Middle tobacco leaves from variable geographic regions (Sangzhi, Fenghuang and Jingzhou) and regions with different altitudes in Xiangxi were used as materials to analyze the physical and chemical characteristics of tobacco leaves. The results showed that the differences in leaf width, rate of water content and thickness of tobacco leaves had reached significant level (P<0.05). Tobacco leaves from Sangzhi district has the longest leaf width (26.52 cm) and the highest rate of water content (18.98%), while the most thick leaf appeared in Jingzhou (96.67 μm). Tobacco leaves from different geographic regions in Xiangxi showed big differences in potassium, chlorine, total sugar, carotenoid and rutin contents, margin of two sugars, the ratio of potassium to chlorine, and the total sugar content increases with the reducing of altitude while the content of carotenoid and rutin increases with the rising of altitude. The difference of leaf width of tobacco leaves at different altitudes had reached significant level (P<0.01), with leaf width at altitudes of 1 000–1 100 m the highest (26.52 cm). The differences in chemical indexes including total nitrogen, nicotine, carotenoid and polyphenols, the ratio of total sugar to nicotine of tobacco leaves at different altitudes had reached significant level, these indexes all reduced with the rising of altitude.