The evidence for an internal hump is somewhat weaker for PA01 than PA14 but we note that our test is conservative, as we have not included data on the effectiveness of either strain at inhibiting find more itself. As both of these values are zero (see Methods), including these values would produce a much more pronounced hump. Table 1 Linear and quadratic regressions of inhibition of clinical isolates by sterile (non heat treated) cell free extract of PA01 and PA14 cultures as function of genetic distance (Figure 2) Source df Value St Error t P-value Multiple R2 AIC PA01 Linear model 0.072 0.059 90.91 Intercept 1 3.27 0.969 3.38 0.0014 Linear term 1 -2.41 1.31 -1.84

0.072 4SC-202 chemical structure Residual SE 53

0.55 PA01 Quadratic model 0.010 0.160 86.94 Intercept 1 -17.00 8.81 -2.08 0.043 Linear term 1 53.94 22.61 2.38 0.021 Quadratic term 1 -38.89 15.58 -2.50 0.016 Residual SE 52 0.53 PA14 Linear JQ-EZ-05 model 0.15 0.044 39.80 Intercept 1 1.99 0.71 2.81 0.0072 Linear term 1 -1.45 0.98 -1.48 0.15 Residual SE 47 0.36 PA14 Quadratic model < 0.0001 0.345 26.08 Intercept 1 -37.51 8.62 -4.35 0.0001 Linear term 1 109.8 24.23 4.53 < 0.0001 Quadratic term 1 -77.88 16.95 -4.59 < 0.0001 Residual SE 46 0.30 To verify that genetic distance correlates with resource use, we measured the metabolic similarity of toxin producing

strains to the clinical isolates using Biolog plates (see Methods). Metabolic profiles become more divergent with increasing genetic distance, as expected, reflected in the significantly Acyl CoA dehydrogenase negative linear relationship observed between Jaccard distance and metabolic correlation between pairs of strains (PA01: slope ± standard error = -0.493 ± 0.213; multiple R2 = 0.098, t ,49 = -2.312, P = 0.025; PA14: slope ± standard error = -0.644 ± 0.208, multiple R2 = 0.164, t 49 = -3.104, P = 0.0032). These results lend support to the idea that genetic distance is linked to ecological divergence. It is further notable that inhibition score peaked at intermediate metabolic similarities for both PA01 and PA14 but was statistically significant only for PA14 (see Additional file 1: Table S1 and Additional file 2: Figure S1; F-ratio test on the fitting of the quadratic term, PA01: F1,48 = 0.176, P = 0.68; PA14: F1,42 = 7.00, P = 0.011). It is not immediately obvious why we detected a significant quadratic relationship between inhibition score and metabolic similarity in one strain but not the other. One possibility is that the Biolog plates we used here, which provide profiles on carbon substrate metabolism, represent one of many possible dimensions along which ecological divergence can proceed.