Figure 4.

Impact of native DGC on interspecies killing and cdi‐GMP levels. A. A. tumefaciens strains with either an empty vector control, pdgcA, patu2091, pdgcC, patu2691, patu1207 and patu5372 were incubated for 14 h on a I medium (pH5.5) with E. coli containing pRL662 which harbours a gentamicin cassette. Bacteria were then resuspended in PBS and plated on LB agar‐containing gentamicin. All experiments are the mean of three independent biological experiments with standard deviation error bars. Statistical significance was determined using students t‐test comparing each strain with the vector control, p < 0.05 *, p < 0.01 ** B. C‐di‐GMP levels were quantified via LC‐MS/MS in strains expressing either an empty vector control, atu5372 or atu2691. Native DGSc were induced with 0.5 mM IPTG for 16 h in 523 rich medium and cells equivalent to an OD₆₀₀ 5 were collected. Samples of interest were compared to a standard curve derived from measurements of known concentrations of pure cdi‐GMP to determine the concentration (in nM) of c‐di‐GMP in the samples. All experiments are the mean of two independent biological experiments with standard deviation error bars.