Single factor combined with orthogonal test was employed to optimize the best technology for fermentation of tobacco leaves by Lactobacillus plantarum in this study. The chemical components, aroma substances and sensory evaluation of fermented tobacco leaves were analyzed. The optimal fermentation technology of tobacco leaves by L. plantarum was achieved when the addition of L. plantarum, glucose syrup and soy protein peptide was 0.80‰, 5.0‰, and 0.05‰ respectively, with fermentation for 8 days. The starch content of tobacco leaves decreased to the most significantly, and 9 kinds of new volatile aroma substances were detected. The content of nicotine decreased by 5.58%. Fermented tobacco leaves from different regions was compared, and it was found that the addition of L. plantarum, carbon and nitrogen sources could effectively improve the smoking quality of tobacco leaves from various regions. The content of starch decreased significantly while that of soluble total sugar and reducing sugar increased significantly (P＜0.05), among which the tobacco leaves from Chongqing changed to the most extent. The total alkaloids and total nitrogen content of tobacco leaves decreased slightly while the sensory evaluation was enhanced significantly (P＜0.05), among which the tobacco leaves from Guizhou improved to the most extent by 1.875 points.