• Korean Journal of Environmental Agriculture
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• v.38 no.4
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• pp.269-272
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• 2019

BACKGROUND: Since 2015, fire blight disease caused by Erwinia amylovora has been devastating apple and pear orchards every year. To quickly block the disease spreading, infected apple and pear trees have been buried in soil. However, concern on the possibility of the pathogen survival urgently requires informative data on the buried host plants. Therefore, this study was conducted to investigate the survival of the pathogen from the buried host plants. METHODS AND RESULTS: Apple trees buried in 42 months ago in a Jecheon site and pear trees buried in 30 months ago in an Anseong site were excavated using an excavator. Plant samples were taken from stems and twigs of the excavated trees. The collected 120 samples were checked for rotting and used for bacterial isolation, using TSA, R2A, and E. amylovora selection media. The purely isolated bacteria were identified based on colony morphology and 16S rDNA sequences. Wood rotting and decay with off smells and discoloring were observed from the samples. A total of 17 genera and 48 species of bacteria were identified but E. amylovora was not detected. CONCLUSION: Our investigation suggests that the survival of E. amylovora doesn't seem possible in the infected hosts which have been buried in soil for at least 30 months. Therefore, the burial control can be considered as a safe method for fire blight disease.

• The Plant Pathology Journal
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• v.39 no.1
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• pp.52-61
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• 2023

Fire blight, caused by Erwinia amylovora, is one of the major bacterial disease of apple and pear, causing enormous economic losses worldwide. Several control measures against E. amylovora have been reported till date, however, none of them have proved to be effective significantly against the pathogen. In this study, mechanisms of the copper-based control agents (CBCAs): copper oxychloride (COCHL), copper oxide (COX), copper hydroxide (CHY), copper sulfate basic (CSB), and tribasic copper sulfate (TCS) and their disease severity reduction efficacy against E. amylovora were analyzed. Bis-1,3-dibutylbarbituric acid trimethine oxonol, carboxyl fluorescein diacetate succinimidyl ester, and 5-cyano-2,3-ditolyl tetrazolium chloride staining were used to check the damage of membrane potential, cytoplasmic pH_in, and respiration of CBCAs-treated E. amylovora, respectively. High disturbance in the membrane potential of E. amylovora was found under COX and COCHL treatments. Similarly, higher significant changes in the inner cytoplasmic pH_in were observed under COX, COCHL, and TCS treatment. CHY and COCHL-treated E. amylovora showed a significant reduction in respiration. In vitro bioassay results revealed that CHY, CSB, and TCS at 2,000 ppm reduced the severity of fire blight both in pre- and post-treatment of CBCAs in immature apple fruits and seedlings. Overall, the most effective CBCAs against E. amylovora could be CHY at 2,000 ppm as its showed inhibition mechanisms and disease severity reduction.

• 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.

• Korean Journal of Agricultural and Forest Meteorology
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• v.25 no.4
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• pp.302-310
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• 2023

In this study, the extent of the impact of rainfall on the outbreak of fire blight during the blossom infection risk period was explored. In the Chungnam province, the outbreak of fire blight disease began in 2015, and changes in the outbreak's scale were most pronounced between 2020 and 2022, significantly escalating from 63 orchards in 2020 to 170 orchards in 2021, before decreasing to 46 orchards in 2022. In 2022, the number of incidence has decreased and the number of canker symptom in branches has also decreased. It was evaluated that the significant decrease of fire blight disease in 2022 was due to the dry weather during the flowering season. In other words, this yearly fluctuation in fire blight outbreaks was correlated with the presence or absence of rainfall and accumulated precipitation during the blossom infection risk period. This trend was observed across all surveyed regions where apples and pears were cultivated. Among the weather conditions influencing the blossom infection risk period, rainfall notably affected the activation of pathogens from over-wintering cankers and flower infections. In particular, precipitation during the initial 3 days of the blossom infection risk warning was confirmed as a decisive factor in determining the outbreak's scale.

• The Plant Pathology Journal
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• v.40 no.3
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• pp.282-289
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• 2024

Fire blight is a bacterial disease caused by Erwinia amylovora. In Korea, fire blight was first reported in 2015 in an orchard. If the infection is confirmed, all trees in the orchard must be removed and the orchard must remain closed for 3 years. Since 2020, if the number of trees infected with fire blight is less than 5% of the total trees in the orchard, only the infected tree and adjacent trees are removed in Korea. Three years after removal, the trees can be replanted after confirming that the orchard soil is free from E. amylovora. In this study, a protocol was established for detecting E. amylovora in soil via selective enrichment, using tryptic soy broth with 0.05% bile salts and 50 ㎍/ml cycloheximide, and real-time polymerase chain reaction. This protocol resulted in a 1,000-times improved detection limit for E. amylovora in soil samples compared to that in unenriched samples. Soil monitoring was performed for orchards where fire blight-infected trees had been removed 3-27 months prior; the selected orchards were monitored every 3 months. Monitoring confirmed that E. amylovora was not present in the soil at any site in any of the orchards. A new detection protocol facilitates the monitoring of E. amylovora in soil and could help permit the replanting of trees in orchards. Also monitoring results provide evidence that trees can be planted earlier.

• Korean Journal of Plant Resources
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• v.36 no.1
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• pp.100-106
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• 2023

In order to control the fire blight disease, all plants within the radius of the diseased orchard were removed in the early stage of the outbreak, or antibiotics control was performed for prevention. Since the beginning of antibiotics use on plants, the potential for development of resistance to antibiotics by the plant pathogen and unintended detrimental effects on the fruit trees and environment has become a problem. The purpose of this study is to determine the degree of phytotoxicity to fruit trees caused by excessive spraying of the fire blight disease disinfectant and to establish basic data for safe disinfectant guide. We analyzed whether damage to the fruit tree and the maximum residual limit of fruit was exceeded when three kinds of the fire blight disease disinfectants were continuously sprayed in excess of the number of safe use during the growing season. There was no phytotoxicity in apple 'Fuji' and pear 'Niitaka', and oxolinic acid was detected beyond the limit of quantitation in 'Fuji' grown without a bag, and the other disinfectants were detected below the maximum residue limit. When these disinfectants are continuously sprayed in excess of the number of safe, phytotoxicity may remain on the fruit. Therefore, it is necessary to observe the prescribed dilution factor and observe the safe frequency and the timing of use.

• Research in Plant Disease
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• v.30 no.2
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• pp.139-147
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• 2024

Erwinia amylovora is a gram-negative plant pathogen that causes fire blight in apple and pear trees, resulting in significant damage worldwide. In this study, we monitored the emergence of fire blight from 2019 to 2023 to determine the emergence patterns and the factors affecting the outbreak of the disease. As a result of the 5-year survey on the emergence of fire blight, a total of 2,029 cases have emerged, mostly in apple trees of 1,378 cases (67.9%) followed by 645 cases (31.8%) in pear trees, and from quince, hawthorn, and mountain ash trees. Fire blight appeared in specific areas of Gyeonggi, Chungnam, Gangwon, and Chungbuk provinces in 2019, but spread to Andong and Yesan in 2021, Muju and Bonghwa in 2023. In 2020 and 2021, there were 744 and 618 cases of fire blight outbreaks, respectively, compared to other years (188-245 cases/year). Notably, 914 of these cases were observed in apple trees from May to July, with 667 cases reported in Chungju and Jecheon. The incidence of fire blight was positively correlated with the daily maximum temperatures and rainy days in January and February, as well as the rainy days in May and June. The average age of the diseased pear trees was 25 years, higher than the 10-year average age of the apple trees. This study provides fundamental information to understand the status and factors affecting the fire blight emergence in Korea. Prevention measures should be established through continuous analysis of the status of fire blight.

• 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.40 no.2
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• pp.160-170
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• 2024

Erwinia amylovora is a plant pathogen that causes fire blight on apples and pears. Bacteriophages, which are viruses that selectively infect specific species of bacteria and are harmless to animal cells, have been considered as biological control agents for the prevention of bacterial pathogens. In this study, we aimed to use bacteriophages that infect E. amylovora as biocontrol agents against fire blight. We isolated bacteriophages Fifi044 and Fifi318 infecting E. amylovora, and characterized their morphology, plaque form, and genetic diversity to use as cocktails for disease control. The stabilities of the two phages were investigated at various temperatures and pH values and under sunlight, and long-term storage experiment was conducted for a year. To evaluate whether the two phages were suitable for use in cocktail form, growth curves of E. amylovora were prepared after treating the bacterial cells with single phages and a phage cocktail. In addition, a disease control test was conducted using immature apples and in vitro cultured apple plantlets to determine the biocontrol effects of the phage cocktail. The two phages were morphologically and genetically different, and highly stable up to 50℃ and pH value from 4 to 10. The phages showed synergistic effect when used as a cocktail in the inhibition of host bacterial growth and the disease control. This study demonstrated that the potential of the phage cocktail as a biocontrol agent for commercial use.

• Research in Plant Disease
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• v.24 no.3
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• pp.182-192
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• 2018

Since fire blight (Erwinia amylovora) firstly broke out at mid-Korea in 2015, it is necessary to investigate potential spread of the invasive pathogen. To speculate environmental factors of fireblight epidemic based on disease triangle, a fire blight predicting program, MARYBLYT, was run with the measured meteorological data in 2014-2017 and the projecting future data under RCP8.5 scenario for 2020-2100. After calculating blossom period of Singo pear from phenology, MARYBLYT was run for blossom blight during the blossom period. MARYBLYT warned "Infection" blossom blight in 2014-15 at Anseong and Cheonan as well as Pyungtak and Asan. In addition, it warned "Infection" in 2016-17 at Naju. More than 80% of Korean areas were covered "Infection" or "High", therefore Korea was suitable for fire blight recently. Blossom blight for 2020-2100 was predicted to be highly fluctuate depending on the year. For 80 years of the future, 20 years were serious with "Infection" covered more than 50% of areas in Korea, whereas 8 years were not serious covered less than 10%. By comparisons between 50% and 10% of the year, temperature and amount of precipitation were significantly different. The results of this study are informative for policy makers to manage the alien pathogen.