Life without a cell membrane: regeneration of protoplasts from disintegrated cells of the marine green alga Bryopsis plumosa.

When the multi-nucleate giant cells of the green alga Bryopsis plumosa (Huds.) Ag. are injured, the protoplasm is extruded from the cells and can generate spontaneously numerous new cells. The cell organelles aggregate rapidly in seawater and become covered with a gelatinous envelope within 15 minutes. A lipid cell membrane is formed inside the envelope within 9 to 12 hours and about 15% of the original cell membrane is recycled to make the membrane of new protoplasts. Cytochemical studies using Nile Red and various enzymes revealed that the primary envelope is initially composed of polysaccharides, and then transformed into a polysaccharide-lipid complex. Fluorescein diacetate staining showed that the primary envelope has some characteristics of a cell membrane including semi-permeability and selective transport of materials. The aggregation of cell organelles appears to be mediated by two kinds of materials, one present in vacuolar sap and the other on the surface of the cell organelles. About a thousand new cells were generated from a single disintegrated branch and 40% of them eventually developed into mature plants.