A Novel Method for Extracting Protoplasts from Large Brown Algae

Protoplasts have been isolated without the application of walldegrading enzymes from three large brown algal species: Macrocystisangustifolia, Ecklonia radiata and Durvillaea potatorum. Thecentral feature of this new protocol is the removal of wall-boundcalcium by substitution with sodium from the isolation medium.The new protocol is specific for cortex and inner meristodermcell walls with highest yields obtained from meristematic oryoung tissue. Protoplasts, extracted with this method, are approximately5–10 µm in diameter with viability estimates rangingfrom 73–86%. Consistent yields of 10⁷ protoplasts g^–1fresh weight have been obtained within 2–3 for all threespecies and this compares favourably with yields achieved usinga conventional enzyme-based system. Key words: Brown algae, protoplasts, alginate, calcium, enzymes