Abstract: Different sulphur application rates were designed to determine the optimum sulphur application rate in southern Anhui tobacco area and its effect on the quality of flue-cured tobacco. The results showed that proper sulphur application could promote the growth and development of tobacco plants, sulphur application increased the sulphur content in all organs of the tobacco plant, and the increment pattern of sulphur content was ranked as root < stem < leaf. The total sulphur content under the same sulphur application rate was ranked as root < stem < leaf. As for leaf, the maximum value of sulphur content was measured in above leaf layer, the minimum was occurred in below leaf layer and the intermediate was measured in middle leaf layer. The total sulphur application rate of flue-cured tobacco should not exceed 45.0 kg/hm2, and appropriate sulphur applied could enhance the leaf sugar accumulation. There was negative effect of sulphur application on the total potassium accumulation, external and economic traits. Excessive application of sulphur decreased smoking quality of flue-cured tobacco. Therefore, adjusted the type of fertilizer in flue-cured tobacco production, reduced the proportion of sulphur content in the fertilizer could enhance the intrinsic quality in southern Anhui province.