The achromobactin biosynthetic pathway is a particularly valuable resource for the study of these enzymes as it relies on the action of all three types of synthetase GDC 0032 research buy [22, 24]. Achromobactin has been shown to be important for virulence in Dickeya dadantii (formerly Erwinia chrysanthemi) [25], and both see more pyoverdine and achromobactin contribute to epiphytic fitness of P. syringae pv. syringae 22d/93 [21], but the contribution of siderophores

to virulence of P. syringae 1448a has not previously been characterized. We therefore examined the roles of both achromobactin and pyoverdine in virulence of P. syringae 1448a, as well as their relative contribution to iron uptake and growth under more precisely defined conditions. Results Identification www.selleckchem.com/TGF-beta.html and in silico characterization of the P. syringae 1448a pyoverdine locus The biosynthesis of pyoverdine has been most extensively studied in P. aeruginosa PAO1 and most, if not all, of the genes required for pyoverdine synthesis in this strain have now been identified [[6, 10, 26]]. Ravel and Cornelis [8] used the PAO1 pyoverdine genetic locus as a blueprint for annotation of the pyoverdine loci from three other fluorescent pseudomonads, including P. syringae pv. tomato DC3000. We adopted a similar strategy to interrogate

the P. syringae 1448a genome, individually BLASTP searching all of the known PAO1 pyoverdine proteins against the P. syringae 1448a sequence database [27]. The genomic organization of pyoverdine genes in P. syringae 1448a is highly similar to the Staurosporine molecular weight P. syringae DC3000 genetic locus presented by Ravel and Cornelis [8], but less similar to that of PAO1 (Figure 1A, Table 1). Given the similarity with the P. syringae DC3000 genetic locus and the excellent earlier analysis of Ravel and Cornelis, we confine our analysis of the non-NRPS genes of P. syringae 1448a to two aspects not previously noted by them. The first concerns the only PAO1 gene that clearly lacks an ortholog in P. syringae, pvdF, which encodes an enzyme required for generating the N5-formyl-N5-hydroxyornithine residues that are present in the PAO1 (but not P. syringae) pyoverdine side chain. Instead,

P. syringae 1448a contains a gene (Pspph1922; marked * in Figure 1A) that is 37% identical at a predicted protein level to the syrP gene of Pseudomonas syringae pv. syringae. Originally mis-annotated as a putative regulatory gene, SyrP has subsequently been shown to be an aspartate hydroxylase that is required for synthesis of the NRPS-derived phytotoxin syringomycin [28]. On this basis we propose that Pspph1922 very likely catalyzes β-hydroxylation of two hydroxyaspartate residues expected to be present in the P. syringae 1448a pyoverdine side chain (Figure 1B), with equivalent iron-chelating roles to the N5-formyl-N5-hydroxyornithine residues of PAO1 pyoverdine. We also note that P. syringae 1448a contains two orthologs of the PAO1 ferripyoverdine receptor gene fpvA.