• Title/Summary/Keyword: Xanthomonas campestris pv. vesicatoria.

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Homologous Expression and T3SS-Dependent Secretion of TAP-Tagged Xo2276 in Xanthomonas oryzae pv. oryzae Induced by Rice Leaf Extract and Its Direct In Vitro Recognition of Putative Target DNA Sequence

  • Kim, Seunghwan;Nguyen, Thi-Dieu-Hanh;Lee, Joohee;Hong, Myoung-Ki;Pham, Tan-Viet;Ahn, Yeh-Jin;Lee, Byoung-Moo;Han, Ye Sun;Kim, Dong-Eun;Kim, Jeong-Gu;Kang, Lin-Woo
    • Journal of Microbiology and Biotechnology
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    • v.23 no.1
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    • pp.22-28
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    • 2013
  • Xo2276 is a putative transcription activator-like effector (TALE) in Xanthomonas oryzae pv. oryzae (Xoo). Xo2276 was expressed with a TAP-tag at the C-terminus in Xoo cells to enable quantitative analysis of protein expression and secretion. Nearly all TAP-tagged Xo2276 existed in an insoluble form; addition of rice leaf extracts from a Xoosusceptible rice cultivar, Milyang23, significantly stimulated secretion of TAP-tagged Xo2276 into the medium. In a T3SS-defective Xoo mutant strain, secretion of TAPtagged Xo2276 was blocked. Xo2276 is a Xoo ortholog of Xanthomonas campestris pv. vesicatoria (Xcv) AvrBs3 and contains a conserved DNA-binding domain (DBD), which includes 19.5 tandem repeats of 34 amino acids. Xo2276- DBD was expressed in E. coli and purified. Direct in vitro recognition of Xo2276-DBD on a putative target DNA sequence was confirmed using an electrophoretic mobility shift assay. This is the first study measuring the homologous expression and secretion of Xo2276 in vitro using rice leaf extract and its direct in vitro binding to the specific target DNA sequence.

Evaluation of Early Generation of Crosses for Incorporation of Resistance to Bacterial Spot into Sweet Pepper (감미종(甘味種) 고추에 더뎅이병(病) 저항성(抵抗性)을 도입(導入)하기 위한 교잡(交雜) 초기세대(初期世代) 검정(檢定))

  • Jeong, Ho Jeong;Kim, Byung Soo;Shon, Eun Young
    • Current Research on Agriculture and Life Sciences
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    • v.12
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    • pp.23-28
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    • 1994
  • A leading sweet pepper cultivar, Keystone Resistant Giant #3, was crossed with a line with resistance to Xanthomonas campestris pv. vesicatoria, PI271322, for incorporation of the resistance and to study the inheritance of resistance to the disease. Seedlings of parents, $F_1$, $F_2$ and backcross populations of a cross between Keystone Resistant Giant #3 and PI271322 were inoculated with race 1 and race 3 of Xanthamonas campestris pv. vesicatoria by infiltrating bacterial suspension into abaxial side of leaves. PI271322 was carrying a gene ($Bs_3$) for hypersensitive resistance to race 1 of X. c. pv. vesicatoria and also a component of non-hypersensitive resistance to race 3 inheriting in a quantitative mode. Correlation coefficient between disease indices to race 1 and race 3 of non-hypersensitive plants in $F_2$ population was highly significant, thus indicating that a component of non-hypersensitive general resistance in PI271322 acted consistently to both races. Individual plants with resistance to both race 1 and race 3 of X. c. pv. vesicatoria were selected and a breeding program for incorporation of the resistance is continued.

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Biodiversity and Phylogenetic Analysis of Streptomyces Collected from Bamboo Forest Soil (대나무 산림토양으로부터 수집한 Streptomyces 속 방선균의 계통학적 다양성)

  • Lee, Hyo-Jin;Whang, Kyung-Sook
    • Korean Journal of Microbiology
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    • v.46 no.3
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    • pp.262-269
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    • 2010
  • To investigate a quantitative evaluation of the actinobacteria, we have collected samples from various kinds of bamboo forest soil. Each different layers contained $2.7{\times}10^6-2.7{\times}10^8$ CFU/g of actinobacteria which was the highest in litter layers of Sasa boreali forest soil. We obtained 330 actinobacteria from different layers of bamboo forest soil; litter (100 strains), humus (70 strains), and rhizosphere soil (160 strains). Based on the colony morphology (aerial mycelium, substrate mycelium, and soluble pigment), isolates were divided into thirty-six groups and we selected 50 representative isolates. 16S rRNA gene sequence analysis showed Streptomyces was major actinobacteria (94%) and they were categorized as cluster I (2 strains), II (35 strains), III (6 strains), and IV (7 strains), respectively. The diversity index of 50 Streptomyces collected from the bamboo forest soil was calculated with the Shannon-Wiener method. Bamboo litter showed higher diversity index level of 3.33 than that of humus and rhizosphere soil. Also, antibiotic activities of our isolates were investigated against Botrytis cinerea, Xanthomonas campestris, Xanthomonas axonopodis pv. vesicatoria, and Bacillus cereus and found in 74, 16, 25, and 24 strains, respectively.

Methylome Analysis of Two Xanthomonas spp. Using Single-Molecule Real-Time Sequencing

  • Seong, Hoon Je;Park, Hye-Jee;Hong, Eunji;Lee, Sung Chul;Sul, Woo Jun;Han, Sang-Wook
    • The Plant Pathology Journal
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    • v.32 no.6
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    • pp.500-507
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    • 2016
  • Single-molecule real-time (SMRT) sequencing allows identification of methylated DNA bases and methylation patterns/motifs at the genome level. Using SMRT sequencing, diverse bacterial methylomes including those of Helicobacter pylori, Lactobacillus spp., and Escherichia coli have been determined, and previously unreported DNA methylation motifs have been identified. However, the methylomes of Xanthomonas species, which belong to the most important plant pathogenic bacterial genus, have not been documented. Here, we report the methylomes of Xanthomonas axonopodis pv. glycines (Xag) strain 8ra and X. campestris pv. vesicatoria (Xcv) strain 85-10. We identified $N^6$-methyladenine (6mA) and $N^4$-methylcytosine (4mC) modification in both genomes. In addition, we assigned putative DNA methylation motifs including previously unreported methylation motifs via REBASE and MotifMaker, and compared methylation patterns in both species. Although Xag and Xcv belong to the same genus, their methylation patterns were dramatically different. The number of 4mC DNA bases in Xag (66,682) was significantly higher (29 fold) than in Xcv (2,321). In contrast, the number of 6mA DNA bases (4,147) in Xag was comparable to the number in Xcv (5,491). Strikingly, there were no common or shared motifs in the 10 most frequently methylated motifs of both strains, indicating they possess unique species- or strain-specific methylation motifs. Among the 20 most frequent motifs from both strains, for 9 motifs at least 1% of the methylated bases were located in putative promoter regions. Methylome analysis by SMRT sequencing technology is the first step toward understanding the biology and functions of DNA methylation in this genus.

Copper Resistance and Race Distribution of Xanthomonas campestris pv. vesicatoria on Pepper in Korea (한국에서의 더뎅이병균의 구리 저항성과 레이스 분포)

  • 이승돈;조용섭
    • Korean Journal Plant Pathology
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    • v.12 no.2
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    • pp.150-155
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    • 1996
  • 1993년과 1994년에 걸쳐 한국의 고추 재배 지역에서 분리한 고추 더뎅이병균은 66개의 균주중 24개가 race 1이었으며, 나머지는 42균주가 race 3이었다. Race 2균주는 발견되지 않았다. 총 66개의 균주중 8%가 200 $\mu\textrm{g}$/ml의 황산구리에 저항성을 보였으며, 100 $\mu\textrm{g}$/ml의 streptomycin sulfate에는 단지 한 균주만이 저항성을 나타냈다. Race 1균주중 21%가 구리에 저항성을 보였으나 race 3균주는 모두 구리에 감수성을 나타냈다. 모든 감수성 균주는 구리 농도 1 $\mu\textrm{g}$/ml에서 죽거나 아주 낮은 수준만이 생존하였으나 저항성 균주는 128 $\mu\textrm{g}$/ml에서도 생존하였다.

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Occurrence and Distribution of Bacterial Canker of Red Pepper Caused by Clavibacter michiganensis subsp. michiganensis (Clavibacter michiganensis subsp. michiganensis에 의한 고추 궤양병)

  • 이승돈
    • Plant Disease and Agriculture
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    • v.5 no.2
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    • pp.105-110
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    • 1999
  • Bacterial leaf spot by Xanthomonas campestris pv. vesicatoria has been known to cause serious problem in red pepper in Korea. However recent survey showed that most smptoms in the leaves were mixed with two different symptoms one was leaf spot and the other was canker. bacteria isolated from canker were identified as Clavibacter michiganensis subsp. michiganensis on the basis of biochemical and physiological characteristics. The causal bacteria were non-motile rod-shaped and Gram-positive. The lesions on pepper leaves appeared at first as small blisters or pimple-like white spots which enlarged in size at a later stage. The centers of some of the spots became necrotic and brown and were surrounded by a white halo. Pathogenicity tests were performed on pepper cv. Alchan seedling by spraying of bacterial suspension. During 1997 and 1998 total 17% of 527 fields surveyed were infected by C. michiganensis subsp. michiganensis. The canker of red pepper caused by C. michiganensis subsp. michiganensis was first identified in this study in Korea, and new name "gueyangbyung" was tentatively given to the disease.

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Elevated CO2 and Temperature Effects on the Incidence of Four Major Chili Pepper Diseases

  • Shin, Jeong-Wook;Yun, Sung-Chul
    • The Plant Pathology Journal
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    • v.26 no.2
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    • pp.178-184
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    • 2010
  • Four major diseases of chili pepper including two fungal diseases, anthracnose (Colletotrichum acutatum) and Phytophthora blight (Phytophthora capsici), and two bacterial diseases, bacterial wilt (Ralstonia solanacearum) and bacterial spot (Xanthomonas campestris pv. vesicatoria), were investigated under future climate-change condition treatments in growth chambers. Treatments with elevated $CO_2$ and temperature were maintained at $720ppm{\pm}20ppm$ $CO_2$ and $30^{\circ}C{\pm}0.5^{\circ}C$, whereas ambient conditions were maintained at $420ppm{\pm}20ppm$ $CO_2$ and $25^{\circ}C{\pm}0.5^{\circ}C$. Pepper seedlings or fruits were infected with each pathogen, and then the disease progress was evaluated in the growth chambers. According to paired t-test analyses, bacterial wilt and spot diseases significantly increased by 24% (p=0.008) and 25% (p=0.016), respectively, with elevated $CO_2$ and temperature conditions. On the other hand, neither Phytophthora blight (p=0.906) nor anthracnose (p=0.125) was statistically significant. The elevated $CO_2$ and temperature accelerated the progress of bacterial wilt by two days and bacterial spot by one day compared to the ambient treatment. Temperature regime studies of the diseases without changes in $CO_2$ confirmed that the accelerated bacterial disease progress was mainly due to the increased temperature rather than the elevated $CO_2$ conditions.

Functional Characterization of PR-1 Protein, β-1,3-Glucanase and Chitinase Genes During Defense Response to Biotic and Abiotic Stresses in Capsicum annuum

  • Hong, Jeum-Kyu;Hwang, Byung-Kook
    • The Plant Pathology Journal
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    • v.21 no.3
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    • pp.195-206
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    • 2005
  • Spatial and temporal expression of pathogenesis-related (PR) gene and proteins has been recognized as inducible defense response in pepper plants. Gene expression and/or protein accumulation of PR-1, $\beta-1,3-glucanase$ and chitinase was predominantly found in pepper plants during the inoculations by Xanthomonas campestris pv. vesicatoria, Phytophthora capsici and Colletotrichum coccodes. PR-1 and chitinase genes were also induced in pepper plants in response to environmental stresses, such as high salinity and drought. PR-1 and chitinase gene expressions by biotic and abiotic stresses were regulated by their own promoter regions containing several stress-related cis-acting elements. Overexpression of pepper PR-1 or chitinase genes in heterogeneous transgenic plants showed enhanced disease resistance as well as environmental stress tolerances. In this review, we focused on the putative function of pepper PR-1, $\beta-1,3-glucanase$ and chitinase proteins and/or genes at the biochemical, molecular and cytological aspects.

Isolation and In vitro and In vivo Antifungal Activity of Phenylacetic acid Produced by Micromonospora aurantiaca Strain JK-1

  • Kim, Hyo-Jin;Hwang, In-Sun;Kim, Beom-Seok;Hwang, Byung-Kook
    • The Plant Pathology Journal
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    • v.22 no.1
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    • pp.75-89
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    • 2006
  • The actinomycete strain JK-1 that showed strong inhibitory activity against some plant pathogenic fungi and oomycetes was isolated from Jung-bal Mountain in Ko-yang, Korea. The strain JK-1 produced spores singly borne on sporophores and the spores were spherical and 0.9-1.2 11m in diameter. The cell wall of the strain JK-1 contained meso-diaminopimelic acid. The actinomycete strain JK-1 was identified as the genus Micromonospora based on the morphological, physiological, biochemical and chemotaxonomic characteristics. From the 168 rDNA analysis, the strain JK-1 was assigned to M aurantiaca. The antibiotic MA-1 was purified from the culture broth of M aurantiaca JK-1 using various purification procedures, such as Diaion HP20 chromatography, C18 flash column chromatography, silica gel flash column chromatography and Sephadex LH-20 column chromatography. $^{1}H-$, $^{13}C-NMR$ and EI mass spectral analysis of the antibiotic MA-1 revealed that the antibiotic MA-1 is identical to phenylacetic acid. Phenylacetic acid showed in vitro inhibitory effects against fungal and oomycete pathogens Alternaria mali, Botrytis cinerea, Magnaporthe grisea, Phytophthora capsici and yeast Saccharomyces cerevisiae at < 100 $\mug$ $ml^{-1}$. In addition, phenylacetic, acid completely inhibited the growth of Sclerotinia sclerotiorum, Bacillus subtilis, Candida albicans, Xanthomonas campestris pv. vesicatoria at < $\mug$ $ml^{-1}$. Phenylacetic acid strongly inhibited conidial germination and hyphal growth of M grisea and C. orbiculare. Phenylacetic acid showed significantly high levels of inhibitory' effect against rice blast and cucumber anthracnose diseases at 250 $\mug$ $ml^{-1}$. The control efficacies of phenylacetic acid against the two diseases were similar to those of commercial compounds tricyclazole, iprobenfos and chlorothalonil .n the greenhouse.

Growth and Fruit Characteristics of Selections from Local Cultivars of Pepper (고추 주요 재래종 선발계통의 생육 및 과실 특성)

  • Kown, Young Seok;Kim, Byung Soo;Jeong, Ho Jeong
    • Current Research on Agriculture and Life Sciences
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    • v.10
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    • pp.117-123
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    • 1992
  • Twenty and 24 lines selected from Korean local cultivar, Kalmi and Punggak, respectively, were grown in the University experiment farm to select promising lines for recurrent parent in a backcross breeding program for incorporation of resistance to Phytophthora capsici and Xanthomonas campestris pv vesicatoria into the Korean local cultivars. All the selections of 'Kalmi' and 'Punggak' were relatively late in maturity and the range of the variation was wider in the selections of 'Kalmi' than in those of 'Punggak'. Many of the selections from 'Kalmi' were vigorous, high in sugar content, and tolerant to virus in the field. Many of the selections from 'Punggak' were bearing large fruits with thick pericarp and high milling percentage. Line 6, 9, 14, 18 of 'Kalmi' and line 14, 16, 18 of 'Punggak' were selected as promising lines for recurrent parent in the backcross program.

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