Proton Motive Potential Difference Across the Plasma Membrane of Plant Cells: a Comparison of Cultured Cells and Protoplasts From Italian Ryegrass (Lolium multiflorum Lam.) Endosperm

Abstract

The electrochemical proton gradient (ΔμH⁺) was measured across the plasma membranes of ryegrass (Lolium multiflorum Lam.) endosperm cells grown in suspension culture. The pH gradient (ΔpH) and the transmembrane electrical potential (ΔΨ) of cells and protoplasts were compared. The pH gradient (ΔpH) was measured from the distribution of acetylsalicylic acid and 5,5-dimethyloxazolidine and the membrane potential (ΔΨ) was calculated from the distribution of the lipophilic tetraphenylphosphonium cation (TPP⁺) between the cells and medium. The protoplasts and cells maintained a similar ΔpH. At the external pH of the growth medium (pH 5.5), the cytoplasmic pH was about 7. Under these conditions the ΔΨ was -60 mV (negative inside) for cells and -46 mV for protoplasts. ΔpH and ΔΨ have values which vary reciprocally depending on the external pH. The main contributor to ΔμH⁺ at pH 4.0 is ΔpH whereas, at pH 7.0, ΔΨ is the main component. The external K⁺ concentration influences the cytoplasmic pH significantly only at the higher external pH values.