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The isolation of leaf protoplasts from the submerged aquatic angiosperm Potamogeton lucens L.
Author(s) -
STAAL M.,
PRINS H. B. A.,
HARMELEN M.,
HELDER R. J.
Publication year - 1988
Publication title -
plant, cell and environment
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.646
H-Index - 200
eISSN - 1365-3040
pISSN - 0140-7791
DOI - 10.1111/j.1365-3040.1988.tb01155.x
Subject(s) - protoplast , botany , biology , photosynthesis , potamogetonaceae , potamogeton , aquatic plant , macrophyte , ecology
. 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 HCO 3 − for photosynthesis. 14 CO 2 fixation experiments at pH 6.0 and 8.5 suggest that these isolated protoplasts can also use bicarbonate as a carbon source.