Mechanisms of chloride partitioning in the leaves of salt-stressed Sorghum bicolor L.

Chloride transport in sheath and blade tissue and the cellular distribution of Cl^- were investigated in an attempt to determine the physiological basis of the preferential accumulation of Cl^- in sheaths of salt-stressed sorghum ( Sorghum bicolor L.). Import and export of ³⁶Cl^- in leaf sheaths and blades of intact sorghum were followed over a 2 week period. X-ray microanalysis of frozen-hydrated bulk tissue samples was used to determine the accumulation of Cl^- and other elements in the vacuoles of sheath and blade cells.
Sheath tissue accumulated Cl^- despite a relatively high Cl^- turnover rate. Chloride was shown to accumulate in most cell types of the sheath, particularly in adaxial epidermal cells. After an initial increase in the concentration of Cl^-, blade tissue regulated Cl^- levels within certain limits. Chloride levels in blades were greater in the abaxial and adaxial epidermal cells than in other cell types. The epidermal cells of blades accumulated Cl^- to approximately the same concentration as sheath epidermal cells. The Cl^- concentration in the photosynthetically active mesophyll and bundle sheath cells, however, remained low.
Thus, the partitioning of Cl^- previously observed in the leaves of salinized sorghum apparently results from the ability of bundle sheath and mesophyll cells to maintain concentrations of Cl^- at lower levels than do epidermal cells. In addition, the relatively large sheath parenchyma cells tend to serve as reservoirs for the storage of Cl^-.     

K/Na selectivity at the plasmalemma of cortical root cells and preferential release of sodium to the xylem vessels in roots of Atriplex hortensis

Using excised roots of Atriplex hortensis L., cv. Gelbe Gartenmelde, the uptake, accumulation and xylem transport of K⁺ and Na⁺ have been measured. Influx as well as xylem transport proved to discriminate little between K⁺ and Na⁺, when considered in relation to the external solution. Both K⁺ and Na⁺ inhibited the uptake and xylem transport of each other to about the same degree. Measurements of intracel-lular Na⁺ fluxes by means of compartment analysis indicated that the low degree of K/Na discrimination during uptake was due to low influx selectivity. Moreover, K⁺/Na⁺ exchange at the plasmalemma was not very efficient in Atriplex roots. In order to establish the basis of the low K/Na discrimination in xylem transport, the rates of K⁺ and Na⁺ transport were related to the cytoplasmic K⁺ and Na⁺ concentrations to yield the selectivity ratio of transport, S(transport) = (φ_cx(K) × [Na⁺]_c)/(φ_cx(Na) × [K⁺]_c). Under all conditions this ratio was far below one indicating that Na⁺ was favoured during xylem release in excised roots of Atriplex at low external concentrations. The implications of this discrimination in favour of Na+ are discussed with respect to salt tolerance of A. hortensis .     

Ionic regulation and Na+,K+-ATPase activity in gills and kidney of the Antarctic aglomerular cod icefish exposed to dilute sea water

When Notothenia neglecta was exposed to diluted, half strength, sea water for 6 h or 10 days, serum concentrations of Cl^-, Na⁺, K⁺ and Mg²⁺ did not differ from those of sea water controls. This indicates that the fish were capable of both short- and long-term regulation. Renal Na⁺,K⁺-ATPase activity decreased after a 6 h exposure to diluted sea water, but there were no differences between diluted sea water and controls after 10 days of exposure.     

Salt tolerance of the reed plant Phragmites communis

Reed plants ( Phragmites communis Trinius) were grown at NaCl concentrations up to 500 m M and their growth, mineral contents and leaf blade osmotic potential were determined. Addition of NaCl up to 300 m M did not affect growth significantly. Sucrose, Cl^-and Na⁺ concentrations in the shoots increased with the salinity of the medium and the shoot water content decreased. K⁺ always contributed most to the leaf osmotic potential. Even in the presence of 250 m M NaCl in the rooting medium, the leaf blade contained only 50 mM Na⁺, suggesting that the plants have an efficient mechanism for Na⁺ exclusion. ²²Na⁺ uptake experiments suggested that the retranslo-cation of absorbed Na⁺ from shoots to the rooting medium lowered the uptake of Na⁺.     

Comparative studies of inorganic substances in the blood of fishes from the Scagerac Sea

The concentrations of the main plasma inorganic electrolytes Na⁺, K⁺, Ca²⁺, Mg²⁺, Cl^- and and PO₄^3- have been determined for different orders of marine fishes. For Na⁺ and Cl^- a typical decrease was found when passing from cyclostomes, holocephalans and elasmobranchs to teleosts. The concentrations of K⁺, Ca²⁺ and Mg²⁺ showed a similar trend except that there was a rise in the teleost group, which showed a large range of variation for these three ions. In the case of PO₄^3- no significant differences between groups were found.     

Impairment of Na+,K+-ATPase activity following enterotoxigenic Campylobacter jejuni infection: changes in Na+, Cl− and 3-O-methyl-D-glucose transport in vitro, in rat ileum

Abstract Unidirectional fluxes of Na⁺, Cl⁻ and 3-O-methyl-D-glucose (3-MG) were measured in vitro across Campylobacter jejuni live culture-infected and control rat ileal short-circuited tissues by the Using Chamber technique. Net secretion of Na⁺ and enhanced secretion of Cl⁻ ions was observed in the infected animals ( P < 0.001, n =6) as compared to the net absorption of Na⁺ and marginal secretion of Cl⁻ ions in the control animals. There was a significant decrease in the mucosal-to-serosal fluxes of 3-MG in C. jejuni -infected rat ileum. The specific Na⁺,K⁺-ATPase activity when measured biochemically in the membrane-rich fraction of enterocytes was found to be significantly lower (58%) in the infected group as compared to the control group ( P < 0.001). Our results therefore suggest that infection with an enterotoxigenic C. jejuni inhibits the Na⁺,K⁺-ATPase activity in rat enterocytes. The impairment of Na⁺,K⁺-ATPase activity thus appears to induce a secondary change in Na⁺,Cl⁻ and 3-MG transport in vitro in rat ileum.     

Control of Na+ and K+ transport in Spergularia marina I. Transpiration effects

In this paper we begin our study of factors controlling Na⁺ and K⁺ uptake in the halophyte Spergularia marina (L.) Griseb., with emphasis on plants growing at moderate salinity (0.2x sea water). The involvement of transpiration was considered first because of its potential to account for much or all of the transport of ions, and particularly of Na⁺, to the shoot under these growth conditions. Transpiration was constant with time through most of the light period, quickly dropping to 6% of the day time rate at night. ²²Na⁺ uptake, on the other hand, showed much less day/night variation, and relative transport to the shoot was constant. After establishing that transpiration was linearly related to leaf weight, possible transpiration effects were further considered as correlations between leaf weight and transport to the shoot. Under constant, day-time conditions, with linear effects of time and plant size removed, total transport of ²²Na⁺ to the shoot (per plant) was not correlated to leaf weight. A similar result was found when transport was expressed per gram of root, and when partitioning of total label to the shoot was considered. Finally, the correlation was considered between leaf weight and a Na⁺/K⁺ enrichment factor defined as the Na⁺/K⁺ ratio in the leaves divided by that in the roots. This correlation was also insignificant. The results indicate that analysis of control of Na⁺ and K⁺ uptake and transport in this experimental system need not consider effects of transpiration.     

Sucrose and ouabain effects on the kinetic properties of a membrane-bound (Na++ K++ Mg2+) ATPase in sugar beet roots

Kinetic studies of a microsomal (Na⁺+ K⁺+ Mg²⁺)ATPase from sugar beet roots ( Beta vulgaris L. cv. Monohill) show that sucrose influences the MgATPase in different ways depending on the presence of K⁺ and/or Na⁺ 1) In the presence of the substrate MgATP and Na⁺ the effect of sucrose follows simple Michaelis-Menten kinetics. 2) In the presence of substrate together with K⁺ or (K⁺+ Na⁺), sucrose has little effect on the ATPase activity. 3) In the presence of Na⁺, onabain acts as an uncompetitive inhibitor with respect to MgATP. 4) In the presence of K⁺ or (K⁺+ Na⁺), the inhibition by ouabain is somewhat depressed and shows non-linearity when 1/v is plotted versus 1/MgATP. 5) Sucrose and Na⁺ activate in a competitive way, so that a successive increase of the Na⁺ level decreases the activation by sucrose. Both K_m and V-values are thereby changed. 6) The sucrose activation in the presence of Na⁺ is also influenced by ouabain. It is, therefore, suggested that Na⁺ may regulate the interference between the Na⁺/K⁺ pump and a sucrose sensitive system.     

K+ status and ABA affect both exudation rate and hydraulic conductivity in sunflower roots

Twenty‐day‐old sunflower plants ( Helianthus annuus L. cv. Sun‐Gro 380) grown in nutrient solutions with different KCl levels were used to study the effects of K⁺ status of the root and of abcisic acid (ABA) on the exudation rate (J_v), the hydraulic conductivity of the root (L_p), the fluxes of exuded K⁺ and Na⁺ (J_K and J_Na), and the gradient of osmotic pressure between the xylem and the external medium. J_v and L_p increased in direct proportion to the K⁺ starvation of the root. Also addition of ABA (4 µ M ) at the onset of exudation in the external medium made J_v and L_p rise, and this effect also increased with the degree of K⁺ starvation. Similarly, K⁺ starvation and ABA promoted both the flux of exuded Na⁺ and the accumulation of Na⁺ in the root. We suggest that ABA acts as a regulating signal for the radial transport of water across the root, and that potassium may be an effector of this mechanism.     

Glutamate Induces a Calcineurin-Mediated Dephosphorylation of Na+,K+-ATPase that Results in Its Activation in Cerebellar Neurons in Culture

Abstract: In primary cultures of cerebellar neurons glutamate neurotoxicity is mainly mediated by activation of the NMDA receptor, which allows the entry of Ca²⁺ and Na⁺ into the neuron. To maintain Na⁺ homeostasis, the excess Na⁺ entering through the ion channel should be removed by Na⁺,K⁺-ATPase. It is shown that incubation of primary cultured cerebellar neurons with glutamate resulted in activation of the Na⁺,K⁺-ATPase. The effect was rapid, peaking between 5 and 15 min (85% activation), and was maintained for at least 2 h. Glutamate-induced activation of Na⁺,K⁺-ATPase was dose dependent: It was appreciable (37%) at 0.1 µ M and peaked (85%) at 100 µ M . The increase in Na⁺,K⁺-ATPase activity by glutamate was prevented by MK-801, indicating that it is mediated by activation of the NMDA receptor. Activation of the ATPase was reversed by phorbol 12-myristate 13-acetate, an activator of protein kinase C, indicating that activation of Na⁺,K⁺-ATPase is due to decreased phosphorylation by protein kinase C. W-7 or cyclosporin, both inhibitors of calcineurin, prevented the activation of Na⁺,K⁺-ATPase by glutamate. These results suggest that activation of NMDA receptors leads to activation of calcineurin, which dephosphorylates an amino acid residue of the Na⁺,K⁺-ATPase that was previously phosphorylated by protein kinase C. This dephosphorylation leads to activation of Na⁺,K⁺-ATPase.     

Possible Involvement of K^＋/Na^＋ in Assessing the Seed Vigor Index

More substances leaked from a higher-vigor seed sample than from a lower-vigor sample. This indicates that, in some cases, electric conductivity does not represent seed vigor level very well, especially for high-vigor seeds. Results from germination, germination index, leachate conductivity, and the ratio of K^＋/Na^＋ from three-seed lots of Chinese cabbage （Brassica pekinensis （Louv.） Rupr） showed that K^＋/Na^＋ correlated well with germination and germination index. The ability of K^＋/Na^＋ to indicate well changes in vigor was further supported by investigation in soybean （Glycine max （L.） Merr.） seeds and another cultivar of Chinese cabbage seeds. Thus, seed leakage of K^＋/Na^＋ can accurately indicate seed vigor, whereas the conductivity test failed to do so. Furthermore, K^＋/Na^＋ showed up bigger quantitative differences in vigor level than did the conductivity test. This findings provide a more sensitive and accurate index for the assessment of seed vigor. The mechanisms of Na^＋ and K^＋ ion transport are also discussed.     

Screening plants for salt tolerance by measuring K+ flux: a case study for barley

Development of salt-tolerant genotypes is central both to remediation of salinity-affected land and to meet increasing global food demand, which has been driving expansion of cropping into marginal areas. The bottleneck of any breeding programme is the lack of a reliable screening technique. This study tested the hypothesis that the ability of plants to retain K⁺ under saline conditions is central to their salt tolerance. Using seven barley cultivars contrasting in salt tolerance (CM72, Numar, ZUG293, ZUG95, Franklin, Gairdner, ZUG403), a comprehensive study was undertaken of whole-plant (growth rate, biomass, net CO₂ assimilation, chlorophyll fluorescence, root and leaf elemental and water content) and cellular (net fluxes of H⁺, K⁺, Na⁺ and Ca²⁺) responses to various concentrations of NaCl (20–320 m m ). Na⁺ selective microelectrodes were found to be unsuitable for screening purposes because of non-ideal selectivity of the commercially available Na⁺ LIX. At the same time, our results show very strong negative correlation between the magnitude of K⁺ efflux from the root and salt tolerance of a particular cultivar. K⁺ efflux from the mature root zone of intact 3-day-old seedlings following 40 min pretreatment with 80 m m NaCl was found to be a reliable screening indicator for salinity tolerance in barley. As a faster and more cost-effective alternative to microelectrode measurements, a procedure was developed enabling rapid screening of large numbers of seedlings, based on amount of K⁺ leaked from plant roots after exposure to NaCl.     

Sodium ion transport in Neurospora crassa

Na⁺ influx and efflux in Neurospora crassa RL21a can be studied separately to calculate net Na⁺ movements. In the absence of external K⁺, Na⁺ influx was independent of the K⁺ content of the cells, but when K⁺ was present, the inhibition of Na⁺ influx by external K⁺ was higher the higher the K⁺ content. Efflux depended on the K⁺ and Na⁺ content, and on the history of the cells. Efflux was higher the higher the Na⁺ and K⁺ contents, and, in low-K⁺ cells, the efflux was also higher in cells grown in the presence of Na⁺ than when Na⁺ was given to cells grown in the absence of Na⁺. Addition of K⁺ to cells in steady state with external Na⁺ resulted in a net Na⁺-loss. In cells grown without Na⁺ this loss was a consequence of the inhibition of Na⁺ influx. In Na⁺-grown cells, addition of K⁺ inhibited Na⁺ influx and increased Na⁺ efflux.     

An effect of Na+ on K+ uptake in intact seedlings of Zea mays

Models for the regulation of K⁺ uptake in higher plant roots have become more complex as studies have moved from the level of excised low-salt roots to that of intact plants grown under fully autotrophic conditions. In this paper we suggest that some of the differences between the conditions are qualitative, possibly requiring fundamental changes to the model, rather than simply quantitative.
The uptake of K⁺ by low-salt roots of Zea mays L. [(A619 x Oh 43) x A632], was independent of Na⁺ concentration over a wide range. However, independence of Na⁺ was not the case in plants grown on complete nutrient medium in the light: inclusion of Na⁺ in the uptake medium enhanced K⁺ uptake. In the presence of Na⁺, K⁺ uptake rates were similar in whole plants with high root K⁺ contents to rates in excised or intact, low-salt roots.     

Brain (Na+, K+)-ATPase and Noradrenergic Activity: Effects of Hyperinnervation and Denervation on High-Affinity Ouabain Binding

Abstract: To examine the role of nerve-specific (Na⁺, K⁺)-ATPase in chronic changes in noradrenergic activity, we examined the effects of noradrenergic denervation and hyperinnervation on p -nitrophenylphosphatase activity and on total and nerve-specific ouabain binding. High-affinity and erythrosin B-sensitive binding were compared as measurements of nerve-specific binding. Hyperinnervation and denervation was produced in cerebellum and cerebral cortex, respectively, by 6-hydroxydopamine lesions of the dorsal noradrenergic bundle. Hyperinnervation increased, and denervation decreased, enzyme activity, high-affinity ouabain inhibition, and erythrosin B-sensitive ouabain binding. As (Nat⁺, K⁺)-ATPase has a major role in the regulation of neural excitability and energy metabolism, and the ouabain binding site has been shown to have endogenous ligands, these changes in (Na⁺, K⁺)-ATPase may be important in the long-term regulation of neuron function by norepinephrine.     

Rootstock-mediated changes in xylem ionic and hormonal status are correlated with delayed leaf senescence, and increased leaf area and crop productivity in salinized tomato

Tomato crop productivity under salinity can be improved by grafting cultivars onto salt-tolerant wild relatives, thus mediating the supply of root-derived ionic and hormonal factors that regulate leaf area and senescence. A tomato cultivar was grafted onto rootstocks from a population of recombinant inbred lines (RILs) derived from a Solanum lycopersicum  ×  Solanum cheesmaniae cross and cultivated under moderate salinity (75 m m NaCl). Concentrations of Na⁺, K⁺ and several phytohormones [abscisic acid (ABA); the cytokinins (CKs) zeatin, Z; zeatin riboside, ZR; and the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC)] were analysed in leaf xylem sap in graft combinations of contrasting vigour. Scion leaf area correlated with photosystem II (PSII) efficiency ( F _v/ F _m) and determined fruit productivity. Xylem K⁺ (but not Na⁺), K⁺/Na⁺, the active CK Z, the ratio with its storage form Z/ZR and especially the ratio between CKs and ACC (Z/ACC and Z + ZR/ACC) were positively loaded into the first principal component (PC) determining both leaf growth and PSII efficiency. In contrast, the ratio ACC/ABA was negatively correlated with leaf biomass. Although the underlying physiological mechanisms by which rootstocks mediate leaf area or chlorophyll fluorescence (and thus influence tomato salt tolerance) seem complex, a putative potassium–CK interaction involved in regulating both processes merits further attention.     

Salinity response of a freshwater charophyte, Chara vulgaris

Abstract. Chara vulgaris L. growing in an oligohaline lake was adapted to laboratory conditions and subjected to long-term salinity treatments ranging from 0 to 350 mol m ³ NaCl added to the lake water (40–680 mosmol kg ¹). Osmotic potential and concentration of the main osmotically active solutes (K⁺, Na⁺, Mg²⁺, Cl and sucrose) in the vacuolar sap of the central internodal cells were estimated. C. vulgaris did regulate turgor but incompletely. Turgor decreased from 335 mosmol kg ¹ under control conditions to 52–111 mosmol kg ¹ at 350 mol m ³ NaCl. The enhancement of πⁱ was achieved by increase in both ions and sucrose. Sterile and fertile plants differed in their response to osmotic stress. In sterile plants, the ions accounted for about 87% of the vacuolar osmotic potential. The increase of πⁱ under osmotic stress was exclusively due to an accumulation of Na⁺ and Cl^-. In fertile plants, sucrose accounted for about 35% of πⁱ and ions for about 51% Under osmotic stress, sucrose content increased together with the ionic content of Na⁺ and Cl^-.     

Promoting effect of fusicoccin on Na+ efflux in barley roots: evidence for a Na+−H+ antiport

Abstract. The effect of fusicoccin (FC) on the K⁺stimulated Na⁺ efflux in root cells of Na⁺ loaded barley roots was studied. FC (0.02 mM) stimulated Na⁺ efflux in the presence of K⁺ and its effect was synergistic with that of K⁺, in a similar way as its effect on proton extrusion. Decreasing the pH of the elution medium promoted Na⁺ efflux and partially replaced the effect of FC. As FC is known to increase the electrochemical proton gradient at the plasmalemma level, these results are consistent with the hypothesis that Na⁺ is extruded in exchange for H⁺. A further support to this view came from the finding that Na⁺ efflux was also promoted by a lipophilic cation, tributylbenzylammonium (TBBA ⁺), which stimulates H ⁺ extrusion and is generally accepted not to enter the cells by means of the same carrier as K ⁺.     

Effect of high external NaCl concentrations on ion transport within the shoot of Lupinus albus. I. Ions in xylem sap

Abstract. Xylem sap was collected from individual leaves of intact transpiring lupin plants exposed to increasing concentrations of NaCl by applying pneumatic pressure to the roots. Concentrations of Na⁺ and Cl⁻ in the xylem sap increased linearly with increases in the external NaCl concentration, averaging about 10% of the external concentration. Concentrations of K⁺ and NO₃⁻, the other major inorganic ions in the sap, were constant at about 2.5 and 1.5 mol m⁻³, respectively. There was no preferential direction of Na ⁺ or Cl⁻ to either young or old leaves: leaves of all ages received xylem sap having similar concentrations of Na⁺ and Cl⁻, and transpiration rates (per unit leaf area) were also similar for all leaves. Plants exposed to 120–160 mol m⁻³ NaCl rapidly developed injury of oldest leaves; when this occurred, the Na⁺ concentration in the leaflet midrib sap had increased to about 40 mol m⁻³ and the total solute concentration to 130 osmol m⁻³. This suggests that uptake of salts from the transpiration stream had fallen behind the rate of delivery to the leaf and that salts were building up in the apoplast.     

Puccinellia tenuiflora maintains a low Na+ level under salinity by limiting unidirectional Na+ influx resulting in a high selectivity for K+ over Na+

Puccinellia tenuiflora is a useful monocotyledonous halophyte that might be used for improving salt tolerance of cereals. This current work has shown that P. tenuiflora has stronger selectivity for K⁺ over Na⁺ allowing it to maintain significantly lower tissue Na⁺ and higher K⁺ concentration than that of wheat under short- or long-term NaCl treatments. To assess the relative contribution of Na⁺ efflux and influx to net Na⁺ accumulation, unidirectional ²²Na⁺ fluxes in roots were carried out. It was firstly found that unidirectional ²²Na⁺ influx into root of P. tenuiflora was significantly lower (by 31–37%) than in wheat under 100 and 150 m m NaCl. P. tenuiflora had lower unidirectional Na⁺ efflux than wheat; the ratio of efflux to influx was similar between the two species. Leaf secretion of P. tenuiflora was also estimated, and found the loss of Na⁺ content from leaves to account for only 0.0006% of the whole plant Na⁺ content over 33 d of NaCl treatments. Therefore, it is proposed that neither unidirectional Na⁺ efflux of roots nor salt secretion by leaves, but restricting unidirectional Na⁺ influx into roots with a strong selectivity for K⁺ over Na⁺ seems likely to contribute to the salt tolerance of P. tenuiflora .