The invention discloses a domestication method of strawberry tissue culture plants. The method includes steps including cover opening domestication of the tissue culture plants, re-domestication in agreenhouse, root cleaning, cultivation and management. The tissue culture plants are domesticated twice before transplantation to improve the adaptiblity, a more suitable cultivation condition is provided for domestication, and via the method, domesticated strawberry is high in survival rate, short in rejuvenation period, fast in growth and improved in quality; and the method is easy to master, adomestication is easy to construct, the cost is lower, repeated use is allowed, and the greenhouse can be used to plant other crops after domestication.