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Bistable expression of the  Salmonella enterica  pathogenicity island 1 (SPI-1) and the flagellar network (Flag) has been described previously. In this study, simultaneous monitoring of OFF and ON states in SPI-1 and in the flagellar regulon reveals independent switching, with concomitant formation of four subpopulations: SPI-1 OFF  Flag OFF , SPI-1 OFF  Flag ON , SPI-1 ON  Flag OFF , and SPI-1 ON  Flag ON . Invasion assays upon cell sorting show that none of the four subpopulations is highly invasive, thus raising the possibility that Flag OFF  cells might contribute to optimal invasion as previously proposed for SPI-1 OFF  cells. Time lapse microscopy observation indicates that expression of the flagellar regulon contributes to the growth impairment previously described in SPI-1 ON  cells. As a consequence, growth resumption in SPI-1 ON  Flag ON  cells requires switching to both SPI-1 OFF  and Flag OFF  states.

Single Cell Analysis of Bistable Expression of Pathogenicity Island 1 and the Flagellar Regulon in Salmonella enterica