Environmental disturbances such as drought can impact soil health and the resistance (ability to withstand environmental stress) and resilience (ability to recover functional and structural integrity after stress) of soil microbial functional activities. A paucity of information exists on the impact of drought on soil microbiome and how soil biological systems respond to and demonstrate resilience to drought stress. To address this, we conducted a systematic review and meta-analysis (using only laboratory studies) to assess the response of soil microbial biomass and respiration to drought stress across agriculture, forest, and grassland ecosystems. The meta-analysis revealed an overall negative response of microbial biomass in resistance (−31.6 %) and resilience (−0.3 %) to drought, suggesting a decrease in soil microbial biomass content. Soil microbial respiration also showed a negative response in resistance to drought stress indicating a decrease in soil microbial respiration in agriculture (−17.5 %), forest (−64.0 %), and grassland (−65.5 %) ecosystems. However, it showed a positive response in resilience to drought, suggesting an effective recovery in microbial respiration post-drought. Soil organic carbon (SOC), clay content, and pH were the main regulating factors of the responses of soil microbial biomass and respiration to drought. In agriculture ecosystem, soil pH was primarily correlated with soil microbial respiration resistance and resilience to drought, potentially influenced by frequent land preparation and fertilizer applications, while in forest ecosystem SOC, clay content, and pH significantly impacted microbial biomass and respiration resistance and resilience. In grassland ecosystem, SOC was strongly associated with biomass resilience to drought. The impact of drought stress on soil microbiome showed different patterns in natural and agriculture ecosystems, and the magnitude of microbial functional responses regulated by soil intrinsic properties. This study highlighted the importance of understanding the role of soil properties in shaping microbial responses to drought stress for better ecosystem management.