The invention discloses a domestication cultivation method of bletilla striata tissue culture seedlings. The domestication cultivation method comprises the following steps: A, preparing a substrate, wherein the substrate comprises mixtures of pine nut shells, humus soil and sheep manure and mixtures of sawdust, chicken manure, bean cakes and sesame oil residues; B, digging a culture pool in a greenhouse, C, successively spreading the mixtures of the pine nut shells, humus soil and sheep manure and the mixtures of the sawdust, chicken manure, bean cakes and sesame oil residues in the culture pool; D, selecting the bletilla striata tissue culture seedlings, and then cleaning, carrying out disinfection and drying; E, planting the tissue culture seedlings obtained in the step D on the substrate in the culture pool, wherein the row spacing is 2 cm to 4 cm, and the plant spacing is 1 cm to 2 cm; and F, managing: controlling the temperature in the greenhouse to be within 22 to 25 DEG C, the humidity to be within 60% to 70%, the illumination intensity to be within 2500 LX to 3000 LX, and the illumination time for 10 h to 13 h daily; regularly watering; and transplanting in fields after 3 months. The bletilla striata tissue culture seedlings are domesticated by using the method, so that good growth of the tissue culture seedlings can be ensured, the ratio for meeting requirements of field transplantation is greatly increased, and the qualified rate of the tissue culture seedlings is improved.