Tropical soil microorganisms are major recyclers of biosphere organic carbon. However, the link of tropical microorganisms to the two primary carbon inputs (roots and litter) remains unclear. Here, we studied the effects of excluding living roots and litter on microorganisms in leaf litter and topsoil in rainforests and plantations in Sumatra, Indonesia. Unexpectedly, excluding living roots strongly decreased microbial biomass and basal respiration in litter but not in soil, indicating that tropical trees prioritize mining for nutrients in litter layer rather than mineral soil. Contrary to litter, soil microbial communities were predominantly influenced by long-term factors related to land-use history. Litter removal neither significantly affected microbial biomass nor community structure in soil, suggesting compartmentalized carbon processing between litter and soil. Our study sheds new light on fundamental-ecosystem processes in the tropics and calls for greater consideration of the litter layer and for including root-derived resources in global carbon cycling models.