The effects of wood vinegar (WVG) on microbial activity and communities of activated sludge were investigated in a sequencing batch reactor (SBR) process. Results showed that the optimal WVG concentration was 4 mu L/L when the pollutants removal efficiency and microbial activity were promoted by a WVG dilution factor of 1000. WVG could reduce the increase in microbial species richness, which led to a more notable variety of microbial species diversity. The enhanced microbial activity and communities were addressed to the promotion of 7 main classes of microbes in Proteobacteria, Bacteroidetes, Acidobacteria, and Nitrospirae phyla. The growth of ammonia-oxidizing bacteria (AOB), nitrite-oxidizing bacteria (NOB), and main genera of denitrifying bacteria (DNB), phosphorus-accumulating organisms (PAOs), and glycogen-accumulating organisms (GAOs) could be promoted by WVG, which improved the sewage treatment effectiveness in a SBR.