• Microbiology and Biotechnology Letters
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• v.19 no.1
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• pp.14-20
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• 1991

- The purpose of this work is to investigate the effects of various substrates on biosynthesis of ethylene by the Kudzu strain of Pseudomonas syn'ngae pv. Phaseolicola causing halo blight. In the intact cell of P. sym'ngue, optimal condition for ethylene production was achieved at p1-I 7.5 and $30^{\circ}C$ for 9 to 10 hours of culture. Ethylene was most effectively produced from amino acids such as Asn, Gln, Asp ans Glu, compared to those of various kinds of sugars. While ethylene production from $\alpha$-ketoglutarate ($\alpha$-KG) was gradually increased throughout 51 hours incubation period tested. Ethylene production derived from citrate, $\alpha$-KG and oxalacetate as well as a few amino acids was further enhanced by the addition of histidine or arginine. In cell-free ethylene-forming system, ethylene was most effectively produced from $\alpha$-KG, compared to those from citrate, oxalacetate, Glu, Arg, or Asp, at 0.5 mM among the range from 0.25 mM to 5 mM. Anlinooxyacetate, an inhibitor of a pyridoxal phosphate-linked enzyme, completely inhibited ethylene evolution derived from Glu but not affect that derived from $\alpha$-KG. The results obtained in this work suggest that $\alpha$-KG might be a direct precursor of ethylene production in this organism than any other substrates tested.

• The Plant Pathology Journal
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• v.22 no.2
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• pp.119-124
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• 2006

Kudzu vine(Pueraria montana var. lobata) is an invasive climbing woody vine that envelops trees and shrubs, pressing physically and shutting out sunlight, which needs to be controlled. Kudzu vine pathogens were surveyed as a way to seek its biocontrol agents in 2002. Occurrence of a bacterial halo blight disease of kudzu vine was observed at several localities in Korea including Euiwang and Suwon in Gyeonggi Province, Daejon, and Gochang and Buan in Jeonbuk Province. Symptoms of brown to black spots with a surrounding yellowish halo appeared from June and lasted till the rainy season without much expansion, but accompanying often leaf blight and defoliation. Isolated bacteria were identified as Pseudomonas syringae pv. phaseolicola based on physiological and cultural characteristics, Biolog, fatty acid and 16S rDNA sequencing analyses. In artificial inoculation test, these bacteria produced the same halo spot symptoms on kudzu vine and bean plants. They also induced hypersensitive responses (HR) on tobacco, tomato, and chili pepper leaves. This is the first report of a bacterial disease of kudzu vine in Korea, and the bacterial pathogen can be used as a biocontrol agent against the pest plant.

• Microbiology and Biotechnology Letters
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• v.22 no.5
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• pp.485-490
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• 1994

A restriction endonuclease, PsyI, has been isolated from Pseudomonas syringae pv. pha- seolicola, and its catalytic properties have been studied. This enzyme was purified through strepto- mycin sulfate and ammonium sulfate fractionation, phosphocellulose Pll, DEAE-cellulose, hydroxy- apatite and Sephadex G-100 column chromatography. It's molecular weight was about 50,000 dalton as determined by 7.5% polyacrylamide gel electrophoresis containing 0.1% SDS. In catalytic proper- ties, PsyI shows stable at wide ranges of pH between 7.0 and 10.0, of temperature between 30$\circ$C and 37$\circ$C, and its thermal stability is between 25$\circ$C, and 45$\circ$C, at the presence Of 10 mM MgCl$_{2}$-PsyI essentially require Na salt for enzyme reaction, is rather inhibited in the high Na salt concent- ration. The presence of 2-mercaptoethanol is absolutely required for the enzyme activity. This endonuclease, PsyI was determined to be an isoschizomer of SalI from the results of the restriction mapping and DNA sequencing.

• The Plant Pathology Journal
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• v.15 no.3
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• pp.137-143
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• 1999

A novel chromosomal gene tagging technique using a specific fragment of the fatty acid desaturase-like open reading frame (des-like ORF) from the tox-argK gene cluster of Pseudomonas syringae pv. phaseolicola was developed to identify Xanthomonas spp.released into the environment as biocontrol agents. X. campestris pv. convolvuli FB-635, a pathogen of Convolvulus arvensis L., (bindweed), was chosen as the organism in which to develop and test the system. A 0.52 kb DES fragment amplified from P. syringae pv. phaseolicola C-199 was inserted into pGX15, a cosmid clone containing a 10.3 kb Eco RI-HindIII fragment derived from the xanthomonadin biosynthetic gene cluster contained in plasmid pIG102, to create a pigG::DES insertion. The 10.8 kb EcoRI-BamHI fragment carrying the pigG:: DES insertion was cloned into pLAFR3 to generate pLXP22. pLXP22 was then conjugated into X. campestris pv. convolvuli FB-635 and the pigG::DES insertion integrated into the bacterial chromosome by marker exchange. Rifampicin resistant, tetracycline sensitive, starch hydrolyzing, white colonies were used to differentiate the marked strain from yellow pigmented wild-type ones. PCR primers specific for the unique DES fragment were used for direct detection of the marked strain. Result showed the marked strain could be detected at very low levels even in the presence of high levels of other closely related or competitive bacteria. This PCR-based DES-tagging system provides a rapid and specific tool for directly monitoring the dispersal and persistence of Xanthomonas spp.released into the environment.