FIG 1.

Adaptive laboratory evolution (ALE) experiment of plasmid-cured P. putida S12 to increasing concentrations of toluene. (A) Experimental design of ALE. ALE was performed on three plasmid-cured P. putida S12 strains (S12-06, S12-10, and S12-22). In the ALE experiment, LB medium (yellow) was used as the growth medium with the addition of an increasing toluene concentration of 0.05% (vol/vol) every cycle (gray). (B) Plasmid-cured P. putida S12 regained the ability to grow in high toluene concentrations. The solvent tolerance phenotype of ALE-derived strains was tested by observing strain growth on LB medium with 10% (vol/vol) toluene within 48 h. Single and double asterisks indicate the control and sample strains, respectively, that were taken for whole-genome sequencing. This experiment was performed with three biological replicates, and error bars indicate standard deviations.