• Korean Journal Plant Pathology
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• v.10 no.1
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• pp.13-17
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• 1994

A semiselective agar medium (XCO) was developed for the isolation of bacterial blight pathogen, Xanthomonas campestris pv. oryzae, from rice seed. The medium contained yeast extract 1.0 g, peptone 2.0 g, sucrose 5.0 g, sodium glutamate 1.0 g, FeSO4.7H2O 0.05 g, Fe.EDTA 1 mg, cephalexin 20 mg, Evan blue (0.1%) 1.5 ml, bromcresol purple (0.1%) 2.5ml, cycloheximide 100 mg and agar 15.0 g per liter. Colonies of X. c. pv. oryzae were 4~5 mm in diameter, smooth, round, blue (darker center) and convex after 6 days incubation at 28$^{\circ}C$. The recovery of 6 isolates of X. c. pv. oryzae on the XOC medium ranged from 81% to 120% (mean 98.2%) in comparison to Wakimoto's medium. The number of saprophytic bacteria from 10 rice seed lots on XCO medium was reduced to 70.4% of that on Wakimoto's medium. The recovery of X. c. pv. oryzae added to rice seed on XOC medium ranged from 67% to 87% (mean 75.6%) of that on Wakimoto's medium.

• Microbiology and Biotechnology Letters
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• v.32 no.3
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• pp.256-264
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• 2004

A new primer set was developed for the detection and identification of Xanthomonas oryzae pv. oryzae, the bacterial leaf blight (BLB) pathogen in rice plant. The nucleotide sequence of hpaA gene was determined from X. o. pv. oryzae str. KACC10331, and the sequence information was used to design primers for the application of the polymerase chain reaction (PCR). The nucleotide sequence of hpaA from X. o. pv. oryzae str. KACC 10331 was aligned with those of X. campestris pv. vesicatoria, X. campestris pv. campestris, X. axonopodis pv. citri, and X. axonopodis pv. glycines. Based on these results, a primer set(XOF and XOR) was designed for the specific detection of hpaA in X. o. pv. oryzae. The length of PCR products amplified using the primer set was 534-bp. The PCR product was detected from only X. o. pv. oryzae among other Xanthomonas strains and reference bacteria. This product was used to confirm the conservation of hpaA among Xanthomonas strains by Southern-blotting. Furthermore, PCR amplification with XOF and XOR was used to detect the pathogen in an artificially infected leaf. The sensitivity of PCR detection in the pure culture suspension was also determined. This PCR-based detection methods will be a useful method for the detection and identification of X. o. pv. oryzae as well as disease forecasting.

• Korean Journal of Plant Resources
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• v.12 no.2
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• pp.152-160
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• 1999

Bacterial population density on soybean leaves was $10^2~10^5CFU/cm^2$. Bacterial population density was increased by progress of plant growth stage. Population density of soybean sprout rotting bacteria on soybean leaves was $0~10^3CFU/cm^2$. Population density of soybean sprouts rotting bacteria was related to cultivating area, but not related to plant growth stage. Cultivar and population density of soybean sprout rotting bacteria were less corelated, and varied by plant growth stages and plant parts. Erwina cypripedii, E. carotovora subsp. carotovora, Xanthomonas campestris pv. glycines, Staphylococcus sp., and Micrococcus sp. were identified as pathogenic bacteria causing soybean sprout rot. In generally population density of E. cypripedii, E. carotovora subsp. carotovora, Micrococcus sp., and X. campestris pv. glycines were high.

• Korean Journal Plant Pathology
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• v.13 no.3
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• pp.179-183
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• 1997

We have screened hypersensitive responses of 18 cultivars of Nicotiana tabacum to Xanthomonas campestris pv. campestris. TC500 cultivar produced the most strong hypersensitive response (HR) to Xanthomonas campestris pv. campestris. By NTG mutagenesis, nonhypersensitive mutants (XHN 514-774, XHN 620-831) were generated, which does not induce hypersensitive response on tobacco leaves (Nicotiana tabacum cv. TC500). Also nonpathogenic mutant (XPN 1001), which does not incite any of the black rot symptoms on leaves was generated. We observed that HR mutants were still pathogenic on cabbage leaves producing black rot symptoms and nonpathogenic mutant induced HR in tobacco leaves. The inplanta growth of wild type and HR mutants were examined for up to 120 hrs after inoculation : population of wild type strain increased to $10^{8}$ in 24hrs, but rapidly declined thereafter; HR-mutants increased to more than $10^{6}$ in 48 hrs after inoculation but subsequently stabilized and slowly decreased. We observed that wild type and these mutans produced similar amounts of degradative enzymes such as protease, pectate lyase, cellulase and amylase.

• Korean Journal Plant Pathology
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• v.11 no.1
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• pp.53-60
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• 1995

Inoculation with the compatible strain Ds 1 of Xanthomonas campestris pv. vesicatoria caused brownish ad water-soaked lesions, but incompatible strain Bv5-4a produced hypersensitive symptoms with local necrosis on tomato (cv. Kwangyang) leaves. Bacterial populations of the compatible strains Ds 1 propagated more greatly than the incompatible strain Bv5-4a at the frist onset, but no differences were observed 5 days after inoculation. The bacterial infection induced the synthesis and accumulation of soluble proteins in tomato leaves, especially in the incompatible interaction. Native-polyacrylamide gel electrophoresis distinguished the soluble proteins in the tomato leaves infected by the compatible or incompatible strains. A protein of low molecular weight occurred only in the incompatible interaction. Some pathogenesis-related (PR) proteins, especially the 15, 18, 23, 26 and 54 kDa proteins, were detected only in the infected tomato leaves. In the two-dimensional electrophoresis, some proteins with different molecular weights (Mr. 21∼29 kDa) and the pI 8∼9 appeared more distinctly only in the incompatible interaction. These data suggest that the de novo synthesis of some PR proteins in tomato may be significant in defense against X. c. pv. vesicatoria.

• Korean Journal Plant Pathology
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• v.14 no.2
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• pp.125-129
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• 1998

Phytopathogenic bacteria causing soft-rot many vegetables; extracellular enzymes produced by them, pectate lyase(Pel) is important pathogenicity facotrs which cause tissue maceration and cell death. Ten of seventeen plant pathogenic bacteria showed weak Pel activity, four of them showed low Pel activity and Erwinia acrotovora subsp. carotovora, E. chrysanthemi, Pseudomonas marginalis and Xanthomonas campestris pv. campestris showed high Pel activity in the polygalacturonate yeast extract agar (PAY) plate. High Pel activity of the four bacteria species produced the highest Pel activity when pectin or polygalacturonic acid (PGA) was added to minimal salts (MS) medium. Pel activity of the four bacterial species was the highest at 2$0^{\circ}C$ among different temperature conditions. The rate and amount of maceration of potato tuber tissue were highest at 2$0^{\circ}C$ in E. carotovora subsp. carotovora, E. chrysanthemi and P. marginalis, while those were the highest at $25^{\circ}C$ in X. campestris pv. campetris.

• Korean Journal Plant Pathology
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• v.13 no.1
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• pp.5-12
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• 1997

Genetic diversity of forty-four strains of Xanthomonas campestris pv. vesicatoria from diverse geographic origins was investigated using random amplified polymorphic DNA (RAPD) of genomic DNA. One hundred and thirty-seven amplified fragments were produced by polymerase chain reaction with a set of 14 random primers, and the sizes of amplified DNA fragments ranged approximately from 0.3 to 3.2 kb. Cluster analysis of genetic similarity among the strains generated the dendrogram that clearly separated all strains from each other. The 44 strains of X. campestris pv. vesicatoria were classified into 4 major genomic DNA RAPD groups and 15 subgroups at the genetic similarity of 0.60 and 0.92, respectively. The strains from foreign countries formed discrete subgroups, but the United States strain 87-77 clustered closely with some of Korean strains together. Thirty-nine Korean strains were classified into 11 subgroups, and especially Masan strain Ms93-1 clustered distinctly far from the other Korean strains. RAPD polymorphism suggests strongly the occurrence of genetic differentiation of X. campestris pv. vesicatoria and the existence of genetically distinctive subgroups among the populations in Korea.

• Research in Plant Disease
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• v.12 no.2
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• pp.134-138
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• 2006

A new bacterial disease of broccoli (Brassica oleracea var. italica) was observed on field-grown plants in Pyungchang during 2003 and 2004. Seedling infections first appeared as a blackening along the margins of the cotyledon. Cotyledon shriveled and dropped off. Infected seedlings were stunted and yellowed and eventually died. The disease was easily recognized by the presence of yellow, V-shaped, or U-shaped areas extending inward from margin of the leaf. As the disease progressed, the yellow lesions turned brown and the tissues died. Isolations made from diseased leaves on yeast extract dextrose calcium carbonate agar yielded nearly pure cultures of a yellow-pigmented bacterium typical of a xanthomonad. Two bacterial strains were purified and used for further tests. Pathogenicity of strains was confirmed on 3-week-old crucifer (cabbage, Chinese cabbage, kale, radish and broccoli) plants cut by scissors with bacterial suspensions containing $10^8 cfu/ml$ of phosphate buffered saline. The Biolog and fatty acid analyses and 16S rDNA sequencing of two strains (SL4797 and SL4800) from broccoli black rot showed that they could be identified as X. campestris pv. campestris because of their high similarity to the tester strain (X. campestris pv. campestris NCPPB528) with a match probability of 100%. This is the first report of black rot of broccoli in Korea.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.71.1-71
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• 2003

Most major plant pathogenic bacteria in Korea belong to Xanthomonas spp.. Xanthomonas oryzae pv. oryzae is a major pathogen in rice, X. campestris pv. vesicatoria in pepper, X. axonopodis pv. giycines in soybean, X. campestris pv. campestris in cabbage, and X. axonoposid pv. citri in tangerin. Host specificity of the bacterial pathogen depends on the avirulence gene in the pathogen and the corresponding resistance gene in host plants. Many avirulence genes in bacteiral pathogen located on the native plasmids. However, the presence of the native plasmids in Xanthomonas spp. was not investigated well. In order to study the host specificity, we isolated native plasmids from Xanthomonas spp. and compared those plasmids each other, The presence of the native plasmids and the characteristics of the plasmids depended on the bacterial strains. In the X. axonopodis pv. glycines, most strains carried native plasmids but some strains did not. Some strains carry about 60 kb native plasmids including 3 different aviurlence genes. We will discuss the characteristics of the native plasmids isolated from the Xanthomonas spp.

• The Plant Pathology Journal
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• v.15 no.1
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• pp.57-61
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• 1999

Xanthomonas campestris pv. glycines causes bacterial pustule disease on susceptible soybean leaves and produces a bacteriocin, named glycinecinA, against most xanthomonads including Xanthomonas campestris pv. vesicatoria. One of the 5 isolated DNA regions responsible for bacteriocin production, a 1.7 kb DNA region for the glycinecinA gene, was used as a probe to detect the presence of the homolog DNA in other bacterial strains. Among 55 bacterial strains tested, only X. campestris pv. glycines showed the positive signal with glycinecinA DNA. Two oligomers, heu2 and heu4, derived from a glycinecinA DNA were used to carry out the polymerase chain reaction (PCR) analysis with chromosomal DNA from 55 different bacterial strains including 24 different strains of X. campestris pv. glycines, 9 different pathovars of xanthomonads, and other 22 bacterial strains of different genus and species. By separation of the PCR products on agarose gel, a 0.86 kb DNA fragment was specifically detected when X. campestris pv. glycines was present in the amplification assay. The 0.86 kb fragment was not amplified when DNA from other bacteria was used for the assay. Southern analysis with glycinecinA DNA showed that the PCR signal was obtained with X. campestris pv. glycines isolates from various geographic regions and soybean cultivars. Therefore, the 1.7 kb DNA region for the glycinecinA gene can be used for the pathovar-specific probe for the DNA hybridization and the primers heu2 and heu4 can be used for the pathovar-specific primers for the PCR analysis to detect X. campestris pv. glycines.