Abstract: In order to achieve the goal of soil improvement in tobacco growing cinnamon soil, field experiments were conducted in tobacco-planting soil of Jiaxian county, Henan province to determine the effects of mixed application of peanut shell biochar and sesame cake fertilizer, bio-organic fertilizer, animal and plant organic fertilizer on nutrients and microbial functional diversity and microbe quantity. The results showed that, compared with chemical fertilizer, mixed application of biochar and organic fertilizers increased contents of soil alkali hydrolysable nitrogen, phosphorus, potassium and organic carbon. The best effects were observed in the bio-organic fertilizer treatments, which increased the contents of soil alkali hydrolysable nitrogen, phosphorus, potassium and organic carbon by 28.18%, 54.81%, 46.72% and 31.12% respectively, compared with CK. The metabolic activities of microorganisms were increased by mixed application of biochar and three organic fertilizers, which improved the utilization of phenolic acid carbon source and the utilization of carbohydrate carbon sources, compared with CK. In addition, mixed application of biochar and organic fertilizers reduced the population of fungi, increased the population of soil bacteria, actinomyces, bacteria/fungi and actinomycetes/fungi, and bio-organic fertilizers increased the population of soil bacteria and actinomyces by 192.45% and 45.77% respectively, compared with CK. The results of this study indicated that mixed application of biochar and organic fertilizers not only increased the contents of cinnamon soil available nutrients and organic carbon, but also increased functional diversity of microbial communities and microbial community structure of the soil and the bio-organic fertilizers had the optimal effects.