Microfluorimetric quantitation of cellulose biosynthesis by plant protoplasts using Calcofluor White

A microfluorimetric procedure, using Calcofluor White, has been developed for the measurement of cellulose biosynthesis by cultured protoplasts of tobacco (Nicotiana tabacum L. cv. Xanthi nc). The procedure was compared to a conventional method for cellulose estimation, that employing the anthrone reagent following exhaustive extraction of the developing cell walls. The results indicate that the amount of fluorescence emitted following Calcofluor White treatment is a specific measurement of cell wall cellulose levels. The procedure possesses the twin advantages of ease of manipulation and of greatly enhanced sensitivity (in the picogram range) as compared to other methods.