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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 g 1(-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.

Growth of Micro-organisms in Gel-stabilized Two-dimensional Diffusion Gradient Systems