Characteristics of transmembrane proton transport in the cells of Lupinus polyphyllus

Abstract

The aim of this work was to examine H+-ATPase hydrolytic activity and the stability of transmembrane electrochemical gradients in membrane vesicles isolated from seedlings and leaf cells of an invasive Washington lupine (Lupinus polyphyllus Lindl.) and compare them with non-invasive yellow lupine (Lupinus luteus L.). Temperatures of 25 and 30°C, keeping in mind possible climate warming, were used. For harder stress conditions, short term cold treatment (−8°C in vivo) was used. Plasmalemma-, tonoplast-, and endoplasmic reticulum-enriched membrane fractions were obtained from a sucrose density gradient and identified. Differences in ATPase hydrolytic activity were significant only between lupine species and were more obvious in plasmalemma-enriched fractions. Preincubation of seedlings and leaves at −8°C for 15 min to 24 h showed that microsomic fraction membranes of invasive lupine were more stable (according to Na+-diffusive potential) at low temperature compared to yellow lupine ones. The level of transmembrane electrochemical potential, mainly evoked by ATP-dependent active proton transport, was almost equal in both lupine species. Supposedly, the cells of invasive lupine are able to maintain transmembrane electrochemical potential by the employment of lower hydrolytic activity of H+-ATPase, thus saving energy for growth.