Abstract: ObjectiveThe present study established in vitro culture system of Camellia nitidissima through callus ap-proach to provide theoretical and practical support for rapid reproduction and germplasm resources preservation. MethodAt seed germination stage, sand was used instead of medium for aseptic germination. Shoot tip, stem and leaf of mature plant were used as explants. Callus induction,callus differentiation and rooting culture were conducted in WPM media sup-plemented with various hormones at different densities. ResultThe mature seeds were peeled, and their endosperms were removed. Endosperm-removed embryos were buried in the sand wetted by sterile liquid. In such condition, average germi-nation rate was 95.40%, average germination time was 19.10 d. The suitable culture medium for callus induction of cotyle-don and hypocotyl was WPM+2.00 mg/L 6-BA+0.50 mg/L 2,4-D. The suitable culture medium for callus differentiation was WPM+2.00 mg/L 6-BA+0.10 mg/L NAA, and differentiation coefficient was 4.56. The optimum culture medium for rooting culture was 1/2WPM+0.50-1.00 mg/L IBA,and rooting rate was 45.40%-54.00%,rooting time was 19.80-20.90 d. Conclu-sionThe in vitro culture system of C. nitidissima is as follows:callus is induced by hypocotyl, then callus differentiate into whole plant.