Organic materials as the important carbon and nitrogen sources for soil microorganisms play crucial roles in regulating rhizosphere microbial community, nutrient cycling and improving crop yield and quality. Under the same amount of nitrogen input, a field experiment was conducted to examine the effects of different organic materials (rice straw, rape green manure and cake fertilizer, compared to the conventional chemical fertilizers as the control) on enzyme activities, rhizosphere microbial functional diversity and yield and quality of tobacco leaves in order to provide scientific information on proper use of organic materials to improve tobacco leaf quality. The results showed that the addition of rice straw and rape greens significantly enhanced urease activity, while had no significant effect on hydrogen peroxide activity in tobacco rhizosphere. Biolog analysis revealed that the addition of rice straw and rape greens significantly increased AWCD values and functional diversity indices in tobacco rhizosphere at the late growth stage. The principal component analysis indicated that the differences in carbon sources utilized by rhizosphere microbes were an important factor in the variation of microbial community in tobacco rhizosphere, in which sugars had the greatest effect. In addition, the addition of rice straw and rape greens significantly decreased reducing sugar content and sugar to nicotine ratio, improved the proportion of mid to high-grade leaves and thus increased the production value of tobacco leaves by 29.4% and 19.8%, respectively, as compared with the control. It is indicated that the proper use of organic materials as partial replacement of chemical fertilizers, can effectively improve the microbial environment of tobacco rhizosphere and tobacco leaf quality, thus can be recommended as an effective cultivation practice to high-quality tobacco leaf production.