Abstract: The callus induction was carried out with the leaves of aseptic seedling of tobacco E'yan 1,and the optimal hormone proportion in culture medium was screened out,and the system of tobacco cell suspension culture was established preliminarily.The results showed that MS was the optimal basic medium for callus induction,and the optimal formula was MS+NAA 1.0 mg/L+6-BA 0.5 mg/L;while the optimal formula for culture medium of callus was MS+2,4-D 0.1 mg/L+NAA 1.5 mg/L+6-BA 0.3 mg/L.On the 6th day of callus culturing,the logarithm growth period started;and on the 14th day,the dry weight increased to 12.536 times of its initial dry weight.The system of tobacco cell suspension culture was established based on this formula.