Effect of Sudden Salt Stress on Ion Fluxes in Intact Wheat Suspension Cells

Although salinity is one of the major problems limiting agriculturalproduction around the world, the underlying mechanisms of highNaCl perception and tolerance are still poorly understood. Theeffects of different bathing solutions and fusicoccin (FC),a known activator of plasma membrane ATPase, on plasma membranepotential (E_m) and net fluxes of Na⁺, K⁺and H⁺were studied inwheat suspension cells (Triticum aestivum) in response to differentNaCl treatments. E_mof cells in Murashige and Skoog (MS) mediumwas less negative than in cells exposed to a medium containing10 mM KCl + 0.1 m M CaCl₂(KSM) and to a basic salt medium (BSM),containing 1 m M KCl and 0.1 m M CaCl₂. Multiphasic Na⁺accumulationin cells was observed, peaking at 13 min after addition of 120m M NaCl to MS medium. This time scale was in good agreementwith net Na⁺flux changes measured non-invasively by moving ion-selectivemicroelectrodes (the MIFE system). When 120 m M NaCl was addedto all media studied, a quick rise of Na⁺influx was reversedwithin the first 20 min. In both 120 and 20 m M NaCl treatmentsin MS medium, net Na⁺efflux was observed, indicating that activeNa⁺transporters function in the plant cell response to saltstress. Lower external K⁺concentrations (KSM and BSM) and FCpre-treatment caused shifts in Na⁺fluxes towards net influxat 120 m M NaCl stress. Copyright 2000 Annals of Botany Company Sodium, potassium, proton, membrane potential, fusicoccin, salt stress, wheat, Triticum aestivum     

Ionic Relations of Garden Orache, A triplex hortensis L.: Growth and Ion Distribution at Moderate Salinity and the Function of Bladder Hairs

The growth of garden orache, A triplex hortensis was studiedunder conditions of mild NaCl or Na₂SO₄ salinity. Growth, drymatter production and leaf size were substantially stimulatedat 10 mM and 50 mM Na⁺ salts. Increased growth, however, appearedto be due to a K⁺-sparing effect of Na⁺ rather than to salinityper se. The distribution of K⁺ and Na⁺ in the plant revealeda remarkable preference for K+ in the roots and the hypocotyl.In the shoot the K/Na ratio decreased strongly with leaf age.However, the inverse changes in K⁺ and Na⁺ content with leafage were dependent on the presence of bladder hairs, which removedalmost all of the Na⁺ from the young leaf lamina. Measurementsof net fluxes of K⁺ and Na⁺ into roots and shoots of growingAtriplex plants showed a higher K/Na selectivity of the netion flux to the root compared to the shoot. With increasingsalinity the selectivity ratio S_{K, Na}^* of net ion fluxes tothe roots and to the shoots was increased. The data suggestthat recirculation of K⁺ from leaves to roots is an importantlink in establishing the K/Na selectivity in A. hortensis plants.The importance of K⁺ recirculation and phloem transport forsalt tolerance is discussed. Key words: Atriplex hortensis, Salinity, Potassium, Sodium, K⁺ retranslocation, Bladder hairs, Growth stimulation     

Inhibition of Nitrate Assimilation in Roots in the Presence of Ammonium: The Moderating Influence of Potassium

Experiments were conducted to investigate the effect of concentrationof NH₄⁺ in nutrient solution on root assimilation of NO₃^–and to determine whether the NH₄⁺NO₃^– interaction wasmodified in the presence of K⁺. Dark-grown, detopped corn seedlings(cv. Pioneer 3369A) were exposed for 8 h to 0.15 mM Ca(NO₃)₂and varying concentrations of (NH₄)₂SO₄ in the absence or presenceof 0.15 mM K₂SO₄. The accelerated phase of NO₃^– uptakeappeared most sensitive to restriction by additions of 0.15mM (NH₄)₂SO₄. In the absence of K⁺, the restriction increasedonly slightly even when solution (NH₄)₂SO₄, was increased from0.15 mM to 12.5 mM which was accompanied by an increase of NH₄⁺in the tissue from about 7.0 to 35 µmol g^–1 fr.wt. of root. Increasing concentrations of solution NH₄⁺ progressivelyinhibited net K+ uptake. At the highest solution NH₄⁺ concentrations,there was an initial net efflux of K⁺ and no net influx occurredduring the treatment period. The severity of the NH₄)SO₄ restrictionof NO₃^– uptake was moderated considerably in the presenceof K⁺ as long as a net influx of K⁺ occurred. However, net influxof K⁺ was not associated with alteration of NH₄⁺ uptake, assimilation,or accumulation in the root tissue. The lack of correlationbetween the severity of restriction of NO₃^– uptake andendogenous NHJ suggested the restriction resulted from an effectexerted by exogenous NH₄⁺ which tended to saturate at lowersolution NHJ concentrations or by inhibitory factors generatedduring assimilation of NH₄⁺. Several mechanisms were postulatedto account for the moderating influence of K⁺. In all experiments,root NO₃^– reduction was restricted by the presence ofambient NH₄⁺. The quantitative decreases in reduction tendedto be less than decreases in NO₃^– uptake and therefore,could result from inhibition solely of uptake with subsequentlimitation in availability of substrate for the reduction process,but the possibility of a direct effect on reduction could notbe excluded.     

K+-Na+ Exchange and Selectivity in Barley Root Cells: Effect of Na+ on the Na+ Fluxes

Using the compartmental analysis the unidirectional Na⁺ fluxesin cortical cells of barley roots, the cytoplasmic and vacuolarNa⁺ contents Q_c and Q_v, and the trans-root Na⁺ transport R'have been studied as a function of the external Na⁺ concentration.Using the re-elution technique the effect of low K⁺ concentrationson the plasmalemma efflux _co of Na⁺ (K+-Na+ exchange) and onR' was investigated at different Na+ concentrations and correspondinglydifferent values of the cytoplasmic sodium content Qc. The relationof the K+-dependent Na+ efflux coK+-dep to Qc or to the cytoplasmicNa+ concentration obeyed Michaelis-Menten kinetics. This isconsistent with a linkage of co, K+-dep to K+ influx by a K⁺-Na⁺exchange system. The apparent Km corresponded to a cytoplasmicNa⁺ concentration of 28 mM at 0·2 mM K⁺ and about 0·2mM Na⁺ in the external solution. 0·2 mM K⁺ stimulatedthe plasma-lemma efflux of Na⁺ and inhibited Na⁺ transport selectivelyeven in the presence of 10 mM Na⁺ in the external medium showingthe high efficiency of the K⁺-Na⁺ exchange system. However,_co, K⁺-dep was inhibited at 10 mM Na¹ compared to lower Na¹concentrations suggesting some competition of Na¹ with K¹ atthe external site of the exchange system. The effect of theNa+ concentration on Na¹ influx _oc is discussed with respectto kinetic models of uuptake.     

Effects of Na2SO4, K2SO4 and KCl on Growth and Ion Uptake of Callus Cultures of Vaccinium corymbosum L. cv. Blue Crop

Non-selected and Na₂SO-, K₂SO₄- or KCl-selected callus culturesof Vaccinium corymbosum L. cv. Blue Crop were grown on mediasupplemented with 0, 25 and 50 mM Na₂SO₄ (non-selected and Na₂SO(-selectedonly), 0, 25 and 50mMK₂SO₄ (non-selected and K₂SO₄-selectedonly) or 0, 50 and 100 mM KCl (non-selected and KCl-selectedonly). On all media, growth of selected callus (on a fresh-weightor dry-weight basis) was greater than that of non-selected callus,and selected callus grew optimally on the level and type ofsalt on which it was selected. Selected callus was friable andmaintained a higher f. wt:d. wt ratio. Tissue water potentialin selected callus was more negative than in non-selected callus. Flame photometry and chloridometry showed Na⁺, K⁺ and Cl^–accumulated in callus to concentrations equal to or greaterthan the initial concentration in the medium. Turbidometry showedthat tissue SO₄^2- concentration was lower than the concentrationin the medium. In most cases selected callus accumulated moreNa⁺, K^sup, SO₄^2– or Cl^– than non-selected callus.Vacuolar ion concentration was measured by electronprobe X-raymicroanalysis, and on most media selected callus had highervacuolar ion concentrations than non-selected callus. SO₄^2–and Cl^– were accumulated in the vacuoles at concentrationshigher than the external medium, but vacuolar Na⁺ concentrationdid not reach external concentration on Na₂SO₄ and on potassiumsalts was maintained between 12 and 17 mM. Vacuolar K⁺ concentration(approx. 142–191 mM on no salt) decreased on Na₂SO₄ andincreased on K₂SO₄ and KCl. There was no precise correlation between total or specific ionaccumulation (Na⁺, K⁺, SO₄^2– and Cl^– and fresh-weightyield. Results suggest that selection results in adaptationin response to decreased water potential of the medium. Vaccinium corymbosum, blueberry, electronprobe X-ray microanalysis, callus, in vitro selection, salt tolerance, KCl, K₂SO₄, Na₂SO₄     

Effect of K+, its Counter Anion, and pH on Sodium Efflux from Barley Root Tips

Sodium efflux from ²²Na⁺-loaded root tips root tips of Hordeumvulgare L. was markedly increased by replacing 10mM Na₂SO₄ inthe washing solution by K₂SO₄ with the same electrical conductivity.This increase was inhibited by both an uncoupler and an inhibitorof oxidative phosphorylation but not by ouabain. Potassium ionsdid not enhance Na⁺ efflux in the presence of a rapidly absorbedcounter anion, such as Cl^–, instead of . Efflux of ²²Na⁺ could also be enhanced by a low pH in theabsence of K⁺; this was prevented by uncouplers, but not byan inhibitor of the mitochondrial ATPase. It seems that K⁺ indirectly enhances Na⁺ efflux. It is suggestedthat metabolic K⁺ uptake in excess of the counter anion resultsin a proton gradient across the plasmalemma (acid outside) inducingH⁺/Na⁺ antiport.     

Ion Imbalance in Capsicum annum scarbrous diminutive a Wilty Mutant of pepper: II.RUBIDIUM FLUX

Root tips of the wilty pepper mutant scarbrous diminutive accumulateless rubidium than those of the normal genotype. This phenomenonwas evident in root tips excised from plants maintained for2 d in CaSO₄ solution (low salt plants), especially in the lowerexternal concentration range (0.1– 1.0 mM) of RbCl. Theefflux rate of Rb⁺ from mutant root tips was twice as high asin normal root tips. These results indicate that the ability of the mutant rootsto absorb and accumulate Rb⁺ and K⁺ is impaired. This defectcould be a consequence of either an impaired Na⁺/K⁺ carriersystem, or increased leakiness of mutant membranes, or both. The fact that the normal roots can accumulate Rb⁺ much fasterthan mutant roots supports the first alternative, i.e. thatthe high affinity carrier system was impaired in the mutantroots. However, the higher efflux rate of Rb⁺ from the mutantroots suggests that membrane leakiness was also affected.     

Cytosolic potassium controls CFTR deactivation in human sweat duct

Absorptive epithelial cells must admit large quantities of salt (NaCl) during the transport process. How these cells avoid swelling to protect functional integrity in the face of massive salt influx is a fundamental, unresolved problem. A special preparation of the human sweat duct provides critical insights into this crucial issue. We now show that negative feedback control of apical salt influx by regulating the cystic fibrosis transmembrane conductance regulator (CFTR) Cl^– channel activity is key to this protection. As part of this control process, we report a new physiological role of K⁺ in intracellular signaling and provide the first direct evidence of acute in vivo regulation of CFTR dephosphorylation activity. We show that cytosolic K⁺ concentration ([K⁺]_c) declines as a function of increasing cellular NaCl content at the onset of absorptive activity. Declining [K⁺]_c cause parallel deactivation of CFTR by dephosphorylation, thereby limiting apical influx of Cl^– (and its co-ion Na⁺) until [K⁺]_c is stabilized. We surmise that [K⁺]_c stabilizes when Na⁺ influx decreases to a level equal to its efflux through the basolateral Na⁺-K⁺ pump thereby preventing disruptive changes in cell volume. electrolytes; phosphatases; protein kinase A; cystic fibrosis transmembrane conductance regulator; epithelial Na⁺ channel     

Transport of Methylammonium and Ammonium Ions by Elodea densa

Excised leaves of Elodea densa rapidly absorb methylamine¹ fromdilute solutions (up to 2.0 mM). The influx isotherm is hyperbolic,with a K_? of approximately 160 µM. Influx is reduced followingtransfer of leaves from light to darkness, and at low temperature.Low concentrations of ammonia reduce the influx greatly, apparentlyby competition between NH⁺₄ and CH₃NH⁺₃, but K⁺ and Na⁺ havelittle effect, nor has removal of Cl^–. Influx is veryinsensitive to external pH over the range 5.0 to 9.0, with usuallya small increase between pH 9.0 and 10.0. When leaves are pretreatedin solutions containing nitrogenous compounds subsequent influxcan be decreased (by ammonia), unchanged (by methylamine) oreven increased (by arginine, proline and imidazol). Influx of methylamine and ammonia lowers influx of K⁺ (Rb⁺)and of Cl and increases efflux of K⁺ into solutions initiallyfree of K⁺. Fluxes of Ca⁺⁺ are not affected and there is netefflux of H⁺ into unbuffered solutions. The results show that uptake of methylamine and ammonia underthese conditions is primarily by transport (uniport) of CH3NHJand NHJ and that diffusion of CH₃NH⁺₃ and NH⁺₃ is insignificant.In Elodea, unlike some of the plants that have been previouslystudied, maintenance of charge-balance during transport of CH₃NH⁺₃and NH⁺₃ appears to involve accumulation of organic acid anions.     

Internal Factors Regulating Nitrate and Chloride Influx in Plant Cells

The primary factor determining the observed decrease in activeC1^– influx during salt accumulation in carrot and barleyroot cells has been shown to be the concentration of C1^–+ NO₃^– in the vacuole. The relationship between C1^– influx and the vacuolar concentrationsof various substances was examined after the tissues had accumulatedions from various salt solutions. After accumulating K⁺ malate,C1^– influx was not reduced, but after accumulating C1^–or NO₃^– salts, C1^– influx was reduced by up to 90per cent. Considering all treatments, C1^– influx was notcorrelated with the vacuolar concentration of K⁺, Na⁺, (K⁺+Na⁺),reducing sugars, malate, C1^–, or NO₃^–, nor withthe cellular osmotic pressure. The correlation coefficient betweenCl^– influx and log (C1^– + NO₃^– concentrationin the vacuole) was highly significant, and accounted for allthe variation in C1^– influx in this experiment. Net NO₃^– influx is similarly reduced by a high C1^–concentration in the vacuole. External Cl^– and NO₃^–have quantitatively different, apparently competitive, effectson C1^– influx. These differ from the apparently negative-feedbackeffects of C1^– and NO₃^– in the vacuole, which arequantitatively similar. Decreasing the internal hydrostatic pressure by raising theexternal osmotic pressure increased active K⁺ influx in Valoniaventricosa, but had no effect on C1^– or K⁺ influx in carrotor maize root cells. Cl^– influx is not related to thereducing sugar concentration during ageing drifts in excisedcarrot root tissue. Acetazolamide did not inhibit C1^– influx to carrot tissue. The implications of this type of negative feedback regulation,and the relationship between C1^– and NO₃^– transportare discussed.     

Effect of NaCI Salinity on Flows and Partitioning of C, N, and Mineral Ions in Whole Plants of White Lupin, Lupinus albusL.

Plants of Lupinus albus L., cv. Ultra, were grown hydroponicallywith NO₃^–-nutrition for 51 d under control (0.05 mol m^–3Na⁺ and 10 mol m^–3 Cl^–) and saline (40 mol m^–3NaCI) conditions. Plants were harvested 41 and 51 d after germinationand analysed for content and net increment of C, N and the mineralcations K⁺, Na⁺, Mg²⁺, and Ca²⁺ and the anions Cl^–, NOJ,malate, phosphate, and SO₄^2–. Roots, stem interaodes,petioles and leaflets were analysed separately. During the studyperiod net photosynthesis, respiratory losses of CO₂ from shootand root and the composition of the spontaneously bleeding phloemsap and the root pressure xylem exudate were also determined.Using molar ratios of C over N in the transport fluids, incrementsof C and N, and photosynthetic gains as well as respiratorylosses of C, the net flows of C and N in the xylem and phloemwere then calculated as in earlier studies (Pate, Layzell andMcNeill, 1979a). Knowing the carbon flows, the ratios of ionto carbon in the phloem sap, and ion increments in individualorgans, net flows of K⁺, Na⁺, and Cl^– over the study periodwere also calculated. Salt stress led to a general decrease of all partial componentsof C and N partitioning indicating that inhibitions were notdue to specific effects of NaCI salinity on photosynthesis oron NO₃ uptake. However, there were differences between variouslyaged organs, and net phloem export of nitrogenous compoundsfrom ageing leaves was substantially enhanced under saline conditions.In addition, NO₃^–reduction in the roots was specificallyinhibited. Uptake and xylem transport of K⁺ was more severelyinhibited than photosynthetic carbon gain or NO₃^– uptakeby the root. K⁺ transport in the phloem was even more severelyrestricted under saline conditions. Na⁺ and Cl^– flowsand uptake, on the other hand, were substantially increasedin the presence of salt and, in particular, there were thenmassive flows of Na^– in the phloem. The results are discussedin relation to the causes of salt sensitivity of Lupinus albus.The data suggest that both a restriction of K⁺ supply and astrongly increased phloem translocation of Na⁺ contribute tothe adverse effects of salt in this species. Restriction ofK⁺ supply occurs by diminished K⁺ uptake and even more by reducedK⁺ cycling within the plant. Key words: Lupinus albus, salt stress, phloem transport, xylem transport, partitioning, carbon, nitrogen, K⁺, Na⁺, CI^–     

Varietal differences in sodium uptake in barley cultivars exposed to soil salinity or salt spray

Barley varieties are known to differ in the extent of Na⁺ andCl^– accumulation in leaves when grown in saline soil orhydroponic culture. In particular, the cv. Chevron accumulatesmore Na⁺ than the more salt-tolerant cv. CM67, and has lowerleaf K⁺ concentrations. When salt was applied as a spray tothe leaves, CM67 accumulated more Na+ than Chevron, and theselection Sinis 27 (from a landrace collected on the Sinis Peninsulaof Sardinia) accumulated more Na⁺ than Sinis 28. In some casesleaf K⁺ concentrations decreased in response to high concentrationsof salt sprayed on to the leaves. Accumulation of Na⁺ was greaterin the 4th leaf than in the flag leaf. Added CaCI₂ had oppositeeffects when added to the salt applied to the soil or to thesaline spray. In the soil, CaCI₂ reduced Na⁺ uptake; appliedto the leaf it increased Na⁺ uptake. Pre-wetting the leavesbefore the salt spray, or washing the leaves with non-salinewater 1 h after the salt spray, reduced the uptake of Na⁺ andCl^–. It is clear that tolerance to salt applied as saltspray or in the soil are different characteristics. Key words: Barley, salt, foliar uptake, calcium     

The Role of the Stem in the Partitioning of Na+ and K+ in Salt-Treated Barley

Hordeum vulgare cv. California Mariout was grown for 50 d insand culture at 100 mol m^–3 NaCl. Xylem sap was collectedthrough incisions at the base of individual leaves along thestem axis by applying pressure to the root system. K⁺ concentrationsin the xylem sap reaching individual leaves increased towardsthe apex, while concentrations of Na⁺, NO₃^–, and Cl^–declined. Phloem exudate was obtained by collecting into Li₂EDTAfrom the base of excised leaves. K/Na ratios of phloem exudatesincreased from older to younger leaves. K/Na ratios in xylem sap and phloem exudate were combined withchanges in ion content between two harvests (38 and 45 d aftergermination) and the direction of phloem export from individualleaves, to construct an empirical model of K⁺ and Na⁺ net flowswithin the xylem and phloem of the whole plant. This model indicatesthat in old leaves, phloem export of K⁺ greatly exceeded xylemimport. In contrast, Na⁺ export was small compared to importand Na⁺ once imported was retained within the leaf. The direction of export strongly depended on leaf age. Old,basal leaves preferentially supplied the root, and most of theK⁺ retranslocated to the roots was transferred to the xylemand subsequently became available to the shoot. Upper leavesexported to the apex. Young organs were supplied by xylem andphloem, with the xylem preferentially delivering Na⁺ , and thephloem most of the K⁺ . For the young ear, which was still coveredby the sheath of the flag leaf, our calculation predicts phloemimport of ions to such an extent that the surplus must havebeen removed by an outward flow in the xylem. Within the culm,indications for specific transfers of K⁺ and Na⁺ between xylemand phloem and release or absorption of these ions by the tissuewere obtained. The sum of these processes in stem internodes and leaves ledto a non-uniform distribution of Na⁺ and K⁺ within the shoot,Na⁺ being retained in old leaves and basal stem internodes,and K⁺ being available for growth and expansion of young tissues. Key words: Hordeum vulgare L., K⁺, Na⁺, stem, salt stress     

Dependence of Na+-K+ pump current-voltage relationship on intracellular Na+, K+, and Cs+ in rabbit cardiac myocytes

To examine effects of cytosolicNa⁺, K⁺, and Cs⁺ on the voltagedependence of the Na⁺-K⁺ pump, we measuredNa⁺-K⁺ pump current (I_p)of ventricular myocytes voltage-clamped at potentials(V_m) from 100 to +60 mV. Superfusates weredesigned to eliminate voltage dependence at extracellular pump sites.The cytosolic compartment of myocytes was perfused with patch pipette solutions with a Na⁺ concentration ([Na]_pip)of 80 mM and a K⁺ concentration from 0 to 80 mM or withsolutions containing Na⁺ in concentrations from 0.1 to 100 mM and K⁺ in a concentration of either 0 or 80 mM. When[Na]_pip was 80 mM, K⁺ in pipette solutionshad a voltage-dependent inhibitory effect on I_pand induced a negative slope of theI_p-V_m relationship. Cs⁺ in pipette solutions had an effect onI_p qualitatively similar to that ofK⁺. Increases in I_p with increasesin [Na]_pip were voltage dependent. The dielectriccoefficient derived from[Na]_pip-I_p relationships at thedifferent test potentials was 0.15 when pipette solutions included 80 mM K⁺ and 0.06 when pipette solutions were K⁺ free.

     

Functional characterization of the neuronal-specific K-Cl cotransporter: implications for [K+]o regulation

The neuronal K-Cl cotransporter isoform (KCC2) was functionallyexpressed in human embryonic kidney (HEK-293) cell lines. Two stablytransfected HEK-293 cell lines were prepared: one expressing anepitope-tagged KCC2 (KCC2-22T) and another expressing theunaltered KCC2 (KCC2-9). The KCC2-22T cells produced aglycoprotein of ~150 kDa that was absent from HEK-293 control cells.The ⁸⁶Rb influx in both cell lineswas significantly greater than untransfected control HEK-293 cells. TheKCC2-9 cells displayed a constitutively active⁸⁶Rb influx that could beincreased further by 1 mMN-ethylmaleimide (NEM) but not by cellswelling. Both furosemide [inhibition constant (K_i) ~25µM] and bumetanide (K_i~55 µM) inhibited the NEM-stimulated ⁸⁶Rb influx in the KCC2-9cells. This diuretic-sensitive⁸⁶Rb influx in theKCC2-9 cells, operationally defined as KCC2 mediated, required external Clbut not external Na⁺ and exhibiteda high apparent affinity for externalRb⁺(K⁺)[Michaelis constant(K_m) = 5.2 ± 0.9 (SE) mM; n = 5] but alow apparent affinity for externalCl(K_m >50 mM). Onthe basis of thermodynamic considerations as well as the unique kineticproperties of the KCC2 isoform, it is hypothesized that KCC2 may servea dual function in neurons: 1) themaintenance of low intracellularCl concentration so as toallow Cl influx vialigand-gated Cl channelsand 2) the buffering of externalK⁺ concentration([K⁺]_o) in the brain.

     

Shoot-Dependent Regulation of Sodium and Potassium Fluxes in Roots of Whole Barley Seedlings

Unidirectional fluxes and the cytoplasmic and vacuolar contentsof potassium and sodium in root cells of intact barley seedlings(Hordeum vulgare L., cv. Villa) were determined by use of compartmentalanalysis. In addition, the net vacuolar accumulation J_cv andthe xylem transport ø_cx of K⁺ and Na⁺ were measured.Both of these data were needed for the evaluation of the effluxdata. Fluxes and compartmental contents of K⁺ and Na⁺ were comparableto data obtained with excised roots. The effect of the shoot-to-rootratio—as varied by partial excision of the seedlings seminalroots—on the fluxes and contents was investigated. Highershoot-to-root ratios induced an increase in xylem transport,in plasmalemma influx, and also in the cytoplasmic content ofK⁺ and Na⁺. With potassium the plasmalemma efflux was almostunaltered while the tonoplast fluxes and vacuolar content weredecreased (in presence of Na⁺). With sodium, on the other hand,the plasmalemma efflux and the tonoplast fluxes were also increasedin the plants having one root and a high shoot-to-root ratio.These changes occurred even under conditions of low humidity,when transpiration was low and guttation occurred. The latterwas also increased at the high shoot-to-root ratio. The observedchanges could be due to a relieved feedback control of ion fluxesby the shoot and mediated in part by a relatively higher supplyof photosynthates in the plants having one root In addition,hormonal signals were suggested to participate. In particulara possibly decreased level of cytokinins in the plants havingonly one root could contribute to the signal. The observed changesappear to be responses of the plant to an alteration that canoccur under natural conditions when the root system is damaged.     

Sodium gradient-dependent transport of magnesium in rat ventricular myocytes

Cytoplasmic concentration of Mg²⁺([Mg²⁺]_i) was measured with a fluorescentindicator furaptra in ventricular myocytes enzymatically dissociatedfrom rat hearts (25°C). To study Mg²⁺ transport acrossthe cell membrane, cells were treated with ionomycin inCa²⁺-free (0.1 mM EGTA) and high-Mg²⁺ (10 mM)conditions to facilitate passive Mg²⁺ influx. Rate of riseof [Mg²⁺]_i due to the net Mg²⁺influx was significantly smaller in the presence of 130 mMextracellular Na⁺ than in its absence. We also tested theextracellular Na⁺ dependence of the net Mg²⁺efflux from cells loaded with Mg²⁺. After[Mg²⁺]_i was raised by ionomycin and highMg²⁺ to the level 0.5-0.6 mM above the basal value(~0.7 mM), washout of ionomycin and lowering extracellular[Mg²⁺] to 1.2 mM caused rapid decline of[Mg²⁺]_i in the presence of 140 mMNa⁺. This net efflux of Mg²⁺ was completelyinhibited by withdrawal of extracellular Na⁺ and waslargely attenuated by imipramine, a known inhibitor of Na⁺/Mg²⁺ exchange, with 50% inhibition at 79 µM. The relation between the rate of net Mg²⁺ efflux andextracellular Na⁺ concentration([Na⁺]_o) had a Hill coefficient of 2 and[Na⁺]_o at half-maximal rate of 82 mM. Theseresults demonstrate the presence of Na⁺ gradient-dependentMg²⁺ transport, which is consistent withNa⁺/Mg²⁺ exchange, in cardiac myocytes.

     

Enhancing effects of transition metals on the salt taste responses of single fibers of the frog glossopharyngeal nerve: specificity of and similarities among Ca2+, Mg2+ and Na+ taste responses

Fibers of the frog glossopharyngeal nerve (water fibers) thatare sensitive to water also respond to CaCl₂, MgCl₂ and NaCl.In the present study, interaction among cations (Ca²⁺, Mg²⁺and Na⁺) on taste cell membrane in frogs was studied using transitionmetals (NiCl₂, CoCl₂ and MnCl₂), which themselves are barelyeffective in producing neural response at concentrations below5 mM. Unitary discharges from single water fibers were recordedfrom fungiform papillae with suction electrode. Transition metalions (0.05–5.0 mM) had exclusively enhancing effects onthe responses to 50 mM Ca²⁺, 100 mM Mg²⁺ and 500 mM Na⁺. Theeffects of transition metal ions were always reversible. Therank order of effectiveness of transition metals at 1 mM inthe enhancement of the responses to 50 mM CaCl₂, 100 mM MgCl₂and 500 mM NaCl was NiCl₂ > CoCl² > MnCl₂. The concentrationof transition metal ions effective to enhance salt responsewas almost the same among Ca²⁺, Mg²⁺ and Na⁺ responses. Theresults suggest that a common mechanism is involved in the enhancementof Ca²⁺, Mg²⁺ and Na⁺ taste responses. The enhanced Mg²⁺ responseand the enhanced Na⁺ response were greatly inhibited by theaddition of Ca²⁺ ions, and the enhanced Ca²⁺ response was inhibitedby the addition of Mg²⁺ or Na⁺ ions, suggesting that competitiveantagonism occurs between Ca²⁺ and Mg²⁺ ions and between Ca²⁺and Na⁺ ions in the presence of Ni²⁺ ions. Ni²⁺ ions had a dualeffect on the Ca²⁺ response induced by low concentration (0.1mM) of CaCl₂: enhancement at lower concentrations (0.02–0.1mM) of NiCl₂ and inhibition at higher concentrations (0.5–5mM)of NiCl₂. The present results suggest that transition metalions do not affect the receptor-antagonist complex, but affectonly the receptor-agonist complex.     

Measurements of Ion Concentrations and Fluxes in Dunaliella parva

Measurements of K⁺, Na⁺, and Cl^– were made on a halotolerantstrain of Dunaliella growing at 500 mM NaCl, 25 ?C, and a relativelylow light intensity (6000 Lx). Much effort was spent in searchingfor a means of measuring the extracellular volume of fluid trappedbetween the cells of centrifuged pellets. All of the sugarstried as markers were rejected because they were found to bedigested in the cell suspension. The most suitable marker wasfound to be [¹⁴C]polyethylene glycol² (mol. wt. 4000); althoughthis substance was apparently adsorbed to the cell exterior,it was found possible to correct for adsorption and then obtaina reasonable figure for the trapped fluid. The final concentrationsof cell K⁺ and Na⁺ were 128 ? 53 mM and 131 ? 117 mM respectively.Cl^– balanced the sum of K⁺ Na⁺. Influxes of ²²Na⁺, ⁴²K⁺,and ³⁶C1^– were measured in cells in which the ions werein the steady state. Averages of 610 and 6.6 nmol m^–2s^–1 were obtained for Na⁺ and K⁺ respectively. Cl^–influx was divided into 2 phases with values of 1540 and 178mmol m^–2 s^–1. The faster influx was considered tobe across the outer cell membrane. The membrane responsiblefor the slower influx has not been identified. By comparingvalues of the potential difference calculated from the Nernstand Goldman equations, it was concluded that Na⁺ and K⁺ areprobably controlled by active mechanisms, whereas cell Cl^–is likely to be at thermodynamic equilibrium with the medium.     

Potassium Transport in Non-Growing Corn Root Tissue as Affected by IAA and GA3

Experiments were done to determine if the spontaneous recoveryof non-growing segments of corn root (Zea mays L.) from excisioninjury is dependent on auxin. Washing the segments with 5 runindoleacetic acid (IAA) for 2 to 4 hours gave a small but significantincrease in K⁺ (⁸⁶Rb) influx, used here as a parameter reflectingrecovery of electrogenie H⁺-efflux pumping. This promotive effectwas obtained only after an hour of washing, and was sustainedby 100 nm gibberellic acid (GA₃). Any early responses to auxinwere obscured by an adverse reaction of the root cells to externalIAA which resulted in a transitory inhibition of H⁺ pumpingand K⁺ influx. Pretreatment of excised root tips with 10 µM IAA in thegrinding medium protected a plasmalemma-enriched fraction ofthe microsomes during isolation, giving increased uncoupler-sensitiveATPase activity. Non-growing root tissue thus shows three responses to auxin:an adverse reaction at the outer surface of the plasmalemmawhich blocks H⁺ pumping; a protective or restorative effecton the H⁺-ATPase; an increased capacity for K⁺ influx duringthe developmental phase of washing, which is augmented by thepresence of GA₃. (Received March 31, 1986; Accepted September 8, 1986)