In-situ determination of intracellular concentrations of K+ in barley (Hordeum vulgare L. cv. Kara) using the K+-binding fluorescent dye benzofuran isophthalate

The tetra[acetoxymethyl] ester of the K⁺-binding fluorescent dye benzofuran isophthalate (PBFI-AM) was used to determine changes in intracellular potassium (K⁺) concentrations and to measure net transport of K⁺ in barley (Hordeum vulgare L. cv. Kara) root and leaf protoplasts. When this dye binds to free K⁺ inside the cytoplasm, the fluorescence intensity ratio 340/380 nm increases in direct relation to the K⁺ concentration. Because of a delay in the uptake of dye into the vacuoles, it is possible to determine K⁺ concentrations in the vacuoles and transport of K⁺ from the cytoplasm into the vacuole. The uptake of PBFI-AM in root and leaf protoplasts of barley differed in the absence or presence of external K⁺ and was faster at pH 5.5 than at pH 7.0. The fluorescence intensity of the dye was stable for at least 20 h when the protoplasts were kept at 4°C. In the presence of nigericin, the fluorescence intensity of both cells and protoplasts was linearly related to the external concentration of K⁺ (up to 100 mM).     

Epidermal solute concentrations and osmolality in barley leaves studied at the single-cell level

The solute relations of the upper epidermis of the third leaf of barley (Hordeum vulgare L. cv. Klaxon) were studied by analysing vacuolar saps extracted from individual cells. Their osmolality (nanolitre osmometry) and the concentrations of K, Na, Ca, Cl, P, S (energy dispersive X-ray analysis) and NO ₃ ^– (microfluorometry) were measured. All of the osmotically important solutes were accounted for. These were K⁺, NO ₃ ^– , Cl^–, and Ca²⁺. The concentration of each solute varied along the leaf blade and changed with leaf age. Calcium in particular increased during leaf ageing, exceeding concentrations of 50 mM. Plants starved of Ca²⁺ during this period accumulated epidermal K⁺ instead of Ca²⁺. Leaf ageing was accompanied by an increase in epidermal osmolalities by about 100 mosmol · kg^–1. When compared to the bulk leaf extract, epidermal cell extracts exhibited significantly higher concentrations of NO ₃ ^– , Cl^– and Ca²⁺, similar concentrations of K⁺ and Na⁺, and lower concentrations of P. In plants subjected to various levels of NaCl stress (up to 200 mM), epidermal concentrations of Cl^– always exceeded those of the bulk extract, while Na⁺ concentrations were similar. Epidermal cells osmotically adjusted to the increase in the external salt concentration.Abbreviations EDX analysis energy dispersive X-ray analysis We wish to thank Paul Richardson, Jeremy Pritchard, Peter Hinde, Eirion Owen and Andrew Davies (Banger) for their helpfull discussion and technical advice. This work was financed by a grant (UR5/ 521) from the Agricultural and Food Research Council.     

The influence of potassium content on the kinetics of potassium influx into excised ryegrass and barley roots

The influx of K⁺ into excised roots of barley (Hordeum vulgare L.) and ryegrass (Lolium multiflorum L.) previously grown with or without K⁺ was measured in K⁺ solutions ranging in concentration from 0.01 to 50 mM. In both species the K⁺ influx was lower in the roots with high K⁺ content. The extent of reduction by high internal [K⁺] decreased with external concentration above 1 mM. These results support the contention that at high external concentrations passive diffusion makes significant contributions to observed fluxes.     

Alpha-aminoisobutyric acid uptake in primary cultures of astrocytes

Homotypically pure cultures of rat brain astrocytes were used to examine some aspects of non-neuronal A-system (alanine preferring) amino acid uptake. The Asystem specific probe, alpha-aminoisobutyric acid is transported rapidly, and a steady state distribution ratio of 9–25 is reached after 30 minute incubations. Kinetic estimates derived from uptake progress curves indicated aK _m of 1.35 mM and aV _max of 133 nmol/min/mg protein. Uptake is reduced in the absence of either Na⁺ or K⁺. Elevations in extracellular K⁺, a putative metabolic modulator of neuroglia, did not affect uptake.     

Effects of abscisic acid on sequestration and exchange of Na+ by barley roots

Excised Na⁺-starved barley roots were suspended in solutions of Na⁺ in combination with NO ₃ ^- , Cl^-, and SO ₄ ^2- , and effects of the added phytohormone, abscisic acid (ABA), to the medium were determined. Abscisic acid increased the rate of Na⁺ (²²Na⁺) accumulation and the amount of Na⁺ deposited in the vacuoles. These stimulating effects of ABA were modified by anions following the sequence NO ₃ ^- >Cl^->SO ₄ ^2- . Testing whether the magnitude of the pH gradient across the plasmalemma of the cells of the root cortex affects rates of Na⁺ accumulation and their dependence upon ABA, we observed that, in the pH range from 4 to 8, the ABA-induced stimulation was strongest at pH 5.8, and least at pH 4. Changes in pH during the experiment caused changes in the rates of Na⁺ accumulation in agreement with experiments performed at constant pH values. Simultaneously with ABA-enhanced accumulation, loss of Na⁺ occurred. Loss of Na⁺ was strongest at pH 4 and was affected by anions, being greatest with SO ₄ ^2- and following the sequence SO ₄ ^2- >Cl^->NO ₃ ^- . On the basis of the finding that initial acceleration of uptake as well as loss of Na⁺ depended on the pH of the medium we suggest that, in barley roots, ABA stimulates an exchange of Na⁺ for H⁺ at the plasmalemma of the cortical cells. The results indicate that ABA-stimulated expulsion of Na⁺, in combination with ABA-stimulated sequestration in the vacuoles, constitutes one of the mechanisms which enable barley plants to tolerate higher than normal levels of Na⁺.Abbreviations ABA abscisic acid - FW fresh weight     

Acetylcholine-Induced Na+ influx in the mouse lacrimal gland acinar cells: Demonstration of multiple Na+ transport mechanisms by intracellular Na+ activity measurements

Summary In the isolated, superfused mouse lacrimal gland, intracellular Na⁺ activities (aNa _i ) of the acinar cells were directly measured with double-barreled Na⁺-selective microelectrodes. In the nonstimulated conditionaNa _i was 6.5±0.5 mM and membrane potential (V _m ) was –38.9±0.4 mV. Addition of 1 mM ouabain or superfusion with a K⁺-free solution slightly depolarized the membrane and caused a gradual increase inaNa _i . Stimulation with acetylcholine (ACh, 1 M) caused a membrane hyperpolarization by about 20 mV and an increase inaNa _i by about 9 mM in 5 min. The presence of amiloride (0.1 mM) reduced the ACh-induced increase inaNa _i by approximately 50%, without affectingV _m and input resistance in both nonstimulated and ACh-stimulated conditions. Acid loading the acinar cells by an addition/withdrawal of 20 mM NH₄Cl or by replacement of Tris⁺-buffer saline solution with HCO ₃ ^– /CO₂-buffered solution increasedaNa _i by a few mM. Superfusion with a Cl^–-free NO ₃ ^– solution or 1 mM furosemide or 0.5 mM bumetanide-containing solution had little effect on the restingaNa _i levels, however, it reduced the ACh-induced increase inaNa _i by about 30%. Elimination of metabolite anions (glutamate, fumarate and pyruvate) from the superfusate reduced both the restingaNa _i and the ACh-induced increase inaNa _i .The present results suggest the presence of multiple Na⁺ entry mechanisms activated by ACh, namely, Na⁺/H⁺ exchange, Na-K-Cl cotransport and organic substrate-coupled Na⁺ transport mechanisms.     

Mesurement of longitudinal ion profiles in single roots of Hordeum and Atriplex by use of flameless atomic absorption spectroscopy

Summary A method is described by which the Na⁺ and K⁺ content in 0.5 mm sections of single roots of Hordeum distichon L. and Atriplex hortensis L. can be determined by use of flameless atomic absorption spectroscopy. By this method the longitudinal profiles of K⁺ and Na⁺ along low salt roots and roots which had been equilibrated with or grown in K⁺-free 1 mM Na⁺-solution were determined. The profiles reveal that high K⁺/Na⁺ ratios in the cytoplasm are maintained also in K⁺-free solutions. In solutions containing 1 mM Na⁺ a high K⁺/Na⁺ selectivity was found to be dependent on sufficient aeration. From the ion profiles the cytoplasmic (110 mM) and vacuolar (20 mM) K⁺ concentration in low salt barley roots—values which are unobtainable by compartmental analysis—could be estimated.     

Electrophysiology of phagocytic membranes: intracellular K+ activity and K+ equilibrium potential in macrophage polykaryons

The role of K⁺ as current carrier during the slow membrane hyperpolarizations (SH) elicited by iontophoretic Ca²⁺ injections into macrophage polykaryons is studied. The intracellular K⁺ activity (a_K) and the K⁺ equilibrium potential (E_K) are measured using ion-sensitive microelectrodes. The mean value of a_K is 84 ± 5 mM in a culture medium containing 5.3 mM K⁺, but increases to 100 ± 8 mM when the extracellular K⁺ concentration is raised to 30.3 mM. Under the same conditions the values of E_K obtained from the Nernst equation are −81 ± 2 mV and −40 ± 2 mV, respectively. The reversal potentials (E_R) of the SH are calculated from changes observed in transmembrane potential and input resistance, according to an equivalent model based only on passive ionic fluxes. The mean E_R values obtained are −74 ± 8 mV in the presence of low K⁺ concentration and −37 ± 3 mV for the high K⁺ medium. These values are significantly smaller than the estimated E_K for the corresponding situations. Evidence for the existence of an electrogenic (Na⁺ + K⁺)-ATPase activity is also presented. The evidence indicates that an increase in the membrane potassium permeability can account for about 90% of the total permeability change occurring during the SH.     

Potassium transport in red blood cells of frog Rana temporaria: demonstration of a K−Cl cotransport

Pathways of K⁺ movement across the erythrocyte membrane of frog Rana temporaria were studied using ⁸⁶Rb as a tracer. The K⁺ influx was significantly blocked by 0.1 mmol·l^-1 ouabain (by 30%) and 1 mmol·l^-1 furosemide (by 56%) in the red cells incubated in saline at physiological K⁺ concentration (2.7 mmol·l^-1). Ouabain and furosemide had an additive effect on K⁺ transport in frog red cells. The ouabain-sensitive and furosemide-sensitive components of K⁺ influx saturated as f(K⁺)_e with apparent K _m values for external K _e ⁺ concentration of 0.96±0.11 and 4.6±0.5 mmol·l^-1 and V _max of 0.89±0.04 and 2.8±0.4 mmol·l cells^-1·h^-1, respectively. The residual ouabain-furosemide-resistant component was also a saturable function of K _e ⁺ medium concentration. Total K⁺ influx was significantly reduced when frog erythrocytes were incubated in NO _- ³ medium. Furosemide did not affect K⁺ transport in frog red cells in NO ₃ ^- media. At the same K _e ⁺ concentration the ouabain-furosemide-insensitive K⁺ influx in Cl^- medium was significantly greater than that in NO _- ³ medium. We found no inhibitory effect of 1 mmol·l^-1 furosemide on Na⁺ influx in frog red cells in Cl^- medium. K⁺ loss from the frog erythrocytes in a K⁺-free medium was significantly reduced (mean 58%) after replacement of Cl^- with NO _- ³ . Furosemide (0.5 mmol·l^-1) did not produce any significant reduction in the K⁺ loss in both media. The Cl^--dependent component of K⁺ loss from frog red cells was 5.7±1.2 mmol·l^-1·h^-1. These results indicate that about two-thirds of the total K⁺ influx in frog erythrocytes is mediated by a K–Cl cotransport which is only partially blocked by furosemide.Abbreviations DMSO dimethyl sulphoxide - K _e ⁺ external concentration of K⁺ - K _m apparent Michaelis constant for external - K⁺ K _e ⁺ at V _max/2 - RBC red blood cell(s) - V _max maximal velocity of the unidirectional K⁺ influx - TRIS tris(hydroxymethyl)aminomethane     

Effect of nitrogen stress and abscisic acid on nitrate absorption and transport in barley and tomato

The potential of barley (Hordeum vulgare L.) and tomato (Lycopersicon esculentum Mill.) roots for net NO ₃ ^- absorption increased two-to five fold within 2 d of being deprived of NO ₃ ^- supply. Nitrogen-starved barley roots continued to maintain a high potential for NO ₃ ^- absorption, whereas NO ₃ ^- absorption by tomato roots declined below control levels after 10 d of N starvation. When placed in a 0.2 mM NO ₃ ^- solution, roots of both species transported more NO ₃ ^- and total solutes to the xylem after 2 d of N starvation than did N-sufficient controls. However, replenishment of root NO ₃ ^- stores took precedence over NO ₃ ^- transport to the xylem. Consequently, as N stress became more severe, transport of NO ₃ ^- and total solutes to the xylem declined, relative to controls. Nitrogen stress caused an increase in hydraulic conductance (L _p) and exudate volume (J _v) in barley but decrased these parameters in tomato. Nitrogen stress had no significant effect upon abscisic acid (ABA) levels in roots of barley or flacca (a low-ABA mutant) tomato, but prevented an agerelated decline in ABA in wild-type tomato roots. Applied ABA had the same effect upon barley and upon the wild type and flacca tomatoes: L _p and J _v were increased, but NO ₃ ^- absorption and NO ₃ ^- flux to the xylem were either unaffected or sometimes inhibited. We conclude that ABA is not directly involved in the normal changes in NO ₃ ^- absorption and transport that occur with N stress in barley and tomato, because (1) the root ABA level was either unaffected by N stress (barley and flacca tomato) or changed, after the greatest changes in NO ₃ ^- absorption and transport and L _p had been observed (wild-type tomato); (2) changes in NO ₃ ^- absorption/transport characteristics either did not respond to applied ABA, or, if they did, they changed in the direction opposite to that predicted from changes in root ABA with N stress; and (3) the flacca tomato (which produces very little ABA in response to N stress) responded to N stress with very similar changes in NO ₃ ^- transport to those observed in the wild type.Abbreviation and symbols ABA abscisic acid - J_v exudate volume - L_p root hydraulic conductance     

Ion permeation and rectification in ATP-sensitive channels from insulin-secreting cells (RINm5F): Effects of K+, Na+ and Mg2+

Summary Patch-clamp techniques were used to study the permeability to ions of an ATP-sensitive channel in membranes from the pancreatic B-cell line (RINm5F). With patches in the outside-out configuration, theI-V curves for different Na⁺–K⁺ mixtures in the bath and 140 mM K⁺ in the pipette were almost linear, and crossed the zero-current axis at voltages that indicated a variable permeability ratio. When K⁺ was added symmetrically, the plot of the conductancevs. K⁺ activity exhibited saturation, with aG _max of about 160 pS and a half-maximal activity of 216 mM. TheI-V behavior for different K⁺–Na⁺ mixtures in the bath could be accurately described with a model based on Eyring theory, assuming two sites and one-ion occupancy. For K⁺, the dissociation constants (K_K) of the two sites were 290 and 850 mM, the lower value pertaining to the site close to the intracellular medium. In experiments with inside-out patches, both Na⁺ and Mg²⁺, when present in the bath, induced a voltagedependent block of the outward current. Fitting the data with the model suggested that for these ions only one of the two sites binds significantly, the corresponding dissociation constants being (mM): 46 for Na⁺ and 34 for Mg²⁺. Blocking by Na⁺ and Mg²⁺ may account for the low outward current seen in intact cells. This hypothesis is consistent with the observation that such current is further reduced by addition of 2,4-DNP, since metabolism inhibitors are expected to lower the ATP level, thereby liberating Mg²⁺ from the Mg²⁺-ATP complex, as well as inducing accumulation of Na⁺ by decreasing the rate of the Na⁺–K⁺ pump.     

Electrophysiology of the marine diatom Coscinodiscus wailesii. III. Uptake of nitrate and ammonium

Electrophysiological investigations have been carried out to study the uptake of nitrate and ammonium in the marine diatom Coscinodiscus wailesii. Double-barrelled glass microelectrodes were used to record the responses of voltage (V) and resistance (R) between the inserted electrode tip and the outside upon exposure to external nitrate or ammonium. Significant responses were found to be restricted to certain periods following nitrogen-starvation, but consistent with the requirements of nitrogen uptake under physiological conditions. The response of V upon exposure to M concentrations of nitrate was a delayed and reversible depolarization which decreased in amplitude upon repetitive exposures in the time range of a minute. For the ionic conditions of natural sea-water (about 500 mM Na⁺ and pH 8), these effects are thermodynamically consistent with a sodium-nitrate symport mechanism and inconsistent with the familiar proton-nitrate symport mechanism in glycophytic cells of vascular plants and fresh water algae (<1 mM Na⁺, pH <6). Replacement of extracellular K⁺ in artificial sea-water (normal, 10 mM K⁺) by ammonium caused no significant V response in non-starved cells; however, depolarizations were observed in N-starved cells, indicating an increase of the permeability ratio Pammonium/Pk from about 1.0 to about 1.2 under N-starvation.     

Ability of leaf mesophyll to retain potassium correlates with salinity tolerance in wheat and barley

This work investigated the importance of the ability of leaf mesophyll cells to control K⁺ flux across the plasma membrane as a trait conferring tissue tolerance mechanism in plants grown under saline conditions. Four wheat (Triticum aestivum and Triticum turgidum) and four barley (Hordeum vulgare) genotypes contrasting in their salinity tolerance were grown under glasshouse conditions. Seven to 10‐day‐old leaves were excised, and net K⁺ and H⁺ fluxes were measured from either epidermal or mesophyll cells upon acute 100 mM treatment (mimicking plant failure to restrict Na⁺ delivery to the shoot) using non‐invasive microelectrode ion flux estimation (the MIFE) system. To enable net ion flux measurements from leaf epidermal cells, removal of epicuticular waxes was trialed with organic solvents. A series of methodological experiments was conducted to test the efficiency of different methods of wax removal, and the impact of experimental procedures on cell viability, in order to optimize the method. A strong positive correlation was found between plants' ability to retain K⁺ in salt‐treated leaves and their salinity tolerance, in both wheat and especially barley. The observed effects were related to the ionic but not osmotic component of salt stress. Pharmacological experiments have suggested that voltage‐gated K⁺‐permeable channels mediate K⁺ retention in leaf mesophyll upon elevated NaCl levels in the apoplast. It is concluded that MIFE measurements of NaCl‐induced K⁺ fluxes from leaf mesophyll may be used as an efficient screening tool for breeding in cereals for salinity tissue tolerance.     

Pyruvate carboxylase from rainbow trout liver

Summary A method is described for the partial purification of pyruvate carboxylase from rainbow trout liver. The enzyme has a pH optimum of about 8.0, possesses an absolute requirement for activation by acetylCoA, and prefers MgATP over other nucleoside triphosphates. K⁺ causes a decrease in the apparentK _m for HCO ₃ ^– . AcetylCoA activation shows positive cooperativity withK _a=0.072 mM andn _H=1.78 at pH 7.7, 2.5 mM free Mg²⁺, 100 mM K⁺, and saturating concentrations of substrates. A high acetylCoA concentration causes a decrease in the apparentK _m values for MgATP and HCO ₃ ^– and a biphasic double reciprocal plot with pyruvate as the varied substrate. MgADP and AMP are competitive inhibitors with respect to MgATP. The enzyme shows a U-type response to the adenylate energy charge and retains considerable activity throughout a wide range of energy charge values. It is proposed that intramitochondrial acetylCoA concentration and the adenylate energy charge control the rate of pyruvate carboxylation in vivo.Abbreviations DTT dithiothreitol - PMSF phenylmethylsulfonylfluoride     

Ethylene and nitric oxide are involved in maintaining ion homeostasis in <Emphasis Type="Italic">Arabidopsis</Emphasis> callus under salt stress

In the present study, the role of ethylene in nitric oxide (NO)-mediated protection by modulating ion homeostasis in Arabidopsis callus under salt stress was investigated. Results showed that the ethylene-insensitive mutant etr1-3 was more sensitive to salt stress than the wild type (WT). Under 100 mM NaCl, etr1-3 callus displayed a greater electrolyte leakage and Na⁺/K⁺ ratio but a lower plasma membrane (PM) H⁺-ATPase activity compared to WT callus. Application of exogenous 1-aminocyclopropane-1-carboxylic acid (ACC, an ethylene precursor) or sodium nitroprusside (SNP, a NO donor) alleviated NaCl-induced injury by maintaining a lower Na⁺/K⁺ ratio and an increased PM H⁺-ATPase activity in WT callus but not in etr1-3 callus. The SNP actions in NaCl stress were attenuated by a specific NO scavenger or an ethylene biosynthesis inhibitor in WT callus. Under 100 mM NaCl, the NO accumulation and ethylene emission appeared at early time, and NO production greatly stimulated ethylene emission in WT callus. In addition, ethylene induced the expression of PM H⁺-ATPase genes under salt stress. The recovery experiment showed that NaCl-induced injury was reversible, as signaled by the similar recovery of Na⁺/K⁺ ratio and PM H⁺-ATPase activity in WT callus. Taken together, the results indicate that ethylene and NO cooperate in stimulating PM H⁺-ATPase activity to modulate ion homeostasis for salt tolerance, and ethylene may be a part of the downstream signal molecular in NO action.     

The regulation of K+ influx in excised barley roots

The electrochemical potential differences for potassium, between excised barley (Hordeum vulgare L.) roots and external media containing 0.05 mM KCl+0.5 mM CaSO₄, were determined over a 4-h period during which initially low-K⁺ roots accumulated K⁺ by pretreatment in 50 mM KCl plus 0.5 mM CaCl₂. This pretreatment resulted in increased internal [K⁺], decreased K⁺ influx (as measured from 0.05 mM KCl+0.5 mM CaSO₄) and decreased values of . These observations indicate that the decline of K⁺ influx associated with increased internal K⁺ concentration cannot be accounted for by passive adjustment to the electrochemical gradient for this ion.     

Relation between permeability to potassium and sodium ions and fusicoccin-stimulated hydrogen-ion efflux in barley roots

Summary Measurements are described of fusicoccin (FC)-stimulated H⁺ efflux in barley (Hordeum vulgare L.) roots when K⁺ and Na⁺ concentrations were varied. In low-salt roots H⁺ efflux was stimulated in both 5 mM KCl and NaCl. In salt-saturated roots H⁺ efflux was stimulated more effectively in KCl than in NaCl solution. The stimulation of H⁺ efflux thus is parallel with the selectivity of these different root preparations for K⁺ and Na⁺ and with estimates of permeability ratios (P _Na/P _K) determined from electrical measurements. It is suggested that the results support electrogenic coupling between FC-stimulated H⁺ efflux and cation uptake.     

ROLE OF INORGANIC IONS IN CONTROLLING SEDIMENTATION RATE OF A MARINE CENTRIC DIATOM DITYLUM BRIGHTWELLI1,2

The settling rates and intracellular levels of K⁺, Na⁺, Cl^-, Mg²⁺ and Ca²⁺ were measured in Ditylum bright-welli (West) Grunow, grown axenically in an enriched seawater medium at 20 C at 4,000 lx on an 8:16 LD schedule. Cells at the end of the dark period have high Na⁺ (118 mM), low K⁺ (64 mM) and low Cl^- (117 mM) relative to levels at the end of the light period when K⁺ (126 mM) and Cl^- (154 mM) are high and Na⁺ (101 mM) is low. There is no significant change in Mg²⁺ (16–18 mM) or Ca²⁺ (3–4 mM) with time. The net result of the ion changes during the light period is to increase cell density by about 3.4 mg ml^-1. This change can account for the increase in settling rate of ca. 0.3 day^-1 during the same interval. The density of the cell contents, calculated from observed ion concentrations, is 15–18 mg.ml^-1 less than that of the seawater medium. The ion and settling rate changes are light-dependent and do not persist in the dark or under constant light (ca. 850 lx), but cells do exhibit a free-running circadian rhythm in cell division under continuous dim illumination. The cell vacuole expands during the light period and contracts during the dark, apparently in response to the net ion fluxes. D. brightwelli appears to regulate its density by active ion selectivity accompanied by trans-vacuolar water movement.     

Substitution of Na+ by K+ in Tissues and Root Vacuoles of Barley (Hordeum vulgare L. cv. Aramir)

The aim of this paper was to investigate whether Na⁺ accumulated in the vacuoles of barley roots can be replaced by K⁺, or else remain irreversibly trapped.Barley seedlings (Hordeum vulgare L. cv. Aramir) were grown for 2 days on nutrient solutions and then salinized over 24 h by a linear increase in NaCl concentration (50 mol · m⁻³). NaCl was replaced by an equimolar concentration of KCl and 1 day later the KCl concentration was gradually decreased over 23 hours. After this the plants were grown for 2 further days on basic nutrient solution. Plant material was harvested at each experimental step in order to measure plant growth and K⁺ and Na⁺ concentrations in vacuoles and tissues.The results from quantitated, energy dispersive X-ray analyses on frozen, hydrated vacuoles of the rhizodermls and 3 subsequent cortical cell rows, point to a proportional replacement of K⁺ by Na⁺ after NaCl stress. A replacement of Na⁺ by K⁺ was clearly found in the rhizodermal vacuoles after KCl treatment. Flame emission spectrometric data from corresponding root segments support these results.