Interestingly, a prophage element found in the identical spot (be

Interestingly, a prophage element found in the identical spot (between mutS and cinA) in the genome of P. fluorescens SBW25 http://​www.​sanger.​ac.​uk/​Projects/​P_​fluorescens has a similar overall organization but contains a P2-like bacteriophage tail cluster (orf5 through orf18) similar to that in phage CTX (Fig. 1), thus resembling another class of phage tail-like bacteriocins, the R-type pyocins of P. aeruginosa [19]. Furthermore, a homologous region from P. fluorescens Pf0-1 (CP000094) contains

both the lambda-like and P2-like tail clusters (Fig. 1), making it similar to the hybrid R2/F2 pyocin locus from P. aeruginosa PAO1 [19]. The differences in organization of the putative phage tail-like pyocins among these prophages clearly indicate that the corresponding loci are subject to extensive recombination, with a possible recombination hotspot between two highly conserved DNA segments comprised of the phage repressor (prtR) and holin click here (hol) genes, and the endolysin (lys) gene (Fig. 1). In strains Pf-5 and Q8r1-96, the putative prophage ABT-263 01-like pyocins are integrated between mutS and the cinA-recA-recX genes (Fig. 1), which suggests that these elements might be activated this website during the SOS response, as is the putative prophage gene cluster integrated into the mutS/cinA region of P. fluorescens DC206 [21]. The mutS/cinA region

is syntenic in several Gram-negative bacteria [22], and comparisons reveal that prophage 01-like elements occupy the same site in the genomes of P. fluorescens Pf0-1, P. fluorescens SBW25, and P. entomophila L48 [23], whereas unrelated prophages reside upstream of cinA in P. putida F1 (GenBank CP000712) and P. syringae pv. tomato DC3000 [24]. The latter strain, as well as P. putida KT2440 [25], carry SfV-like bacteriophage tail assembly clusters elsewhere in the genome. The putative F- and R-pyocins appear to be ubiquitously distributed among

strains of P. fluorescens as illustrated by a screening experiment PDK4 (Fig. 4) in which genomic DNA of different biocontrol strains was hybridized to probes targeting the lambda-like and P2-like bacteriophage tail gene clusters of Q8r1-96 and SBW25, respectively. The F- and R-pyocin-specific probes each strongly hybridized to DNA from 12 of 34 P. fluorescens strains, while the remaining 22 strains tested positive with both probes. Figure 4 Southern hybridization of DNA from 34 strains of P. fluorescens with probes targeting F-pyocin- and R-pyocin-like bacteriophage tail assembly genes. Total genomic DNA from each strain was digested with EcoRI and PstI restriction endonucleases, separated by electrophoresis in a 0.8% agarose gel, and transferred onto a BrightStar-Plus nylon membrane. The blots were hybridized with biotin-labeled probes prepared from P. fluorescens strains Q8r1-96 (A) and SBW25 (B) targeting the SfV-like (A) and CTX-like (B) bacteriophage tail assembly genes, respectively.

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