• The Plant Pathology Journal
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• v.37 no.2
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• pp.137-151
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• 2021

The present study describes the bacterial blight of walnut, caused by Xanthomonas arboricola pv. juglandis (Xaj) in the northern Gyeongbuk province, Korea. Disease symptoms that appear very similar to anthracnose symptoms were observed in walnut trees in June 2016. Pathogens were isolated from disease infected leaves, fruits, shoots, bud, flower bud of walnut, and cultured onto yeast dextrose carbonate agar plates. Isolated bacteria with bacterial blight symptoms were characterized for their nutrient utilization profiles using Biolog GN2 and Vitek 2. In addition, isolates were subjected to physiological, biochemical, and morphological characterizations. Furthermore, isolates were identified using 16S rDNA sequence analysis, and multi-locus sequence analysis using atpD, dnaK, efp, and rpoD. To confirm pathogenicity, leaves, fruits, and stems of 3-year-old walnut plants were inoculated with bacterial pathogen suspensions as a foliar spray. One week after inoculation, the gray spots on leaves and yellow halos around the spots were developed. Fruits and stems showed browning symptoms. The pathogen Xaj was re-isolated from all symptomatic tissues to fulfill Koch's postulates, while symptoms were not appeared on control plants. On the other hand, the symptoms were very similar to the symptoms of anthracnose caused by Colletotrichum gloeosporioides. When walnut plants were inoculated with combined pathogens of Xaj and C. gloeosporioides, disease symptoms were greater in comparison with when inoculated alone. Xaj population size was more in the month of April than March due to their dormancy in March, and sensitive to antibiotics such as oxytetracycline and streptomycin, while resistant to copper sulfate.

• Research in Plant Disease
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• v.28 no.3
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• pp.152-161
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• 2022

Several types of chemical bactericides have been used to control fire blight. However, their excessive usage leads to environmental deterioration. Therefore, several researchers have analyzed antagonistic microorganisms as promising, effective, and safe biological control agents (BCAs). The primary aim of this study was to screen for potential antagonistic bacteria that suppress Erwinia amylovora. Among the 45 isolates studied, 5 strains showed the largest inhibition zone against E. amylovora. 16S rRNA gene sequencing identified them as Bacillus amyloliquefaciens (KPB 15), B. stratosphericus (KPB 21), B. altitudinis (KPB 25), B. safensis (KPB 31), and B. subtilis (KPB 39). KPB 25 and 31 reduced the lesion size of fire blight by 50% in immature apple fruits, and did not show antagonism against each other. Therefore, KPB 25 and 31 were selected to develop an antagonistic mixture against fire blight. Although the mixture with KPB 25 and 31 showed a slightly increased ability to reduce lesion size on immature fruits, they did not exhibit a synergistic effect in reducing E. amylovora population compared to each strain alone. Nevertheless, we have identified these two strains as useful and novel BCAs against fire blight with additional benefits safety and potential in developing a mixture without loss of their activity, owing to the absence of antagonism against each other.

• The Plant Pathology Journal
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• v.23 no.3
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• pp.203-209
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• 2007

Infection structures were observed using a fluorescence microscope at the penetration sites on the leaves of potato plants pre-inoculated with the bacterial strains Pseudomonas putida TRL2-3, Micrococcus luteus TRK2-2, and Flexibacteraceae bacterium MRL412, which mediated an induced systemic resistance on potato plants against late blight disease caused by Phytophthora infestans. In order to compare the infection structures on the leaves expressing systemic acquired resistance, the leaves of potato plants pre-treated with DL-3-amino butyric acid (BABA) were also observed after challenge inoculation with the same pathogen. The infection structures were investigated. The total number of germination and appressorium formation of P. infestans were counted. Furthermore, the frequencies of fluorescent epidermal cells at the penetration sites, which indicate a defense response of plant cell, were estimated. There were no differences on the germination rates of the fungal cysts among the untreated control, BABA pre-treated, and bacterial strains pre-inoculated plants. However, appressorium formation was slightly decreased on the leaves of BABA pre-treated plants compared to those of untreated as well as bacterial strains pre-inoculated plants. Furthermore, the frequencies of fluorescent cells of BABA pre-treated and bacterial strains pre-inoculated were higher than that of untreated plants, indicating an active defense reaction of the host cells against the fungal attack. On the other hand, the pre-treatment with BABA caused a stronger fluorescent of epidermal cells at the penetration sites compared to the pre-inoculation with the bacterial strains. Interestingly, the frequency of fluorescent cells by BABA, however, was lower than that by the bacterial strains. Based on the results it is suggested that the infection structures showing resistance reaction on the leaves of potato plants were different between by pre-inoculation with bacterial strains and by pre-treatment with BABA against the late blight pathogen.

• Research in Plant Disease
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• v.24 no.1
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• pp.41-51
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• 2018

Since fire blight disease on apple and pear was produced in Korea in 2015, there were no registered chemicals to control against this disease. Instead, several antibacterial chemicals that were registered for other bacterial diseases such as soft rot and bacterial spot have been authorized by Rural Development Administration (RDA). However, these chemicals are not tested efficacy for fire blight disease except damage by those treatments on apple and pear in Korea. Thus, we evaluated efficiency using in vitro and in planta assays of antibacterial chemicals such as antibiotics and copper compounds including kasugamycin, oxytetracycline, oxolinic acid and streptomycin, and copper hydroxide, copper sulfate, oxine copper and tribasic copper sulfate, respectively. We also tested two kinds of biological agents. As expected, significant antibacterial effect was observed in vitro test of both antibiotics and copper-based chemicals. In planta test based on disease severity including ooze and water-soaked formation on immature pears, bacterial populations on blooms, and blight lesion formation in artificially inoculated shoots, kasugamycin, oxytetracycline and streptomycin have been shown the most efficiency among tested antibiotics. Four copper-based chemicals tested in this study, control effects are little bit lower than agricultural antibiotics but they seem to be available to use in terms of winter season. Biocontrol agents were also shown possibility to treat in eco-friendly farms. In addition, there are no antibiotic resistance genes in Korean isolates against antibiotics, which were selected for suppression of fire blight in this study.

• The Plant Pathology Journal
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• v.23 no.2
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• pp.109-112
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• 2007

This study was to examine the efficacy of a root-dipping application of antagonistic bacterial strains for the control of Phytophthora blight of pepper caused by P. capcisi, and to evaluate their plant growth-promoting effects in the field in 2005 and 2006. The candidate antagonistic rhizobacterial strains CCR04, CCR80, GSE09, ISE13, and ISE14 were treated by dipping plant roots with bacterial suspensions prior to transplanting. The candidate rhizobacterial strains CCR04, CCR80, GSE09, and ISE14 significantly (P=0.05) reduced the disease incidence and the area under the disease progress curves when compared to buffer-treated controls in at least a year test. The metalaxy l(fungicide-treated control) resulted in one of the lowest disease incidences among the treatments in both years. Moreover, the strains CCR04, CCR80, GSE09, and ISE13 significantly (P=0.05) increased the fruit weights and/or numbers of peppers in at least a year test compared to the buffer-treated controls. These results suggest that the antagonistic rhizobacterial strains CCR04, CCR80, and GSE09 could be efficient biocontrol agents by controlling Phytophthora blight of pepper and promoting the plant growth when treated with root-dipping at transplanting.

• The Plant Pathology Journal
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• v.29 no.4
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• pp.418-426
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• 2013

Pseudomonas isolates from different crop plants were screened for in vitro growth inhibition of Phytophthora capsici and production of biosurfactant. Two in vivo experiments were performed to determine the efficacy of selected Pseudomonas strains against Phytophthora blight of pepper by comparing two fungicide treatments [acibenzolar-S-methyl (ASM) and ASM + mefenoxam]. Bacterial isolates were applied by soil drenching ($1{\times}10^9$ cells/ml), ASM ($0.1{\mu}g$ a.i./ml) and ASM + mefenoxam (0.2 mg product/ml) were applied by foliar spraying, and P. capsici inoculum was incorporated into the pot soil three days after treatments. In the first experiment, four Pseudomonas strains resulted in significant reduction from 48.4 to 61.3% in Phytophthora blight severity. In the second experiment, bacterial treatments combining with olive oil (5 mL per plant) significantly enhanced biological control activity, resulting in a reduction of disease level ranging from 56.8 to 81.1%. ASM + mefenoxam was the most effective treatment while ASM alone was less effective in both bioassays. These results indicate that our Pseudomonas fluorescens strains (6L10, 6ba6 and 3ss9) that have biosurfactant-producing abilities are effective against P. capsici on pepper, and enhanced disease suppression could be achieved when they were used in combination with olive oil.

• Proceedings of the Korean Society of Crop Science Conference
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• 1990.05a
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• pp.48-49
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• 1990

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2002.11a
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• pp.44.3-45
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• 2002

• The Plant Pathology Journal
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• v.28 no.4
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• pp.364-372
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• 2012

Xanthomonas oryzae pv. oryzae causing bacterial leaf blight disease in rice produces and secretes Hpa1 protein that belongs to harpin protein family. Previously it was reported that Hpa1 induced defense responses when it was produced in tobacco. In this study, we expressed hpa1 gene in rice and Arabidopsis to examine the effects of Hpa1 expression on disease resistance to both fungal and bacterial pathogens. Expression of hpa1 gene in rice enhanced disease resistance to both X. oryzae pv. oryzae and Magnaporthe grisea. Interestingly, individual transgenic rice plants could be divided into four groups, depending on responses to both pathogens. hpa1 expression in Arabidopsis also enhanced disease resistance to both Botrytis cineria and Xanthomonas campestris pv. campestris. To examine genes that are up-regulated in the transgenic rice plants after inoculation with X. oryzae pv. oryzae, known defense-related genes were assessed, and also microarray analysis with the Rice 5 K DNA chip was performed. Interestingly, expression of OsACS1 gene, which was found as the gene that showed the highest induction, was induced earlier and stronger than that in the wild type plant. These results indicate that hpa1 expression in the diverse plant species, including monocot and dicot, can enhance disease resistance to both fungal and bacterial plant pathogens.

• Journal of Microbiology and Biotechnology
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• v.24 no.6
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• pp.732-739
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• 2014

We describe the development of a polymerase chain reaction method for the rapid, precise, and specific detection of the Xanthomonas oryzae pv. oryzae (Xoo) K3a race, the bacterial blight pathogen of rice. The specific primer set was designed to amplify a genomic locus derived from an amplified fragment length polymorphism specific for the K3a race. The 1,024 bp amplicon was generated from the DNA of 13 isolates of Xoo K3a races out of 119 isolates of other races, pathovars, and Xanthomonas species. The assay does not require isolated bacterial cells or DNA extraction. Moreover, the pathogen was quickly detected in rice leaf 2 days after inoculation with bacteria and at a distance of 8 cm from the rice leaf 5 days later. The results suggest that this PCR-based assay will be a useful and powerful tool for the detection and identification of the Xoo K3a race in rice plants as well as for early diagnosis of infection in paddy fields.