Isolation of protoplasts and vacuoles from storage tissue of red beet

A fast and efficient method is described for the isolation of protoplasts and vacuoles from storage tissue of Beta vulgaris L. The viability of the isolated protoplasts is indicated by the development within a few hours of plasma strands with active cyclosis as well as by transport activity.The effect of aging of the tissue on the yield and on the properties of the protoplasts is examined. Adding 1 millimolar dithiothreitol to the water during the aging of the tissue prevents a change of the cell wall structure which otherwise reduces the protoplast yield within 2 days to zero. When protoplasts are isolated from tissue after different times of aging, they show an increase in transport activity which parallels that in the intact tissue.