The invention belongs to the technical field of tissue culture, and particularly relates to a treatment method for relieving dormancy of terminal buds of peony tissue culture seedlings. The method comprises the following steps of collecting explants, inducing germination, carrying out multiplication culture, inducing rooting, inducing terminal buds, breaking dormancy and the like. The inventor believes that the main reasons of low transplanting survival rate of existing peony tissue culture seedlings are as follows: firstly, after induced roots of the tissue culture seedlings grow, terminal bud dormancy phenomenon exists, and growth of overground parts is stopped; and secondly, although the rooting rate effect of the existing peony tissue culture seedling is improved to a certain extent, a root system is not developed, and along with the dormancy of the terminal buds and the reduction of nutritional requirements, the growth vitality of the root system after rooting is further weakened, and the survival rate after transplanting is lower. Aiming at the problems, on one hand, the inventor improves the rooting induction effect by optimizing related culture media and culture conditions, on the other hand, terminal bud dormancy is relieved and the growth vigor is improved by regulating and controlling the environment, so that root system germination and growth are promoted, and the transplanting survival rate is finally improved.