Measuring the Osmotic Water Permeability of the Plant Protoplast Plasma Membrane: Implication of the Nonosmotic Volume

Starting from the original theoretical descriptions of osmotically induced water volume flow in membrane systems, a convenient procedure to determine the osmotic water permeability coefficient (P (os)) and the relative nonosmotic volume (beta) of individual protoplasts is presented. Measurements performed on protoplasts prepared from pollen grains and pollen tubes of Lilium longiflorum cv. Thunb. and from mesophyll cells of Nicotiana tabacum L. and Arabidopsis thaliana revealed low values for the osmotic water permeability coefficient in the range 5-20 microm.s(-1) with significant differences in P (os), depending on whether beta is considered or not. The value of beta was determined using two different methods: by interpolation from Boyle-van't Hoff plots or by fitting a solution of the theoretical equation for water volume flow to the whole volume transients measured during osmotic swelling. The values determined with the second method were less affected by the heterogeneity of the protoplast samples and were around 30% of the respective isoosmotic protoplast volume. It is therefore important to consider nonosmotic volume in the calculation of P (os) as plant protoplasts behave as nonideal osmometers.