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     

Vacuolar Na/K Exchange, its Occurrence in Root Cells of Hordeum, Atriplex and Zea and its Significance for K/Na Discrimination in Roots

Using excised low-salt roots of barley and Atriplex hortenslsthe transport of endogenous potassium through the xylem vesselswas studied It was enhanced by nitrate and additionally by sodiumions which apparently replaced vacuolar potassium which wasthen available in the symplasm of root cells for transport tothe shoot Vacuolar Na/K exchange also has been investigatedby measurements of longitudinal ion profiles in single rootsof both species. In Atriplex roots a change in the externalsolution from K⁺ to Na⁺ induced an exchange of vacuolar K⁺ forNa⁺, in particular in the subapical root tissues and led toincreased K⁺ transport and loss of K⁺ from the cortex. In inverseexperiments a change from Na⁺ to K⁺ did not induce an exchangeof vacuolar Na⁺; merely in meristematic tissues Na⁺—apparentlyfrom the cytoplasm—was extruded in exchange for K⁺. Inroots of barley seedlings without caryopsis, as in excised roots,a massive exchange of K⁺ for Na⁺ was observed in the continuouspresence of external 1.0 mM Na and 0.2 mM K. This exchange alsowas attributed to the vacuole and was most pronounced in theyoung subapical tissues. It did not occur, however, in the correspondingtissues in roots of fully intact barley seedlings. In these,the young tissues retained a relatively high K/Na ratio alsoin their vacuoles. Similarly, contrasting results were obtainedwith intact and excised roots of Zea mays L. Based on theseresults a scheme of the events that lead to selective cationuptake in intact barley roots is proposed. In this scheme acrucial factor of selectivity is sufficient phloem recirculationof K⁺ by the aid of which K⁺ rich cortical cells are formednear the root tip. When matured these cells are suggested tomaintain a high cytoplasmic K/Na ratio due to K⁺ dependent sodiumextrusion at the plasmalemma and due to recovery of vacuolarK⁺ by Na/K exchange across the tonoplast. Key words: Potassium/Sodium selectivity, Vacuolar exchange, Xylem transport, Hordeum, Zea, Atriplex     

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     

Effect of Sodium on Phosphate Uptake in Unicellular and Filamentous Cyanobacteria

The effect of Na⁺ on phosphate uptake was studied in four strainsof cyanobacteria: Synechococcus PCC 7942, Gloeothece PCC 6501,Phormidium sp. and Chlorogloeopsis PCC 6912. Phosphate uptakewas stimulated by Na⁺ in all cases. Li⁺ and K⁺ acted as partialanalogues for Na⁺. Half-saturation [K_1/2(Na⁺)] of phosphateuptake was reached with Na⁺ concentrations ranging from 317µM in Chlorogloeopsis to 659 µM in Phormidium. Theconcentration of phosphate required to reach half-saturationof phosphate uptake [K_1/2(P_i)]was not changed by the presenceof Na⁺. (Received April 11, 1994; Accepted July 5, 1994)     

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.     

Sodium-Stimulation of Phosphate Uptake in the Cyanobacterium Anabaena PCC 7119

Anabaena PCC 7119 showed higher rates of phosphate uptake whencells were under P-starvation. Phosphate uptake was energy-dependentas indicated the decrease observed when assays were performedin the dark or in the presence of inhibitors of photosyntheticelectron transport, energy transfer and adenosine triphosphataseactivity. Phosphate uptake was stimulated by Na⁺ both in P-sufficientcells and P-starved cells. Li⁺ and K⁺ acted as partial analoguesfor Na⁺. The Na⁺-stimulation of phosphate uptake followed Michaelis-Mentenkinetics, half-saturation (K) of phosphate uptake was reachedwith a Na⁺ concentration of 212 µM. The absence of Na⁺reduced the rates of phosphate uptake at all phosphate concentrationsassayed (1–20 µM). The maximum uptake rates (V_max)decreased from 658 nmol P (mg dry wt)^-1 h^-1 in the presenceof Na⁺ to 149 nmol P (mg dry wt)^-1 h^-1 in the absence of Na⁺.The absence of Na⁺ did not change significantly the concentrationof phosphate required to reach half-saturation (K) (3.01 µMin the presence of Na⁺ vs 3.21 µM in the absence of Na⁺).In the presence of Na⁺ the rate of phosphate uptake was affectedby the pH; optimal rates were observed at pH 8. In the absenceof Na⁺ phosphate uptake was not affected by the pH; low rateswere observed in all cases. Monensin, an ionophore which collapsesNa⁺-gradients, reduced the rate of phosphate uptake in Na⁺-supplementedcells. These results indicated the existence of a Na⁺-dependentphosphate uptake in Anabaena PCC 7119. (Received September 8, 1992; Accepted November 17, 1992)     

Effects of Fusicoccin and Abscisic Acid on Sugar and lon Transport from Plant Glands

Fusicoccin (FC) inhibited net excretion of Cl^– by theglands of the pitchers of the carnivorous plant Nepenthes hookeriana;of Na⁺ and Cl^– by the salt glands of the halophytes Limoniumvulgare and L. pectinatum and of K⁺ in the nectar of Acer platanoidesflowers. It had no effect on K⁺ elimination with nectar of Impatienswalleriana (extrafloral nectaries) and Abutilon striatum. Abscisicacid (ABA) stimulated net excretion of K⁺ and Cl^– in Nepenthesand of Na⁺ and Cl^– in Limonium but had no effects on K⁺in nectar. Thus, FC and ABA had opposing effects on ion excretionby the salt eliminating glands of Limonium and Nepenthes. Bothcompounds, however, had similar effects on sugar secretion ofnectary glands which was either inhibited or unaffected by FCand ABA. It is suggested that the effects of FC and ABA on ion excretionby gland cells could be reconciled with literature showing FC-stimulationand possible ABA-inhibition of proton pumps at the plasmalemmaof plant cells. Nepenthes hookeriana, Limonium vulgare, Limonium pectinatum, Acer platanoides, salt-glands, nectaries, excretion, fusicoccin, abscisic acid, proton pump     

Na+/H+ Antiporter in Tonoplast Vesicles from Rice Roots

The Na⁺/H ⁺ antiporter in vacuolar membranes transports Na⁺from the cytoplasm to vacuoles using a pH gradient generatedby proton pumps; it is considered to be related to salinitytolerance. Rice (Oryza sativa L.) is a salt-sensitive crop whosevacuolar antiporter is unknown. The vacuolar pH of rice roots,determined by ³¹P-nuclear magnetic resonance (NMR), increasedfrom 5.34 to 5.58 in response to 0.1 M NaCl treatment. Transportof protons into the tonoplast vesicles from rice roots was fluorometricallymeasured. Efflux of protons was accelerated by the additionof Na⁺. Furthermore, the influx of ²²Na⁺ into the tonoplastvesicles was accelerated by a pH gradient generated by proton-translocatingadenosine 5'-triphosphatase (H⁺-ATPase) and proton-translocatinginorganic pyro-phosphatase (H⁺-PPase). We concluded that thisNa⁺/H⁺antiporter functioned as a Na⁺ transporter in the vacuolarmembranes. The antiporter had a K_m of 10 mM for Na⁺ and wascompetitively inhibited by amiloride and its analogues. TheKⁱ values for 5-(N-methyl-N-isobutyl)-amiloride (MIA), 5-(N-ethyl-N-isopropyI)-amiloride(EIPA), and 5-(N, N-hexamethylene)-amiloride (HMA) were 2.2,5.9, and 2.9 µ M, respectively. Unlike barley, a salt-tolerantcrop, NaCl treatment did not activate the antiporter in riceroots. The amount of antiporter in the vacuolar membranes maybe one of the most important factors determining salt tolerance. ¹This work was supported by a grant from Bio-Media Project ofthe Japanese Ministry of Agriculture, Forestry and Fisheries(BMP96-III-1).     

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     

Further Studies of Electro-Osmosis in Nitella in Relation to Pores in Membranes

By measuring the number of water molecules per ion which weremoved electro-osmotically through cells of Nitella translucensand N. flexilis it has been shown that a significant differenceexisted between samples of these species in 1965. In each speciesthe electro-osmotic efficiency was greater with Na⁺ than withK⁺⁺. Also 10^-4 to 10^-5 M IAA tended to decrease electro-osmoticefficiencies while IAA, after 30 min. treatment, produced asignificant increase in water flow into the treated end of aliving cell. Calculations based on this work suggest that about10⁸–10⁹ pore sites per cm² exist on the surface membranesfor Na⁺ or K⁺ ion transport.     

Alloxan-induced diabetes reduces sarcolemmal Na+-K+ pump function in rabbit ventricular myocytes

The effect of diabetes on sarcolemmal Na⁺-K⁺ pump function is important for our understanding of heart disease associated with diabetes and design of its treatment. We induced diabetes characterized by hyperglycemia but no other major metabolic disturbances in rabbits. Ventricular myocytes isolated from diabetic rabbits and controls were voltage clamped and internally perfused with the whole cell patch-clamp technique. Electrogenic Na⁺-K⁺ pump current (I_p, arising from the 3:2 Na⁺-to-K⁺ exchange ratio) was identified as the shift in holding current induced by Na⁺-K⁺ pump blockade with 100 µmol/l ouabain in most experiments. There was no effect of diabetes on I_p recorded when myocytes were perfused with pipette solutions containing 80 mmol/l Na⁺ to nearly saturate intracellular Na⁺-K⁺ pump sites. However, diabetes was associated with a significant decrease in I_p measured when pipette solutions contained 10 mmol/l Na⁺. The decrease was independent of membrane voltage but dependent on the intracellular concentration of K⁺. There was no effect of diabetes on the sensitivity of I_p to extracellular K⁺. Pump inhibition was abolished by restoration of euglycemia or by in vivo angiotensin II receptor blockade with losartan. We conclude that diabetes induces sarcolemmal Na⁺-K⁺ pump inhibition that can be reversed with pharmacological intervention. sodium transport; insulin; angiotensin II; cardiomyopathy; hyperglycemia     

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     

Transepithelial resistance can be regulated by the intestinal brush-border Na(+)/H(+) exchanger NHE3

Initiation of intestinal Na⁺-glucose cotransport results intransient cell swelling and sustained increases in tight junction permeability. Since Na⁺/H⁺ exchange has beenimplicated in volume regulation after physiological cell swelling, wehypothesized that Na⁺/H⁺ exchange might also berequired for Na⁺-glucose cotransport-dependent tightjunction regulation. In Caco-2 monolayers with activeNa⁺-glucose cotransport, inhibition ofNa⁺/H⁺ exchange with 200 µM5-(N,N-dimethyl)- amiloride induced 36 ± 2% increases in transepithelial resistance (TER). Evaluation using multiple Na⁺/H⁺ exchange inhibitors showed thatinhibition of the Na⁺/H⁺ exchanger 3 (NHE3)isoform was most closely related to TER increases. TER increases due toNHE3 inhibition were related to cytoplasmic acidification becausecytoplasmic alkalinization with 5 mM NH₄Cl prevented bothcytoplasmic acidification and TER increases. However, NHE3 inhibitiondid not affect TER when Na⁺-glucose cotransport wasinhibited. Myosin II regulatory light chain (MLC) phosphorylationdecreased up to 43 ± 5% after inhibition ofNa⁺/H⁺ exchange, similar to previous studiesthat associate decreased MLC phosphorylation with increased TER afterinhibition of Na⁺-glucose cotransport. However, NHE3inhibitors did not diminish Na⁺-glucose cotransport. Thesedata demonstrate that inhibition of NHE3 results in decreased MLCphosphorylation and increased TER and suggest that NHE3 may participatein the signaling pathway of Na⁺-glucosecotransport-dependent tight junction regulation.

     

Ion and Glycerol Concentrations in 12 Isolates of Dunaliella

Ginzburg, M., and Ginzburg, B. Z., 1985. Ion and glycerol concentrationsin 12 isolates of Dunaliella.—J. exp. Bot. 36: 1064–1074. Twelve isolates of Dunaliella with average cell volumes rangingfrom 50 to 1400x10^–18 m³ were grown in batch culture at0.5 M or 2.0 M NaCl. Glycerol and ions (Na⁺, K⁺, Mg²⁺, CI^–,phosphate) were measured in log-phase cultures. The contentsof Mg²⁺, K⁺ and phosphate per cell were found to be a functionof cell-volume. Cell glycerol, Na⁺ and Cl^– were functionsof cell-volume and of the NaCl concentration in the medium.Solute concentrations were calculated from the measured cell-volumesand from the ³H₂O content of pellets corrected for intercellularspace using Blue Dextran. Cell glycerol was found to accountfor about one-half of the expected osmolarity, the remainderbeing largely accounted for by Na⁺ and CI^–. Key words: —Dunaliella, isolates, glycerol, ion concentrations     

Direct evidence of Na+/Ca2+ exchange in squid rhabdomeric membranes

Na⁺/Ca²⁺exchange has been investigated in squid(Loligopealei) rhabdomeric membranes.Ca²⁺-containing vesicles have beenprepared from purified rhabdomeric membranes by extrusion throughpolycarbonate filters of 1-µm pore size. After removal of externalCa²⁺, up to 90% of the entrappedCa²⁺ could be specificallyreleased by the addition of Na⁺;this finding indicates that most of the vesicles containedNa⁺/Ca²⁺exchanger. The Na⁺-inducedCa²⁺ efflux had a half-maximumvalue (K_1/2) of~44 mM and a Hill coefficient of ~1.7. The maximalNa⁺-inducedCa²⁺ efflux was ~0.6 nmolCa²⁺ · s¹ · mgprotein¹. SimilarNa⁺-inducedCa²⁺ effluxes were measured ifK⁺ was replaced withLi⁺ orCs⁺. Vesicles loaded withCa²⁺ byNa⁺/Ca²⁺exchange also released this Ca²⁺byNa⁺/Ca²⁺exchange, suggesting thatNa⁺/Ca²⁺exchange operated in both forward and reverse modes. Limited proteolysis by trypsin resulted in a rate ofCa²⁺ efflux enhanced byapproximately fivefold when efflux was activated with 95 mM NaCl. For vesicles subjected to limited proteolysis by trypsin,Na⁺/Ca²⁺exchange was characterized by aK_1/2 of ~25 mMand a Hill coefficient of 1.6. For these vesicles, the maximalNa⁺-inducedCa²⁺ efflux was about twice asgreat as in control vesicles. We conclude thatNa⁺/Ca²⁺exchange proteins localized in rhabdomeric membranes mediate Ca²⁺ extrusion in squid photoreceptors.     

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     

K+Nutrition and Na+Toxicity: The Basis of Cellular K+/Na+Ratios

The capacity of plants to maintain a high cytosolic K⁺/Na⁺ratiois likely to be one of the key determinants of plant salt tolerance.Important progress has been made in recent years regarding theidentification and characterization of genes and transportersthat contribute to the cytosolic K⁺/Na⁺ratio. For K⁺uptake,K⁺efflux and K⁺translocation to the shoot, genes have been isolatedthat encode K⁺uptake and K⁺release ion channels and K⁺carriersthat are coupled to either a H⁺or Na⁺gradient. Although thepicture is less clear for the movement of Na⁺, one pathway,in the form of non-selective ion channels, is likely to playa role in Na⁺uptake, whereas Na⁺efflux and compartmentationare likely to be mediated by H⁺-coupled antiport. In addition,several proteins have been characterized that play prominentroles in the regulation of K⁺and/or Na⁺fluxes. In this BotanicalBriefing we will discuss the functions and interactions of thesegenes and transporters in the broader context of K⁺nutritionand Na⁺toxicity. Copyright 1999 Annals of Botany Company Salinty, K⁺/N⁺ratio, transporter, membrane.     

The Effect of Fusicoccin and Monovalent Cations on the Viability of Wheat Seeds

Fusicoccin (FC), K⁺ and Na⁺ restore the viability of aged wheatseeds and isolated wheat embryos. We studied the effect of FCand K⁺ on the evolution of the proton extrusion activity andnet K⁺ uptake during the early phases of germination of wheatembryos isolated from unaged and aged seeds. Our data show thatin this material, germination is also characterized by the activationof the proton-extruding activity and by the leakage and subsequentre-absorption of K⁺. The percentage of germination appearedto be correlated with the amount of proton extrusion and K⁺net uptake. In aged embryos the positive effects of FC and K⁺on germination were correlated with a marked increase in bothproton extrusion and K⁺ net uptake. These findings suggest that:(a) in wheat seeds, germination is also correlated to the activationof a mechanism linked to H⁺/K⁺ exchange; (b) the damage of thismechanism is, under our conditions, responsible for the lossof viability.     

Osmotic and ionic regulation in Nitella

When the osmotic value of an internodal cell of Nitella flexiliswas modified by the method of transcellular osmosis, the normalosmotic value was chiefly restored by the release or absorptionof K⁺. The release or uptake of Na⁺ was observed only when themodification of osmotic value was significant. Both the uptakeand release of K⁺ were linearly dependent on the degree of modificationof the osmotic value. The effectiveness of alkali metal cationsin restoring the osmotic value in cells of lower osmotic valueswas in the order K⁺>Rb⁺>Na⁺, Cs⁺>Li⁺. The absorptionof K⁺ by cells of lower osmotic values depended strongly ontemperature, while the release of K⁺ from cells of higher osmoticvalues did not. To clarify whether the Nitella cell regulates the osmotic valueor regulates the concentration of K⁺ in the vacuole, the cellsap was exchanged for artificial cell saps whose osmotic valuesand ionic concentrations were varied independent of each other.It was shown that in Nitella two regulating mechanisms are operating,one which regulates the osmotic value of the cell sap irrespectiveof the level of vacuolar K⁺ (0.1–140 mM) and another whichregulates the vacuolar K⁺-level when it is abnormaly high (>160mM). Both mechanisms are assumed to operate in order to keepthe concentration of K⁺ in the cytoplasm at a constant level.The presence of Na⁺ (0–100 mM) and Ca²⁺ (5–40 mM)did not affect the movement of K⁺ during osmoregulation. ¹Present address: Sanki Engineering Limited, Nagaokakyo, Kyoto,Japan. (Received December 19, 1973; )