in a field orchard, in which it showed a significant control efficacy, which was not statistically different with that of a commercial acaricide. BZA also had significant antibacterial activities against three species of plant pathogenic bacteria when it was added to the bacterial cultures, in which it showed the highest inhibitory activity against a bacterial wilt-causing pathogen, Ralstonia solanacearum. The bacterial pathogen caused significant disease symptom to young potato plants. However, BZA significantly suppressed the disease occurrence. This study suggests that BZA can be used to develop a novel crop protectant to control mite and bacterial pathogen.

• Mycobiology
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• v.33 no.1
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• pp.35-40
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• 2005

Selected isolates of Pseudomonas fluorescens (Pf4-92 and PfRsC5) and P. aeruginosa (PaRsG18 and PaRsG27) were examined for growth promotion and induced systemic resistance against Fusarium wilt of chickpea. Significant increase in plant height was observed in Pseudomonas treated plants. However, plant growth was inhibited when isolates of Pseudomonas were used in combination with Fusarium oxysporum f. sp. ciceri (FocRs1). It was also observed that the Pseudomonas spp. was colonized in root of chickpea and significantly suppressed the disease in greenhouse condition. Rock wool bioassay technique was used to study the effect of iron availability on the induction of systemic resistance to Fusarium wilt of chickpea mediated by the Pseudomonas spp. All the isolates of Pseudomonas spp. showed greater disease control in the induced systemic resistance (ISR) bioassay when iron availability in the nutrient solution was low. High performance liquid chromatography (HPLC) analysis indicated that an the bacterial isolates produced more salicylic acid (SA) at low iron ($10\;{\mu}M$ EDDHA) than high iron availability ($10\;{\mu}Fe^{3+}$ EDDHA). Except PaRsG27, all the three isolates produced more pseudobactin at low iron than high iron availability.

• The Plant Pathology Journal
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• v.36 no.4
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• pp.355-363
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• 2020

Bacterial traits for virulence of Ralstonia solanacearum causing lethal wilt in plants were extensively studied but are not yet fully understood. Other than the known virulence factors of Ralstonia solanacearum, this study aimed to identify the novel gene(s) contributing to bacterial virulence of R. solanacearum. Among the transposon-inserted mutants that were previously generated, we selected mutant SL341F12 strain produced exopolysaccharide equivalent to wild type strain but showed reduced virulence compared to wild type. In this mutant, a transposon was found to disrupt the murI gene encoding glutamate racemase which converts L-glutamate to D-glutamate. SL341F12 lost its motility, and its virulence in the tomato plant was markedly diminished compared to that of the wild type. The altered phenotypes of SL341F12 were restored by introducing a full-length murI gene. The expression of genes required for flagella assembly was significantly reduced in SL341F12 compared to that of the wild type or complemented strain, indicating that the loss of bacterial motility in the mutant was due to reduced flagella assembly. A dramatic reduction of the mutant population compared to its wild type was apparent in planta (i.e., root) than its wild type but not in soil and rhizosphere. This may contribute to the impaired virulence in the mutant strain. Accordingly, we concluded that murI in R. solanacearum may be involved in controlling flagella assembly and consequently, the mutation affects bacterial motility and virulence.

• Journal of Microbiology and Biotechnology
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• v.31 no.9
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• pp.1241-1255
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• 2021

This study was carried out to explore a non-chemical strategy for enhancing productivity by employing some antagonistic rhizobacteria. One hundred eighteen bacterial isolates were obtained from the rhizospheric zone of various crop fields of Gangwon-do, Korea, and screened for antifungal activity against Fusarium wilt (Fusarium oxysporum f. sp. lactucae) in lettuce crop under in vitro and in vivo conditions. In broth-based dual culture assay, fourteen bacterial isolates showed significant inhibition of mycelial growth of F. oxysporium f. sp. lactucae. All of the antagonistic isolates were further characterized for the antagonistic traits under in vitro conditions. The isolates were identified on the basis of biochemical characteristics and confirmed at their species level by 16S rRNA gene sequencing analysis. Arthrobacter sulfonivorans, Bacillus siamensis, Bacillus amyloliquefaciens, Pseudomonas proteolytica, four Paenibacillus peoriae strains, and Bacillus subtilis were identified from the biochemical characterization and 16S rRNA gene sequencing analysis. The isolates EN21 and EN23 showed significant decrease in disease severity on lettuce compared to infected control and other bacterial treatments under greenhouse conditions. Two bacterial isolates, EN4 and EN21, were evaluated to assess their disease reduction and growth promotion in lettuce in field conditions. The consortium of EN4 and EN21 showed significant enhancement of growth on lettuce by suppressing disease caused by F. oxysporum f. sp. lactucae respectively. This study clearly indicates that the promising isolates, EN4 (P. proteolytica) and EN21 (Bacillus siamensis), can be commercialized and used as biofertilizer and/or biopesticide for sustainable crop production.

• Research in Plant Disease
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• v.18 no.4
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• pp.361-369
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• 2012

A population density model for bacterial wilt, which is caused by Ralstonia solanacearum, in hot pepper was developed to estimate the primary infection date after overwintering in the field. We developed the model mechansitically to predict reproduction of the pathogen and pathogensis on seedlings of the host. The model estimates the pathogen's populations both in the soil and in the host. In order to quantify environmental infection factors, various temperatures and initial population densities were determined for wilt symptoms on the seedlings of hot pepper in a chamber. Once, the pathogens living in soil multiply up to 400 cells/g of soil, they can infect successfully in the host. Primary infection in a host was supposed to be started when the population of the pathogen were over $10^9$ cells/g of root tissue. The estimated primary infection dates of bacterial wilt in 2011 in Korea were mostly mid-July or late-July which were 10-15 days earlier than those in 2010. Two kinds of meterological data, synoptic observation and field measurements from paddy field and orchard in Kyunggi, were operated the model for comparing the result dates. About 1-3 days were earlier from field data than from synoptic observation.

• The Korean Journal of Pesticide Science
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• v.13 no.1
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• pp.45-53
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• 2009

An antagonistic bacterial strain KLF01 was isolated from rhizosphere of tomato and identified to be Lactobacillus sp. by biochemical and genetic analysis. This strain showed antagonism against the used plant pathogenic bacteria like Ralstonia solanacearum, (bacterial wilt), Xanthomonas axonopodis pv. citri, (Citrus canker), Xanthomonas campestris pv. vesicatoria (Bacterial spot), Eriwinia pyrifoliae (Shoot-blight) and Eriwinia carotovora subsp. carotovora group (Potato scab) through agar well diffusion method. In planta test done by drench application of strain KLF01 $(4{\times}10^8 cfu/ml)$ into the experimental plot containing tomato (Solanum lycopersicum L.) cultivar 'Lokkusanmaru' and red pepper (Capsicum annuum L.) cultivar 'Buja' plants, in pot test post-inoculated with the plant pathogenic bacteria, R. solanacearum significantly reduced the disease severity, compared to the non-treated plants.

• The Korean Journal of Mycology
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• v.43 no.3
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• pp.185-190
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• 2015

Water extract from spent mushroom substrate of Hericium erinaceus inhibited the mycelial growth of seven strain of tomato pathogenic fungi including Phytophthora capsici and the growth of Ralstonia solanacearum. Control efficacy of tomato bacterial wilt by treatment of 33.3% and 50% water extract from spent mushroom substrate of Hericium erinaceus was showed 58.3%, 83.3%, respectively.

• Journal of Microbiology and Biotechnology
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• v.17 no.11
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• pp.1765-1771
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• 2007

A polymerase chain reaction (PCR)-based method was developed to detect the DNA of Ralstonia solanacearum, the causal agent of bacterial wilt in various crop plants. One pair of primers (RALSF and RALSR), designed using cytochrome c1 signal peptide sequences specific to R. solanacearum, produced a PCR product of 932 bp from 13 isolates of R. solanacearum from several countries. The primer specificity was then tested using DNA from 21 isolates of Ralstonia, Pseudomonas, Burkholderia, Xanthomonas, and Fusarium oxysporum f. sp. dianthi. The specificity of the cytochrome c1 signal peptide sequences in R. solanacearum was further confirmed by a DNA-dot blot analysis. Moreover, the primer pair was able to detect the pathogen in artificially inoculated soil and tomato plants. Therefore, the present results indicate that the primer pair can be effectively used for the detection of R. solanacearum in soil and host plants.

• Research in Plant Disease
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• v.22 no.4
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• pp.236-242
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• 2016

Bacterial wilt caused by Ralstonia solanacearum is one of the most devastating diseases in major Solanaceae crops. The pathogen is easily disseminated and survives for many years in plant farming system. Although chemicals are applied to control the disease, they are of limited efficacy and cause several problems. Therefore, the use of phage therapy has been suggested to control the disease as a biological agent. In this study, we discovered bacteriophages lysing diverse Ralstonia isolates from plant and soil samples obtained from the potato cultivated field in Jeju. Three times repeated pickings of plaques resulted in obtaining 173 single phages showing diverse spectrum of host-specificity. With the results, 12 core phages were selected and dendrogram was generated. Genetic diversity of the selected phages was also confirmed by AFLP (Amplified Fragment of Length Polymorphism) fingerprinting. The stability of the phages was investigated in various temperatures and various conditions of pH in vitro. The phages were stable at $16^{\circ}C-44^{\circ}C$ and pH 6-10. Morphological characterization of the phages revealed they were all classified into the Podoviridae, but had diverse head sizes. The results of this research will contribute to control the disease and further researches regarding genetic and molecular aspects will facilitate understanding phage and bacteria interaction.