Taguchi Fumiko, Ogawa Yujiro, Takeuchi Kasumi, Suzuki Tomoko, Toyoda Kazuhiro, Shiraishi Tomonori, Ichinose Yuki
The Graduate School of Natural Science and Technology, Okayama University, Tsushima-naka 1-1-1, Okayama 700-8530, Japan.
J Bacteriol. 2006 Dec;188(24):8376-84. doi: 10.1128/JB.00763-06. Epub 2006 Oct 6.
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 (Deltaorf3 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 Deltaorf3 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 Deltaorf3 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 Deltaorf3 mutant. The phenotypes of the Deltaorf3 mutant and an AHL synthesis (DeltapsyI) mutant were similar, although the mutant-specific characteristics were more extreme in the Deltaorf3 mutant. The swarming motility of the Deltaorf3 mutant was greater than that of the DeltapsyI mutant. This was attributed to the synergistic effects of the overproduction of biosurfactants and/or alternative fatty acid metabolism in the Deltaorf3 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.
丁香假单胞菌烟草致病变种6605拥有一个参与鞭毛蛋白糖基化的遗传区域。该区域由三个开放阅读框组成:orf1、orf2和orf3。我们之前的研究表明,orf1和orf2编码糖基转移酶;另一方面,orf3在鞭毛蛋白的翻译后修饰中不起作用。尽管Orf3的功能仍不清楚,但orf3缺失突变体(Deltaorf3突变体)对烟草植株的毒力有所降低。Orf3与脂肪酸延长循环中的3-氧代酰基-(酰基载体蛋白)合酶III具有显著的同源性。Deltaorf3突变体形成群体感应分子酰基高丝氨酸内酯(AHLs)的能力显著降低,这表明Orf3是AHL合成所必需的。与野生型菌株相比,Deltaorf3突变体的群体游动性、生物表面活性剂产生以及对H2O2和抗生素的耐受性均增强。对接种在烟草叶片表面的细菌进行扫描电子显微镜观察发现,Deltaorf3突变体的细胞周围几乎没有细胞外聚合物基质。Deltaorf3突变体和AHL合成(DeltapsyI)突变体的表型相似,尽管突变体特异性特征在Deltaorf3突变体中更为极端。Deltaorf3突变体的群体游动性大于DeltapsyI突变体。这归因于Deltaorf3突变体中生物表面活性剂的过量产生和/或脂肪酸代谢改变的协同作用。此外,铁和生物表面活性剂的量似乎参与了丁香假单胞菌烟草致病变种6605群体感应调控下的生物膜形成。