The K326 variety of fluecured tobacco was selected as experimental material, which was field planted in Yunnan Province, Fujian province and Henan province of China. The middle leaves (leaf 11) of tobacco were collected four times during the mature leaf stage. The δ¹³C, total organic carbon, total nitrogen, LMA, photosynthetic pigment were investigated to compare different ecologic area fluecured tobacco smokes and the δ¹³C value distribution with the physiological and ecological adaptation. Important findings: (1) three ecology tobacco area δ¹³C value, total carbon, C/N, LMA, Chlorophyll a, Chlorophyll results showed Yunnan > Fujian > Henan；total nitrogen results showed Henan> Yunnan > Fujian；Chlorophyll b results showed Yunnan > Henan> Fujian; Content of carotenoid results showed Henan> Fujian> Yunnan. Among them δ¹³C value, total carbon, C/N, Carotenoids were similar in Yunnan and Fujian. (2) The correlation analysis suggested that δ¹³C value with total carbon were positive correlated of Yunnan, negative correlated of Fujian and Henan. In the three ecological areas, δ¹³C value and total nitrogen were negatively correlated; δ¹³C value with photosynthetic pigment was positive correlated in Yunnan and Henan, was negatively correlated in Fujian. That δ¹³C value and the smoke alkali, the nitrogen, the potassium, the chlorine assumes the negative correlation, with total sugar and reducing sugar have positive correlation. (3) Yunnan fluecured tobacco had good fragrance, medium irritation, moderate strength and the best chemical constituents’ coordination; Henan fluecured tobacco had a higher amount of aroma with heavier irritation, while Fujian fluecured tobacco showed a worse characteristic in amount of aroma and chemical constituents’ coordination. In conclusion the δ¹³C value was associated in physiological features and quality evaluation of tobacco. The study found using δ¹³C value, physiological features and quality evaluation could be used to identify fluecured tobacco aroma type and quality characteristics from different ecological tobacco areas.