【Objective】This study was trying to find out a proper application rate of specialized vinasse organic manure for flue-cured tobacco in Guizhou karst area.【Method】A field experiment was designed and laid out to have 5 treatments. i.e. CK (3 t•hm^-2 of conventional vinasse organic manure), T1 (3 t•hm^-2 of specialized vinasse organic manure), T2 (3.6 t•hm^-2 of specialized vinasse organic manure), T3 (3.6 t•hm^-2 of specialized vinasse organic manure, + 300 kg•hm^-2 of potassium fertilizer, and T4 (1 t•hm^-2 of specialized vinasse organic manure + 2 t•hm^-2 of chemical fertilizer), and carried out for analysis and comparison of effects of the five treatments on ecology and yield of flue-cured tobacco.【Result】Results show that compared with CK, Treatments T1, T2 and T4 improved the crop in plant height, stem circumference and leaf area to a varying degree; Treatments T1, T2 and T3 did in root fresh weight, root dry weight and root volume; Treatment T4 increased rhizosphere indexes significantly；Treatment T1 and T2 reduced the incidence of black mites by 49.8%; Treatment T1, T3 and T4 lowered the incidence of climate spot disease by 62.5%, 37.5%, and 25%, respectively; and Treatment T2, T3 and T4 got free of mosaic disease. In terms of production of flue-cured tobacco, Treatment T4 was the highest in yield and followed by Treatment T1 and T2, reaching 1623 kg•hm^-2 and 1434 kg•hm^-2, respectively, and Treatments T1~T4 increased by 10.4%~25.0% over CK. The contents of nicotine, total nitrogen, total sugar, reducing sugar, phosphorus, potassium and chloride ions in tobacco leaves of all the tested treatments varied within the suitable range of high-quality tobacco leaves. Application of specialized vinasse organic manure reduced the content of nicotine, total nitrogen and phosphorus, and kept the potassium ion content in a proper range that made internal quality of the flue-cured tobacco gradually reasonable. 【Conclusion】In summary, the use of the specialized vinasse organic manure in addition to chemical fertilizer can improve growth and development as well as quality of the flue-cured tobacco.