• Korean Journal of Agricultural Science
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• v.50 no.4
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• pp.723-734
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• 2023

Erwinia pyrifoliae, which causes black shoot blight in apple and pear trees, was first identified in Korea in 1995. Extensive measures are typically used to control the disease by eradicating trees in diagnosed orchards, owing to the detrimental impact of the disease on apple and pear production. However, despite governmental efforts, the disease has continuously spread. In this study, we analyzed the current status of the black shoot blight occurrence in apple and pear orchards between 1995 to 2022. Our findings reveal that over the past 28 years, black shoot blight has occurred in 26 cities and districts across five Korean provinces. The affected regions are predominantly concentrated in the northern part of Korea, including the Gangwon and Gyeonggi provinces. Furthermore, black shoot blight has gradually expanded to the northern provincial regions of Chungbuk, Chungnam, and Gyeongbuk, which are centrally situated in Korea. Furthermore, the occurrence pattern of black shoot blight differed between apple and pear orchards; in apple orchards, black shoot blight occurred consistently each year, with a sudden increase in cases in 2020; however, in pear orchards, it has considerably decreased since 2007. To the best of our knowledge, this is the first comprehensive report on the occurrence of black shoot blight in apple and pear trees in 28 years, and the results will provide valuable insights for future disease management strategies.

• The Plant Pathology Journal
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• v.34 no.3
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• pp.191-198
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• 2018

Fast and accurate diagnosis is needed to eradicate and manage economically important and invasive diseases like fire blight. Loop-mediated isothermal amplification (LAMP) is known as the best on-site diagnostic, because it is fast, highly specific to a target, and less sensitive to inhibitors in samples. In this study, LAMP assay that gives more consistent results for on-site diagnosis of fire blight than the previous developed LAMP assays was developed. Primers for new LAMP assay (named as DS-LAMP) were designed from a histidine-tRNA ligase gene (EAMY_RS32025) of E. amylovora CFBP1430 genome. The DS-LAMP amplified DNA (positive detection) only from genomic DNA of E. amylovora strains, not from either E. pyrifoliae (causing black shoot blight) or from Pseudomonas syringae pv. syringae (causing shoot blight on apple trees). The detection limit of DS-LAMP was 10 cells per LAMP reaction, equivalent to $10^4$ cells per ml of the sample extract. DS-LAMP successfully diagnosed the pathogens on four fire-blight infected apple and pear orchards. In addition, it could distinguish black shoot blight from fire blight. The $B{\ddot{u}}hlmann$-LAMP, developed previously for on-site diagnosis of fire blight, did not give consistent results for specificity to E. amylovora and on-site diagnosis; it gave positive reactions to three strains of E. pyrifoliae and two strains of P. syringae pv. syringae. It also, gave positive reactions to some healthy sample extracts. DS-LAMP, developed in this study, would give more accurate on-site diagnosis of fire blight, especially in the Republic of Korea, where fire blight and black shoot blight coexist.

• The Plant Pathology Journal
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• v.39 no.1
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• pp.141-148
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• 2023

Black shoot blight disease caused by Erwinia pyrifoliae has serious impacts on quality and yield in pear production in Korea; therefore, rapid and accurate methods for its detection are needed. However, traditional detection methods require a great deal of time and fail to achieve absolute quantification. In the present study, we developed a droplet digital polymerase chain reaction (ddPCR) method for the detection and absolute quantification of E. pyrifoliae using a pair of species-specific primers. The detection range was 10³-10⁷ copies/ml (DNA templates) and cfu/ml (cell culture templates). This new method exhibited good linearity and repeatability and was validated by absolute quantification of E. pyrifoliae DNA copies from samples of artificially inoculated immature pear fruits. Here, we present the first study of ddPCR assay for the detection and quantification of E. pyrifoliae. This method has potential applications in epidemiology and for the early prediction of black shoot blight outbreaks.

• The Plant Pathology Journal
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• v.36 no.5
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• pp.428-439
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• 2020

Erwinia pyrifoliae is a Gram-negative bacterial plant pathogen that causes black shoot blight in apple and pear. Although earlier studies reported the genome comparison of Erwinia species, E. pyrifoliae strains for such analysis were isolated in 1996. In 2014, the strain E. pyrifoliae EpK1/15 was newly isolated in the apple tree showing black shoot blight in South Korea. This study aimed to better understand the similarities and differences caused by natural variations at the genomic level between newly isolated E. pyrifoliae EpK1/15 and the strain Ep1/96, which were isolated almost 20 years apart. Several comparative genomic analyses were conducted, and Clusters of Orthologous Groups of proteins (COG) database was used to classify functional annotation for each strain. E. pyrifoliae EpK1/15 had similarities with the Ep1/96 strain in stress-related genes, Tn3 transposase of insertion sequences, type III secretion systems, and small RNAs. The most remarkable difference to emerge from this comparison was that although the draft genome of E. pyrifoliae EpK1/15 was almost conserved, Epk1/15 strain had at least three sorts of structural variations in functional annotation according to COG database; chromosome inversion, translocation, and duplication. These results indicate that E. pyrifoliae species has gone natural variations within almost 20 years at the genomic level, and we can trace their similarities and differences with comparative genomic analysis.

• Plant Disease and Agriculture
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• v.5 no.1
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• pp.46-49
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• 1999

Bacterial blight occurred on the flowers, leaves, twigs and branches of apple trees (Malus domestica Borkh) in Chunchon, Kangwon-Do. Infected flowers and leaves turned brownish black, and they were fallen down or remained hanging in the trees. Under humid conditions during postblossom period, the casual bacterium spreaded along twigs and killed them. However, killing of branches was very rare. The symptoms on apple trees were very similar to symptoms of fire blight. But the causal organism isolated from the lesions was identified as Pseudomonas syringae based on physiological and chemical characteristics. This is the first described bacterium that causes apple shoot blight in Korea.

• The Plant Pathology Journal
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• v.38 no.3
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• pp.194-202
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• 2022

Erwinia amylovora and Erwinia pyrifoliae cause fire blight and black-shoot blight, respectively, in apples and pears. E. pyrifoliae is less pathogenic and has a narrower host range than that of E. amylovora. Fire blight and black-shoot blight exhibit similar symptoms, making it difficult to distinguish one bacterial disease from the other. Molecular tools that differentiate fire blight from black-shoot blight could guide in the implementation of appropriate management strategies to control both diseases. In this study, a primer set was developed to detect and distinguish E. amylovora from E. pyrifoliae by conventional polymerase chain reaction (PCR). The primers produced amplicons of different sizes that were specific to each bacterial species. PCR products from E. amylovora and E. pyrifoliae cells at concentrations of 10⁴ cfu/ml and 10⁷ cfu/ml, respectively, were amplified, which demonstrated sufficient primer detection sensitivity. This primer set provides a simple molecular tool to distinguish between two types of bacterial diseases with similar symptoms.

• The Plant Pathology Journal
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• v.38 no.4
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• pp.355-365
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• 2022

Erwinia amylovora and E. pyrifoliae are the causative agents of destructive diseases in both apple and pear trees viz. fire blight and black shoot blight, respectively. Since the introduction of fire blight in Korea in 2015, the occurrence of both pathogens has been independently reported. The co-incidence of these diseases is highly probable given the co-existence of their pathogenic bacteria in the same trees or orchards in a city/district. Hence, this study evaluated whether both diseases occurred in neighboring orchards and whether they occurred together in a single orchard. The competition and virulence of the two pathogens was compared using growth rates in vitro and in planta. Importantly, E amylovora showed significantly higher colony numbers than E. pyrifoliae when they were co-cultured in liquid media and co-inoculated into immature apple fruits and seedlings. In a comparison of the usage of major carbon sources, which are abundant in immature apple fruits and seedlings, E. amylovora also showed better growth rates than E. pyrifoliae. In virulence assays, including motility and a hypersensitive response (HR), E. amylovora demonstrated a larger diameter of travel from the inoculation site than E. pyrifoliae in both swarming and swimming motilities. E. amylovora elicited a HR in tobacco leaves when diluted from 1:1 to 1:16 but E. pyrifoliae does not elicit a HR when diluted at 1:16. Therefore, E. amylovora was concluded to have a greater competitive fitness than E. pyrifoliae.

• 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.

• The Plant Pathology Journal
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• v.34 no.5
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• pp.445-450
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• 2018

Bacteriophages, bacteria-infecting viruses, have been recently reconsidered as a biological control tool for preventing bacterial pathogens. Erwinia amylovora and E. pyrifoliae cause fire blight and black shoot blight disease in apple and pear, respectively. In this study, the bacteriophage phiEaP-8 was isolated from apple orchard soil and could efficiently and specifically kill both E. amylovora and E. pyrifoliae. This bacteriophage belongs to the Podoviridae family. Whole genome analysis revealed that phiEaP-8 carries a 75,929 bp genomic DNA with 78 coding sequences and 5 tRNA genes. Genome comparison showed that phiEaP-8 has only 85% identity to known bacteriophages at the DNA level. PhiEaP-8 retained lytic activity up to $50^{\circ}C$, within a pH range from 5 to 10, and under 365 nm UV light. Based on these characteristics, the bacteriophage phiEaP-8 is novel and carries potential to control both E. amylovora and E. pyrifoliae in apple and pear.

• Research in Plant Disease
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• v.22 no.4
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• pp.269-275
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• 2016

Black shoot blight disease caused by Erwinia pyrifoliae have damaged economic loss to apple and pear growers until now since it was firstly reported in 1995 in Korea. This study was performed to reduce economic loss by mandatory eradication of all infected trees in case of more 10% disease incidence per orchard as official control. It also aims to set up effective management protocol for this disease by examining how far bacterial pathogen is present from the border of symptomatic and asymptomatic regions in infected apple twigs. Colony-PCR using isolated bacterial cells instead of genomic DNA was used to identify bacterial pathogen, EpSPF/EpSPR primer designed in enterobacterial repetitive intergenic consensus (ERIC) region was selected as specific for E. pyrifoliae. As results of monitoring of this disease during April to October in 2014-2015 by colony-PCR, occurrence of this disease was frequent from mid-May to early-July, when daily average temperature was around $25^{\circ}C$. Moreover, bacterial cells were continuously detected only in symptomatic regions and also asymptomatic regions of less than 20 cm from symptomatic regions. Therefore, we concluded that pruning of infected twigs at the region of more than 20 cm from symptomatic regions might be effective to manage black shoot blight disease in apple trees.