• Research in Plant Disease
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• v.13 no.2
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• pp.88-92
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• 2007

A total of 41 Pseudomonas syringae pv. syringae, the causal agent of bacterial blossom blight, were isolated from kiwifruit plants in Korea. Among them, two strains showing streptomycin resistance were examined to investigate the structure of resistant determinants by PCR and nucleotide sequence analysis. PCR results suggested that the streptomycin resistance is mediated by strA-strB genes carried on Tn5393a. Insertion sequences, IS6100 and IS1133, which were located within or downstream of tnpR gene in Xanthomonas campestris and Erwinia amylovora were not found. Nucleotide sequences of strA-strB were 100% identical with Tn5393a. Two stretomycin resistant strains had three plasmids. Southern blot hybridization using strA-strB probe indicated that the resistant genes were carried on a 100kb plasmid.

• Research in Plant Disease
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• v.28 no.4
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• pp.221-230
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• 2022

The fire blight caused by Erwinia amylovora (Ea) was first reported in 2015 in Korea, and the disease has rapidly spread to 22 regions until 2021. In Korea, all host plants in the apple and pear orchards where fire blight occurred should be eliminated and buried by the Plant Protection Act. To prevent the spread of the disease, all burial sites were prohibited from planting the new host plants for the next three years. To confirm the eradication efficiency of Ea and the reoccurrence of fire blight, the surveillance facilities were established on three burial sites from 2019 to 2020 in Anseong-si, Gyeonggi-do, and Chungju-si, Chungcheongbuk-do. As host plants, five apple trees of fire blight-susceptible cultivar 'Fuji', were planted in each facility. All facilities were enclosed with fences and nets and equipped with two CCTVs, motion sensors, and several other sensors for recording weather conditions to monitor the environment of the sentinel plants in real-time. The sentinel plants were checked for the reoccurrence of fire blight routinely. Suspicious plant parts were collected and analyzed for Ea detection by loop-mediated isothermal amplification polymerase chain reaction and conventional polymerase chain reaction. Until November 2022, Ea has not been detected in all sentinel plants. These results might support that the burial control of infected plants in soil works efficiently to remove Ea and support the possibility to shorten the prohibition period of host plant establishment in the burial sites.

• 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.

• Parasites, Hosts and Diseases
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• v.37 no.4
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• pp.257-263
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• 1999

Identification of the genes responsible for the recovery of virulence in brain-passaged Acanthamoeba culbertsoni was attempted via mRNA differential display polymerase chain reaction (mRNA DD-PCR) analysis. In order to identify the regulatory changes in transcription of the virulence related genes by the brain passages, mRNA DD-PCR was performed which enabled the display of differentially transcribed mRNAs after the brain passages. Through mRNA DD-PCR analysis. 96 brain-passaged amoeba specific amplicons were observed and were screened to identify the amplicons that failed to amplify in the non-brain-passaged amoeba mRNAs. Out of the 96 brain-passaged amoeba specific amplicons, 12 turned out to be amplified only from the brain-passaged amoeba mRNAs by DNA slot blot hybridization. The clone, A289C, amplified with an arbitrary primer of UBC ＃289 and the oligo dT$_{11}$-C primer, revealed the highest homology (49.8％) to the amino acid sequences of UPD-galactose lipid transferase of Erwinia amylovora, which is known to act as an important virulence factor. The deduced amino acid sequences of an insert DNA in clone A289C were also revealed to be similar to cpsD, which is the essential gene for the expression of type III capsule in group B streptococcus. Upregulated expression of clone A289C was verified by RNA slot blot hybridization. Similar hydrophobicity values were also observed between A289C (at residues 47-66) and the AmsG gene of E. amylovora (at residues 286-305: transmembrane domains). This result suggested that the insert of clone A289C might play the same function as galactosyl transferase controlled by the AmsG gene in E. amylovora.a.

• Herbal Formula Science
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• v.19 no.2
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• pp.83-92
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• 2011

Objectives : This study was carried out to moniter microbial flora on freeze-dried herbal powder and identify isolated bacteria. Methods : We measured the total number of bacteria and fungi in 29 herbal powder which had made according to the guideline of KFDA. For the identification, we observed microscopic properties and carried out polymerase chain reaction(PCR). The purified DNA was analyzed by DNA sequencer. Results : Among the 29 herbal powders, the fungi were detected only one sample as unacceptable range of total aerobic bacteria. Isolated bacteria were identified as Bacillus cereus, B. subtilis, B. megaterium, B. licheniformis, Erwinia tasmaniensis, E. amylovora, and Pantoea agglomerans by 16S rDNA analysis. E. tasmaniensis was observed 20 herbal samples. Conclusions : According to above results, further studies for the effective sterilization of low herbal materials should be needed.

• Microbiology and Biotechnology Letters
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• v.51 no.4
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• pp.538-541
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• 2023

In this study, we present the draft genome of Aureobasidium pullulans strain MHAU2101, which is the first strain to effectively control fire blight caused by Erwinia amylovora in Korea. The genome of strain MHAU2101 was composed of 28,669,322 base pairs, with a C+G content of 50.4%. The assembly comprised 17 contigs and had 99.22% completeness. The results of this study will be a valuable resource for future research on the biocontrol mechanism of A. pullulans strain MHAU2101.

• 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.

• Molecules and Cells
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• v.25 no.1
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• pp.30-42
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• 2008

The disease-specific (dsp) region and the hypersensitive response and pathogenicity (hrp) genes, including the hrpW, $hrpN_{Ep}$, and hrpC operons have previously been sequenced in Erwinia pyrifoliae WT3 [Shrestha et al. (2005a)]. In this study, the remaining hrp genes, including the hrpC, hrpA, hrpS, hrpXY, hrpL and hrpJ operons, were determined. The hrp genes cluster (ca. 38 kb) was comprised of eight transcriptional units and contained nine hrc (hrp conserved) genes. The genetic organization of the hrp/hrc genes and their orientation for the transcriptions were also similar to and collinear with those of E. amylovora, showing ${\geq}80%$ homologies. However, ORFU1 and ORFU2 of unknown functions, present between the hrpA and hrpS operons of E. amylovora, were absent in E. pyrifoliae. To determine the HR active domains, several proteins were prepared from truncated fragments of the N-terminal and the C-terminal regions of $HrpN_{Ep}$ protein of E. pyrifoliae. The proteins prepared from the N-terminal region elicited HR, but not from those of the C-terminal region indicating that HR active domains are located in only N-terminal region of the $HrpN_{Ep}$ protein. Two synthetic oligopeptides produced HR on tobacco confirming presence of two HR active domains in the $HrpN_{Ep}$. The HR positive N-terminal fragment ($HN{\Delta}C187$) was further narrowed down by deleting C-terminal amino acids and internal amino acids to investigate whether amino acid insertion region have role in faster and stronger HR activity in $HrpN_{Ep}$ than $HrpN_{Ea}$. The $HrpN_{Ep}$ mutant proteins $HN{\Delta}C187$ (D1AIR), $HN{\Delta}C187$ (D2AIR) and $HN{\Delta}C187$ (DM41) retained similar HR activation to that of wild-type $HrpN_{Ep}$. However, the $HrpN_{Ep}$ mutant protein $HN{\Delta}C187$ (D3AIR) lacking third amino acid insertion region (102 to 113 aa) reduced HR when compared to that of wild-type $HrpN_{Ep}$. Reduction in HR elicitation could not be observed when single amino acids at different positions were substituted at third amino acids insertion region. But, substitution of amino acids at L103R, L106K and L110R showed reduction in HR activity on tobacco suggesting their importance in activation of HR faster in the $HrpN_{Ep}$ although it requires further detailed analysis.

• 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.

• The Plant Pathology Journal
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• v.22 no.4
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• pp.375-380
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• 2006

In the survey from 1992 to 2000, twenty-eight parasitic diseases were observed in major apple producing areas in Korea. The predominant apple diseases were white rot(Botryosphaeria dothidea), Marssonina blotch(Marssonina mali), Valsa canker(Valsa ceratosperma), Alternaria leaf spot(Alternaria mali), and bitter rot(Collectotrichum gloeosporioides and C. acutatum). Apple scab that reappeared in 1990 after disappearance for 15 years was disappeared again since 1997. A viroid disease(caused by apple scar skin viroid) was newly found in this survey. The five diseases, fire blight(Erwinia amylovora), black rot(Botryosphaeria obtusa), scab(Cladosporium carpophilum), Monochaetia twig blight(Monochaetia sp.), and brown leaf spot(Hendersonia mali), which had once described in 1928 but no further reports on their occurrence, were not found in this survey. However, blossom blight(Monilinia mali), brown rot(Monilinia fructigena), and pink rot(Trichothecium roseum), which did not occur on apple after mid 1970s, were found in this survey.