Abstract
Abstract. A method for isolating protoplasts from leaves of the submerged aquatic angiosperm Potamogeton lucens L. is described. The protoplasts are produced enzymatically from leaf strips using 1.5% (w/v) Cellulysin, 0.3% (w/v) Macerozyme R10, and 2.5% (v/v) β‐glucuronidase at 27°C in the dark. Subsequently the protoplasts are purified on a discontinuous Ficoll gradient. The yield obtained is approximately 20% of the starting material on a chlorophyll basis. The viability is high, namely more than 90% as estimated with Evans Blue. Cells of the intact leaves of P. lucens can use HCO3− for photosynthesis. 14CO2 fixation experiments at pH 6.0 and 8.5 suggest that these isolated protoplasts can also use bicarbonate as a carbon source.
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