Abstract: Stem segments of Anoectochilus roxburghii (wall.) lindl. Huaye were used as the explants to induce bud clumps. Varied medium (i.e., MS, B₅ or improved MS) and phytohormone (i.e., TDZ, 6-BA, KT or IBA) were applied for the tissue regeneration experiment of an orthogonal design. It was found the major factors affecting the bud multiplication to be basal medium, KT, 6-BA and TDZ. The optimal multiplication was achieved by using the improved MS+6-BA with additions of 2.0 mg·L^-1+KT 0.5 mg·L^-1+NAA 0.1 mg·L^-1+sugar 30.0 g·L^-1+agar powder 3.0 g·L^-1, and carrageenan 3.6 g·L^-1. An averaged propagation coefficient of 4.58 in 60 days was obtained. For rooting, the improved MS+6-BA with added 0.3 mg ·L^-1+AC 0.5 g·L^-1+sugar 20.0 g·L^-1+agar powder 3.6 g·L^-1 and carrageenan 3.6 g·L^-1, rendered a 100.0% rate in 60 days.