The invention discloses a method for testing cold resistance of tobacco in the seedling stage, which includes the steps: firstly, raising seedlings by floating; secondly, transplanting the tobacco seedlings; thirdly, domesticating each seedling in a greenhouse for 2-3 days when each seedling grows 6-8 true leaves; fourthly, performing low-temperature stress treatment at 4 DEG C-8 DEG C, and recording morphological response characteristics of the seedlings; fifthly, allowing the seedlings to resume growing at the normal temperature, namely restoring the temperature and light conditions of greenhouse domestication after low-temperature stress treatment, and recording recovery growth conditions of the seedlings; and sixthly, evaluating cold resistance of tobacco in the seedling stage, namelyperforming comprehensive evaluation according to the morphological response characteristics and the normal-temperature recovery growth conditions of the tobacco seedlings subjected to low-temperaturestress treatment, and dividing cold resisting degree into a low-temperature sensitive type, low cold resistance type and high cold resistance type. The method is objective in results and high in accuracy. The basis of cold resistance evaluation is intuitive, simple and clear. The method is easy in operation, highly repeatable, and applicable to guiding safety breeding of cold-resistant tobacco and safety production of tobacco leaves.