Complete genome sequence and comparative analysis of the metabolically versatile <i>Pseudomonas putida</i> KT2440

Nelson, Karen E.; Weinel, Christian; Paulsen, Ian T.; Dodson, Robert J.; Hilbert, Helmut; Santos, Vítor A. P. Martins dos; Fouts, Derrick E.; Gill, Steven R.; Pop, Mihai; Holmes, Megan E.; Brinkac, Lauren; Beanan, Maureen J.; DeBoy, Robert T.; Daugherty, Sean C.; Kolonay, James F.; Madupu, Ramana; Nelson, William; White, Owen; Peterson, Jeremy; Khouri, Hoda; Hance, Ioana R.; Lee, P. Chris; Holtzapple, Erik; Scanlan, David J.; Tran, Kevin; Moazzez, A.; Utterback, Teresa; Rizzo, Michael; Lee, K.; Kosack, Daniel S.; Moestl, D.; Wedler, Holger; Lauber, Jürgen; Stjepandić, Diana; Hoheisel, Jörg D.; Straetz, M.; Heim, Sabina; Kiewitz, Claudia; Eisen, Jonathan A.; Timmis, Kenneth N.; Düsterhöft, Andreas; Tümmler, Burkhard; Fraser, Claire M.

doi:10.1046/j.1462-2920.2002.00366.x

Cited by 1,198 publications

(703 citation statements)

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“…Cell-cell communication has been recently shown to take place during bacterial colonization of the tomato rhizosphere, and the production of AHLs in rhizosphere populations has been reported (43). In the soil and rhizospherecolonizing bacterium P. putida KT2440, no production of AHLs has been detected to date, and there was no genetic evidence that this microorganism possesses a quorum-sensing system (32). The results presented here indicate that KT2440 does respond to population density via a still-unidentified signaling mechanism.…”

Section: Discussionmentioning

confidence: 99%

Cell Density-Dependent Gene Contributes to Efficient Seed Colonization by Pseudomonas putida KT2440

Espinosa‐Urgel

Ramos

2004

Appl Environ Microbiol

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We have characterized the expression pattern of a gene, ddcA, involved in initial colonization of corn seeds by Pseudomonas putida KT2440. The ddcA gene codes for a putative membrane polypeptide belonging to a family of conserved proteins of unknown function. Members of this family are widespread among prokaryotes and include the products of a Salmonella enterica serovar Typhimurium gene expressed during invasion of macrophages and psiE, an Escherichia coli phosphate starvation-inducible gene. Although its specific role is undetermined, the presence of ddcA in multicopy restored the seed adhesion capacity of a KT2440 ddcA mutant. Expression of ddcA is growth phase regulated, being maximal at the beginning of stationary phase. It is independent of RpoS, nutrient depletion, or phosphate starvation, and it is not the result of changes in the medium pH during growth. Expression of ddcA is directly dependent on cell density, being also stimulated by the addition of conditioned medium and of seed exudates. This is the first evidence suggesting the existence of a quorum-sensing system in P. putida KT2440. The potential implication of such a signaling process in seed adhesion and colonization by the bacterium is discussed.

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Section: Discussionmentioning

confidence: 99%

Cell Density-Dependent Gene Contributes to Efficient Seed Colonization by Pseudomonas putida KT2440

Espinosa‐Urgel

Ramos

2004

Appl Environ Microbiol

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“…The resulting strain, EU9, did show increased attachment, but only at early time points; further biofilm development and detachment kinetics were similar to those of the wild-type strain (Fig. 3), indicating that the hyperadherent character of ibi-626 is not solely due to the disruption of olpA, and that an additional mutation might be present.Complete sequencing of ibi-626 reveals a point mutation that affects lapGThe genome of mutant ibi-626 was fully sequenced and compared with the reference genome of KT2440 (Nelson et al, 2002) to define potential mutations that could contribute to the phenotype of this strain. Several apparent discrepancies with respect to the reference sequence were checked by PCR amplification and sequencing of the corresponding region in ibi-626 and our laboratory KT2440 strain.…”

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Selection of hyperadherent mutants in Pseudomonas putida biofilms

et al. 2011

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A number of genetic determinants required for bacterial colonization of solid surfaces and biofilm formation have been identified in different micro-organisms. There are fewer accounts of mutations that favour the transition to a sessile mode of life. Here we report the isolation of random transposon Pseudomonas putida KT2440 mutants showing increased biofilm formation, and the detailed characterization of one of them. This mutant exhibits a complex phenotype, including altered colony morphology, increased production of extracellular polymeric substances and enhanced swarming motility, along with the formation of denser and more complex biofilms than the parental strain. Sequence analysis revealed that the pleiotropic phenotype exhibited by the mutant resulted from the accumulation of two mutations: a transposon insertion, which disrupted a predicted outer membrane lipoprotein, and a point mutation in lapG, a gene involved in the turnover of the large adhesin LapA. The contribution of each alteration to the phenotype and the possibility that prolonged sessile growth results in the selection of hyperadherent mutants are discussed. INTRODUCTIONThe development of a multicellular community associated with a surface and surrounded by an exopolymeric matrix, referred to as a biofilm, is common to a variety of bacteria under different environmental conditions. Biofilm formation has received increasing attention due to its importance in medicine, since biofilm populations are considered relevant to chronic infection, and are more resistant to the action of antibiotics and biocidals than planktonic populations (Anderson & O'Toole, 2008;Høiby et al., 2010). Biofilm development is also relevant in industrial settings and in the design of bioreactors (Nicolella et al., 2000;Singh et al., 2006). Genetic determinants that play a role in biofilm formation have been unravelled in different bacterial species, and environmental and cellular signals that influence this process have also been described. These include iron availability, quorum sensing, and the intracellular secondary messenger cyclic di-GMP (Banin et al., 2006;Patriquin et al., 2008;Ueda & Wood, 2009;Coenye, 2010). Changes in the levels of cyclic di-GMP correlate with phenotypic changes associated with virulence, motility, colony morphology, production of exopolysaccharides, and the transition between planktonic and sessile lifestyles (Hengge, 2009; Römling & Simm, 2009, and references therein). Changes in the expression of different functions are also associated with one or the other lifestyle; in Pseudomonas aeruginosa and Pseudomonas putida, for example, differential expression of flagellar components has been observed when comparing planktonic and biofilm populations, and even during the various stages of biofilm development (Sauer & Camper, 2001;Sauer et al., 2002;Toutain et al., 2007). In the former organism, swarming motility and surface attachment are inversely regulated through a pathway that involves BifA, a cyclic-di-GMP phosphodiesterase, and the membra...

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“…A homology search revealed that orf3 is highly homologous to the putative 3-oxoacyl-(acyl carrier protein [ACP]) synthase III (␤-ketoacyl-ACP synthase III) genes of Escherichia coli, Salmonella enterica serovar Typhimurium, and Pseudomonas putida strain KT2440 (24,36). It has been reported that a 3-oxoacyl-ACP synthase III called FabH is one of the enzymes in the type II fatty acid synthesis system (20).…”

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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

et al. 2006

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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 (⌬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 ⌬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 H 2 O 2 and antibiotics were enhanced in the ⌬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 ⌬orf3 mutant. The phenotypes of the ⌬orf3 mutant and an AHL synthesis (⌬psyI) mutant were similar, although the mutant-specific characteristics were more extreme in the ⌬orf3 mutant. The swarming motility of the ⌬orf3 mutant was greater than that of the ⌬psyI mutant. This was attributed to the synergistic effects of the overproduction of biosurfactants and/or alternative fatty acid metabolism in the ⌬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.Pseudomonas syringae pv. tabaci 6605, an phytopathogenic bacterial isolate, causes wildfire disease on host tobacco plants and induces a hypersensitive reaction (HR) on nonhost plants.In previous studies, we demonstrated that flagellin, a component of the flagellar filament, is a major elicitor of HR by P. syringae pv. tabaci 6605 (30, 35). Flagellin of P. syringae pv. tabaci 6605 induces HR on nonhost plants but not on its host tobacco plant. Although the deduced amino acid sequence of P. syringae pv. glycinea race 4 flagellin (FliC) is identical to that of P. syringae pv. tabaci 6605 flagellin, the flagellin of P. syringae pv. glycinea does induce HR on tobacco plants, suggesting that posttranslational modification of flagellin determines the specificity to induce HR (34). We recently reported that genes existing upstream of fliC, which encodes flagellin protein, are involved in the glycosylation of flagellin in these two pathovars (33, 36). There are three open reading frames, namely, orf1, orf2, and orf3, in the glycosylation islands of P. syringae pv. tabaci and pv. glycinea (Fig. 1). Because the proteins encoded by orf1 and orf2 showed homology to a putative glycosyltransferase and the molecular mass of the flagellin of each orf1 and orf2 deletion mutant (⌬orf1 and ⌬orf2 mutants) was decreased, these genes are thought to encode the glycosyltransferases. I...

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Complete genome sequence and comparative analysis of the metabolically versatile Pseudomonas putida KT2440

Cited by 1,198 publications

References 56 publications

Cell Density-Dependent Gene Contributes to Efficient Seed Colonization by Pseudomonas putida KT2440

Cell Density-Dependent Gene Contributes to Efficient Seed Colonization by Pseudomonas putida KT2440

Selection of hyperadherent mutants in Pseudomonas putida biofilms

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

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