In saline soils under semi-arid climate, low matric and osmotic potential are the main stressors for microbes. But little is known about the impact of water potential (sum of matric and osmotic potential) and substrate composition on microbial activity and biomass in field collected saline soils. Three sandy loam soils with electrical conductivity of the saturated soil extract (ECe) 3.8, 11 and 21 dS m(-1) (hereafter referred to EC3.8, EC11 and EC21) were kept at optimal water content for 14 days. After this pre-incubation, the soils were either left at optimal water content or dried to achieve water potentials of -2.33, -2.82, -3.04 and -4.04 MPa. Then, the soils were amended with 20 g kg(-1) pea or wheat residue to increase nutrient supply. Carbon dioxide emission was measured over 14 days; microbial biomass C was measured at the end of the experiment. Cumulative respiration decreased with decreasing water potential and was significantly (P < 0.05) lower in soils at water potential -4 MPa than in soils at optimal water content. The effect of residue type on the response of cumulative respiration was inconsistent; with residue type having no effect in the saline soils (EC11 and EC21) whereas in the non-saline soil (EC3.8), the decrease in respiration with decreasing water potential was less with wheat than with pea residue. At a given water potential, the absolute and relative (in percentage of optimal water content) cumulative respiration was lower in the saline soils than in the non-saline soil. This can be explained by the lower osmotic potential and the smaller microbial biomass in the saline soils. However, even at a similar osmotic potential, cumulative respiration was higher in the non-saline soil. It can be concluded that high salt concentrations in the soil solution strongly reduce microbial activity even if the water content is relatively high. The stronger relative decrease in microbial activity in the saline soils at a given osmotic potential compared to the non-saline soil suggests that the small biomass in saline soils is less able to tolerate low osmotic potential. Hence, drying of soil will have a stronger negative effect on microbial activity in saline than in non-saline soils.