Abstract: To study the response of flue-cured tobacco to bio-organic fertilizer, and provide references for flue- cured tobacco production, the effect of bio- organic fertilizer on flue- cured tobacco physiologicalbiochemical indexes, yield and quality was analyzed by different bio-organic fertilizer application tests. The results showed that the combined application of bio- organic fertilizer increased roots volume and net photosynthetic rate of flue-cured tobacco leaves, promoted the growth of flue-cured tobacco, increased firstclass tobacco leaves ratio, yield and quality, and out-put value; the yield of flue-cured tobacco increased by 9.35%-11.2% and reached 2159.1 kg/hm2 and 2361.0 kg/hm2, the out-put value increased by 2.51%-15.2% and reached 55057.1 yuan/hm2 and 61858.2 yuan/hm2, the ratio of first-class tobacco leaves increased by 0.95%-3.63%; economic benefit increased by 2.14%-14.5% under the same other production costs; economic character index (ECI ) and chemical components usability index (CCUI ) of combined bio- organic fertilizer application treatment were better than those of CK. Therefore, reasonably combined application of bio-organic fertilizer has positive effect on flue-cured tobacco production.