Isolation and cultivation of protoplasts of Caloglossa leprieurii.

Protoplasts, as important media in genetic engineering, had been isolated from some genera of seaweeds in the beginning of this decade. Using a mixed enzyme solution composed of 3% Sea snail enzyme, 3% Cellulase Onozuka R-10, 1% Macerozyme and 0.5% Pectinase, viable protoplasts were isolated from Caloglossa leprieurii (Mont) J. Ag. (Ceramiales, Rhodophyta). The viability of protoplast was determined by its exclusion of Evans Blue stain. The absence of cell walls were observed in the protoplasts, and were determined by Calcofluor White, reduced osmolarity, electron microscopic examination and self-fusion. About half of the protoplasts survived and divided at a high rate when they were cultivated in a thin liquid layer of modified medium No.5 (MES as basic medium, supplemented with MS medium, vitamins and 1ppm IAA, 1.5 ppm KT). Regenerated plantlet appeared after 45 days. Tetrasporangia appeared in the blade of some regenerated plants after 4 months. Due to its properties of Being large, polynucleal and regenerative, protoplasts of C. leprieurii offer a good prospect in the field of cell fusion and genetic operation of seaweeds.