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Microbial communities are important to ecosystem function and sensitive to hydrological dynamics. However, we lack predictable knowledge about how soil microorganisms respond to water table drawdown in different depths. This research used a high-throughput sequencing method to determine the responses of prokaryotic communities to the changes of water table and depth on Zoige peatlands. Our results showed that water table drawdown reduced alpha diversity indices (observed species, Shannon diversity and Chao1 richness) of prokaryotic communities. Intriguingly, the reduction of diversity varied in different depths, and was statistically significant in intermediate layers (20-30 cm and 50-60 cm), but not in the surface (0-10 cm) or deep layer (90-100 cm). In deeper layers there was greater relative abundance of most anaerobic microorganisms (e.g. Chloroflexi, Planctomyctes and NC10), but lesser amounts of most aerobes (e.g. Proteobacteria and Acidobacteria). However, the vertical distribution of prokaryotic microbiota along the depth gradient was altered by water table drawdown, mainly by enriching oligotrophs (e.g. Acidobcteria) over copiotrophs (e.g. Bacteriodetes). In addition, we found that the most important soil parameters influencing community structure were soil pH, total organic carbon and total nitrogen. Our study illuminates that the variations of prokaryotic communities caused by water table drawdown are depth-dependent, and that water table drawdown leads to predictive changes of microbiota in peatlands.