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A better understanding of soil microbial processes is required to improve the synchrony between nutrient release from plant residues and crop demand. Phospholipid fatty acid analysis was used to investigate the effect of two crop rotations (continuous maize and maize–crotalaria rotation) and P fertilization (0 and 50 kg Pha⁻¹yr⁻¹, applied as triple superphosphate) on microbial community composition in a highly weathered soil from western Kenya. Microbial substrate use in soils from the field experiment was compared in incubation experiments. Higher levels of soil organic matter and microbial biomass in the maize–crotalaria rotation were connected with higher total amounts of phospholipid fatty acids and an increase in the relative abundances of indicators for fungi and gram-negative bacteria. P fertilization changed the community profile only within the continuous maize treatment. The decomposition of glucose …