Na+, K+ and Cl- in Xylem Sap Flowing to Shoots of NaCl-Treated Barley

Munns, R. 1985. Na⁺, K⁺ and Cl^– in xylem sap flowing toshoots of NaCl-treated barley.—J. exp. Bot. 36: 1032–1042. Na⁺, Cl^– and K⁺ concentrations were measured in xylemsap obtained by applying pressure to the roots of decapitatedbarley plants grown at external [NaCl] of 0, 25, 50, 100, 150and 200 mol m^–3. For any given NaCl treatment, ion concentrationsin the xylem sap were hyperbolically related to the flux ofwater. Ion concentrations in sap collected at very low volumefluxes (without applied pressure) were 5–10 times higherthan in sap collected at moderate fluxes (under pressure). Fora given moderate volume flux, Na+ concentration in the xylemsap, [Na⁺]_x, was only 4.0 mol m^–3 at external [NaCl] of25–150 mol m^–3, and increased to 7.0 mol m^–3at 200 mol m^–3. [Cl-]x showed a similar pattern. Thisshows there would be little difference in the rate of uptaketo the shoot of plants at 25–150 mol m^–3 externalNaCl and indicates little change even at 200 mol m^-3 NaCl becausetranspiration rates would be much lower. Thus the reduced growthof the shoot of plants at high NaCl concentrations is not dueto higher uptake rates of Na⁺ or Cl^–. The fluxes of Na⁺, Cl^– and K^– increased non-linearlywith increasing volume flux indicating little movement of saltin the apoplast. The flux of K⁺ increased even when [K⁺]_x wasgreater than external [K⁺], indicating that membrane transportprocesses modify the K⁺ concentration in the transpiration streamas it flows through the root system. Key words: -Xylem sap, Na⁺, K⁺, Cl^– fluxes, salinity, barley     

Nitrate Inhibition of Chloride Influx in Barley: Implications for a Proposed Chloride Homeostat

Net accumulation of Cl^– by intact barley plants was virtuallyeliminated in roots and reduced by 40% in shoots when externalmedia (0.5 mol m^–3 CaSO₄ plus 0–5 mol m^–3KCI) were supplemented with 0.25 mol m^– Ca(NO₃)₂. Plasmalemma³⁶Cl^– influx (_oc) was shown to be insensitive to externalNO₃^- in plants which had previously been grown in solutionslacking ^–3, but _oc became sensitive to NO₃^-after a lagperiod of 3–6 h. Kinetic analyses revealed that the inhibitionof 36C1^– influx by external NO₃^- was complex. At 0.25mol m^–3 NO₃^- the V_max for Cl^– influx was reducedby greater than 50%, with insignificant effects upon K_m. At0.5 mol m^–3 NO₃^- there was no further effect upon V_maxbut K_m for influx increased from 38±5 mmol m^–3to 116±26 mmol m^–3. By contrast, Cl^– effluxwas found to be insensitive to external NO₃^-. A model for theregulation of Cl^– influx is proposed which involves bothnegative feedback effects from vacuolar NO₃^- +Cl^–) concentrationand (external) NO₃^- inhibition of Cl^– influx at the plasmalemma.These combined effects serve to discriminate against Cl^–accumulation, favouring NO₃^- accumulation, when the latter ionis available. Such observations are inconsistent with recentproposals for the existence of bona fide homeostats for chlorideaccumulation in higher plants. Key words: Nitrate inhibition, Chloride influx, Barley     

Further Characteristics of Salt-Dependent Bicarbonate Use by the Seagrass Zostera muelleri

Millhouse, J. and Strother, S. 1987. Further characteristicsof salt-dependent bicarbonate use by the seagrass Zostera muelleri.—J.exp. Bot. 38: 1055–1068. The contribution of HCO^–₃to photosynthetic O₂ evolutionin the seagrass Zostera muelleri Irmisch ex Aschers. increasedwith increasing salinity of the bathing seawater when the inorganiccarbon concentration was kept constant. K_1/2 (seawater salts)for HCO^–₃ -dependent photosynthesis was 66% of seawatersalinity. Both short- and long-term pretreatment at low salinitiesstimulated photosynthesis in full strength seawater. Twentyfour hours pre-incubation of seagrass plants in 3·0 molm^–3 NaHCO₃ resulted in increased photosynthesis at allsalinities, apparently due to stimulation of HCO^–₃ use(K_1/2 (seawater salts) = 26%). V_max (HCO^–₃) was not affectedby low salinity pretreatment. The kinetics of HCO^–₃ stimulationby the major seawater cations was investigated. Ca²⁺ was themost effective cation with the highest V_max (HCO^–₃) andwith K_1/2(Ca²⁺) = 14 mol m^–3. Mg²⁺ was also very effectiveat less than 50 mol m^–3 but higher concentrations wereinhibitory. This inhibition cannot be accounted for solely byprecipitation of MgCO₃. Na⁺ and K⁺ were both capable of stimulatingHCO^–₃ use. Stimulation was in two distinct parts. Up to500 mol m^–3, both citrate and chloride salts gave similarresults (K_1/2(Na⁺) 81 mol m^–3, V_max(HCO^–₃) 0·26µmol O₂ mg^–1 chl min^–1), but use of citratesalts above 500 mol m^–2 caused a second stimulation ofHCO^–₃ use (K_1/2(Na⁺) 830 mol m^–3, V_max(HCO^–₃)0·68 µmol O₂ mg^–1 chl min^–1). V_max(HCO^–₃)for the second-phase Na⁺ or K⁺ stimulation was of the same orderas for Ca²⁺-stimulated HCO^–₃ use. To further characterizesalt-dependent HCO^–₃ use, the sensitivity of photosynthesisto Tris and TES buffers was investigated. The effects of Trisappear to be due to the action of Tris⁺ causing stimulationof HCO^–₃ -dependent photosynthesis in the absence of salt,but inhibition of HCO^–₃ use in saline media. TES has noeffect on photosynthesis. External carbonic anhydrase, althoughimplicated in salt-dependent HCO^–₃ use in Z. muelleri,could not be detected in whole leaves. Key words: Zostera muelleri, HCO^–₃ use, salinity     

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^–     

Potassium Transport in Enteromorpha intestinalis (L.) Link: MEASUREMENT OF INTRACELLULAR K+, EXCHANGE FLUXES AND THERMODYNAMIC ANALYSIS

Potassium transport has been studied in the marine euryhalinealga, Enteromorpha intestimlis cultured in seawater and in low-salinitymedium (Artificial Cape Banks Spring Water, ACBSW; 25·5mol m^–3 Cl^–, 20·4 mol m^–3 Na⁺, 0·5mol m^–3 K⁺). K⁺ fluxes were measured using ⁴²K⁺ and ⁸⁶Rb⁺although ⁸⁶Rb⁺ does not act as an efficient K⁺ analogue in thisplant. ⁴²K⁺ experiments on seawater plants typically exhibiteda single protoplasmic exchange phase whereas ⁸⁶Rb⁺ exhibitedtwo exchange phases. Compartmental analysis of ⁸⁶Rb⁺ effluxexperiments on seawater-grown Enteromorpha plants were usedto deduce the intracellular partition of K⁺ between the cytoplasm(279±38 mMolal) and vacuole (405±68 mMolal). Theplasmalemma K⁺ flux in plants in seawater was greater in thelight than in the dark (563±108 nmol m^–2 s^–1versus 389±66·7 nmol m^–2 s^–1). Inlow-salinity plants, separate cytoplasmic and vacuolar exchangephases were apparent. Analysis of ⁴²K⁺ efflux experiments onlow-salinity plants yielded a cytoplasmic K⁺ of 222±38mMolal and a vacuolar K⁺ of 82±11 mMolal. The plasmalemmaand tonoplast flux was 23±4·5 nmol m^–2 s^–1. The Nernst equation showed that, although K⁺ was close to electrochemicalequilibrium, active accumulation of K⁺ across the plasmalemmaoccurred in plants in seawater and ACBSW both in the light anddark. K⁺ was also actively transported inwards across the tonoplastin low-salinity plants. The electrochemical potential for K⁺across the plasmalemma ranged from 2·41±0·60kJ mol^–1 in plants grown in seawater in the light to 5·79±0·87kJ mol^–1 for plants in ACBSW in the light. Although K⁺is close to electrochemical equilibrium, the flux of K⁺ in plantsin both seawater and ACBSW media is high, hence the power consumptionof K⁺ transport is high. The permeability of K⁺ (P_K⁺) was significantlyhigher in the light than in the dark in plants in seawater (about7·0 versus 2·5 nm s^–1) but in plants inlow-salinity (ACBSW) medium the permeability was independentof light (about 12 nm s^–1). The energy requirements ofactive K⁺ transport by ATP-dependent pumps is discussed. Key words: Enteromorpha, Potassium transport, Ionic relations, Saltwater, Low salinity, Thermodynamics     

Ontogenetic Changes in the Chemical Composition of Ricinus communis Grown with NO-3 or NH+4 as N Source

Ricinus communis L. var. Gibsonii was grown in Long Ashton nutrientmedium with either 12mol m^–3 NO^–₃ or 8.0 mol m^–3NH⁺₄ as N source. Two plants from each N treatment were harvestedtwice a week and analysed for C, N, P, S, NO^–₃, SO^2–₄Cl^–K⁺Na⁺, Ca²⁺ Mg²⁺ and ash alkalinity. Statistical analysis of thedata showed that the effect of age and N source was differentfor the chemical variables analysed. Thus [Na⁺] was unaffectedby age or N source, and for both N sources [Mg²⁺] started atthe same level and decreased at the same rate as the plantsmatured. With NH⁺₄ as N source, [SO^2–₄] was higher thanwith NO^–₃, but did not alter with age. The concentrations,in mmol g^–1 dry wt, of C, organic N, K⁺ and Ca²⁺ weredifferent for the two N sources, but the levels of these variablesaltered with age in the same way for both N sources; i.e. therewas no age x N interaction. In the case of P, NO^–₃, Cl^– and COO^–, however,age-related variations were different for the two N sources.It is concluded, inter alia, that [Na⁺] is determined by external[Na⁺] alone, and that K⁺, Ca²⁺ and Cl^– are the inorganicions actively involved in charge balance during ion uptake bythe roots. Key words: Ontogeny, Chemical composition, Plant nutrition     

Salinity-induced Malate Accumulation in Chara

Ion absorption by Chara corallina from solutions containingpredominantly KC1 or RbCl at up to 100 mol m^–3 resultedin accumulation of salts and turgor regulation. Turgor regulationdid not occur in solutions containing Na⁺ or Li⁺salts. Duringion absorption from various salts of K⁺ and Rb⁺ vacuolar cationconcentration exceeded Cl^– concentration. This differencewas shown to be balanced by the synthesis and accumulation ofmalate. Vacuolar malate concentration reached 48 mol m₃^–,with accumulation occurring at rates of up to 0.45 mol m^–3h^–1. Malate accumulation was inhibited by low externalpH and was dependent upon external HCO₃^– concentration.The synthesis of malic acid and its subsequent dissociationimposed a severe acid load on the cell. Biophysical regulationof cellular pH was achieved by a H⁺efflux at a rate of about40 nmol m^–2 s^–1from the cell. The results presentedargue against cytoplasmic Cl^–, HCO₃^– or pH regulatingmalate accumulation in Chara and it is suggested that malatetransport across the tonoplast may regulate malate accumulation. Key words: Malate, Chara corallina, pH regulation, salinity     

External Potassium Supply is not Required for Root Growth in Saline Conditions: Experiments with Ricinus communis L. Grown in a Reciprocal Split-Root System

Ricinus communis L. (castor bean) plants were grown in the absence(control) and in the presence of 100molm^–3NaCl with areciprocal split-root system, in which K⁺ was supplied to oneand NO₃^– to the other part of the root system. In theseplants shoot and, to a lesser extent, total root growth wereinhibited compared to plants with non-split roots. Without andwith NaCl, growth of roots receiving NO₃^– but noK⁺ (‘minusK/plus N-roots’) was substantially more vigorous thanunder the reverse conditions (‘plus K/minus N-roots¹).100mol m^–3 NaCl inhibited growth of minus K/plus N-roots¹to the same extent as that of non-split roots, indicating thatexternally supplied K⁺ was not required for root growth undersaline conditions. In growth media without added K⁺ the rootdepleted the external low K ⁺ levels resulting from chemicalsdown to a minimum value C_mln (1.0 to 1.4 mmol m^–3); inthe presence of 100 mol m^–3 NaCl, C_min, was higher (10–18mmol m^–3) and resulted from an initial net loss of K ⁺.C_min, was pH-dependent The distribution of K⁺, Na⁺ and Mg²⁺along the root was measured. In meristematic root tissues, K⁺ concentrations were scarcely affected by external K⁺ or byNaCl, where Na ⁺ concentrations were low, but somewhat elevatedat low external K+ and/or high NaCl. In differentiated, vacuolatedtissues K ⁺ concentrations were low and Na⁺ concentrations high,if K ⁺ was not supplied externally and/or NaCl was present.The longitudinal distribution of ions within the root was usedto estimate cytoplasmic and vacuolar ion concentrations. Thesedata showed a narrow homoeostasis of cytoplasmic K+ concentrations(100–140 mol m^–3) independent of external K ⁺ supplyeven in the presence of 100 mol m ^–3 NaCl. CytoplasmicNa ⁺ concentrations were maintained at remarkably low levels.Hence, external K⁺ concentrations above C_min, were not requiredfor maintaining K/Na selectivity, i.e. for controlling Na⁺ entry.The results are discussed with regard to mechanisms of K/Naselectivity and to the importance of phloem import of K⁺ forsalt tolerance of roots and for cytoplasmic K⁺ homoeostasis. Key words: Ricinus communis, nitrate, potassium, root (split-root), salt tolerance, phloem transport     

Potassium-Ammonium Uptake Interactions in Tobacco Seedlings

Short-term (< 12 h) uptake experiments were conducted with6–7-week-old tobacco (Nicotiana tabacum L. cv. Ky 14)seedlings to determine absorption interactions between K⁺ andNH₄⁺. At equal solution concentrations (0.5 mol m^–3) netK⁺ uptake was inhibited 30–35% by NH₄⁺ and NH₄⁺ uptakewas decreased 9–24%. Removal of NH₄⁺ resulted in completerecovery in K⁺ uptake rate, but NH⁴⁺ uptake rate did not recoverwhen K⁺ was removed. In both cases, inhibition of the uptakerate of one cation saturated as the concentration of the othercation was increased up to 0.5 mol m^–3. The relative effectof K⁺-NH₄⁺ interactions was not altered when Cl- was replacedwith SO₄^2–, but the magnitudes of the uptake rates wereless in the absence of Cl-. The V_max for NH₄⁺ uptake was reducedfrom 128 to 105 µmol g^–1 dry wt. h^–1 in thepresence of 0.5 mol m^–3 K⁺ and the K_m for NH₄⁺ doubledfrom 12 to 27 mmol m^–3 in the presence of K⁺. The resultsof these K⁺-NH₄⁺ experiments are interpreted as mixed-noncompetitiveinteractions. However, an enhanced efflux of K⁺ coupled to NH₄⁺influx via an antiporter cannot be ruled out as contributingto the decrease in net K⁺ uptake. Key words: Nicotiana tabacum, K⁺, NH₄⁺, Uptake interactions     

Nitrogen-13 Studies of Nitrate Fluxes in Barley Roots: II. EFFECT OF PLANT N-STATUS ON THE KINETIC PARAMETERS OF NITRATE INFLUX

Influx of nitrate into the roots of intact barley plants wasfollowed over periods of 1–15 min using nitrogen-13 asa tracer. Based on measurements taken over 15 min from a rangeof external nitrate concentrations (0·2–250 mmolm^–3), the kinetic parameters of influx, I_max and K_m, werecalculated. Compared with plants grown in the presence of nitrate throughout,plants that had been starved of N for 3 d showed a significantlygreater value ofI_max for ¹³N-nitrate influx (by a factor of1·4–1·8), but a similar value of K_m (12–14mmol m^–3). Pre-treating N-starved plants with nitratefor about 5 h further increased the subsequent rate of ¹³N-nitrateinflux, but had little effect in the unstarved controls. Allowingfor this induction of additional nitrate transport, the differencein rates of nitrate influx in control and N-starved plants wassufficient to account for the previously-observed differencein net uptake by the two groups of plants. In barley plants grown without any exposure to nitrate, butwith ammonium as N-source, both I_max and K_m for subsequent ¹³N-nitrateinflux were significantly decreased (by about one-half) comparedwith the corresponding nitrate-grown controls. The importance of changes in the rate of influx in the regulationof net uptake of nitrate is discussed. Key words: Ion transport, nitrate, influx, kinetic parameters, N-deficiency     

Response of Wheat Seedlings to Low O2 Concentrations in Nutrient Solution: II. K +/Na+ SELECTIVITY OF ROOT TISSUES9

This paper reports the effects of low O₂ concentration (0–01,0–055, and 0.115mol m^–3) in nutrient solutions onK⁺/Na⁺ selectivity of growing and mature root tissues of 6-to 8-d-old, intact, wheat (Triticum aestivum cv. Gamenya) seedlings. Increases in anaerobic catabolism and decreases in O₂ uptake,K⁺ uptake and K⁺/Na⁺ selectivity were all more pronounced and/oroccurred at higher external O₂ concentrations in the apex (0–2mm) than in the expanding tissues (2–4 mm); these growingtissues were, in turn, more affected than the expanded tissuesof the roots (4–12 mm). Selectivity for K⁺ over Na⁺ in roots and shoots was particularlysensitive to O₂ deficiency. For example, in apical tissues (0–2mm) K ⁺ /Na⁺ selectivity was already reduced at 0.115 mol m^–3O₂, yet at this O₂ concentration there was no effect on eithergrowth or (K⁺/Na⁺) uptake. Upon transfer from 0.01 to 0.26 mol m^–3 O₂, a detailedstudy of the 12 mm root tips showed that 70% of these tips regainedhigh (K⁺ + Na⁺) concentrations and K⁺/^Na+ ratios. In contrast,there was no recovery in the remaining 30% of the 12 mm roottips. Net K⁺ transport to the shoots during the period afterre-aeration was negative for the population as a whole. Theseverity of these effects supports the view that the root tipsand the stele were more susceptible to O2 deficiency than wasthe cortex of the fully-developed root tissues. Key words: Hypoxia, K⁺/Na⁺ selectivity, expanded and expanding tissues     

Potassium Transport Across the Membranes of Chara: I. THE RELATIONSHIP BETWEEN RADIOACTIVE TRACER INFLUX AND ELECTRICAL CONDUCTANCE

Smith, J. R., Smith, F. A. and Walker, N. A. 1987. Potassiumtransport across the membrane of Chara. I. The relationshipbetween radioactive tracer influx and electrical conductance.—J.exp. Bot. 38:731–751. The ⁴²K influx () and the electrical conductance (G_m) were measured simultaneously for the ‘membrane’of internodal cells of Chara australis as a function of theexternal [KCl] (K?. In bathing solutions of pH = 5?0, progressively increased from 20?5to 430?60 nmol m^–2 s^–1 and G_m increased from 0?36?0?02to 3?8?0?8 S m^–2 when K? was increased from 0?1 to 10mol m^–3. The resting membrane potential difference (p.d.)was approximately -135 mV for low K? and approached the expectedNernst equilibrium p.d. for K⁺ ions when K? > 1?0 mol m^–3.Measurements of ³⁶Cl influx suggested that the ⁴²K influx waspredominantly electrogenic. The equivalent Goldman permeabilityto K⁺ ions (P_k) was approximately 20–30 nm s^–1 anddid not vary significantly with increasing K?. The equivalentconductance attributable to the electrogenic transport of K⁺ ions was calculated from assuming passive, independent diffusionof K⁺ ions and the ratio was found to be typically close to one. It was also found that themagnitudes of and G_m measuredsimultaneously for each individual cell were also well correlatedfor K? 1?0 mol m^–3, and that the slope of the line ofbest fit was close to one. For each K? it was found that theconductance not attributable to K⁺ translocation and presumablyassociated primarily with the transport of protons or theirequivalents was typically 0?2–0?5 Sm^–2. For K? >1?0 mol m^–3 the results indicated that the transport ofK⁺ ions was essentially independent, i.e. there was no evidencefor flux interactions. The results also indicated that the equivalentconductance derived from the measured ⁴²K influx could usefullyindicate the fraction of the electrical conductance attributableto the translocation of K⁺ ions. Key words: Potassium, conductance, influx     

Effect of Divalent Cations on Na+ Permeability of Chara corallina and Freshwater Grown Chara buckellii

The effect of elevated Na⁺ concentration on Na⁺ permeability(P_Na) and Na⁺ influx in the presence of two levels of externaldivalent cations was determined in Chara corallina and freshwater-culturedChara buckellii. When Na⁺ in the medium was increased from 1.0to 70 mol m^–3, Na⁺ influx increased in both species ifCa²⁺ was low (0.1 mol m–3). If Ca²⁺ was increased to 7.0mol m^–3 when Na⁺ was increased, Na⁺ influx remained atthe low control level in C. corallina, and showed only a temporaryincrease in C. buckellii. Mg²⁺ was a better substitute for Ca²⁺in C. buckellii than in C. corallina. Na⁺ permeability data suggest that when the external Ca²⁺ concentrationis low, P_Na does not increase in the presence of elevated NaCl;the increase in Na⁺ influx appears to be due to the increasein external Na⁺ concentration alone. Ca^{2 +} supplementation appearsto decrease P_Na whereas supplemental Mg²⁺ has no effect. Na⁺ effluxes were computed from previously determined net fluxesand the influxes. It was found that for both species, fluxesin both directions were stimulated in response to all experimentaltreatments, but Na⁺ influx always exceeded efflux. This resultedin net Na⁺ accumulation in the vacuoles of both species. The results are discussed with reference to net flux and electrophysiologicaldata obtained previously under identical conditions, as wellas the comparative salinity tolerance of both species and theNa⁺/divalent cation ratio. Key words: Na⁺ influx, Na⁺ tolerance, membrane potential, permeability, Chara     

Diurnal K+ and Anion Transport in Phaseolus Pulvinus

Diurnal movement of Phaseolus leaf is caused by deformationof the laminar pulvinus located at the joint of the leaf bladeand the petiole. The plants were cultured in solutions withvarious ion compositions, and changes of K⁺, Na⁺, Ca²⁺, Mg²⁺,Cl^–, NO₃– and P₁ concentrations both in the upperand lower parts of the laminar pulvinus were measured. Culturein 10 mM KCl solution caused an increase in K⁺ and Cl^–concentrations both in the upper and lower parts without anysignificant change in the concentration of NO₃^–; culturein 10 mM KNO₃ solution caused an increase in K⁺ and NO₃^–concentration without any significant change in the concentrationof Cl^–; and culture in 10 mM KH₂PO₄ solution caused anincrease in K⁺ and P₁ concentrations without any significantchange in the concentrations of NO₃- and Cl^–. K⁺ moved from the upper to lower parts or from the lower toupper parts diurnally in all plants cultured in any solutionmentioned above. The main inorganic anion that accompanied thisK⁺ movement was Cl^– in KCl solution, and NO₃^– inKNO₃ solution. When the seedlings were cultured in distilledwater or in KH₂PO₄ solution, neither Cl^– NO₃^– norP₁ accompanied this K⁺ movement. In these cases, mainly H⁺ and/ororganic anions are supposed to move in exchange for and/or incombination with K⁺ movement. (Received November 8, 1982; Accepted June 13, 1983)     

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.     

Chloride Transport in Chara: I. KINETICS AND CURRENT-VOLTAGE CURVES FOR A PROBABLE PROTON SYMPORT

Chara cells show an inward positive electric current acrossthe plasmalemma when exposed to Cl^– under voltage-clampconditions. The rapid rise of this current suggests that itis directly associated with the inward transport of Cl^–.The dependence of the current on Cl^– concentration showssaturation, the data fitting the Michaelis-Menten equation withV_m up to 100 nmol m^–2 s^–1 (for Cl^–starvedcells) with K_M 10–20 µM, and with some allowancefor an unstirred layer of water adjacent to the membrane. Theeffects on the current of clamp potential, illumination, withdrawalof alkali metal cations, and addition of amine were also investigated.These results suggest that the mechanism is the symport of 2H⁺ with each Cl^–, and that the actions of light, externalK⁺, and amine in stimulating Cl^–, influx are indirect.     

The Ionic Relations of Acetabularia mediterranea

The concentrations of K⁺, Na⁺, and Cl^– in the cytoplasmand the vacuole of Acetabularia mediterranea have been measured,as have the vacuolar concentrations of SO₄^–– andoxalate. The electrical potential difference between externalsolution, and vacuole and cytoplasm has been measured. The resultsindicate that Cl^– and SO₄^–– are probably transportedactively into the cell, and that active transport of Na⁺ isoutwards. The results for K⁺ are equivocal. The fluxes of K⁺,Na⁺, Cl^–, and S0₄^–– into the cell and theeffluxes of Na⁺ and Cl^– have been determined. The Cl^–fluxes are extremely large. In all cases the plasmalemma isthe rate-limiting membrane for ion movement. A technique isdescribed for the preparation of large, completely viable cellfragments containing only cytoplasm, with no vacuole.     

Possible reasons for relative salt stress tolerance in nodules of white lupin cv. Multolupa

The effects of different NaCl concentrations on the growth andnitrogen fixation activity of white lupin (Lupinus albus [L.])was studied over a 6 d period. Plant growth parameters, photosynthesisand shoot respiration were unaffected by NaCl concentrationsup to 150 mol m^–3. However, nitrogenase activity decreasedwith increased NaCl concentration up to 100 mol m_–3, whilstthe O₂ diffusion resistance increased with 100 mol m^–3NaCl, but showed no further change when 150 mol m^–3 NaClwas applied for 6 d. Increases in NaCl concentration decreasednodular starch content while increasing sucrose content, suggestingan osmotic regulation. These changes were associated with a77% decrease in sucrose synthase activity. The effect on theO₂ diffusion resistance was paralleled by changes in glycoproteincontent of the nodules, as determined by immunogold localizationand ELISA. X-ray microanalysis studies of nodules showed that,following a 6 d exposure to 150 mol m^–3 NaCl, Na⁺ ionswere largely excluded from the infected zone, whilst only lowlevels of Cl^- ions penetrated into this region. Na⁺ entry intoroots and leaves was also at a low level. Leghaemoglobin contentdecreased with saline stress, as did superoxide dismutase; whichdecreased by 36% following exposure to 100 mol m^–3 saltfor 6 d. These results are discussed in relation to the relativesalt tolerance of the Multolupa/ISLU-16 symbiosis. Key words: Salt stress, nodules, nitrogen fixation, oxygen diffusion, carbohydrates, Lupinus albus     

Uptake of Imidazole and its Effects on the Intracellular pH and Ionic Relations of Chara corallina

Ammonia (pK_a 9.25) and methylamine (pK_a, 10.65) increase cytoplasmicpH and stimulate Cl^– influx in Chara corallina, theseeffects being associated with influx of the amine cations ona specific porter. The weak base imidazole (pK_a 6.96) has similareffects but diffuses passively into the cell both as an unionizedbase and as a cation. When the external pH is greater than 6.0influx of the unionized species predominates. Imidazole accumulates to high concentrations in the vacuole,where it is protonated. Cytoplasmic pH and vacuolar pH riseby only 0.2–0.3 units, suggesting a large balancing protoninflux across the plasma membrane. Balance of electric chargeis partially maintained by net efflux of K⁺ and net influx ofCl^–. Calculation of vacuolar concentrations of imidazole(from (¹⁴C] imidazole uptake, assuming that there is no metabolism)plus K⁺ and Na⁺ indicates an excess of cations over inorganicanions (Cl^–). However, although the osmotic potentialof the cells increases, also indicating increased solute concentrations,the increase is less than that predicted by the calculated ionicconcentrations. This discrepancy remains to be resolved. Becausethe osmotic potential also increases when imidazole is absorbedfrom Cl^–-free solutions it is likely that maintenanceof charge-balance can also involve synthesis and vacuolar storageof organic or amino acids. Key words: Imidazole, potassium, intracellular pH, membrane transport, Chara     

Influence of Age of Culture and Light Intensity on Solute Concentrations in Two Dunaliella Strains

Two strains of Dunaliella, one halotolerant and one halophilic,were grown in batch culture at NaCl concentrations varying from500 mol m^–3 to 3000 mol m^–3. Measurements were madeof the following solutes: glycerol, Na⁺, K⁺, Mg²⁺, Cl^–,phosphate in the cells of logarithmic and of stationary-phasecultures. The method used was to separate the cells from thebulk of the medium by differential density centrifugation. Soluteconcentrations were calculated using Blue Dextran as a markerfor extracellular space. It was found that in log-phase cells,glycerol accounted for one-half to two-thirds of the total cellsolute concentration, the remainder being largely accountedfor by Na⁺ and Cl^–. In the stationary phase glycerol felland Na⁺, plus Cl^–, rose. Light intensity was found toaffect cell volume and solute content. The means whereby soluteconcentrations are controlled is discussed. Key words: Osmoregulation, Ion concentrations, Dunaliella