Identification of a transport mechanism for NH4+ in the symbiosome membrane of pea root nodules.

摘要

Symbiosome membrane vesicles, facing bacteroid-side-out, were purified from pea (Pisum sativum) cv. Solara root nodules and used to study NH4+ transport across the membrane by recording vesicle uptake of the NH4+ analogue [14C]methylamine (MA). Membrane potentials (DELTApsi) were imposed on the vesicles using K+ concentration gradients and valinomycin, and the size of the imposed DELTApsi was determined by measuring vesicle uptake of [14C]tetraphenylphosphonium. Vesicle uptake of MA was driven by anegative DELTApsi and was stimulated by a low extravesicular pH. Protonophore-induced collapse of the pH gradient indicated that uptake of MA was not related to the presence of a pH gradient. The MA-uptake mechanism appeared to have a large capacity fortransport, and saturation was not observed at MA concentrations in the range of 25 鍹 to 150 mM. MA uptake could be inhibited by NH4+, which indicates that NH4+ and MA compete for the same uptake mechanism. The observed fluxes suggest that voltage-driven channels are operating in the symbiosome membrane and that these are capable of transporting NH4+ at high rates from the bacteroid side of the membrane to the plant cytosol. It is concluded that the pH of the symbiosome space is likely to be involved in regulation of the flux.