Abstract: This study attempts the establishment of the tissue culturing system using Dendrobium nobile seeds as the starting materials. It is found that the MS medium supplemented with 1.5 mg/L 6-BA+0.5 mg/L NAA+2% sucrose is the most suitable for the induction of protocorm, showing an induction rate of 91.67% after being cultured for 15 days. Adding 0.25 mg/L NAA and 3% sucrose to the MS medium with 1.5 mg/L 6-BA is more suitable to form adventitious seedlings with dark green sturdy leaves, and the addition of 0.1 mg/L NAA and 2% sucrose is suitable for root emergency and growth. The transgenic system for Dendrobium nobile tissues is also studied. It is found that compared with GV1301 strain, the Agrobacterium tumefaciens EHA105 strain can be used to obtain higher transformation rates. The rate of transformation using root segments, root tips and tissue-cultured seedlings as the starting materials reaches the highest when they are soaked with EHA105 for 30 minutes and then co-cultured for 3 days, but the rate of transformation using nodes with axillary buds reaches the highest if co-cultured for 5 days. The results of the present study can provide meaningful techniques for further research on Dendrobium nobile and the breeding of Dendrobium varieties.