The isoelectric points of isolated cutides from Citrus aurantium L. Transport properties of membranes depend strongly on the nature and density of charges fixed to the membrane matrix (6,11). Plant cuticles are biological polymer membranes of heterogeneous composition (12). Their transport properties are of great biological and practical importance and interest, as is evident from an extensive literature dealing with cuticular transpiration (21), leaching, uptake of nutrients, growth regulators, herbicides, fungicides, and insecticides applied to the foliage (9).Early indirect evidence suggested that cuticles may function as weak acid ion exchangers. Sorption of organic ions such as dyes, 2,4-D, and naphthaleneacetic acid was found to be pH-dependent; sorption of anions decreased while sorption of cations increased with increasing pH (2,5,14,16). When sorption of anionic and cationic dyes was plotted against pH, the curves had a point of inflection between pH 2.8 and 3.2 (2), which was believed to represent the pK of the cuticle. Further, surfaces of intact leaves selectively bound divalent over monovalent ions (10). From studies concerning the effect of pH on cuticular transpiration, Hartel (4) properties of isolated tomato fruit cuticular membranes directly and quantitatively. Three dissociable groups in the pH ranges 3 to 6 (0.2 meq g-'), 6 to 9 (0.3 meq g-'), and 9 to 12 (0.55 meq g-') were observed. The first two groups were tentatively identified as -COOH groups and the third as phenolic -OH group. The cuticles contained 0.65% (w/w) nitrogen, and above pH 9 more OH-ions were consumed in titration than Ca2+ bound, indicating titration of some basic groups, possibly -NH3+. Cuticles of apple fruits had previously been shown to contain small amounts of protein (8).Thus, there is some evidence that cuticles are bipolar polyelectrolytes, but quantitative data are lacking. In this paper, we present quantitative data on the isoelectric point, on permselectivity of cuticles to ions, and on the amounts of positive and negative charges fixed to the matrix of various isolated cuticles. (18); (c) immersion at room temperature in a solution of ZnC12 in concentrated HCI (1 g in 1.7 ml) (7). Cellular debris still adhering to the cuticles after isolation was removed with a jet of water, exchangeable cations were removed by agitating the cuticles at room temperature in 1 N HCI for 10 to 20 min (three changes) followed by washing with deionized H20 to remove sorbed HCI. Cuticular waxes were removed from a portion of the pear and Citrus leaf cuticular membranes by successive extraction at room temperature with methanol (three changes), 1 part methanol and 1 part chloroform (three changes), and chloroform (five changes) for 10 min each. All cuticular membranes were air-dried and stored until used. In the following, the method of isolation (enz., oxalate, ZnCl2/HCI) and the removal of waxes (-wax) will be stated after the name of the species from which the cuticle was isolated.
MATERIALS AND METHODSMembrane Potential...