Abstract: In this study, TMV resistant characteristics of 20 tobacco cultivars were preliminarily identified and classified based on analyzing the seed germination and seedling growth. Then, Yunyan 201 was chosen as tolerant cultivar and MSK326 was chosen as the sensitive one to be contaminated by TMV virus and then the seedling growth was investigted. The physiological and biochemical indexes were also tested. The seeds of those two cultivars were soaked with different reagents to compare the tolerant effects of TMV, which showed that soaking with reagents could increase the TMV tolerance of tobacco seeds and seedlings. In summary, using 1 g/L thiophanate methyl solution could significantly reduce the morbidity rate and morbidity index of MSK326, and increase its chlorophyll a and b content and peroxidase activity.