Fluxes of Sodium and Potassium in Acetabularia mediterranea

Sodium efflux in Acetabularia mediterranea occurs against agradient of electrochemical potential and is a light-stimulated,temperature-sensitive process; it is not sensitive to the uncouplerCCCP. Sodium influx is stimulated in CCCP and at low temperature.Potassium influx is temperature- and uncoupler-sensitive, butis not light-stimulated. Tracer K efflux shows complex kinetics,which cannot be explained by any arrangement of intracellularcompartments; it appears to be stimulated at low temperatureand is insensitive to light and uncouplers. There is no evidencefor any chemical linkage between fluxes of Na⁺, K⁺, or Cl^–.It is concluded that Na^– efflux at the plasmalemma isan active process, but no consistent explanation can be advancedto account for the results of K⁺ flux measurements.     

Effects of Light/Dark on Anion Fluxes in Isolated Guard Cells of Commelina communis L.

The effects of light/dark on anion fluxes in isolated guardcells of Commelina communis L. have been studied, using ⁸²Brand ³⁶Cl. Transfer of open guard cells from light to dark hasno effect on the ⁸²Br influx, but produces a marked transientstimulation of ⁸²Br or ³⁶Cl efflux, similar to the effect ofsuch transfer on the ⁸⁶Rb fluxes, and to the effects on both⁸⁶Rb and ⁸²Br fluxes of adding ABA. On return of guard cellsto light, after the transient, there is a further reductionin Cl/Br efflux. It is argued that control of a specific processof ion extrusion is important in regulating the ability of guardcells to stay open. In three out of four batches of steady-statetissue labelled with ⁸²Br, the plasmalemma fluxes were highenough, relative to the tonoplast fluxes, for the efflux kineticsto be separable into two exponential components, allowing estimationof bromide contents in cytoplasm and vacuole (Q_c and Q_v), andfluxes at plasmalemma and tonoplast. With opening in light,Q_c increased by 3.9 ± 0.4 pmol mm^–2 µm^–1and Qy by 5.2 ± 0.6 pmol mm^–2 µm^–1(change in content per mm² of epidermis perµm change inaperture). Using rough estimates for the volumes of cytoplasmand vacuole these figures suggest that at 6.1 µm in thedark the concentrations were about 63 mol m^–3 in the cytoplasmand 35 mol m^–3 in the vacuole, rising to about 185 molm^–3 in the cytoplasm and 125 mol m^–3 in the vacuole,at 16.7 µm aperture in light. Neither increase can providean adequate increase in salt concentration to account for theosmotic change required, and some solute other than potassiumsalt must also be involved. In one experiment with 82Br andin the only experiment with 36Cl the plasmalemma flux was lower,and not high enough relative to the tonoplast flux to allowseparation of two phases in the efflux curves, and calculationof cytoplasmic and vacuolar contents and fluxes. The effectsof transfer from light to dark were, nevertheless, similar inboth types of tissue. Key words: Commelina communis L., Light/dark effects, Anion fluxes, Guard cells     

The Effects of Fusicoccin on Anion Fluxes in Isolated Guard Cells of Commelina communis L.

Clint, G. M. 1987. The effects of fusicoccin on anion fluxesin isolated guard cells of Commelina communis L.—J. exp.BoL 38: 863–876. The effects of 3?10^–2 mol m^–3 fusicoccin (FC) onbromide fluxes and contents in isolated guard cells of Commelinacommunis L. have been studied using K⁸²Br at pH 3?9 and pH 6?7.At pH 3?9 FC caused a reduction in both the influx and the effluxof ⁸²Br, whereas at pH 6?7 FC had no effect on the influx butcaused a transient increase in the efflux of ⁸²Br. There wasno obvious change in bromide content with FC treatment at eitherpH. The behaviour of the anion fluxes in response to FC suggeststhat FC does not act solely via a hyperpolarization at the plasmalemma.A redistribution of bromide between the intracellular compartmentssuggests that anion flux from the cytoplasm to the vacuole maybe stimulated by FC at pH 3?9. The failure of guard cells toincrease their anion content on treatment with FC despite anincrease in stomatal aperture and in cation content suggeststhat in FC-induced stomatal opening excess cation is balancedby organic acid synthesis within the guard cell. Key words: Fusicoccin, guard cells, ion fluxes, Commelina communis     

An Examination, by Compartmental Flux Analysis, of the Development of Sodium and Chloride Absorption Capacities in Beetroot Disks

Using beetroot, Beta vulgaris L. var. Avon Early, grown in radioactivelylabelled nutrient solutions, concentrations and fluxes of Na⁺and Cl^– were estimated for cells of freshly cut storageroot disks, by compartmental analysis of radioisotope elutionmeasurements. These values were compared with results obtainedin a similar manner from beetroot grown in non-labelled nutrientsolution and loaded instead during an aging period in ²²Na-or ³⁶Cl- labelled 1 mM NaCl solution. In accord with the generallyaccepted, but never properly tested, view, it was found thatnet Na⁺ influx followed from a reduction in efflux with agingand net Cl^– uptake depended on a marked increase in influx.However, both these important changes took place at the tonoplast,and, although aging led to a reduction of plasmalemma fluxes,at no time was entry of Cl^– into the cytoplasm, or lossof Na⁺ from the cytoplasm, significantly restricted.     

Utilization of the Vibrating Probe and Ion-Selective Microelectrode Techniques to Investigate Electrophysiological Responses to Wounding in Pea Roots

This paper examines the ionic composition of wound-induced electricalcurrents in higher plant tissue, using two non-injurious electrophysiologicaltechniques. By simultaneous recording of K⁺, H⁺ , and Ca²⁺ ionfluxes with extracellular ion-selective microelectrodes, wehave determined that a Ca²⁺ influx (2.4 µA cm^–2),a small H⁺ influx (0.17 µA cm^–2) and a large K⁺efflux (16 µA cm^–2) occur immediately after woundingin roots of Pisum sativum L. var. Greenfeast. Using an extracellularvibrating probe at the wound site, net ion currents of 26 µAcm^–2 were measured 5 min after wounding. In a more concentratedbathing medium (1/4 rather than 1/16 strength Hoagland's solution),net ion currents of 59 µA cm^–2 were measured, andthese would appear to be the largest extracellular currentsthat have been measured in plants. We made a quantitative comparisonof the summed ion fluxes with the net ion currents and thisrevealed that ion fluxes, in addition to those measured here,occur after wounding. Key words: Wounding, ion flux, electric current, calcium, potassium     

Control of Plasma Membrane Cl- Fluxes in Chara corallina by External Cl- and Light

The efflux of Cl^– at the plasma membrane of Chara wasstudied in relation to two treatments known to affect the flux:that of removal of external Cl^– and of light. It is shownthat although removal of external Cl^– results in a rapidreduction in Cl^– efflux (consistent with a direct effectof external Cl^– on the transport system) the magnitudeof this reduction in the dark is greater than the measured darkinflux. Therefore, in the dark at least, it is proposed that1:1 exchange diffusion cannot account for the trans-stimulationof efflux by external Cl^–. Light induces an inhibition of efflux and a concomitant stimulationof Cl^– influx at 20 °C, but at 10 °C the responsesto light of the two fluxes can be separated temporally. It istherefore suggested that the fluxes are not reciprocally dependenton the same factor which mediates in the light response. Furtherconsiderations show that it is unlikely that the cytoplasmicCl^– concentration mediates in the light response of eitherflux, but that changes in cytoplasmic pH may do so.     

Light-Induced H+ Accumulation in the Vacuole of Nitellopsis obtusa

The effects of light on the pH in the vacuole and the electricpotential difference across the plasmalemma and the tonoplastof Nitellopsis obtusa were investigated by means of conventionaland H⁺-specific glass or antimony microelectrodes. Illuminationis found to bring about a decrease in the pH of the vacuolarsap by 0.1–0.5 units concomitant with a depolarizationof the cell. The light-induced changes of the potential differenceand the vacuolar pH depend in different ways on the pH of theexternal medium (pH_o). At pH_o 9.0 cells exhibit great light-inducedpotential changes (up to 100 mV), but only small pH changesof the vacuolar sap. At neutral or slightly acidic pH_o valuesthe amplitude of the light-induced pH changes in the vacuoleincreases up to 0.3–0.5 pH units, but the amplitudes ofthe potential changes at the plasmalemma are relatively small.At pH_o 9.0 a transient acidification of the medium is observedupon illumination whereas at lower pH values light-induced alkalinizationwas only seen. Transfer of the cells from pH_o 9.0 to pH_o 7.5results in a cell hyperpolarization by 60–80 mV and adecrease of the vacuolar pH by 0.4–0.5 units under lightconditions but has no significant effect on the potential andthe vacuolar pH in the darkness. It is proposed that mechanismsof active H⁺ extrusion from the cytoplasm are located both inthe plasmalemma and the tonoplast. The observed acidificationin the vacuole appears to be determined by a light-induced increaseof the concentration of H⁺ in the cytoplasm. The H⁺ conductionof the plasmalemma seems to increase on illumination. The patternof the light-induced H⁺ fluxes across the tonoplast and theplasmalemma depends crucially on the extent of the light-inducedchanges in the H⁺ conductance and on the electrochemical gradientfor H⁺ at the plasmalemma.     

Na+ fluxes in Chara under salt stress

The influx and efflux of Na⁺ across the plasma membrane of Characorallina and Chara longifolia were examined under mild saltstress conditions. Na⁺ influx was found to be rapid in bothspecies with the freely exchangeable cytoplasmic Na⁺ cominginto isotopic equilibrium with external ²²Na⁺ within 1 h ofexposure to isotope. Cytoplasmlc Na⁺ concentration and Na⁺ influxwere greater in C. corallina than in C. longifolla under thesame conditions. Na⁺ influx across the tonoplast was much lowerthan the flux across the plasma membrane. Na⁺ efflux was stimulatedat pH 5 relative to pH 7 by 218% in C. coralllna and 320% inC. longifolia. In both species externally applied Li⁺ inhibitedNa⁺ efflux at pH 5 but not at pH 7. Na⁺ etflux was not significantlyinhibited by amiloride. Key words: Na⁺ influx, Na⁺ efflux, Na⁺/H⁺ antiport, Chara     

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.     

Effects of Cations on the Cytoplasmic pH of Chara corallina

Smith, F. A. and Gibson, J.–L. 1985. Effects of cationson the cytoplasmic pH of Chara corallina.—J.exp. Bot.36: 1331–1340 Removal of external Ca²⁺ from cells of Chara corallina lowersthe cytoplasmic pH, as determined by the intracellular distributionof the weak acid 5,5–dimethyloxazolidine2–,4–dione(DM0), when the external pH is below about 60. This effect isreversed, at least partially, by addition of the following cationsto Ca²⁺-free solutions: tetraethylammonium (TEA⁺) and Na⁺ at5 or 10 mol m^-3, Li⁺ and Cs⁺ (10 mol m^-3), or Mg²⁺, Mn²⁺ andLa³⁺ (02 or 05 mol m^-3). Under the same conditions, increasesin pH sometimes, but not always, occur in the presence of 10mol m^-3 K⁺ or Rb⁺ The results are discussed in relation to the major transportprocesses that determine pH and the electric potential differenceacross the plasma membrane, namely fluxes of H⁺ and of K⁺. Thesimplest explanation of the effects of the various cations testedin this study is that they primarily affect pHi_c via changesin influx of H⁺ but direct effects on the H⁺ pump or on K⁺ fluxesmay also be involved Key words: Chara corallina, cytoplasmic pH, cations, H⁺transport     

Ion Fluxes in 'Isolated' Guard Cells of Commelina communis L.

Ion fluxes have been measured in ‘isolated’ guardcells of Commelina communis L. using ⁸⁶RbCl and K⁸²Br, in epidermalstrips in which all cells other than guard cells have been killedby treatment at low pH. To avoid problems of slow free spaceexchange most fluxes have been measured at pH 3.9, at whichstomata open well in K(Rb) Cl(Br) and are stable for many hours.At pH 3.9 the intracellular ⁸⁶Rb exchanged as a single compartmentwith a half-time of 2–3 h, independent of external concentration(C_o). The influx of ⁸⁶Rb rose with concentration, to a V_maxof about 23 pmol mm^–2 h^–1. The efflux curve of ⁸²Brcould be well fitted by two exponential terms, with half-timesof 38 min (independent of C_o), and 5–35 h (falling withincreasing C_o). Bromide contents of cytoplasm and vacuole (Q_cand Q_v), and fluxes at plasmalemma and tonoplast, were calculatedfrom the efflux kinetics. Over C_o 20–60 mM, as the apertureincreased from 7 µm to 17 µm, the tonoplast flux(0.5–11.5 pmol mm^–2h^–1) was always much lessthan the plasmalemma flux (7–77 pmol mm^–2 h^–1).Q_c and Q_v both increased with aperture. The increase in Q_c of10.3 pmol mm^–2 µm^–1 is adequate to accountfor the osmotic changes required to change the aperture, aspreviously estimated. However, the change in vacuolar contentof only 5.9 pmol mm^–2 µm^–1 is much too smallto account for the osmotic changes required, or to balance thecytoplasmic changes. It appears therefore that increasing KBroutside not only increases the cytoplasmic salt content, andthe Br flux at the tonoplast, but also stimulates the vacuolaraccumulation of some other solute.     

H+ tolerance of Chara Internodal Cells and Apparent Net Influx of H+ in Weakly Acidic Solutions: Implication of the Net Flux of H+ as a Minor Component among the Total Net Flux of Ions across the Intact Cell Membrane of Chara

Precise measurements of the net flux of protons in Chara internodalcells were made with a recently designed high-resolution pH-meter.Survival of intact Chara internodal cells in artificial pondwater (APW) that contained HC1 at various concentrations wasalso examined. The apparent net flux of H⁺ was inward and muchsmaller than that reported so far. In APW at pH 4.005, a valuehigher than the extracellular pH expected from the values ofH⁺ efflux reported to date, all of the intact Chara internodalcells died within a day. With reference to the data on the circadianflow of ions in the pulvinus of Phaseolus [Kiyosawa (1979) PlantCell Physiol. 20: 1621–1634, Hosokawa and Kiyosawa (1983)Plant Cell Physiol. 24: 1065–1072] and ionic regulationin Chara L-cells [Kiyosawa and Okihara (1988) Plant Cell Physiol.29: 9–19], a discussion is presented of the prossiblyminor contribution of the net flux of H⁺ in the generation ofthe electrical membrane potential. Regulation of the net fluxof H⁺ in weakly acidic APW is also discussed. (Received September 4, 1989; Accepted January 25, 1990)     

Inhibition of Cl- Channel Activation in Chara corallina Membrane by Lanthanum Ion

We examined a role of Ca²⁺ in the activation of the two majorion channels, i.e., Cl^– and K⁺ channels at the excitationof the characean plasmalemma. The current-voltage relation (I-Vcurve) of the Chara membrane was compared under the ramp voltageclamp condition before and after external application of 20µMof La³⁺ (a Ca²⁺ channel blocker). The transient inward currentcomponent, which is carried mainly by the efflux of Cl^–,disappeared almost completely in about 30 min with La³⁺ treatment.On the other hand, no effect was observed on the late largeoutward current, which is mainly carried by the efflux of K⁺in a large depolarization region (less negative than –50mV). These results suggest that the Cl^– channel in theChara plasmalemma is activated by Ca²⁺ influx, while the K⁺channel is simply activated by depolarization. (Received April 7, 1986; Accepted June 6, 1986)     

Biophysical and Biochemical Regulation of Cytoplasmic pH in Chara corallina during Acid Loads

The contribution of membrane transport to regulation of cytoplasmicpH in Chara corallina has been measured during proton-loadingby uptake of butyric acid. In the short-term (i.e. up to 20min) uptake of butyric acid is not affected by removal of externalK⁺, Na⁺ or Cl^– but over longer periods uptake is decreased(by 20–50% in different experiments) in the absence ofexternal Na⁺ or, sometimes, K⁺. Influxes of both Na⁺ and K⁺increase temporarily after addition of butyrate, Na⁺ immediatelyand K⁺ after a lag. Effects on Cl^– influx are small butCl^– efflux increases enormously after a short lag. Anapproximate comparison of internal butyrate with changes inthe concentration of K⁺, Na⁺, and Cl^– suggests that initially(i.e. for a few min) cytoplasmic pH is determined by bufferingand possibly by some decarboxylation of organic acids (biochemicalpH regulation), and that biophysical pH regulation involvingefflux of H⁺ balanced by influxes of K⁺, Na⁺ and especiallyefflux of Cl^– progressively becomes dominant. When butyric acid is washed out of the cells, cytoplasmic pHis restored completely or partially (depending on the butyrateconcentration used) and this is independent of the presenceor absence of external Cl^–. Where Cl^– is present,its influx is relatively small. It is suggested that cytoplasmicpH is then controlled biochemically, involving the synthesisof an (unidentified) organic acid and the accumulation of acidicanions in place of butyurate lost from the cell. During thesecond application of butyrate, net Cl^– efflux is small:it is suggested that control of cytoplasmic pH then involvesdecarboxylation of the organic acid anions. The questions of the source of Cl^– lost from the cell(cytoplasm or vacuole) and of possible cytoplasmic swellingassociated with the accumulation of butyrate are discussed. Key words: Chara corallina, butyric acid, cytoplasmic pH, membrane transport     

Potassium Transport in Enteromorpha intestinalis (L.) Link: II. EFFECTS OF MEDIUM COMPOSITION AND METABOLIC INHIBITORS

In springwater (25.5 mol m^–3 Cl^–, 20.4 mol m^–3Na⁺, 0.14 mol m^–3 K⁺) Enteromorpha intestinalis couldnot survive for more than a few weeks unless provided with 0.5mol m^–3 K⁺ in the medium or alternatively exposed to seawaterfor 1 day per week. Maintenance of a cytoplasmic K⁺ level ofabout 200 mol m^–3 is critical for the maintenance of normalmetabolic activity. Net gains of intracellular K⁺ occurred whenthe plants were transferred from low-salinity to seawater; converselylarge net losses occurred when plants were transferred fromseawater to springwater. These two processes were not simplythe reverse of one another; net gain of K⁺ involved a largeincrease in the tracer flux both into and out of the cell butnet loss of K⁺ virtually halted the tracer flux into the cell.Any injury incurred by rapid salinity changes was short-lived;plants were rapidly able to adjust intracellular [K₁.K⁺). K⁺(orto some extent Rb⁺) was found to be necessary in the effluxmedium for ⁴²K⁺ exchange to occur. The osmotic concentrationof the medium was also important but extracellular Na⁺ and Cl^–concentrationswere not critical. K⁺ influx and efflux in both springwaterand seawater were largely independent of light and were sensitivein varying degrees to a range of common metabolic inhibitorsand uncouplers. The results are best explained by the presenceof an active K⁺ influx, generated by an ATP-dependent K⁺ pumpat the plasmalemma. Key words: Enteromorpha, Potassium transport, Salinity changes, Uncouplers, Inhibitors     

Comparison of the Effects of Ammonia and Methylamine on Chloride Transport and Intracellular pH in Chara corallina

Ammonium and methylammonium ions greatly increase the rate ofCl^– transport in Chara corallian. This effect is dependenton the pH of the bathing solution. The amine-stimulated Cl^–influx is small at pH 5·5, increases to a maximum atpH 6·5–7·5, and decreases again as the pHis raised to 8·5. Increased Cl^– influx is accompaniedby an increase in cytoplasmic pH, as calculated from the distributionof DMO. When the external pH lies between 5·5 and 7·3,cytoplasmic pH in the absence of amine is 7·65–7·70,with an increase of 0·15–0·25 in the presenceof amine. As external pH is increased above 7·3, cytoplasmicpH also increases, with progessively less effect of amine. Although the relationship between Cl^– influx and cytoplasmicpH is not simple, the results provide evidence in accord withthe hypothesis that Cl^– transport in Chara involves H⁺—Cl^–symport, or the equivalent OH^–—Cl^– antiport.The possible role of cytoplasmic pH as a factor involved inthe regulation of membrane transport in Chara is discussed.     

Rates of Hydrogen Ion Efflux by Nodulated Legumes Grown in Flowing Solution Culture with Continuous pH Monitoring and Adjustment

The net efflux of H⁺ from lucerne (Medicago saliva L.), redclover (Trifolium pratense L.) and white clover (Trifolium repensL.) growing in flowing solution culture and dependent upon symbioticfixation of atmospheric N, was measured over a 75 d experimentalperiod. Considerable and rapid increases in acidity of the nutrientsolution of up to 1.45 pH units were recorded when the pH wasriot held constant over a 30 h period. There was little differencein H⁺ efflux when solution pH was held constant at 4.75, 5.75or 6.75, but there was an immediate cessation when it was adjustedto 3.75. Differences in the daily net efflux of H⁺ closely followedthe pattern of daily differences in incoming radiation, andthere was also evidence of a diurnal pattern of H⁺ efflux. Althoughthere were initially distinct differences between the speciesin the calculated rate of net H⁺ efflux (µg H⁺ g^–1dry shoot d^–), by day 75 these had diminished. In allspecies, however, the maximum rate of efflux per unit of shootsoccurred during the earlier rapid phases of growth. The measuredefflux of H⁺ was well equated with the plant content of excesscations (as measured by ash alkalinity) and, on average, theratio of acidity produced to N assimilated (expressed as anequivalent) was 0-24. Medicago sativa L., Trifolium pratense L., Trifolium repens L., lucerne, red clover, white clover, acidification, cation/anion balance, flowing solution culture, H⁺ efflux, nitrogen fixation     

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.     

Effects of Some Amino Acid Analogues on the Uptake and Transport of K+ and Ca2+ in Wheat and Mung Bean Seedlings: I. CORTICAL CELL FLUXES AND TRANSPORT TO THE STELE IN EXCISED ROOT SEGMENTS, AS AFFECTED BY p-FLUOROPHENYLALANINE

Effluxes of K⁺ and Ca²⁺ from root segments of both wheat, Triticunaestivum L. cv. Capelle and mung bean, Vigna radiata (L.) Wilczek,were measured in the presence or absence of 20 mol m^–3para-fluorophenylalanine (p-FPA). The results were used to estimatethe compartment contents and transmembrane K⁺ and Ca²⁺ fluxesin root cortex cells. Using the Ussing-Teorell flux equationas the criterion, it was concluded that entry of K⁺ from theoutside solution to the cytoplasm, and from the cytoplasm tothe vacuole were active in both wheat and mung bean. Also, inboth species, Ca²⁺ entered the cytoplasm passively across theplasmalemma and was actively pumped back to the external solution.However, interpretation of the direction of active transportacross the tonoplast depends upon an assumption about Ca²⁺ activityin the cytoplasm. The only qualitative effect of p-FPA was to alter the drivingforce for K⁺ influx, across the plasmalemma in wheat, from anactive to a passive one. Quantitative effects of the analoguewere seen for K⁺ fluxes in both wheat and mung bean and forCa²⁺ fluxes in wheat. The p-FPA reduced transport of K⁺ in bothspecies, while transport of Ca²⁺ was unaffected. The implicationsof these results for the ‘two pump hypothesis’ arediscussed. Key words: Triticum aestivum, Vigna radiata, Two pump hypothesis     

Chloride Transport in Chara corallina and the Electrochemical Potential Difference for Hydrogen Ions

The pH of the cytoplasm of Chara corallina cells has been measuredwith the weak acid 5,5-dimethyloxazolidine-2,4-dione (DM0).Over an external pH range 4·5–9·5 the resultsfit the regression equation pH_cytoplasm=6·28+0·22pH_out. Using measured values of the electric potential difference acrossthe plasmalemma we have calculated the electrochemical potentialdifference across this membrane for H⁺ and Cl^–. Thesedata are used to test the hypothesis that the inward transportof Cl^– is coupled to the inthix of H⁺ or, which comesto the same thing, efflux of OH^–. One-for-one couplingwill not give net Cl^– uptake from solutions with pH greaterthan about 7·2, unless the cytoplasmic Cl^– concentrationis lower than 10 mM, or the pH just outside the membrane islower than that in the bulk solution. It is shown that net Cl^–uptake proceeds from solutions with pH up to 9. The alternative possibility is that Cl^– transport is broughtabout by co-transport of two H⁺ for each Cl^–; this isnot ruled out by the results reported. Such a mechanism mightbe detectable by its electrogenic effect: although such effectshave not been detected, it is shown that they would be smallunder most conditions. Other possible mechanisms are discussed.