• 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.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2021.04a
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• pp.23-23
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• 2021

Apple (Malus×domestica Borkh.; Rosaceae) is an important fruit crop grown mainly in temperate regions of the world. Tissue culture in vitro is a biotechnological technique that has been used to genetically improve cultivars (scions) and rootstocks. This could be important in the production of genetically uniform scions and rootstocks for commercial apple production. In nurseries, apple plants are produced by grafting scions onto rootstocks. The Cornell-Geneva (Geneva® series) breeding program has bred several dwarf rootstocks that are resistant to diseases and pests and are also cold hardy. This study was conducted to determine the optimal medium strength to improve sprouting shoot rate of apical meristem of the apple rootstocks of fire blight resistance. The apple rootstocks apical meristem at size (0.2 mm to 0.3 mm) with axillary buds were cultured on the MS(Murashige & Skoog) medium supplemented with plant growth regulators. The sprouting ratio and growth characteristics was evaluated after eight weeks in vitro culture. The highest rate of bud differentiation and shoot formation were 23.8% and 55.6%, respectively. After 6 weeks, shoots were regenerated from apical meristem, and their growth characteristics was significantly varied on the respective basal medium with different plant growth regulators. Our studies showed that the apple rootstocks the apple rootstocks of fire blight resistance plantlets could be successfully produced from apical meristem differentiated out of young twigs via organogenic regeneration.

• Korean Journal of Agricultural Science
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• v.50 no.2
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• pp.241-247
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• 2023

Erwinia amylovora , which causes fire blight disease on apple and pear trees, is one of the most important phytopathogens because of its devastating impact. Currently, the only way to effectively control fire blight disease is through the use of antibiotics such as streptomycin, kasugamycin, or oxytetracycline. However, problems with the occurrence of resistant strains due to the overuse of antibiotics are constantly being raised. It is therefore necessary to develop novel disease control methods through an advanced understanding of the pathogenesis mechanism of E. amylovora . To better understand the pathogenesis of E. amylovora , we investigated unknown virulence factors by random mutagenesis and screening. Random mutants were generated by Tn5 transposon insertion, and the pathogenicity of the mutants was assessed by inoculation of the mutants on apple fruitlets. A total of 17 avirulent mutants were found through screening of 960 random mutants. Among them, 14 mutants were already reported as non-pathogenic strains, while three mutants, TS3128_M2899 (ΔSUFU ), TS3128_M2939 (ΔwcaG ), and TS3128_M3747 (ΔrecB ), were not reported. Further study of the association between E. amylovora pathogenicity and these 3 novel genes may provide new insight into the development of control methods for fire blight disease.

• The Plant Pathology Journal
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• v.37 no.4
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• pp.396-403
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• 2021

Fire blight disease, caused by Erwinia amylovora, could damage rosaceous plants such as apples, pears, and raspberries. In this study, we designed to understand how E. amylovora affected other bacterial communities on apple rhizosphere; twig and fruit endosphere; and leaf, and fruit episphere. Limited studies on the understanding of the microbial community of apples and changes the community structure by occurrence of the fire blight disease were conducted. As result of these experiments, the infected trees had low species richness and operational taxonomic unit diversity when compared to healthy trees. Rhizospheric bacterial communities were stable regardless of infection. But the communities in endosphere and episphere were significanlty affected by E. amylovora infection. We also found that several metabolic pathways differ significantly between infected and healthy trees. In particular, we observed differences in sugar metabolites. The finding provides that sucrose metabolites are important for colonization of E. amylovora in host tissue. Our results provide fundamental information on the microbial community structures between E. amylovora infected and uninfected trees, which will contribute to developing novel control strategies for the fire blight disease.

• The Plant Pathology Journal
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• v.37 no.4
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• pp.404-412
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• 2021

Despite the plant microbiota plays an important role in plant health, little is known about the potential interactions of the flower microbiota with pathogens. In this study, we investigated the microbial community of apple blossoms when infected with Erwinia amylovora. The long-read sequencing technology, which significantly increased the genome sequence resolution, thus enabling the characterization of fire blight-induced changes in the flower microbial community. Each sample showed a unique microbial community at the species level. Pantoea agglomerans and P. allii were the most predominant bacteria in healthy flowers, whereas E. amylovora comprised more than 90% of the microbial population in diseased flowers. Furthermore, gene function analysis revealed that glucose and xylose metabolism were enriched in diseased flowers. Overall, our results showed that the microbiome of apple blossoms is rich in specific bacteria, and the nutritional composition of flowers is important for the incidence and spread of bacterial disease.

• The Plant Pathology Journal
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• v.38 no.3
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• pp.248-253
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• 2022

Erwinia amylovora is a devastating bacterial plant pathogen that infects Rosaceae including apple and pear and causes fire blight. Bacteriophages have been considered as a biological control agent for preventing bacterial infections of plants. In this study, nine bacteriophages (ΦFifi011, ΦFifi044, ΦFifi051, ΦFifi067, ΦFifi106, ΦFifi287, ΦFifi318, ΦFifi450, and ΦFifi451) were isolated from soil and water samples in seven orchards with fire blight in Korea. The genetic diversity of bacteriophage isolates was confirmed through restriction fragment length polymorphism pattern analysis. Host range of the nine phages was tested against 45 E. amylovora strains and 14 E. pyrifoliae strains and nine other bacterial strains. Among the nine phages, ΦFifi044 and ΦFifi451 infected and lysed E. amylovora only. And the remaining seven phages infected both E. amylovora and E. pyrifoliae. The results suggest that the isolated phages were different from each other and effective to control E. amylovora, providing a basis to develop biological agents and utilizing phage cocktails.

• 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.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.

• 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.

• Research in Plant Disease
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• v.29 no.1
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• pp.79-81
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• 2023

During the nationwide survey of fire blight, the typical shoot blight symptoms were found on Korean mountain ash (Sorbus alnifolia) which was located near an orchard that produced fire blight on pear trees in Eumseong, Korea, May 2021. To identify the causal agent, we progressed isolation from the symptomatic leaves and shoots. Two white and mucoid colonies were isolated into the pure culture. Two isolates were identified as Erwinia amylovora according to the colony-polymerase chain reaction (PCR) with amsB primers and the phylogenetic tree using 16S rRNA sequences. To test of pathogenicity of two isolates, we inoculated immature pear fruits and understock of apple. We observed necrosis and oozes on immature pear fruits and shoot blight resulting in necrosis on apple shoots six days after inoculation. Colonies were recovered from the inoculated pears and apples, and identity was confirmed through colony PCR for amsB genes. To our knowledge, E. amylovora was first reported on Korean mountain ash native to South Korea.