Abstract: In order to screen suitable organic fertilizer species and improve the quality of flue-cured tobacco, in this studyYunyan 87 was used in a field experiment to investigate the effects of different organic fertilizers on biochemical characteristics of flue-cured tobacco planting soil and yield. Four kinds of organic fertilizers (high-carbon base fertilizer, fried sesame, bio-organic fertilizer and decomposed sesame seed cake fertilizer) were applied in combination with a tobacco-specific compound fertilizer separately during different growth periods. The results showed that application of organic fertilizers could increase the content of organic matter, soil microbial functional diversity and soil enzyme activities in the soil. The increase was higher from the treatment with high-carbon base fertilizers at early stages compared with the other treatments while at middle and late stages the treatment of fried sesame had the most significant effect. The quality of flue-cured tobacco was most significantly improved when applied with sesame seeds. The combined application of high-carbon base fertilizers could greatly promote yield and output values of flue-cured tobacco. Overall, it is more suitable to use high-carbon base fertilizers and fried sesame to partially replace chemical fertilizers in Zhoukou tobacco areas.