Abstract:[Background] The continuous cropping and poor fertilization habits in the tobacco-planting areas of Panzhihua have brought serious stress to the nutrient balance and microbial diversity in the soil, leading to declines in the yield and quality of tobacco and seriously affecting the development of the tobacco industry. [Objective] To study the effects of applying rhizobium-inoculated leguminous green manure on the microbial diversity and community composition in the soil cultivated with tobacco for years in Panzhihua and provide a theoretical basis for scientific fertilization and establishment of a reasonable planting pattern of flue-cured tobacco in Panzhihua. [Methods] We collected soil samples from the newly reclaimed tobacco field and the field planted with tobacco for years (both with the application of green manure) in Pingshan Township, Miyi County of Panzhihua. The physical and chemical properties of the soil samples were determined, and the bacterial community composition and diversity were analyzed by high-throughput sequencing. [Results] Inoculation with rhizobia increased the number of nodules and roots of pea seedlings, especially in the newly reclaimed tobacco field. The application of green manure increased the content of organic carbon and inorganic nutrients in soil and reduced soil pH and volume weight. In addition, the application of pea green manure changed the bacterial diversity and community composition in the soil. Specifically, it elevated the Simpson and Shannon indexes of bacteria in the newly reclaimed tobacco field and reduced the relevant diversity indexes in the field cultivated with tobacco for years. The predominant bacterial phyla of the soil samples planted with tobacco for different years included Proteobacteria, Chloroflexi, and Actinobacteria, while the dominant bacterial genera were Acinetobacter, Ochrobactrum, Sphingomonas, and Streptomyces. The redundant analysis showed that soil pH, available potassium (AK), and alkali-hydrolyzed nitrogen (AN) had strong correlations with the microbial community structure (P<0.05). [Conclusion] The application of green manure improves the nutrients and changes the bacterial diversity and community composition of the tobacco-growing soil.