Lake Urmia in north-western Iran is one of the largest hypersaline lakes in the world, and agricultural production in the surrounding area is limited by soil salinity. We investigated the effects of salinity on belowground microbial communities in soils collected from fields of cultivated onions (Allium cepa L.) and lucerne (Medicago sativa L.), and sites with the native halophyte samphire (Salicornia europaea L.). We tested the hypotheses that salinity reduces microbial biomass and changes the structure of the microbial community. The physical and chemical properties of soil samples were analysed, and phospholipid fatty acids were identified as signatures for various microbial groups. We found that the organic carbon (OC) content was the dominant determinant of microbial biomass. We also found linear relationships between OC and the biomass of various groups of organisms across the wide salinity gradient studied. Salinity, on the other hand, caused changes in the microbial fatty acid composition that indicated adaptation to stress and favoured saprotrophic fungi over bacteria, and Gram-negative bacteria over Gram-positive. Principal component analysis showed that salinity variables and microbial stress indices formed one group, and OC and microbial biomass another. The importance of OC for high microbial biomass in severely stressed soils indicates that OC amendment may be used to mitigate salt stress and as a method of managing saline soils.