A Novel Method for Extracting Protoplasts from Large Brown Algae

Abstract

Protoplasts have been isolated without the application of wall degrading enzymes from three large brown algal species: Macrocystis angustifolia, Ecklonia radiata and Durvillaea potatorum. The central feature of this new protocol is the removal of wall-bound calcium by substitution with sodium from the isolation medium. The new protocol is specific for cortex and inner meristoderm cell walls with highest yields obtained from meristematic or young tissue. Protoplasts, extracted with this method, are approximately 5-10 μm in diameter with viability estimates ranging from 73-86%. Consistent yields of 10 7 protoplasts g -1 fresh weight have been obtained within 2-3 h for all three species and this compares favourably with yields achieved using a conventional enzyme-based system