This research aimed to reveal the response characteristics of soil microbial community structure to different degrees of tourism disturbance. To explore the soil microbial community structure’s response mechanism, we set up continuous plots with different interference intensities: high disturbance, middle disturbance, and the control area. We collected 0–10 cm topsoil in all plots and used Illumina MiSeq high-throughput sequencing method to obtain and analyze the response characteristics of soil microbial community composition and structure under different tourism disturbances. These results were then combined with alpha diversity and environmental factors to explore the microbial response mechanism. In the tested soil, Acidobacteria, Chlorocurve, and Proteobacteria were the main bacterial phyla, while Basidiomycota and Ascomycota were the main fungal phyla. Based on the phylum, the relative abundance of the microbial community between the interference groups was compared using a significance test, with significant differences found between the interference groups in the phyla Chloroflexus, GAL15, Rokubacteria, and Blastomonas (p < 0.05). The relative abundance of the dominant phyla in the fungal community was significantly different among the groups (p < 0.05). A principal component analysis of the soil microbial community structure suggested that the soil microbial community structure was significantly different for different interference levels. © 2022 by the authors. Licensee MDPI, Basel, Switzerland.