Abstract: We studied the cryopreservation of germplasm resources of Dioscorea opposita Thunb. by encapsulation-vitrification. D. opposita plantlets of different cultivars were cold-hardened for 1 week at low temperature, then stems with one axillary bud were excised and pre-cultured at low temperature for 3 days. Stems with axillary buds suspended in Murishige and Skoog (MS) inorganic medium supplemented with 3% Na-alginate were placed in MS inorganic medium supplemented with 0.5 mol·L^–1 CaCl₂ solution. Beads were loaded with MS medium co-mixed with 0.2 mol·L^–1 sucrose, 2mol·L^–1 glycerol and 50 g·L^–1 dimethyl sulfoxide (DMSO) for 60 min at 0°C,then were dehydrated with a highly concentrated vitrification solution (30% glycerol+ 15% ethylene glycol+10% DMSO+ 15% polyethylene glycol+0.4 mol·L^–1 sucrose) for 60 min at 0°C and plunged into liquid nitrogen quickly After 24 hours, the beads were rapidly thawed in a water bath at 40°C for 1-3 min. The beads were washed twice with MS medium supplemented with 0.5 mol·L^–1 sucrose, then transferred to regeneration medium (MS+2 mg·L^–1 KT +0.02 mg·L^–1 NAA +3% sucrose+0.5% agar). Stems with one axillary bud could lead to regenerated plantlets with regeneration culture, but the survival rates of cultivars differed: 64.29%, 49.21%, 13.11% and 39.81% for D. opposita 'Tiegun', D. opposita 'Taigu' D. opposita 'Huaiqing No.1' and D. opposita 'No.B', respectively.