Abstract: To investigate the effects of fertilization treatment on the bacterial community structure and functions of tobacco planting soil, MiSeq high-throughput sequencing technology was used to study the changes in soil bacterial community diversity and composition after the application of different fertilization treatments, no fertilizer (CK), organic fertilizer (OM), organic and chemical fertilizer (OM+NPK), and chemical fertilizer (NPK). A PICRUSt analysis was used to analyze the change characteristics of soil bacterial functional gene relative abundance. The results showed that: 1) There was no significant difference in the effects of different fertilization treatments on the diversity and abundance of soil bacterial communities. The dominant phyla were Proteobacteria, Acidobacteria, and Actinobacteria, which accounted for 64.87%-68.51% of the total abundance. All three fertilization treatments increased the relative abundance of Proteobacteria and Actinobacteria and decreased the relative abundance of Acidobacteria, Chloroflexi, and Verrucomicrobia. 2) In total, 6 types of first-level functions and 41 types of second-level functions were obtained, with metabolism being the most prominent function of the bacterial community among the first-level functions. The mixed treatment of organic and chemical fertilizers significantly decreased the relative abundance of functional genes for membrane transport, terpenoid and polyketide metabolism, and increased the relative abundance of functional genes for translation, glycan biosynthesis and metabolism, replication and repair, and cellular motility in bacteria. 3) Soil bacterial community structure was mainly affected by four chemical property indicators, including soil organic carbon, total phosphorus, effective phosphorus, and alkaline dissolved nitrogen, with alkaline dissolved nitrogen and effective phosphorus being the main factors affecting bacterial functions. In conclusion, different fertilization treatments did not significantly affect the soil bacterial diversity α, but they significantly changed the bacterial community composition, thereby affecting the functional gene relative abundance of soil bacteria. Reasonable regulation of fertilizer application types and reduction of chemical fertilizer application ratios can improve the soil microbiological environment of tobacco plantations in central Yunnan.