Protoplast Isolation

Protoplasts are plant cells having their cell wall artificially removed. In this way, they can be used in gene transfer experiments and for the creation of hybrid cells, i.e. cells resulting from the direct fusion of two protoplast cells that might have been derived from entirely different species. In our laboratory we have isolated protoplasts from both mistletoe leaf (Fig. 1D) and callus tissues, at a maximal yield of 100x104 protoplasts/ml, using an enzyme solution of pectinase (0.008-0.016% w/v) and cellulase (0.007-0.014% w/v). The total protoplast yield was higher from calli than leaves, but callus-derived protoplasts were far less viable, probably because older, brownish callus tissues were used for the isolation experiment.

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