Abstract: To explore the effects of three-dimensional film mulching for single tobacco plant on the microorganism quantity in tobacco rhizospheric and non-rhizospheric soils, field experiments were conducted from 2015 to 2016 in Guiyang County, Hunan Province. Three treatments were set:three-dimensional film mulching for single plant, conventional film mulching and without mulching. The numbers of bacteria, actinomycetes and fungi in soil were determined by the dilution plate counting method. The results showed that after tobacco seedling transplanting, the number of soil microorganisms was different at different sampling times. The number of rhizospheric bacteria was significantly different among the three treatments (p < 0.05), with the highest being in three-dimensional film mulching for single plant and the lowest in without mulching. The number of rhizospheric actinomycetes was significantly higher in three-dimensional film mulching for single plant than that in without mulching(p < 0.05), and also higher than that in conventional film mulching (however p < 0.05 only in 2016). The number of rhizospheric fungi in three-dimensional film mulching for single plant and conventional film mulching was significantly higher than that in without mulching(p < 0.05), which was also significantly higher in three-dimensional film mulching for single plant than in conventional film mulching(p < 0.05). The ratio of microorganism quantity between rhizospheric and non-rhizospheric soils was higher in three-dimensional film mulching for single plant than in conventional film mulching and without mulching. In conclusion, three-dimensional film mulching for single plant improves the microorganism quantity in tobacco rhizospheric soil.