Establishment of Protoplast Culture of Solanum sisymbriifolium

The optimal calture conditions were studied for plant regeneration from mesophyll protoplasts of Solanum sisymbriidorium. Aeseptic seedlings of S.sisymbriidorium were used as a meterial for protoplast culture. Many variable protoplasts were isolated by incubating leaf slices in an enzyme solution containing 0.25% Meiserase and 0.05% Macerozyme for 16h at 25℃ without shaking. Protoplast density of 5.0*10[4]/ml in Kao medium containing 5.0 mg/l NAA, 1.0 mg/l BA was optimal for colony formation. Most colonies formed when protoplasts were cultured at 25℃ after initial culture at 30℃ for one week. On the MS agar medium with 1.0mg/l zeatin, 38.5% of protoplast-derived calli dofferentiated shoots. These shoots rooted on 1/2 MS medium with 5.0 mg/l and 2.5g/l gellan gum, anf developed into whole plants