In this study, the mature embryo of Pinus taiwanensis Hayata was used as the experimental material to conduct a preliminary study of in vitro culture and plant regeneration conditions. The results showed that the suitable surface sterilization method for mature embryos of P. taiwanensis Hayata was 75% alcohol treatment for 1 min and 3% sodium hypochlorite for 10 min; the optimal callus induction medium was MS + 1.0 mg·L^-1 6-BA + 0.2 mg·L^-1 NAA; the optimal induction medium for adventitious buds was DCR + 1.0 mg·L^-1 6-BA + 0.05 mg·L^-1 NAA; the suitable induction condition for adventitious bud elongation was DCR + 0.1 mg·L^-1 6-BA + 0.05 mg·L^-1 NAA; the optimal medium for rooting was 1/2 DCR + 2.0 mg·L^-1 IBA + 0.05 mg·L^-1 NAA. The results obtained in this study will be of great significance for the preservation, genetic improvement and high-quality seedling reproduction of P. taiwanensis Hayata germplasms.