Abstract: In order to establish an efficient and rapid system of regeneration, the optimal medium for callus induction from leaves and stem tips in Fragaria nilgerrensis were screened. Compared with 1/2 MS medium, MS medium with 0.4 mg/L TDZ+0.2 mg/L 6-BA was more appropriate for callus induction. Then leaves and stem tips were used as explants to compare the different callus induction rate under four kinds of plant hormones with different concentration combinations, which were designed as orthogonal experiment of L₉(34). The results showed that 2, 4-D has the greatest effect on callus induction of leaves in F. nilgerrensis and the optimal medium was MS +0.4 mg/L TDZ +0.2 mg/L 6-BA +0.15 mg/L 2, 4-D +0.2 mg/L NAA. While NAA has the greatest effect on callus induced from stem tips and the optimal medium was MS +0.2 mg/L TDZ +0.6 mg/L 6-BA +0.15 mg/L 2, 4-D+0.6 mg/L NAA.