commun.). The products of these genes do not have any homologues in the databases. Furthermore, the products of ECA3711, ECA3724 and ECA3730 were detected by Coulthurst et al. (2008) in the secretome of Pa. ECA3724 and ECA3730 are predicted to encode a capsid protein and selleck chemicals llc the major tail tube protein. The presence of these structural components of the

virion in the extracellular medium may suggest that excision of ECA41 from the chromosome is followed by encapsidation. To determine whether these proteins, or any others provided by the prophages, contributed to virulence in Pa, we deleted the entire prophages – both individually and in combination – from the Pa genome, using the limits of the prophage that we had determined experimentally. No differences were detected in the growth rates of TJE101 (ΔECA29), TJE102 (ΔECA41) or TJE103 (ΔECA29, ΔECA41) in PMM or PMB. Culture supernatant samples were taken throughout these growth experiments and the levels of secreted protease, pectate lyase

Cabozantinib and cellulase activities were determined. No changes were observed in the mutants compared with the wild type (data not shown). Swimming motility has previously been shown to be important in Pa potato infections (Mulholland et al., 1993; Evans et al., 2010). Comparison of motility of the prophage deletion strains showed that TJE101 and TJE102 were consistently less motile than the wild type, with a 5–8% reduction in halo size (data Phosphoribosylglycinamide formyltransferase not shown). The decrease, even though small, was statistically significant after multiple biological repetitions (P<0.05, paired t-test) and could result in reduced fitness in the environment. Finally, the ability of the prophage-deficient strains to rot potato tubers was assessed in vivo. The prophage deletion mutants showed a statistically significant reduction in virulence compared with the wild type (Fig. 3) (P<0.05). This result demonstrates that the acquisition of

these prophages has contributed towards the pathogenicity of Pa. Similar to each of the single mutants, the double mutant (TJE103) showed a modest, but statistically significant, reduction in motility and a reduction in virulence in tubers (data not shown). However, due to the intrinsically variable nature of such assays, we were unable to determine whether the impacts of the two mutations were additive. Although the impacts on motility and virulence were not drastic under lab conditions, it is possible that such differences could have significant fitness and survival consequences in the environment and during pathogenesis in the field. The two Pa prophages, ECA29 and ECA41, are likely to be maintained at a metabolic cost to the cell: at 68 kb combined, they represent over 1% of the genome, which must be replicated in each cell cycle. This in itself implies that the prophages may confer a selective advantage on cells that carry them. The results herein demonstrate that these two prophages do contribute to in vivo pathogenicity.