The invention discloses a method for identifying the bacterial wilt resistance of tobacco seedlings. A water retaining agent and pearl stone are mixed uniformly at the ratio of 2:1; the mixture is disinfected and then loaded into a 6-hole cell culture board; tobacco seeds are sowed on the cell culture board, wherein each kind of tobacco seeds is sowed in one hole, and 10-20 tobacco seeds are sowed in each hole; the cell culture board is placed in an axenic culture room to culture the tobacco seeds under the conditions that the temperature environment is 25 DEG C, the illumination time is 16 hours every day, the illumination intensity is 4,000 lux and the relative humidity is 70-80 %; after emergence of seedlings, 5 seedlings are reserved in each hole uniformly; a nutrient solution is prepared from special tobacco seedling raising fertilizers, wherein the concentration of a nitrogen fertilizer reaches 100 ppm; the nutrient solution is top-dressed into the culture board; when the seedlings send forth two true leaves, 1 ml of ralstonia solanacearum suspending liquid is vaccinated into each hole of the cell culture board through a liquid-transferring gun; investigation and recordation are carried out 15 days later. The method provided by the invention is shorter in identification period, can identify 25-day-old seedlings, is not limited by seasons and has high testing stability.