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Journal of Bacteriology, December 2006, p. 8376-8384, Vol. 188, No. 24
0021-9193/06/$08.00+0     doi:10.1128/JB.00763-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

A Homologue of the 3-Oxoacyl-(Acyl Carrier Protein) Synthase III Gene Located in the Glycosylation Island of Pseudomonas syringae pv. tabaci Regulates Virulence Factors via N-Acyl Homoserine Lactone and Fatty Acid Synthesis{triangledown}

Fumiko Taguchi,1 Yujiro Ogawa,1 Kasumi Takeuchi,1,2 Tomoko Suzuki,1 Kazuhiro Toyoda,1 Tomonori Shiraishi,1 and Yuki Ichinose1*

The Graduate School of Natural Science and Technology, Okayama University, Tsushima-naka 1-1-1, Okayama 700-8530, Japan,1 National Institute of Agrobiological Sciences, Kannondai 2-1-2, Tsukuba, Ibaraki 305-8602, Japan2

Received 27 May 2006/ Accepted 15 September 2006

Pseudomonas syringae pv. tabaci 6605 possesses a genetic region involved in flagellin glycosylation. This region is composed of three open reading frames: orf1, orf2, and orf3. Our previous study revealed that orf1 and orf2 encode glycosyltransferases; on the other hand, orf3 has no role in posttranslational modification of flagellin. Although the function of Orf3 remained unclear, an orf3 deletion mutant ({Delta}orf3 mutant) had reduced virulence on tobacco plants. Orf3 shows significant homology to a 3-oxoacyl-(acyl carrier protein) synthase III in the fatty acid elongation cycle. The {Delta}orf3 mutant had a significantly reduced ability to form acyl homoserine lactones (AHLs), which are quorum-sensing molecules, suggesting that Orf3 is required for AHL synthesis. In comparison with the wild-type strain, swarming motility, biosurfactant production, and tolerance to H2O2 and antibiotics were enhanced in the {Delta}orf3 mutant. A scanning electron micrograph of inoculated bacteria on the tobacco leaf surface revealed that there is little extracellular polymeric substance matrix surrounding the cells in the {Delta}orf3 mutant. The phenotypes of the {Delta}orf3 mutant and an AHL synthesis ({Delta}psyI) mutant were similar, although the mutant-specific characteristics were more extreme in the {Delta}orf3 mutant. The swarming motility of the {Delta}orf3 mutant was greater than that of the {Delta}psyI mutant. This was attributed to the synergistic effects of the overproduction of biosurfactants and/or alternative fatty acid metabolism in the {Delta}orf3 mutant. Furthermore, the amounts of iron and biosurfactant seem to be involved in biofilm development under quorum-sensing regulation in P. syringae pv. tabaci 6605.

* Corresponding author. Mailing address: The Graduate School of Natural Science and Technology, Okayama University, Tsushima-naka 1-1-1, Okayama 700-8530, Japan. Phone and fax: (81) 86 251 8308. E-mail: yuki{at}cc.okayama-u.ac.jp.

{triangledown} Published ahead of print on 6 October 2006.

Journal of Bacteriology, December 2006, p. 8376-8384, Vol. 188, No. 24
0021-9193/06/$08.00+0     doi:10.1128/JB.00763-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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