• Title/Summary/Keyword: Xanthomonas

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Cloning of a Chitinase Gene of Xanthomonas sp. Isolated from Soil and its Expression in E. coli. (토양에서 분리된 Xanthomonas sp.의 Chitinase 유전자 cloning과 E.coli에서의 발현)

  • Kim, Ho-Sang;Seong, Ki-Young;Eun, Moo-Young;Hwang, Cher-Won
    • Applied Biological Chemistry
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    • v.41 no.2
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    • pp.125-129
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    • 1998
  • Xanthomonas sp. isolated from soil exhibited cell wall lytic activity of Candida albicans and secreted chitinase in chitin media. Especially, the chitinase activity was induced by chitin and reached a maximum level at 3 days culture in chitin media. We constructed genomic library of Xanthomonas sp. using cosmid vector in E. coli. Oligonucleotide probe was synthesized from the consensus sequence corresponding to chitinase active site, which was derived from the comparison of amino acid sequences of bacterial chitinase genes. Using this oligonucleotide probe, we screened the genomic library. By restriction enzyme mapping of the positive clones, we identified 4 independent clones which may contain the chitinase gene. One of the clones, named pXCH1 (1.2 kb insert), was further analyzed. Northern blot analysis indicated that is transcripts, 1 kb and 0.8 kb, were induced by chitin. When the cloned gene was induced by IPTG in E.coli cell, chitinase activity which was secreted onto culture media was not observed. However, when the cell was disrupted by using sonicator and then centrifuged, the supernatant exhibited chitinase activity. SDS-PAGE of the supernatant indicated that about 35 kDa protein was induced by IPTG. From these results, it was concluded that the cloned DNA was one of the chitinase genes of Xanthomonas sp.

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The Investigation on the Optimum Culture Conditions and the Ice Nucleating Activity of Bacterium Xanthomonas translucens KCTC 2751 (Xanthomonas translucens KCTC 2751의 최적배양과 빙핵 활성 검토)

  • Kim, Young-Mun;Kang, Sung-Il;Jang, Young-Boo;Jun, Byung-Jin;Kong, Jai-Yul
    • Korean Journal of Food Science and Technology
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    • v.38 no.2
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    • pp.249-255
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    • 2006
  • The optimum culture conditions for the ice nucleating activity and the cell growth of Xanthomonas translucens KCTC 2751 were investigated. The optimum initial pH and temperature for the cell growth and the ice nucleating activity were 6.5 and $25^{\circ}C$, respectively. The optimum culture medium for the ice nucleating activity was composed of 1.0% maltose, 1.4% yeast extract, 0.8% digested of gelatin, and 0.03% KCI in distilled water. Freezing operations carried out on distilled water showed that the degrees of supercooling were $-7.90^{\circ}C$ without ice nucleators, $-1.56^{\circ}C$ with silver iodide as a commercial ice nucleator, and $-1.36^{\circ}C$ when Xanthomonas translucens KCTC 2751 were added. During progressive freeze-concentration assays, the addition of Xanthomonas translucens KCTC 2751 led to lower saccharose concentrations in the crystals, while the cells led to higher saccharose concetrations in the concentrated phase.

PCR-Based Sensitive Detection and Identification of Xanthomonas oryzae pv. oryzae (중합효소연쇄 반응에 의한 벼 흰잎마름병균의 특이적 검출)

  • Lee, Byoung-Moo;Park, Young-Jin;Park, Dong-Suk;Kim, Jeong-Gu;Kang, Hee-Wan;Noh, Tae-Hwan;Lee, Gil-Bok;Ahn, Joung-Kuk
    • 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.

Detection of Xanthomonas axonopodis pv. glycines and Survey on Seed Contamination in Soybean Seeds Using PCR Assay (PCR Assay 이용 콩 종자에서 Xanthomonas axonopodis pv. glycines 검출 및 종자오염 조사)

  • Hong, Sung-Jun;Hong, Yeon-Kyu;Lee, Bong-Choon;Lim, Mi-Jung;Yoon, Young-Nam;Hwang, Jae-Bok;Song, Seok-Bo;Park, Sung-Tae
    • Research in Plant Disease
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    • v.13 no.3
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    • pp.145-151
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    • 2007
  • Xanthomonas axonopodis pv. glycines is the causal agent of bacterial pustule of soybean(Glycine max. (L.) Merr), which is one of the most prevalent bacterial diseases in Korea. In this study, Polymerase Chain Reaction (PCR) assay was applied to detect Xanthomonas axonopodis pv. glycines and to survey on seed contamination in 36 soybean cultivars of Korea. And we have to compare PCR assay with dilution-plating assay of detection and identification. We confirmed detection of pathogen from artificial infected seeds and natural Infected seeds using PCR assay. This assay gave results similar to a seed-wash dilution plating assay and proved more effective than classical methods. Results of survey on seed contamination by X. axonopodis pv. glycines from 36 cultivar seeds showed that the pathogen was detected from Pungsan-namulkong, Mallikong, Taekwangkong, Daemangkong, Ajukkarikong using PCR assay. Therefore, The PCR assay provides a sensitive, rapid tool for the specific detection of X. axonopodis pv. glycines in soybean seeds.

Selection of Small Synthetic Antimicrobial Peptides Inhibiting Xanthomonas citri subsp. citri Causing Citrus Canker

  • Choi, Jeahyuk;Park, Euiho;Lee, Se-Weon;Hyun, Jae-Wook;Baek, Kwang-Hyun
    • The Plant Pathology Journal
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    • v.33 no.1
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    • pp.87-94
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    • 2017
  • Citrus canker disease decreases the fruit quality and yield significantly, furthermore, emerging of streptomycin-resistant pathogens threatens the citrus industry seriously because of a lack of proper control agents. Small synthetic antimicrobial peptides (AMPs) could be a promising alternative. Fourteen hexapeptides were selected by using positional scanning of synthetic peptide combinatorial libraries. Each hexapeptide showed different antimicrobial spectrum against Bacillus, Pseudomonas, Xanthomonas, and Candida species. Intriguingly, BHC10 showed bactericidal activity exclusively on Xanthomonas citri subsp. citri (Xcc), while BHC7 was none-active exclusively against two Pseudomonas spp. at concentration of $100{\mu}g/ml$ suggesting potential selectivity constrained in hexapeptide frame. Three hexapeptides, BHC02, 06 and 11, showed bactericidal activities against various Xcc strains at concentration of $10{\mu}g/ml$. When they were co-infiltrated with pathogens into citrus leaves the disease progress was suppressed significantly. Further study would be needed to confirm the actual disease control capacity of the selected hexapeptides.

Shot Hole of Peach and Japanese Plum Caused by Xanthomonas campestris pv. Pruni and Erwinia nigriflens in Korea (Xanthomonascampestris pv. pruni와 Erwinia nigrifluens에 의한 복숭아 및 자두의 세균성구멍병)

  • 최재을;이은정;박영섭
    • Research in Plant Disease
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    • v.6 no.1
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    • pp.10-14
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    • 2000
  • In 1998, bacterial shot hole of peach (Prunus persica) and Japanese plum(Prunus salicina) was found in Naju and Milyang. Five isolates of bacteria isolates from the diseased leaves and fruits of peach and Japanese plum were classified into genus Erwinia and Xanthomonas on diagnostic characteristics. Of five isolates, two were identified as X. campestris pv. pruni, three as E. nigrifluens. E.nigrifluens is the first description of bacteria which causes the disease on peach and Japanese plum in Korea. the symptoms caused by E. nigrifluens were hardly distinguished from those caused by X. campestris pv. pruni. In addition, it was observed that two pathogenic bacteria were isolated from most of naturally infected plants at the same time. from the reason mentioned above, we proposed to use a single common name \"bacterial shot hole of peach and Japanese plum\" for the both bacterial diseases, hereafter.

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PCR-Based Assay for Rapid and Specific Detection of the New Xanthomonas oryzae pv. oryzae K3a Race Using an AFLP-Derived Marker

  • Song, Eun-Sung;Kim, Song-Yi;Noh, Tae-Hwan;Cho, Heejung;Chae, Soo-Cheon;Lee, Byoung-Moo
    • Journal of Microbiology and Biotechnology
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    • v.24 no.6
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    • pp.732-739
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    • 2014
  • We describe the development of a polymerase chain reaction method for the rapid, precise, and specific detection of the Xanthomonas oryzae pv. oryzae (Xoo) K3a race, the bacterial blight pathogen of rice. The specific primer set was designed to amplify a genomic locus derived from an amplified fragment length polymorphism specific for the K3a race. The 1,024 bp amplicon was generated from the DNA of 13 isolates of Xoo K3a races out of 119 isolates of other races, pathovars, and Xanthomonas species. The assay does not require isolated bacterial cells or DNA extraction. Moreover, the pathogen was quickly detected in rice leaf 2 days after inoculation with bacteria and at a distance of 8 cm from the rice leaf 5 days later. The results suggest that this PCR-based assay will be a useful and powerful tool for the detection and identification of the Xoo K3a race in rice plants as well as for early diagnosis of infection in paddy fields.

Rheological Properties of Exopolysaccharide Produced by Xanthomonas sp. EPS-1 (Xanthomonas sp. EPS-1이 생산하는 다당류의 리올로지 특성)

  • Son, Bong-Soo;Park, Seok-Kyu;Kang, Shin-Kwon;Lee, Sang-Won;Sung, Nack-Kie
    • Microbiology and Biotechnology Letters
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    • v.23 no.3
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    • pp.269-274
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    • 1995
  • For the screening of a new functional exopolysaccharide, sugar composition and rheological properties of exopolysaccharide produced from Xanthomonas sp. EPS-1 were investigated. The average molecular weight of exopolysaccharide was determined to be approximately 2.l $\times$ 10$^{6}$ dalton. The new exopolysaccharide EPS-1 was composed of mannose, glucose, galactose and gluco- samine. IR analysis showed that the exopolysaccharide EPS-1 was assumed to be polymer with carbohydrates. NMR analysis showed that exopolysaccharide EPS-1 was presumed to be 4 units of sugar and trace of CH$_{3}$ group. Exopolysaccharide EPS-1 solution showed a characteristic of non-Newtonian fluid properties. At the concentration of 1.0%, the consistency index and the flow behavior index were shown at 10.8352 poise-sec and 0.4419, respectively. All dispersions were pseudoplastic fluids described accurately by Power-law model. Exopolysaccharide EPS-1 was highly viscous at low concentration, with good stability over a wide range of pH 5 to 13. The excellent compatibility of exopolysaccharide EPS-1 was represented with salts such as sodium chloride.

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Bacterial Multiplications and Electrophoretic Patterns of Soluble Proteins in Compatible and Incompatible Interactions of Pepper Leaves with Xanthomonas campestirs pv. vesicatoria (Xanthomonas campestris pv. vesicatoria에 감염된 고추잎의 친화적, 불친화적 반응에서 세균증식과 수용성 단백질의 전기영동 패턴)

  • 이연경;김영진;황병국
    • Korean Journal Plant Pathology
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    • v.10 no.4
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    • pp.305-313
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    • 1994
  • Typically susceptible lesions were developed on pepper (cv. Hanbyul) leaves inoculated with the compatible strains Ds 1 of Xanthomonas campestris pv. vesicatoria. The lesions appeared first water-soaked and then turned yellow with a chlorotic area. In contrast, the leaves inoculated with the incompatible strain 81-23 initially turned yellow and then developed local necrosis. Multiplication of x. c. pv. vesicatoria in pepper leaves also were distinctly different between the two strains. The strain Ds 1 multiplied more greatly than did the strain 81-23 in the infected leaves. X. c. pv. vesicatoria infection of pepper leaves induced the synthesis of soluble proteins, especially more greatly in the compatible than in the incompatible interactions. Some pathogenesis-related (PR) proteins were detected in the intercellular washing fluid (IWF) and extracts of the infected pepper leaves. In particular, the 32 kDa protein on SDS-PAGE gels appeared intensely in the incompatible interaction. In contrast, some proteins with moluecular masses of 65, 71, and 75 kDa disappeared in the infected pepper leaves. Isoelectric focusing could identify the pIs of soluble proteins in infected pepper leaves. The accumulation of the IWF from infected leaves was more conspicuous in the incompatible than the compatible interaction. These results suggest that some extremely acidic and basic proteins were induced and accumulated in the intercellular spaces of infected pepper leaves.

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