Establishment of protoplast culture of Solanum sisymbriifolium

The optimal culture conditions were studied for plant regeneration from mesophyll protoplasts of Solanum sisymbriifolium,. Aseptic seedlings of S. sisymbriifolium were used as a material for protoplast culture. Many viable protoplasts were isolated by incubating leaf slices in an enzyme solution containing 0.25% Meiserase and 0.05% Macerozyme for 16h at 25°C without shaking. Protoplast density of 5.0 × 10⁴/ml in Kao medium containing 5.0mg/l NAA, 1.0mg/1 2,4-D and 1.0mg/1 BA was optimal for colony formation. Most colonies formed when protoplasts were cultured at 25°C after initial culture at 30°C for one week. On the MS agar medium with 1.0mg/1 zeatin, 38.5% of protoplast-derived calli differentiated shoots. These shoots rooted on 1/2 MS medium with 5.0mg/l sucrose and 2.5g/l gellan gum, and developed into whole plants.