The invention provides a Dioscoreae Oppositae 'Qi' tissue culture seedling raising method, which comprises the steps of explant selection and disinfection, callus tissue induction, adventitious bud differentiation, rooting culture, seedling training, transplanting and the like. By taking the nodular stem segments of the potted Dioscoreae Oppositae 'Qi' as explants, after the explants are disinfected by alcohol with volume concentration of 70 percent and sodium hypochlorite with weigh percentage of 2 percent, the callus tissue induction, the adventitious bud differentiation and the rooting culture are conducted on MS+NAA+BA culture mediums, and rooted seedlings are transplanted in mixed mediums for culture, wherein the mixed mediums comprise vegetable garden soil, mountain sand and peat, with the mass ratio of 4:1:1. Compared with traditional Dioscoreae Oppositae 'Qi' seedling raising methods such as a direct root tuber planting method, a cutting breeding method and the like, the Dioscoreae Oppositae 'Qi' tissue culture seedling raising method has the advantages that the genetic stability of the seedlings is good, the breeding cycle is short, the breeding coefficient is high, the operation is simple and the factory production of the seedlings can be realized. By adopting the method, not only can the problems that the quantity of the used seminal roots for the production of the Dioscoreae Oppositae 'Qi' is large, the cost is high and the characters of strain is degenerated be solved, but also technical supports can be provided for the high quality and high yield and the industrialization of the Dioscoreae Oppositae 'Qi' through large-scale seedling breeding.