The experiment was carried out with five flue-cured tobacco varieties to study the changes of growth, chill tolerance, chlorophyll content, root activity, cell membrane permeability, and content of MDA under the condition of 8℃ and 5℃. The results showed that variety Hongda demonstrated the highest tolerance to chill, followed by NC82, RG11, K326 and V2, subsequently. The chilling injury indices had highly significant correlation with MDA content, root activity, chlorophyllous content, respectively, and significant correlation with cell membrane permeability, which had highly significant correlation with MDA content. The physiological indices could be effective in evaluation of flue-cured tobacco resistance to chilling.