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The Szybalski wedge plate technique was modified to generate two-dimensional diffusion gradients for two environmental variables: pH value and NaCl concentration were selected in this study. After experimentation, pH values chosen ranged from 3·9 to 8·1 and salt concentrations from 23 to 79 gl−1. The pH gradients were slightly sigmoidal whilst the salt gradients were approximately linear. The reproducibility of the technique was assessed and found to be satisfactory in replicate growth experiments. A wide range of growth patterns was observed for a large variety of different bacterial types. The growth boundary and the pattern details are sufficiently distinct to suggest that the technique can distinguish between closely related species or strains.
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