Protoplast culture and plantlet regeneration in stock (Matthiola incana R. Br.).

Abstract

Protoplasts were isolated from callus derived from excised cotyledons of stock (Matthiola incana R. Br.), by using an enzyme mixture of 1.5% Meicelase and 0.05% Macerozyme R-10. Modified Murashige-Skoog major elements, Ringe and Nitsch minor elements and modified vitamins supplemented with 0.2mg/l 2, 4-D, 1mg/l NAA, 1mg/l BAP, 8.1% mannitol, 1% glucose and 1% sucrose supported protoplast division at 25°C. The frequency of colony formation 10 days after culture was the highest (17.9% of isolated protoplasts) at protoplast density of 0.5×105/ml. Colonies developed to the callus-stage 2 months after initial culture. When callus pieces were cultured in full strength MS medium supplemented with 1mg/l BAP or 1mg/l zeatin, adventitious buds were induced in 9%-16% of callus explants. The buds elongated with leaves open at 20°C and rooted in medium supplemented with 0.1mg/l or 5mg/l IBA.