• The Plant Pathology Journal
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• v.38 no.5
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• pp.482-489
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• 2022

Fire blight caused by Erwinia amylovora (Ea) is a devastating disease in apple and pear trees. Oxolinic acid (OA), a quinolone family antibiotic that inhibits DNA gyrase, has been employed to control fire blight in South Korea since 2015. The continuous use of this bactericide has resulted in the emergence of OA-resistant strains in bacterial pathogens in other countries. To investigate the occurrence of OA-resistant Ea strains in South Korea, we collected a total of 516 Ea isolates from diseased apple and pear trees in 2020-2021 and assessed their sensitivities to OA. We found that all isolates were susceptible to OA. To explore the possibility of emerging OA-resistant Ea by continuous application of OA, we exposed Ea stains to a range of OA concentrations and constructed OA-resistant mutant strains. Resistance was associated with mutations in the GyrA at codons 81 and 83, which result in glycine to cysteine and serine to arginine amino acid substitutions, respectively. The in vitro growth of the mutants in nutrient media and their virulence in immature apple fruits were lower than those of wild-type. Our results suggest that OA-resistance decreases the fitness of Ea. Future work should clarify the mechanisms by which OA-resistance decreases virulence of this plant pathogen. Continuous monitoring of OA-resistance in Ea is required to maintain the efficacy of this potent bactericide.

• 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 Korean Journal of Mycology
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• v.51 no.4
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• pp.447-460
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• 2023

Fire blight, a plant disease caused by Erwinia amylovora, continues to cause considerable damage to fruit-bearing trees belonging to the Rosaceae family, such as apples and pears. In Korea, to hinder its rapid spread diseased apple and pear trees have been buried since its first occurrence. To determine the safety of this control method, the buried plant materials and adjacent soil have been investigated. In the process of investigating the microbiota of the buried soil, Bisifusarium allantoides, Botryotrichum domesticum, Microascus verrucosus, Paraphoma pye, Phaeosphaeria culmorum, Ramophialophora globispora, Sordaria tomentoalba, and Striaticonidium brachysporum were identified as unrecorded species in Korea. To report the above eight species as Korean unrecorded species, we report their morphological and molecular features in this study.

• The Plant Pathology Journal
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• v.39 no.4
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• pp.397-408
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• 2023

Fire blight disease, caused by Erwinia amylovora, is a devastating affliction in apple cultivation worldwide. Chemical pesticides have exhibited limited effectiveness in controlling the disease, and biological control options for treating fruit trees are limited. Therefore, a relatively large-scale survey is necessary to develop microbial agents for apple trees. Here we collected healthy apple trees from across the country to identify common and core bacterial taxa. We analyzed the endophytic bacterial communities in leaves and twigs and discovered that the twig bacterial communities were more conserved than those in the leaves, regardless of the origin of the sample. This finding indicates that specific endophytic taxa are consistently present in healthy apple trees and may be involved in vital functions such as disease prevention and growth. Furthermore, we compared the community metabolite pathway expression rates of these endophyte communities with those of E. amylovora infected apple trees and discovered that the endophyte communities in healthy apple trees not only had similar community structures but also similar metabolite pathway expression rates. Additionally, Pseudomonas and Methylobacterium-Methylorobrum were the dominant taxa in all healthy apple trees. Our findings provide valuable insights into the potential roles of endophytes in healthy apple trees and inform the development of strategies for enhancing apple growth and resilience. Moreover, the similarity in cluster structure and pathway analysis between healthy orchards was mutually reinforcing, demonstrating the power of microbiome analysis as a tool for identifying factors that contribute to plant health.

• The Korean Journal of Mycology
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• v.50 no.4
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• pp.331-341
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• 2022

This study was conducted to identify yeast species isolated from domestic pear blossom through gene sequencing and analysis of morphological characteristics, and to confirm specific yeast species inhibitory effects toward fire blight in immature apples, pears, and crab apple blossoms. Yeast morphological characteristics were consistent with the known characteristics of Aureobasidium pullulans. Nucleotide sequencing of the D1/D2 region of large-subunit (LSU) 26S ribosomal DNA and the internal transcribed spacer (ITS) region confirmed its identity as A. pullulans (MHAU2101). Inoculation of immature fruits with A. pullulans MHAU2101 before exposure to Erwinia amylovora prevented fire blight symptoms in apples and pears. A. pullulans MHAU2101 treated crab apple blossoms had a significantly lower flower infection rate than untreated blossoms, revealing 64% of the potency of streptomycin. The A. pullulans MHAU2101 treated group also displayed lower E. amylovora density in both pistil and hypanthium compared to the untreated group, especially in the hypanthium. This study confirms that A. pullulans MHAU2101 isolated from domestic pear blossom can effectively suppress the onset of fire blight.

• Research in Plant Disease
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• v.26 no.4
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• pp.222-228
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• 2020

Erwinia amylovora, a causal bacterium of fire blight disease, is registered as a prohibited quarantine pathogen in Korea. To control the disease, the government should diagnose the disease, dig and bury the host trees when fire blight occurs. Fire blight was the first reported in 43 orchards of Anseong, Cheonan, and Jecheon in 2015, and 42.9 ha of host trees were eradicated. However, the disease spread to eleven cities, so that 348 orchards and 260.4 ha of host trees were eradicated until 2019. Fire blight of Asian pear occurred mainly in the southern part of Gyeonggi, and Chungnam province, on average of 29±9.2 orchards per year. And the age of the infected trees were mostly 20-30 years old. In apple trees, the disease occurred mainly in the northern part of Gyeonggi, Gangwon, and Chungbuk province, on average of 41±57.6 orchards per year, increased highly in 2018 and 2019. The age of infected apple trees were under 20 years old. Therefore, because the disease spread rapidly in young apple trees, spraying control agents to the trees in a timely manner and removing infected trees quickly are important to prevent the spread of fire blight in the orchard of immature trees.

• Research in Plant Disease
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• v.29 no.1
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• pp.23-38
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• 2023

The fire blight caused by Erwinia amylovora (Ea) is a devastating disease of Rosaceae plants, including commercially important apple and pear trees. Since the first report in Korea in May 2015, it has been spreading to neighboring regions gradually. Host plants can be infected by pollinators like bees, rainfall accompanied by wind, and cultural practices such as pruning. Many studies have revealed that wild Rosaceae plants such as Cotoneaster spp., Crataegus spp., Pyracantha spp., Prunus spp., and Sorbus spp. can be reservoirs of Ea in nature. However, wild Rosaceae plants in Korea have not been examined yet whether they are susceptible to fire blight. Therefore, the susceptibility to fire blight was examined with 25 species in 10 genera of wild Rosaceae plants, which were collected during 2020-2022, by artificial inoculation. Bacterial suspension (10⁸ cfu/ml) of Ea type strain TS3128 was inoculated artificially in flowers, leaves, stems, and fruits of each plant species, and development of disease symptoms were monitored. Moreover, the presence of Ea bacteria from inoculated samples were checked by conventional polymerase chain reaction. Total 14 species of wild Rosaceae plants showed disease symptoms of fire blight, and Ea bacteria were detected inside of inoculated plant parts. These results suggest that wild Rosaceae plants growing nearby commercial apple and pear orchards in Korea can be Ea reservoirs, and thus they should be monitored regularly to minimize the damage by Ea infection and spreading.

• The Plant Pathology Journal
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• v.19 no.6
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• pp.294-300
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• 2003

Bacterial strains were isolated from diseased samples of shoot blight collected from different pear growing orchards of Chuncheon, Korea from 1995 to 1998. Forty-nine strains showed their pathogenicity on immature fruit and shoot of pear. Microbiological, physiological, and biochemical tests were performed on these pathogenic strains. One strain, designated as WT3 in this study, was selected as a representative strain as it was collected from the first outbreak area in Jichonri, Chuncheon in 1995. Further detailed characterization of the strain WT3 was done by PCR amplification using specific primers described previously for distinguishing Erwinia pyrifoliae from its close pathogen Erwinia amylovora. Based on phenotypical, biochemical, and molecular analyses, strain WT3 was identified as a shoot blight pathogen which was the same as E. pyrifoliae Ep16 previously described by a German group in 1999.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2020.12a
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• pp.48-48
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• 2020

Fire blight, caused by Erwinia amylovora(Burrill), is a destructive disease of apple that damages blossoms, shoots, and woody plant organs. The fire blight disease is a worldwide problem for pome fruit growers because all popular apple cultivars are susceptible to the disease. Recently, fire blight of apple rootstocks has become a serious economic problem in high-density orchard systems in korea. The most commonly used dwarfing root stocks, M.9 and M.26, are highly susceptible to E. amylovora. The objective of the apple rootstock-breeding program has been to develop pomologically excellent rootstocks with resistance to abiotic and biotic stresses, including fire blight. Budagovsky 9 (B.9) apple rootstock is reported to be highly susceptible when inoculated with E. amylovora, although results from multiple trials showed that B.9 is resistant to rootstock blight infection in field plantings. So we tried to collect the apple rootstocks traits of fire blight resistance. The apple genotype Malus Robusta 5 (MR5) represents an ideal donor for fire blight resistance because it was described as resistant to all currently known European strains of the pathogen. The PCR for detecting the MR5 gene using the primers Md_MR5_FL_F/Md_MR5_FL_R. The results of these experiments confirmed some apple rootstocks traits of fire blight resistance showed the MR5. Furthermore, this gene is confirmed to be the resistance determinant of Mr5 as the transformed lines undergo the same gene-for-gene interaction in the host-pathogen relationship MR5-E. amylovora.

• Research in Plant Disease
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• v.23 no.4
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• pp.306-313
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• 2017

Recently fire blight occurred in the Republic of Korea and eradication program for the disease has been executed since then. Specificity and detection sensitivity of the 2 antibody-based diagnostic strips to Korean isolates of Erwinia amylovora (Ea) and their application for on-site diagnosis were evaluated in this study. Ea AgriStrip, a commercial diagnostic kit, and EB strip, developed in this study, reacted positively to the all tested Korean Ea strains and also to most of Erwinia pyrifoliae (Ep) strains causing black shoot blight. They reacted negatively to all Pusedomonas syringae pv. syringae (Pss) strains that cause shoot blight on apple. Detection sensitivity was similar between the 2 strips. For on-site diagnosis, the two strips reacted positively only to the extractions of the fire-blighted samples on all fire blight occurred orchards except one orchard at which on-site diagnosis was carried out at winter time. In addition, they reacted positively to the black-shoot blighted extractions from the black shoot blight occurred apple orchard. These results suggest that both EB strip and Ea AgriStrip would be useful for on-site diagnosis of fire blight in Korea.