The invention discloses a method for improving the transplanting survival rate of a catharanthus roseus tissue culture seeding. The method is characterized by comprising the following steps of: (1) selecting strongly and regularly grown catharanthus roseus aseptic test-tube seedling, opening a cover of a culture flask, adding the suitable amount of distilled water to the culture flask to immerse the root of the catharanthus roseus, and putting the catharanthus roseus seedling in an illumination incubator to culture for 3-4d to finish a seedling hardening operation; (2) taking out the catharanthus roseus aseptic test-tube seedling from the culture flask by a pair of tweezers, flushing a culture medium of the root of the catharanthus roseus aseptic test-tube seedling cleanly by running water, putting the culture medium in a test tube or a culture dish containing a water-culture nutrient solution, putting the test tube or the culture dish in the illumination incubator to continue the seedling hardening operation, and finishing the water culture when a new water root is grown up from the root of the catharanthus roseus plant after the catharanthus roseus plant is cultured for 2-7d; and (3) transplanting the newly grown catharanthus roseus water-culture plant in a transplanting medium containing vermiculite, perlite and turfy to finish the transplanting of the catharanthus roseus tissue culture seeding. The method for improving the transplanting survival rate of the catharanthus roseus tissue culture seeding disclosed by the invention has the advantages of reinforcing the antibacterial ability of the catharanthus roseus tissue culture seeding and remarkably improving the transplanting survival rate of the catharanthus roseus tissue culture seeding.