With Didymodon tectorum collected from the main mine of Baiyun obo as experimental material, we studied the effect of different disinfectants and different concentrations of plant hormone 6BA and IAA, respectively on callus induction and differentiation. It showed that the best disinfectant for D. tectorum gametophyte is 75% alcohol immersion for 30 s and 0.1 g/L mercuric chloride for 90 s. Using Knop medium to culture the upper stemleaf segments of D. tectorum, and a low concentration of 6BA (0.1 mg/L) promotes callus differentiation and gametophyte formation. However, high concentration of 6BA (1.0 mg/L) restrains the callus formation. IAA helps protonemata germinate.