Abstract: The effects of seedling cultivating methods on the biological and physiological characteristics of flue-cured tobacco were studied.The results showed that either nutrition pot or tray heeling-in was able to retard the elongation of taproot, promote the development of side roots, enlarge stem circumference and the volume of root system, significantly increased the chlorophyll content, root vigor and root active absorbing area.The activities of NR, INV, SOD, POD and CAT in seedling were enhanced remarkably, which promoted the metabolism of nitrogen and carbon and the stress resistance of tobacco seedlings.