Plant Regeneration from Mesophyll Protoplasts of Japanese Butterbur (Petasites japonicus Fr. SCHMIDT)

Abstract

Mesophyll protoplasts of butterbur (Fuki) were prepared in the enzyme solution containing 0.l% Pectolyase Y 23, 0.5% Cellulase Onozuka RS, 0.5% Driselase and 0.5M mannitol at 27°C for 120 min with shaking at the rate of 80 str/min. Purified protoplasts were cultured in the medium containing diluted MURASHIGE & SKooG's (MS) salts, vitamins, and various concentration of auxins and cytokinins at 1×105 cells/ml. The culture was kept at 25°C. After about one or one and a half months, cell colonles were transplanted onto the agar medium containing 1/4 MS salts, 1/2 MS vitamins, 1mg/l NAA. 0.5 mg/l BA, 1% sucrose and 0.8% agar. Calluses of 2∼5 mm in diameter were transferred on the agar medium of MS with 3% sucrose and 0.8% agar supplemented with various concentration of NAA and BA to induce shoot. After 6 month-culture, whole plants were obtained.