Calcium can ameliorate Na+ toxicity in plants by decreasing Na+ influx through nonselective cation channels. Here, we show that elevated external [Ca2+] also inhibits Na+-induced K+ efflux through outwardly directed, K+-permeable channels. Noninvasive ion flux measuring and patch-clamp techniques were used to characterize K+ fluxes from Arabidopsis (Arabidopsis thaliana) root mature epidermis and leaf mesophyll under various Ca2+ to Na+ ratios. NaCl-induced K+ efflux was not related to the osmotic component of the salt stress, was inhibited by the K+ channel blocker TEA+, was not mediated by inwardly directed K+ channels (tested in the akt1 mutant), and resulted in a significant decrease in cytosolic K+ content. NaCl-induced K+ efflux was partially inhibited by 1 mm Ca2+ and fully prevented by 10 mm Ca2+. This ameliorative effect was at least partially attributed to a less dramatic NaCl-induced membrane depolarization under high Ca2+ conditions. Patch-clamp experiments (whole-cell mode) have demonstrated that two populations of Ca2+-sensitive K+ efflux channels exist in protoplasts isolated from the mature epidermis of Arabidopsis root and leaf mesophyll cells. The instantaneously activating K+ efflux channels showed weak voltage dependence and insensitivity to external and internal Na+. Another population of K+ efflux channels was slowly activating, steeply rectifying, and highly sensitive to Na+. K+ efflux channels in roots and leaves showed different Ca2+ and Na+ sensitivities, suggesting that these organs may employ different strategies to withstand salinity. Our results suggest an additional mechanism of Ca2+ action on salt toxicity in plants: the amelioration of K+ loss from the cell by regulating (both directly and indirectly) K+ efflux channels.

While the importance of cell type specificity in plant adaptive responses is widely accepted, only a limited number of studies have addressed this issue at the functional level. We have combined electrophysiological, imaging, and biochemical techniques to reveal the physiological mechanisms conferring higher sensitivity of apical root cells to salinity in barley (Hordeum vulgare). We show that salinity application to the root apex arrests root growth in a highly tissue- and treatment-specific manner. Although salinity-induced transient net Na⁺ uptake was about 4-fold higher in the root apex compared with the mature zone, mature root cells accumulated more cytosolic and vacuolar Na⁺, suggesting that the higher sensitivity of apical cells to salt is not related to either enhanced Na⁺ exclusion or sequestration inside the root. Rather, the above differential sensitivity between the two zones originates from a 10-fold difference in K⁺ efflux between the mature zone and the apical region (much poorer in the root apex) of the root. Major factors contributing to this poor K⁺ retention ability are (1) an intrinsically lower H⁺-ATPase activity in the root apex, (2) greater salt-induced membrane depolarization, and (3) a higher reactive oxygen species production under NaCl and a larger density of reactive oxygen species-activated cation currents in the apex. Salinity treatment increased (2- to 5-fold) the content of 10 (out of 25 detected) amino acids in the root apex but not in the mature zone and changed the organic acid and sugar contents. The causal link between the observed changes in the root metabolic profile and the regulation of transporter activity is discussed.

Salinity is a widespread environmental stress that severely limits crop yield worldwide. Cerium oxide nanoparticles (nanoceria) have the unique capability of catalytically reducing levels of stress-induced reactive oxygen species (ROS) including hydroxyl radicals (˙OH) that lack enzymatic scavenging pathways. The underlying mechanisms of how nanoceria ROS scavenging augments plant tolerance to environmental stress are not well understood. Herein, we demonstrate that catalytic ˙OH scavenging by nanoceria in Arabidopsis thaliana leaves significantly improves mesophyll K⁺ retention, a key trait associated with salinity stress tolerance. Leaves with mesophyll cells interfaced with 50 mg L⁻¹ poly(acrylic acid) coated nanoceria (PNC) have significantly higher (P < 0.05) carbon assimilation rates (85%), quantum efficiency of photosystem II (9%), and chlorophyll content (14%) compared to controls after being exposed to 100 mM NaCl for 3 days. PNC infiltrated leaves (PNC-leaves) under salinity stress exhibit lower ROS levels – including hydroxyl radical (41%) and its precursor hydrogen peroxide (44%) – and one fold higher (P < 0.05) cytosolic K⁺ dye intensity in leaf mesophyll cells relative to controls. Non-invasive microelectrode ion flux electrophysiological (MIFE) measurements indicated that PNC-leaves have about three-fold lower NaCl-induced K⁺ efflux from leaf mesophyll cells compared to controls upon exposure to salinity stress. The ROS-activated nonselective cation channels (ROS-NSCC) in the plasma membrane of leaf mesophyll cells were identified as the main ˙OH-inducible K⁺ efflux channels. Long term catalytic scavenging of ˙OH in leaves by PNC enhances plant photosynthetic performance under salinity stress by enabling plasma membrane channels/transporters to coordinately retain higher levels of K⁺ in the leaf mesophyll cell cytosol. PNC augmented plant ROS scavenging provides a key tool for understanding and improving plant tolerance against abiotic stresses such as salinity.

The effects of various Na / Ca ratios on root growth, development, and ion acquisition patterns were studied in hydroponic experiments with barley ( Hordeum vulgare L.) plants. In total, interactions between three different levels of salinity (1, 50 and 100 mM NaCl) and three different levels of Ca2+ (0.1, 1 and 10 mM) were studied (a full factorial experiment). Growth rate and biomass accumulation were significantly lower in salinised roots. In addition to reduction in extension growth, salinity also significantly affected plant developmental processes (for example reduced root hair density and root thickening). Supplemental Ca2+ significantly ameliorated those detrimental effects of salinity. Non-invasive, microelectrode ion-flux (MIFE) measurements showed that the onset of salt stress caused rapid and prolonged efflux of H+, K+ and NH4+ from the root epidermis. This efflux could be significantly reversed, or completely prevented, by the presence of high Ca2+ concentration in the bath solution, even after several days of salt stress. Membrane potential measurements in root epidermal cells showed that high Ca2+ levels in the bath were able to restore (otherwise depolarised) membrane potential back to control level (–120 to –130 mV). At the same time, no significant impact of Ca2+ on net Na+ uptake in plant roots was found. Some limitations of the MIFE technique for study of Na+ uptake kinetics under saline conditions, as well as possible ionic mechanisms underlying the ameliorating Ca2+ effects on ion fluxes in roots of salt-stressed plants, are discussed.