Abstract: In order to study the effects of altitude and varieties and their interactions on polyphenolic contents and compositions in flue-cured tobacco in Weining (2200 m), Nayong (1600 m) and Qixingguan (1000 m) of Bijie, three varieties, including Honghuadajinyuan, Bina1 and K326 were planted on each altitude. The experimentation was designed in random block design of two-factors and three-levels. The results showed that there was a significantly difference in polyphenols content between different varieties of tobacco. The contributing rations of altitude to the content variations of chlorogenic acid, rutin and scopoletin were 2.0%, 74% and 30%, respectively, and that of the variety were 57%, 8.0%, 17%, respectively. Interaction between altitude and variety contributed 41%, 15% and 48%, respectively. Chlorogenic acid content of Honghuadajinyuan was higher at Weining, whereas hlorogenic acid and scopoletin contents of Bina 1 were higher in Qixingguan, and higher chlorogenic acid and scopoletin contents were observed in K326 of Nayong. In order to improve the content of polyphenols in tobacco, suitable varieties should be selected according to altitude.