We studied the effect of no-till (disc seeder), conventional-till (tine scarifier+disc seeder) and rotary-till (rotary hoe+disc seeder) management on soil organic matter (SOM) components, rates of carbon (C) and nitrogen (N) cycling, substrate utilization and microbial community composition. We hypothesized that labile SOM fractions are sensitive to changes in tillage techniques and, in turn mediate any tillage-induced changes in microbial function and composition. A replicated field site was established in May 1998 in the semi-arid agricultural region of Western Australia and soils were collected in September 2004. We found soil pH varied between different tillage techniques as an initial lime application was mixed to deeper soil depths in rotary-till soil than no-till and conventional-till soil. Total-C was greater in surface soil and lower in subsurface soil from no-till and conventional-till plots than from rotary-till plots, but there was no effect of tillage technique on total-C when averaged across soil depths. Light (specific density 0.50) and were greater in no-till and conventional-till soil than rotary-till soil both within, and across soil depths. These soil variables generally increased (r>0.5) with increasing soil pH. Dissolved organic N and gross N mineralization were positively correlated (r>0.90) but neither was affected by tillage techniques. No-till soil had greater utilization of carboxylic acids and lower utilization of amino acids and carbohydrates than conventional-till and rotary-till soil; surface soil also had greater utilization of carboxylic acids than subsurface soil. In turn, substrate utilization differed between soil depths, and between no-till soil and conventional-till and rotary-till soil; these differences were correlated with soil pH, total-N, DOC, LFOM-N and microbial biomass nitrogen (MB-N). Bacterial and fungal biomasses generally decreased with soil depth and were greater in no-till and conventional-till soil than rotary-till soil. Microbial community composition differed between all tillage techniques and soil depths; these differences were correlated with soil textural classes, soil pH, and total, LFOM, DOM and microbial C and N pools. These results indicate that most tillage-induced changes to soil properties were associated with the greater soil disturbance under rotary-till than under no-till or conventional-till management. Our results indicate that tillage-induced changes to soil pH, and LFOM, DOM and microbial biomass pools are likely to be important regulators of the rates of C and N cycling, substrate utilization and microbial community composition in this coarse textured soil.