Isolation of Leaf Protoplasts: Macromolecule Uptake and Growth Substance Response

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Abstract
A simple method, using mixtures of commercially available pectinases and cellulases, is described for the isolation of very large numbers of tobacco leaf protoplasts. These protoplasts have been cultured for several days in suitable media. Pinocytosis was shown to occur when protoplasts were incubated with ferritin. These isolated protoplasts respond dramatically, by progressively bursting, when incubated with growth substances at physiological levels. There was no detectable response to growth substances such as indol-3yl-acetic acid when the anti-auxin transcinnamic acid was also present, indicating that these responses are specifically auxin-induced.

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