Biochar is a potential tool to mitigate climate change by enhancing C sequestration in soils, but its use as a soil amendment to improve soil fertility and crop yields is still a contentious subject. InNorth East (NE) Thailand, biochar has been promoted to restore soil fertility in rubber tree plantations. Despite this, there is scarce information on the impact of biochar application on the soil biota, particularly on microbial communities associatedwith rubber trees. The effects of increasing doses of biochar onmicrobial communities were investigated in a rubber tree plantation in NE Thailand, 28 months after application. Biochar application resulted in increases of soil pH and nutrient contents and also had an impact on both bacterial and fungal communities. Changes in microbial composition and structure were observed although fungal communities weremoremarkedly affected than bacterial communities. The nature and magnitude of the observed changes were strongly related to soil properties (pH, soil moisture and P content),while biochar dose (5, 10 or 20 tons/ha) effect was not significant. Our results highlight the need for additional research for a better understanding of the impact of biochar application on soil microbial communities and further cascading effects on ecosystem functions.