Preparation, culture, and regeneration of protoplasts from filamentous green algae

High yields of viable protoplasts of the filamentous green algae Mougeotia, Stigeoclonium, Ulothrix, and Klebsormidium can routinely be produced by enzymatic digestion of their cell walls. Regenerating cell walls are detectable by electron microscopy within 16 h of transferring the protoplasts from their isolation medium to growth medium supplemented with an osmoticum. All algae except Ulothrix regenerated filaments that were indistinguishable from untreated material. Stigeoclonium protoplasts, as well as regenerating branched filaments, also gave rise to motile cells and clumps of cells.