• 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.

• Research in Plant Disease
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• v.15 no.2
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• pp.83-87
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• 2009

Direct Polymerase Chain Reaction (PCR) method that combines biological and enzymatic amplification of PCR targets was developed for the detection of Xanthomonas axonopodis pv. glycines on soybeen seeds without DNA isolation. Primers Xag F1 and Xag R1 were designed to specifically amplify a 401 bp fragment of the glycinecin A gene of X axonopodis pv. glycines. Xag F1 and Xag R1 were used to carry out the PCR analysis with genomic DNA from 45 different bacterial strains including phylogenetically related bacteria with X axonopodis pv. glycines, and other bacterial strains of different genus and species. The PCR assay using this set of primers were able to detect X axonopodis pv. glycines with DNA concentration as low as 200 fg and $1.8{\times}10^3$ cfu/ml. The Xag was detected from the seed samples incubated for 2 hrs with shaking and the intensity of the band was increase with the incubation time of seeds. The Direct PCR assay method without DNA isolation makes detection of X. axonopodis pv. glycines on soybean seeds easier and more sensitive than other conventional methods. The developed seed assay using direct PCR method will be useful for the specific detection of X. axonopodis pv. glycines in soybean seed samples.

• Journal of Applied Biological Chemistry
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• v.44 no.4
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• pp.173-176
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• 2001

This report concerns the role of indole-3-acetic acid (IAA) in bacterial pustule disease of soybean. Pustule production in soybean leaves caused by Xanthomonas axonopodis pv. glycines was accompanied by a drastic increase in IAA content of host tissues. The phytopathogenic bacterium synthesized IAA in a tryptophan concentration-dependent manner when grown in a defined minimal medium. In complex media, however, the pathogen showed no response to tryptophan feeding, implying that the bacterial biosynthetic machinery of IAA is strictly regulated by nutrient availability of its growth environments. The results may suggest that IAA of bacterial origin and tryptophan of plant origin be involved in the process of pustule symptom development in soybean.

• Research in Plant Disease
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• v.21 no.1
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• pp.40-43
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• 2015

Bacterial pustule of soybean is caused by Xanthomonas axonopodis pv. glycines, one of the most important diseases in soybean. The symptom of bacterial pustule is mainly distributed around leaf veins. However, the reason has not been known. In order to determine pathosystem of bacterial pustule in leaf, soybean leaves were collected and observed using scanning electron microscopy (SEM) and light microscopy. Many hairs were observed at abaxial sides of the leaf, few hairs were observed at tissue around the leaf veins. In addition, unidentified bacterial cells and dusts at the no hair part near veins were observed. In the inoculation assays, the cells of X. axonopodis pv. glycines were observed near leaf veins. The imprint of underside of soybean leaves inoculated with X.axonopodis pv. glycines on PDA showed that the growth of bacteria around veins was observed but no bacterial growth at the part with leaf hairs. Our data demonstrated that soybean leaf hairs play an important role as a physical barrier for structural resistance of soybean against bacterial pustule pathogen.

• Journal of Microbiology and Biotechnology
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• v.18 no.9
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• pp.1500-1509
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• 2008

The hybridization patterns with the avrBs3 gene that is known to determine the recognition of host specificity were used to study the diversity of Xanthomonas axonopodis pv. glycines causing bacterial leaf pustule in soybean. A total of 155 strains were isolated from diverse tissues of soybean cultivars collected in Korea and were classified into six different type strains of OcsF, SL1017, SL1018, SL1045, SL1157, and SL2098 according to the patterns of avrBs3-homologous bands. When these type strains were inoculated on various cultivars, most of the Korean strains mildly induced disease symptoms on the resistant CNS1 cultivars. Unlike other type strains, strain SL2098, which appeared not to contain any avrBs3 homolog, induced only a few pustules on even highly susceptible cultivars. When a plasmid carrying the 3.7-kb avrBs3-homologous gene from strain SL1045 was introduced into SL2098, the transformant could not recover the pathogenicity in susceptible host plants. However, when avrBs3-homologous genes of strain SL1018 were mutated by transposon mutagenesis, one of the mutants in which a 5.2-kb chromosomal band homologous to avrBs3 was disrupted could not induce the hypersensitive response on resistant cultivars such as William82 or CNS2. Our results suggest that the avrBs3 homologs may play important roles in the pathogenicity of Xanthomonas axonopodis pv. glycines and the recognition of soybean cultivars.

• The Plant Pathology Journal
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• v.18 no.3
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• pp.115-120
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• 2002

Xanthomonas axonopodis pv. glycines, the causal agent of bacterial pustule of soybean, induces hypersensitive response (HR) in a non-host plant, hot pepper (Capsicum annuum). A wild-type strain (8ra) and its non-patho-genic mutant (8-13) of X. axonopodis pv. glycines were inoculated into the pepper leaf tissues and their subcellular responses to the bacterial infections were examined by electron microscopy. Intrastructural changes related to HR were found in the leaf tissues infected with 8ra from 8 h after inoculation, characterized by separation of plasmalemma from the cell wall, formation of small vacuoles and vesicles, formation of cell wall apposition, and cellular necrosis. No such responses were observed in the tissues infected with the mutant. In 8ra, the bacterial cells were attached to the cell walls, with the cell wall material dissolved into and appearing to encapsulate the bacterial cells. The bacterial cells later became entirely embedded in the cell wall material. On the other hand, in 8-13, the bacterial cells were usually not attached tightly to the plant cell wall, and no or poor encapsulation of the bacteria by the wall material occurred, although these were encircled by rather loose wall materials at the later stages.

• The Plant Pathology Journal
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• v.17 no.6
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• pp.362.2-362
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• 2001

• The Plant Pathology Journal
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• v.23 no.3
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• pp.193-202
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• 2007

Typical susceptible symptoms of the bacterial pustule disease caused by Xanthomonas axonopodis pv. glycines are pustule and chlorotic haloes that usually occur in leaves of Glycine max plants. The soybean genotype 'PI96188' showed an atypical response to all tested races X. axonopodis pv. glycines, accompanied with necrosis without chlorotic haloes on the underside of the necrotic symptoms. X. axonopodis pv. glycines 8ra grew to levels from 10 to 100 fold lower on PI96188 than on susceptible cultivar Jinjoo1, but 10-fold higher than on the resistant cultivar CNS. The chlorophyll content in PI96188 leaves remained unchanged until 12 days after bacterial infection. Ultrastructural observation showed that the infected leaf cells of PI96188 had intact normal chloroplasts compared to those of the susceptible cultivar Jinjoo1. Chloroplast degradation or the absence of chloroplasts in cells of the infected tissues was observed in Jinjoo1. Senescence-related ACS7 gene was significantly induced in PI96188 compared to those in Jinjoo1 at 2 days after inoculation. While photosynthesis-related rbcS gene showed the dramatic change in Jinjoo1, this gene was constitutively expressed in PI96188. However, expression of the defense-related genes, such as peroxidase and isoflavone synthase in the infected PI96188 leaves was similar to that in Jinjoo1. Together, these results suggest that the novel necrotic symptom in PI96188 is a kind of resistant response different from a typical hypersensitive response in the resistant genotypes.

• 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.

• Journal of Crop Science and Biotechnology
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• v.10 no.3
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• pp.147-158
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• 2007

Xanthomonas axonopodis pv. glycines(Xag) is a pathogen that causes bacterial leaf pustule(BLP) disease in soybeans grown in Korea and the southern United States. Typical and early symptoms of the disease are small, yellow to brown lesions with raised pustules that develop into large necrotic lesions leading to a substantial loss in yield due to premature defoliation. After Xag infects PI 96188, only pustules without chlorotic haloes were observed, indicating the different response to Xag. To identify differentially expressed genes prior to and 24 hr after Xag inoculation to PI 96188 and BLP-resistant SS2-2, an oligonucleotide macroarray was constructed with 100 genes related to disease resistance and metabolism from soybean and Arabidopsis. After cDNAs from each genotype were applied on the oligonucleotide macroarrays with three replicates and dye swapping, 36 and 81 genes were expressed as significantly different between 0 hr and 24 hr in PI 96188 and SS2-2, respectively. Six UniGenes, such as the leucine-rich repeat protein precursor or 14-3-3-like protein, were selected because they down-regulated in PI 96188 and up-regulated in SS2-2 after Xag infection, simultaneously. Using tubulin and cDNA of Jangyeobkong(BLP-susceptible) as controls, the oligonucleotide macroarray data concurred with quantitative real-time RT-PCR(QRT RT-PCR) results in most cases, supporting the accuracy of the oligonucleotide macroarray experiments. Also, QRT RT-PCR data suggested six candidate genes that might be involved in a necrotic response to Xag in PI 96188.