To assess the effect of phloretin on the virulence of Pb1692, we measured the severity of symptoms in two plants, Zantedeschia aethiopica (calla lily) and Solanum tuberosum (potato), as described by Yishay et al. (2008). For this assay, fully expanded young leaves of calla lily and small (25–50 g) potato tubers were taken and surface-sterilized by soaking in 0.5% sodium hypochlorite for 20 min. Then, the samples were washed twice with sterile, double-distilled H2O and air-dried under a hood. For calla lily, leaf discs approximately 20 mm in diameter were excised and transferred to Petri dishes containing MS medium. For potato, whole disinfected tubers were used for the infection assay. Both the leaf discs and the potato tubers were pierced at the center with a sterile tip. Bacteria that had been grown for 16 h, 28°C in liquid LB were diluted to 1 × 108 CFU/mL in sterile, double-distilled H2O with or without the test compound and incubated for 2 h at 28°C on a 150-rpm incubator shaker. After 2 h, leaf discs and potato tubers were inoculated with 10 μL of bacterial suspension (containing 106 CFU) in the presence or absence of phloretin or ciprofloxacin. The inoculated plant material was incubated at 28°C without shaking. Disease severity in calla lily was expressed as the percentage of decayed tissues relative to the total disc area after 15 h. In potato, disease severity was assessed in terms of the percentage of tissue that had rotted after 48 h of exposure. The entire experiment was repeated at least twice with eight replicates for calla lily and four replicates for potato.

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