Abstract: In order to improve tobacco quality and make sustainable development of flue-cured tobacco production, a field experiment was conducted to study the application effects of bio-organic fertilizer on tobacco production. The results showed that the fertilizer could regulate the development and metabolism of tobacco plants, and improve disease resistance. The potassium content was significantly increased and the chemical composition was more coordinate after applying the bio-organic fertilizer. When the bio-organic fertilizer was 20%-40% of total fertilizer applied, the output value increased by 3595.94-4640.49 yuan/ha, the soil enzyme activity was also enhanced, and the bio-organic fertilizer could play an important role in soil improvement.