The strong relationships between plant and soil microbial communities suggest that post-fire vegetation may be a critical driver of the post-fire recovery of the structure and functioning of the soil microbial community. In this study, we conducted an experimental burning and evaluated the effect of the post-fire persistence traits of the vegetation (resprouter and seeder) on the medium-term (3 years after fire) post-fire response of the soil microbial activity in Mediterranean shrublands. The experiment was carried out in a Mediterranean shrubland (Eastern Spain), where four main types of microsites were selected: Bare-soil inter-patch (BS); Resprouter patch (R), Seeder patch (S), and Mixed patch (R+S). For each microsite, we analyzed soil basal respiration, water-soluble carbon, microbial biomass carbon, total organic carbon, and dehydrogenase activity at 0-5 cm soil depth. We also assessed plant cover dynamics. Our results suggest that, in general, fire impacts on soil microbial activity are not long-lasting, with most assessed soil variables being similar between burned and unburned areas three years after the fire. However, while the unburned microsites showed a trend in microbial biomass and activity from lower values in bare soils to higher values in R+S patches, these differences disappeared in the burned area, due to both a slight increase in microbial activity and biomass in bare soils, and the opposite response for soils under R+S patches. Our results highlight the role of the plant persistence trait in the microbial post-fire response of Mediterranean soils.