Abstract: Reductive soil disinfestation (RSD) is an effective and environmental-friendly technology that can overcome the obstacle of soil quality in continuous cropping. To investigate the effects of RSD technology on the quality and microbial community structure of soils before and after tobacco planting, organic bean dregs were applied and RSD treatment was conducted after flooding and plastic film mulching in soils with the serious obstacle of continuous cropping. The changes of chemical properties and microbial community structure in the soils treated by the RSD technology before and after tobacco planting were compared, and the effects of the RSD treatment on tobacco growth vigor and disease resistance were evaluated. The results showed that the RSD treatment before tobacco planting significantly increased the pH value, the contents of NH₄⁺-N and available phosphorus (mass fraction) of the soils. The contents of available phosphorus, available potassium and NH₄⁺-N in the soils treated with the RSD after tobacco planting were significantly higher than those of untreated soils (CK1) as well as the flooded and plastic film mulched soils (CK2). The results of microbial community analysis indicated that the RSD treatment significantly raised the relative abundances of Flavisolibacter, Bacillus, Coprococcus, Podospora and Mortierella, while reduced those of Fusarium, Rhizoctonia and Saitozyma, especially that of bacterial wilt pathogen—Ralstonia. Compared with CK1 and CK2, the RSD treatment promoted the agronomic traits (plant height, number of effective leaves, maximum leaf area) of tobacco plants to some extent and reduced the incidence rate of bacterial wilt by 59.00% and 49.58% respectively. Thereby, the RSD treatment of soil can significantly improve soil microbial community structure and is beneficial to the improvement of soil nutrient as well as tobacco growth and development.