A NOVEL METHOD FOR EXTRACTING PROTOPLASTS FROM LARGE BROWN-ALGAE

Protoplasts have been isolated without the application of wall degradi ng enzymes from three large brown algal species: Macrocystis angustifo lia, Ecklonia radiata and Durvillaea potatorum. The central feature of this new protocol is the removal of wall-bound calcium by substitutio n with sodium from the isolation medium. The new protocol is specific for cortex and inner meristoderm cell walls with highest yields obtain ed from meristematic or young tissue. Protoplasts, extracted with this method, are approximately 5-10 mum in diameter with viability estimat es ranging from 73-86%. Consistent yields of 10(7) protoplasts g-1 fre sh weight have been obtained within 2-3 h for all three species and th is compares favourably with yields achieved using a conventional enzym e-based system.