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Mowing and burning, two common human disturbance methods, have different impacts on the soil fungal and bacterial communities of sandy land ecosystems in semiarid areas. Thus, the effects of long-term mowing and burning on plant and soil microbial communities in the semiarid Horqin Sandy Land are unclear. In this study, the effects of soil microbial community structure from mowing and burning on plant and soil microbial communities over six years were assessed by high-throughput sequencing. First, we found that mowing and burning significantly reduced vegetation coverage (V) (p < 0.05), and burning significantly increased plant Shannon–Wiener diversity (H), Pielou evenness (E), and Simpson dominance (λ) indices (p < 0.05). Second, the two human disturbances markedly affected fungal diversity and abundance more than bacterial diversity and abundance. In fungi, burning significantly reduced beta diversity (p < 0.05) and the relative abundance of Basidiomycota (p < 0.05) but markedly increased the relative abundance of Ascomycota and Mortierellomycota. Third, redundancy analysis (RDA) showed that vegetation coverage was a significant factor in bacterial species distribution (p < 0.05). For fungi, the significant factors were species richness (S) and vegetation coverage (p < 0.01). Taken together, these data suggest that long-term mowing and burning have more of an effect on soil fungal communities than bacteria in a semiarid sandy land, improving our understanding of how terrestrial ecosystems respond to human disturbances and providing guidance for the management and restoration of sandy desertification. • Mowing significantly reduced vegetation coverage. • Burning could change the plant community diversity. • Mowing and burning markedly affected fungal diversity and abundance more than bacteria. • Vegetation coverage was the significant factor in bacteria species distribution. • Fungal community was more easily influenced by plant community diversity. [ABSTRACT FROM AUTHOR]