The antibacterial activities and mechanism of action of acetone extracts from R. rubescens were reported in this paper. The results showed that 80% acetone extracts had both the highest contents of total phenolics and flavonoids. Acetone extracts showed better antibacterial activities against Gram-positive bacterial strains and there were no inhibitory effects found on tested Gram-negative bacteria. In addition, 80% acetone extracts from R. rubescens had relatively higher antibacterial activities with the lowest values of MIC and MBC at 2.5 mg/mL and 5 mg/mL against B. subtilis. The antibacterial mechanism of 80% acetone extracts against Bacillus subtilis might be described as disrupting cell wall, increasing cell membrane permeability, and finally leading to the leakage of cell constituents