Objective The studies on tissue culture and cytological observations of leaf explants of Curculigo orchioides were conducted in order to provide the basis for the rapid propagation of C. orchioides. Methods Young leaf explants of C. orchioides were cultured on MS basal media. Differences in the callus induction and plantlet regeneration rate were observed by different light treatment as well as chemical factors like different phytohormones, casein hydrolysate (CH), and activated charcoal (AC) concentrations. Paraffin method was used to cytological observation. Results For callus induction of leaf explants of C. orchioides, dark treatment gave better results compared to light treatment; among the media tested, the suitable phytohormone combinations were 2.0 mg/L 2, 4-D or 6-BA 1.5 mg/L+2, 4-D 2.5 mg/L, and 300 mg/L CH+0.2% AC was good for plantlet regeneration from leaf explants. The callus from leaf explants mainly originated from midrib. The parenchyma cells near epicuticle of midrib firstly were initiated to division. Then the parenchyma cells of vascular bundle sheath and mesophyll cells on each side of vascular bundle were also divided to form callus. The buds developed on the peripheral parts of the calli, but the roots developed in the regions deep within the calli. Conclusion Tissue culture of young leaf explants of C. orchioides can make the propagation of C. orchioides rapid.