Figure 3.

A large deletion occurs early during the passage of population 4 and provides resistance to the inhibitory agent. (a) SynMap dotplot visualizes the large deletion occurring from 131 to 499 kb in the evolved 4B pMPPla107 as a large shift across the x-axis. The remaining portions of the sequences maintain perfect synteny, indicating that a clean deletion occurred. The x-axis is ancestral pMPPla107 gene order where x_{1, …, N} = gene_{1, …, N} and the y-axis is the line 4B evolved pMPPla107 gene order where y_{1, …, N} = gene_1…N.]. (b) We tested single colony isolates from frozen stocks of the passage line 4 populations at generations 0 and 100 for sensitivity to the inhibitory agent. We found that resistance to the inhibitory agent is present in the generation 0 isolate from population 4. This resistance is maintained in generation 100 (shown) through generation 500 and is the only unique mutation in pMPPla107 other than a synonymous SNP. All overlays were plated after 4 h of growth n KB and spotted with 10 µl of P. stutzeri filter-sterilized supernatants. All images are representative of three biological replicates. Raw image files can be found in electronic supplementary material at Figshare (doi:10.6084/m9.figshare.15072537.v3): DBL408 is electronic supplementary material, figure S4, 4A-0 is electronic supplementary material, figure S5, 4A-100 is electronic supplementary material, figure S6. (Online version in colour.)