3%, and in flooded pots, it was 16.3% (all significantly
different from the null hypothesis), indicating that there was a negative correlation between the competitiveness of the nonmotile mutant and vermiculite water content. Hence, we evaluated the competition for nodulation of all strains in the flooded condition. As shown in Table 2, the behavior of the mutants carrying one flagellum was similar as in field capacity (except LP 6866, which, although occupied 64.3% of nodules, did not deviate significantly from the null hypothesis due to higher experimental variability). As in field capacity, LP 3008 seemed to compete better than LP 3004 against its derivative without a thin flagellum. Meanwhile, the nonmotile double mutants
were again significantly less competitive than in field capacity. Bacterial swimming may be observed in semi-solid agar plates as a colony expansion a this website click here few millimeters below the agar surface, and must not be confused with swarming, which occurs in plates of more concentrated agar where colonies of differentiated cells move on the surface (Harshey, 1994, 2003). Indeed, rhizobia mutants able to produce swimming halos, but swarming colonies were not described (Braeken et al., 2007; Nogales et al., 2010). Our results showing swimming in 0.3% agar indicated that the thin flagellum of B. japonicum is actively used for this motion, because LP 5844 (ΔfliC1-4, producing only the thin
flagellum) formed the widest swimming halo of all mutants. In addition, this strain tumbled more frequently than the wild type. In agreement with our results, Wolfe & Berg (1989) also reported that the swimming halo rate of expansion increases with Urocanase tumble frequency. Thin flagellum derepression in LP 3008 may also cause its faster spread in 0.3% agar; however, it does not explain why the LP 3008 mutant lacking this flagellum still formed wider swimming halos than the corresponding mutant in the LP 3004 background. In 0.3% agar, the consumption of nutrients and release of other chemicals by the rings of bacteria moving inside the medium creates a chemoattractant gradient (Adler, 1966). Thus, the higher chemotaxis of LP 3008 (Althabegoiti et al., 2008) may also contribute to its higher displacement. After characterizing the motility provided by each flagellum, we assessed their roles in the competition for nodulation in vermiculite. Although all mutants moved less than the parental strains in swimming plate assays, they were differently affected in their competitiveness for nodulation, which also depended on the water status of the vermiculite. While mutants lacking the thin flagellum were, in general, more competitive than the parental strains both at field capacity and in the flooded environment, the mutants lacking the thick flagellum were less competitive.