The effects of seed sterilization were studied by different seed sterilization methods. The results showed that sterilization of seeds soaked in aseptic water for 24 hours was better than that without soaking. Under constant temperature, the sterilization effects of three germicides (10%H2O2, 2% sodium hypochlorite and 1‰ HgCl2) on tobacco seeds were studied. The results showed that 10%H2O2 had the greatest sterilization effect. The germ-bearing rate of seeds of K326 treated with 10%H2O2 was 0%. Sterilization of seeds soaked in 75% ethanol before seed sterilization decreased germ-bearing rate. The highest germination rate was observed in treatment with 10%H2O2.