• 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.

• 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.

• The Plant Pathology Journal
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• v.26 no.1
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• pp.63-69
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• 2010

The effect of Acibenzolar-S-methyl (ASM) and Rahnella aquatilis Ra39 against apple fire blight disease caused by Erwinia amylovora were tested as a possible alternative to streptomycin. In vitro studies, no inhibition effect against the pathogen was found when ASM was tested. Under greenhouse conditions, application of R. aquatilis Ra39 with the highly susceptible M26 rootstock resulted in a marked disease suppression. Application of ASM and strain Ra39 caused a high decrease of the disease, 82% and 58% respectively; this was correlated with a reduction of the growth of the pathogen within host plants up to 64% and 49.5% respectively. Further studies in the field under artificial infection condition during full bloom revealed that application of ASM and R. aquatilis Ra39 with Gala variety resulted in a control effect up to 21 and 29% respectively. In physiological studies, enhanced activities of PR-proteins (chitinase and $\beta$-1, 3-glucanase) were detected, which are well known as biochemical markers for systemic acquired resistance. Application of ASM to apple shoots caused the highest chitinase activity followed by strain Ra39. The enzyme activity was increased after 2, 4 and 6 days from application. In addition, ASM-treatment caused the higher $\beta$-1, 3-glucanase activity than strain Ra39. Maximum enzyme activity was recorded after 6 days from application and then decreased after 8 and 10 days from application.

• Research in Plant Disease
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• v.24 no.1
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• pp.41-51
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• 2018

Since fire blight disease on apple and pear was produced in Korea in 2015, there were no registered chemicals to control against this disease. Instead, several antibacterial chemicals that were registered for other bacterial diseases such as soft rot and bacterial spot have been authorized by Rural Development Administration (RDA). However, these chemicals are not tested efficacy for fire blight disease except damage by those treatments on apple and pear in Korea. Thus, we evaluated efficiency using in vitro and in planta assays of antibacterial chemicals such as antibiotics and copper compounds including kasugamycin, oxytetracycline, oxolinic acid and streptomycin, and copper hydroxide, copper sulfate, oxine copper and tribasic copper sulfate, respectively. We also tested two kinds of biological agents. As expected, significant antibacterial effect was observed in vitro test of both antibiotics and copper-based chemicals. In planta test based on disease severity including ooze and water-soaked formation on immature pears, bacterial populations on blooms, and blight lesion formation in artificially inoculated shoots, kasugamycin, oxytetracycline and streptomycin have been shown the most efficiency among tested antibiotics. Four copper-based chemicals tested in this study, control effects are little bit lower than agricultural antibiotics but they seem to be available to use in terms of winter season. Biocontrol agents were also shown possibility to treat in eco-friendly farms. In addition, there are no antibiotic resistance genes in Korean isolates against antibiotics, which were selected for suppression of fire blight in this study.

• Research in Plant Disease
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• v.29 no.3
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• pp.316-320
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• 2023

Recently, the domestic plant disease control policy for fire blight has been implemented partial control in addition to burial control. In this study, an on-site management survey was conducted targeting orchards that implemented partial disease control from 2019 to 2020 in order to find efficient implementation methods for partial disease control. As a result of an investigation into 22 pear and apple orchards in Cheonan and Chungju, 7 orchards were buried. The upper part of the cut infected plants was burned at 16 orchards and covered with plastic vinyl after lime treatment at 6 orchards. The lower stumps of cut infected plants were burned at 7 orchards and covered with plastic vinyl after lime treatment at 15 orchards. There were two orchards where suckers appeared on the stumps even though covers were applied. There was no infection by Erwinia amylovora in the suckers. The conservation condition of lime treatment was good, but warning signs were absent at 6 orchards. Most orchards treated the stumps and surrounding areas with glyphosate-isopropylamine herbicide. The effect of partial control was judged to be safe.

• Korean Journal of Agricultural and Forest Meteorology
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• v.23 no.1
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• pp.15-33
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• 2021

In this study, the possibility of discriminating Fire blight (FB) infection tested using the hyperspectral imagery. The reflectance of healthy and infected leaves and branches was acquired with 5 nm of full width at high maximum (FWHM) and then it was standardized to 10 nm, 25 nm, 50 nm, and 80 nm of FWHM. The standardized samples were divided into training and test sets at ratios of 7:3, 5:5 and 3:7 to find the optimal bands of FWHM by the decision tree analysis. Classification accuracy was evaluated using overall accuracy (OA) and kappa coefficient (KC). The hyperspectral reflectance of infected leaves and branches was significantly lower than those of healthy green, red-edge (RE) and near infrared (NIR) regions. The bands selected for the first node were generally 750 and 800 nm; these were used to identify the infection of leaves and branches, respectively. The accuracy of the classifier was higher in the 7:3 ratio. Four bands with 50 nm of FWHM (450, 650, 750, and 950 nm) might be reasonable because the difference in the recalculated accuracy between 8 bands with 10 nm of FWHM (440, 580, 640, 660, 680, 710, 730, and 740 nm) and 4 bands was only 1.8% for OA and 4.1% for KC, respectively. Finally, adding two bands (550 nm and 800 nm with 25 nm of FWHM) in four bands with 50 nm of FWHM have been proposed to improve the usability of multispectral image sensors with performing various roles in agriculture as well as detecting FB with other combinations of spectral bands.