Abstract: In order to explore the effects of two growth-promoting strain on tobacco growth and rhizosphere soil bacterial community, a pot experiment was conducted by using Yunyan 87 and set up with four groups: sterile water (cotrol), Piriformospora indica (PI), Bacillus subtilis (Tpb55), and P. indica +B. subtilis (PI+Tpb55). The growth index of tobacco plant , rhizosphere soil enzyme activity, bacterial community structure and function were analyzed. Results showed that compared to the control, both P. indica and B. subtilis Tpb55 promoted tobacco growth and increased dry or fresh matter accumulation, and PI+Tpb55 exhibited apparently higher effect than the single PI or Tpb55 (p<0.05). PI, Tpb55, and PI+Tpb55 increased the activities of sucrase, urease, and acid phosphatase in tobacco rhizosphere soil, with the most significant effect (p<0.05) in PI+Tpb55. The above three treatments also increased the number, richness, and diversity of bacterial OTUs in tobacco rhizosphere soil, among which PI+Tpb55 exhibited significant effect, with Chao 1 index increased by 43.9% and Simpson index decreased by 10.1%. After PI+Tpb55 treatment, compared with the control, the relative abundance of Gemmatimonadota, Actinobacteria, and Acidobacteriota increased by 40%, 20.6%, and 90%, respectively. The relative abundance of Gemmatimonas, Vicinamidobacteriaceae, and Haliangium increased by 35.5%, 86.5%, and 135.7%, respectively, up to significant level (p<0.05). Functional prediction of bacterial community indicated that PI, Tpb55 and PI+Tpb55 increased the abundance of functional genes of bacterial community tricarboxylic acid cycle. In conclusion, both the strain P. indica and B. subtilis Tpb55 can promote tobacco growth and improve soil characteristics, with the optimal effect in the synergistic treatment.