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KMID : 0545120030130010110
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Journal of Microbiology and Biotechnology
2003 Volume.13 No. 1 p.110 ~ p.118
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Identification and Characterization of Coronatine-Producing Pseudomonas syringae pv.actinidiae
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HAN, HYO SHIM
KOH, YOUNG JIN/HUR, JAE-SEOUN/JUNG, JAE SUNG
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Abstract
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Pseudomonas syringae pv. actinidiae strains, which cause canker disease in kiwifruit, were collected from kiwifruit orchards in Korea and identified using biochemical and physiological tests. The nucleotide sequences of the 16S rDNA and 16S-23S internally transcribed spacer of the isolates were found to be identical to those of the pathotype strain, Kwl1, of P. syringae pv. actinidiae. Remarkably, no coding sequence for phaseolotoxin biosynthesis or phaseolotoxin-resistant ornithine carbamoytransferase was found by PCR amplification in any of the new Korean isolates of Pseudomonas syringae pv. actinidiae, although this was clearly identified in the control pathotype Kwl1 reference strain. In contrast, three primers sets derived from the coronatine biosynthetic gene cluster and DNA from the Korean strains yielded amplified DNA fragments of the expected size. A sequence analysis of the PCR products revealed that Pseudomonas syringae pv. actinidiae and the Korean strains of pv. actinidiae contained coronafacate ligase genes (cfl) with identical sequenced, whereas their corR genes exhibited 91% sequence similarity. The production of coronatine, instead of phaseolotoxin, by the Korean strains of P. syringae pv. actinidiae was confirmed by a bioassay using reference pathovars known to produce coronatine and phaseolotoxin. The genes for coronatine biosynthesis in the Korean strains of P. syringae pv. actinidiae were found to be present plasmids.
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