In saline environments, plants accumulate Na

With concentrations between 0.1 and 0.2 M, potassium is the most abundant cation in plant cells. The main pool of potassium inside the plant cell is in the vacuole. The function of potassium in this organelle is thought to be purely biophysical; to generate cell turgor to drive cell expansion (1). Under conditions that limit K ϩ availability, the role of K ϩ in the vacuole can be replaced by other ions like Na ϩ , as has been reported to occur under conditions of salt stress (2). Vacuolar concentrations thus vary from high (200 mM) to low (20 mM), suggesting the existence of active K ϩ import and export mechanisms at the vacuolar membrane (3).The role and concentration of K ϩ in other endomembrane organelles in plants are largely unknown. Apart from a specific K ϩ requirement, secondary ion transporters might rely on K ϩ for pH regulation in these organelles. It was shown that apart from the vacuole, V-type H ϩ -transporting ATPase is found in various membranes of the secretory system where vesicle acidification is important for ligand-receptor binding and protein modification, trafficking, and sorting (4, 5). In animal cells, the pH gradient from neutral to acidic along organelles from both the secretory and endocytic pathways is suggested to be under tight control by the operation of secondary ion transporters providing proton leak pathways (6,7,8). Second, solute uptake energized by the pH gradient might be required to generate the osmotic pressure needed for vesicle fusion (5). In view of the abundance of K ϩ in the cell, K ϩ /H ϩ exchangers are likely candidates to be involved in pH and osmoregulation of intracellular compartments as well as active uptake of K ϩ into vacuoles (3) although the biochemical evidence for the existence of such antiporters is scarce (9, 10).In contrast to the limited information available on K ϩ /H ϩ antiporters, many reports have indicated the existence of vacuolar Na ϩ /H ϩ antiporters in plants (11). The first vacuolar Na ϩ /H ϩ exchanger AtNHX1 was identified recently (12), and it was shown that overexpression of this gene in plants enhances salt tolerance (13,14,15). A family of six genes was identified in Arabidopsis (AtNHX1 to AtNHX6) that shows sequence homology to mammalian and yeast NHE or NHX exchangers (16,17). It was demonstrated however that AtNHX1 could catalyze both Na ϩ /H ϩ and K ϩ /H ϩ exchange (14, 18). A similar ion specificity was reported for the human NHE7 isoform. It was shown that this isoform is expressed in the trans-Golgi network, indicating that regulation of pH and ionic composition of intracellular compartments by K ϩ /H ϩ or Na ϩ /H ϩ exchange is an important task of these antiporters (7). This notion was strengthened by the observation that the NHX1 protein is essential for osmotolerance and endosomal protein trafficking in yeast (19,20). Indeed, plant NHX genes were shown not only to be involved in salinity tolerance, but also in vacuolar pH regulation (21,22), and to be induced by NaCl, KCl, and osmotic stress (12,16,(23)(24)(25)(26), or even h...

A tomato (Lycopersicon esculentum Mill. cv. Pera) callus culture tolerant to NaCl was obtained by successive subcultures of NaCl-sensitive calli in medium supplemented with 50 mM NaCl. NaCl-tolerant calli grew better than NaCl-sensitive calli in media supplemented with 50 and 100 mM NaCl. Analysis of callus ion content showed a strong increase in Na+ and Cl- both in NaCl-tolerant and -sensitive calli grown in media containing NaCl for one subculture. Cells from NaCl-tolerant calli showed a higher H+ extrusion activity than those from NaCl-sensitive calli grown for one subculture in the presence of NaCl. The inhibition of H+ extrusion by NaCl-sensitive cells was correlated with an inhibition of microsomal vanadate-sensitive H+-ATPase (EC 3.6.1.35) and ATP-dependent H+ transport, while the stimulation of H+ extrusion by cells tolerant to 50 mM NaCl was correlated with an increase in plasma membrane ATP-dependent H+ transport. The increase of ATP-dependent H+ extrusion in plasma membranes isolated from 50 mM NaCl-tolerant calli was not a result of stimulation of a vanadate-sensitive ATP hydrolytic activity or an increase in passive permeability to H+. Relative to NaCl-sensitive calli, plasma membrane H+-ATPase from calli tolerant to 50 mM NaCl showed a lower Km for Mg2+-ATP. Our results indicate that tolerance of tomato calli to 50 mM NaCl increases the affinity of plasma membrane H+-ATPase for the substrate ATP and stimulates the H+-pumping activity of this enzyme without modifying its phosphohydrolytic activity.

Summary• The effects of salt stress and adaptation on salicylic acid (SA) content and on antioxidant and lipoxygenase (LOX) enzyme activities were studied in tomato ( Lycopersicon esculentum cv. Pera) cells.• NaCl-adapted cells were obtained from calli adapted to 100 m M NaCl by successive subcultures in medium supplemented with 100 m M NaCl. Salt stress treatments consisted of the addition of 100 m M NaCl to cells.• Adapted cells contained a lower concentration of SA than unadapted cells. The lower manganese-containing superoxide dismutase (Mn-SOD) and LOX activities as well as the higher glutathione reductase (GR) and ascorbate peroxidase (APX) activities in adapted cells than in unadapted cells could be correlated with the development of salt adaptation. Salt stress increased APX and LOX activities as well as lipid peroxidation in unadapted cells and increased Mn-SOD activity in both types of cells. Application of 200 µ M SA + 100 m M NaCl inhibited APX activity in both unadapted and adapted cells, induced the Mn-SOD in adapted cells and increased lipid peroxidation in unadapted cells.• Our data indicate that adaptation of tomato cells to NaCl results in a higher tolerance to NaCl-induced oxidative stress and suggest a role for SA in this response.

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