Electrical Potentials and Na, K, and Cl Concentrations in the Vacuole and Cytoplasm of Nitella translucens

The potential differences across the tonoplast and plasmalemmamembranes have been measured in the single cells of Nitellatranslucens, the cells being immersed in an artificial pondwater (composition: NaCl _1.0 mM., KC1 _0.1 mM., CaCl₂, _0.1 mM.).The potential of the cytoplasm is –138 m V with respectto the bathing medium and –18 mV with respect to the vacuole.The concentrations of Na, K, and Cl have been measured in thetwo cell fractions. The concentrations in the flowing cytoplasmare: Na 14 mM., K 119 mM., and Cl 65 mM.; the vacuolar concentrationsare: Na 65 mM., K 75 mM.,and Cl 160 mM. The observed potential differences across the two membranesare compared with the Nernst potentials for all three ions.This analysis shows that all three ions are actively transportedat the plasmalemma: Na is pumped outwards while K and Cl arepumped inwards. At the tonoplast Na is pumped into the vacuolewhile K and Cl are close to electrochemical equilibrium. The inhibitor, ouabain, has no effect on the cell resting potential.     

IONIC COMPOSITION AND ELECTRIC RESPONSE OF LAMPROTHAMNIUM SUCCINCTUM

The concentrations of K⁺, Na⁺ and Cl^– in the cytoplasmof Lamprothamnium succinctum, a brackish water Characeae, areabout 137, 47 and 86 mM respectively. The concentration of K⁺in the cytoplasm is of the same order as that in the cell sap,while the concentrations of Na⁺ and Cl^– are much lowerthan those in the cell sap (i.e., 140 mM Na⁺, 370 mM Cl^–).In the brackish water, in which the plant grows, the internodesis never excitable electrically. However, it acquires excitabilitywhen it is kept in a mannitol-pond water. The action potentialthus elicited is accompanied by a temporary cessation or slowdown of the cytoplasmic streaming. ¹This work was supported by Research Grants from the Ministryof Education of Japan     

X-Ray Microanalysis and Ultrastructural Studies of Cell Compartments of Dunaliella parva

Hajibagheri, M. A., Gilmour, D. J., Collins, J. C. and Flowers,T. J. 1986. X-ray microanalysis and ultrastructural studiesof cell compartments of Dunaliella parva. -J. exp. Bot. 37:1725–1732. Ultrastructural studies of the unicellular green alga Dunaliellaparva showed the presence of cytoplasmic vacuoles. X-ray microanalysiswas performed on sections of cells which had been freeze substitutedin acetone. It was found that the concentrations of both Naand Cl were much higher in the vacuoles than in the cytoplasm.When cells were grown in 0·4 kmol m^–3 NaCl theNa and Cl concentrations in the vacuoles were 349 and 283 molm ^–3 respectively, whilst cytoplasmic Na and Cl concentrationswere 37 and 26 mol m^–3. Corresponding values for cellsgrown in 1·5 kmol m^–3 NaCl were 392 mol m^–3Na and 325 mol m^–3 Cl in the vacuoles and 36 mol m^–3Na and 30 mol m^–3 Cl in the cytoplasm. Immediately afterexposure to an increase in external salinity Na and Q concentrationsincreased in both vacuoles and cytoplasm. The results are discussedwith reference to compartmental models for the ionic relationsof Dunaiiella. Key words: X-ray microanalysis, ultrastructural studies, Dunaliella parva     

Changes in intracellular electrolyte concentrations during apoptosis induced by UV irradiation of human myeloblastic cells

Decreases in the intracellular concentrations of both K⁺ and Cl^– have been implicated in playing a major role in the progression of apoptosis, but little is known about the temporal relationship between decreases in electrolyte concentration and the key events in apoptosis, and there is no information about how such decreases affect different intracellular compartments. Electron probe X-ray microanalysis was used to determine changes in element concentrations (Na, P, Cl, and K) in nucleus, cytoplasm, and mitochondria in U937 cells undergoing UV-induced apoptosis. In all compartments, the initial stages of apoptosis were characterized by decreases in [K] and [Cl]. The largest decreases in these elements were in the mitochondria and occurred before the release of cytochrome c. Initial decreases in [K] and [Cl] also preceded apoptotic changes in the nucleus. In the later stages of apoptosis, the [K] continued to decrease, whereas that of Cl began to increase toward control levels and was accompanied by an increase in [Na]. In the nucleus, these increases coincided with poly(ADP-ribose) polymerase cleavage, chromatin condensation, and DNA laddering. The cytoplasm was the compartment least affected and the pattern of change of Cl was similar to those in other compartments, but the decrease in [K] was not significant until after active caspase-3 was detected. Our results support the concept that normotonic cell shrinkage occurs early in apoptosis, and demonstrate that changes in the intracellular concentrations of K and Cl precede apoptotic changes in the cell compartments studied. sodium; potassium; chloride; cell shrinkage     

X-Ray Micro-Analysis of Cells and Cell Compartments of A triplex spongiosa: I. LEAVES

Atriplex spongiosa was grown in hydroponic culture with additionsof 0, 200, 400 and 600 mM NaCl. Frozen, hydrated cells and cellcompartments of young and mature leaf tissue were analysed byX-ray micro-analysis. Evidence for low K + Na and Cl content,and high K selectivity in the bundle sheath cytoplasm was obtainedfrom data on X-ray count ratios and on total X-ray counts. Vacuolesof the major cell types of the mature leaf had either high Kor high Na and Cl contents when grown in the absence or presenceof NaCl. Comparison of K, Na and Cl content of different cell types inthe mature leaf showed gradients in selectivity for K. relativeto Na between the bundle sheath cells and the bladder cells.In the young expanding leaves salt was sequestered in the numeroussalt bladders on the leaf surface, while the cytoplasm and developingvacuoles of undifferentiated cells contained largely K and littleNa or Cl. The results support general views on the compartmentation ofsalt in plants cells in relation to osmotic or saline stress. Key words: Atriplex spongiosa, X-ray analysis, Salinity, Compartmentation, Leaf     

Cytoplasmic Hydration Triggers a Transient Increase in Cytoplasmic Ca2+ Concentration in Nitella flexilis

Cells of Nitella flexilis were made inexcitable by treatmentwith 10 mM KCl for more than 24 h. A Ca²⁺-sensitive photoproteinaequorin was injected into the cytoplasm of such cells. Forvacuolar per fusion, the central part of an aequorin-loadedcell was immersed in silicone oil, and both cell ends bathedin the perfusion medium were cut off. A large light emissionfrom aequorin was observed when the vacuole was perfused witha hypotonic medium whose osmotic pressure was adjusted to halfof the osmotic pressure of the cell sap. This shows that hydrationof the cytoplasm triggers release of Ca²⁺ from internal stores,since influx of Ca²⁺ from silicone oil is excluded. Hydration of cells was induced in another way. Cells were firstdehydrated by transferring them from 10 mM KCl solution to thatwith 250 mM sorbitol added. This procedure did not affect thecytoplasmic streaming. When cells were rehydrated by transferringthem to 10 mM KCl solution, cytoplasmic streaming was eitherstopped or slowed down in a few seconds. A quick light emissionfrom aequorin was observed in the rehydration, evidence thatcytoplasmic streaming was inhibited by an increase in the cytoplasmicCa²⁺ concentration. Both streaming cessation and aequorin lightemission were observed even in KCl-treated cells which werefurther treated with 5 mM EGTA. Thus, the increase in Ca²⁺ isconcluded to be caused by the release of Ca²⁺ from internalstores. These results support our previous hypothesis [Tazawa et al.(1994) Plant Cell Physiol. 35:63] that, in Nitella flexilis,the increase in the concentration of Ca²⁺ in the cytoplasm whichoccurs on the endoosmotic side of the cell during transcellularosmosis is caused by hydration of the cytoplasm. (Received June 6, 1994; Accepted December 26, 1994)     

The Linkage of Light-stimulated Cl Influx to K and Na Influxes in Hydrodictyon africanum

There are reciprocal stimulations of Cl influx by K and Na,and of K and Na influx by Cl, in the light in Hydrodictyon africanum.The component of the K influx which stimulates, and is stimulatedby, Cl, is independent of the ouabainsensitive mechanism forK influx also found in H. africanum. The concentration dependenceof the cation effects on Cl influx and on the Cl-stimulatedportion of their own influxes are similar. The stimulation withK saturates at about 0.3 mM K; that with Na saturates at about2 mM Na. The Cl-dependent portions of the K and Na influxeshave similar responses to changes in photo-synthetic metabolism(far-red illumination, CDMU, and CCCP) as does the light-stimulatedCl influx. This suggests that Cl influx, and the Cl-stimulatedportions of K and Na influxes are both dependent on photosystem2 of photosynthesis, and are less sensitive to the uncouplerCCCP than is ¹⁴CO₂ fixation or the K-Na pump. It would thusappear that the Cl-dependent portions of the K and Na influxesin the light are linked to the cation-stimulated portion ofthe Cl influx. There is no very great change in the electricalcomponent of the inwardly directed passive driving force oncations under conditions in which Cl is being pumped comparedwith those under which it is not. It is not clear whether suchincrease in this driving force as do occur could account quantitativelyfor the increase in the cation influxes associated with Cl transport,or whether chemical coupling must be invoked. In addition tothe Cl-stimulated portions of the cation influxes, there arealso light-stimulated portions of K and Na influx which areindependent of Cl, not associated with the cation regulatingmechanism, and which seem to have a similar linkage to photosynthesisas does the Cl-K-Na pump. Since the light-stimulated portionof the K efflux appears to be similar to this portion of theK influx, these Cl-independent light-stimulated portions ofK and Na influxes are tentatively related to light-induced changesin cation permeability.     

The Membrane Potential of Nitella translucens

The effects of changing the external concentrations of Na, K,Ca, and Cl on the potentials of the cytoplasm and the vacuolewith respect to the bathing medium of the internodal cells ofNitella translucens have been investigated. The potential differencebetween the vacuole and the cytoplasm is practically unaffectedby the concentration changes. The observed changes of potentialdifference are therefore attributed to the boundary separatingthe cytoplasm from the medium; this boundary is possibly a plasmalemma–cellwall complex. The difference of potential between the cell walland the medium has also been measured and, in the presence ofCa, shown to be markedly sensitive only to the external Ca concentration.The results are divided into two sections: (a) for cells pretreatedin 5 mM NaCl, the subsequent experiments being carried out inCa-free media, and (b) for cells initially immersed in a standardartificial pond water containing the chlorides of Na, K, Ca.With the pretreated cells the external Na/K ratio was variedwith the total NaCl+KCl concentration kept constant at 1.1 mM.The results suggest that over a limited range of concentrationsthe cytoplasm-medium potential difference can be described byan equation similar in form to a Goldman equation but containingonly terms for Na and K, the average value of the permeabilityratio (= P_Na/P_K) being 0.27. In the presence of Ca the effectsof Na and K on the cytoplasm-medium potential difference aregreatly reduced, while the effect of Ca is relatively large.The results cannot be fitted to any form of Goldman equationcontaining terms for the major ions. The possibility of a contributionto the plasmalemma potential from electrogenic pumps is brieflydiscussed. Measurements of the Na and K content of the cytoplasmand the vacuole have been made for the pretreated cells. TheNa concentration in the cytoplasm is 37 mM and in the vacuole73 mM; the K concentration is 93 mM in the cytoplasm and 67mM in the vacuole. The Nernst potentials for both ions are comparedwith the cytoplasm-medium and cytoplasm-vacuole potential differences.This analysis shows that Na is actively transported from thecytoplasm into the medium as well as into the Vacuole; K ispumped into the cytoplasm from the medium but appears to beclose to electrochemical equilibrium across the tonoplast. ThisConfirms previously published work.     

Concentrations and Transport of Solutes in Xylem and Phloem along the Leaf Axis of NaCl-treated Hordeum vulgare

Hordeum vulgare cv. California Mariout was established in sandculture at two different NaCl concentrations (0.5 mol m^–3‘control’ and 100 mol m^–3) in the presenceof 6.5 mol m^–3 K ⁺. Between 16 and 31 d after germination,before stem elongation started, xylem sap was collected by useof a pressure chamber. Collections were made at three differentsites on leaves 1 and 3: at the base of the sheath, at the baseof the blade, i.e. above the ligule, and at the tip of the blade.Phloem sap was collected from leaf 3 at similar sites throughaphid stylets. The concentrations of K ⁺, Na⁺, Mg2⁺ and Ca²⁺were measured. Ion concentrations in xylem sap collected at the base of leaves1 and 3 were identical, indicating there was no preferentialdelivery of specific ions to older leaves. All ion concentrationsin the xylem decreased from the base of the leaf towards thetip; these gradients were remarkably steep for young leaves,indicating high rates of ion uptake from the xylem. The gradientsdecreased with leaf age, but did not disappear completely. In phloem sap, concentrations of K⁺ and total osmolality declinedslightly from the tip to the base of leaves of both controland salt-treated plants. By contrast, Na⁺ concentrations inphloem sap collected from salt-treated plants decreased drasticallyfrom 21 mol m^–3 at the tip to 7.5 mol m^–3 at thebase. Data of K/Na ratios in xylem and phloem sap were used to constructan empirical model of Na⁺ and K⁺ flows within xylem and phloemduring the life cycle of a leaf, indicating recirculation ofNa⁺ within the leaf. Key words: Hordeum vulgare, xylem transport, phloem transport, NaCl-stress     

Photo-induced H+ Transport between Chloroplasts and the Cytoplasm in a Protoplasmic Droplet of Characeae

Svintitskikh, V. A., Andrianov, V. K. and Bulychev, A. A. 1985.Photo-induced H⁺ transport between chloroplasts and the cytoplasmin a protoplasmic droplet of Characeae.—J. exp. Bot. 36:1414–1429. The effects of light on the membrane potential and cytoplasmicpH of isolated droplets of protoplasm from Nitella have beenstudied using microcapillary electrodes and pH-sensitive antimonymicro-electrodes. Illumination of chloroplast-containing dropletscaused a change of the membrane potential with a concomitantacidification of both the cytoplasm and the outer medium, butit had no effect on the electrical resistance of the surfacemembrane. Treatment of protoplasmic droplets with uncouplers(NH₄Cl and CCCP) resulted in a complete inhibition of the light-inducedacidification of the cytoplasm, whereas the energy transferinhibitor DCCD had no effect. A correlation between the formationof a pH gradient across the thylakoid membrane and the acidificationof the cytoplasm was explicable in terms of the assumption ofrestricted spatial communication between the intra-thylakoidvolume and the cytoplasm in intact chloroplast. The photo-inducedacidification of the boundary layer of an external medium wasmarkedly stimulated under the action of inhibitors of H⁺-ATPaseDCCD and DES. These findings suggest that the active extrusionof H⁺ from the cytoplasm into the external medium is not drivenby an ATPase, although H+-conducting channels of membrane ATPaseprovide a pathway for a passive diffusion of protons from outsideinto the cytoplasm Key words: Transport of protons, protoplasmic droplet, intact chloroplasts, Characeae     

Ca Fluxes and Membrane Potentials in Nitella translucens

The concentrations of Ca have been measured in the flowing cytoplasmand the vacuole of the single cells of Nitella translucens,the cells being immersed in an artificial pond Water (composition:NaCl, 1.0 mM; KCl, 0.1 mM; CaCl₂, 0. mM). In the flowing cytoplasmthe total concentration is 8 mM and in the vacuole 12 mM. Measurementsof the electrical potential differences across the plasmalemmaand tonoplast membranes show that the cytoplasm is at a potentialof —134 mV with respect to the bathing medium and —24mV with respect to the vacuole. An attempt has been made tomeasure the tracer fluxes of Ca and it is shown that the cellsare not in flux equilibrium. The influx is 0.046 µµmoles cm^–2 sec^–1; the efflux was too small to measurewith any degree of accuracy. The observed potential differences across both membranes arecompared with the Nernst potentials for Ca. This analysis showsthat Ca is not in electrochemical equilibrium across eithermembrane and that the driving forces on Ca are directed fromthe bathing medium and the vacuole into the cytoplasm. It issuggested that there is no necessity for a metabolically drivenCa pump at the plasmalemma because the low cytoplasmic Ca contentcould be due to the low permeability of the plasmalemma; theGoldman flux equation gives a value of P_Ca = 4.3x10^–8cm sec^–1. A Ca pump at the tonoplast appears to be necessaryto explain the steep electrochemical potential gradient fromthe vacuole to the cytoplasm. The efflux of Ca from the isolated cell wall has been measured.From these measurements it was possible to estimate the concentrationof indiffusible anions in the Donnan Free Space; the value obtainedwas 0.74 equiv. 1.^–1.     

X-Ray Micro-Analysis of Cells and Cell Compartments of Atripiex spongiosa: II. ROOTS

Atripiex spongiosa was grown in hydroponic culture with additionof 0, 50, 200, 400 and 600 mM NaCl. Frozen, hydrated cells ofthe roots were analysed by X-ray micro-analysis. Comparisonswere made of meristematic cells at the root apex and vacuolatedcells 5.0 mm from the apex. High selectivitiy for K relativeto Na was found for the cytoplasm of meristematic cells andthere was little effect of increasing salinity on the ratiosNa/K, Cl/K, Na/P and CI/P. In the cell vacuoles of the cortex,selective uptake of K relative to Na also occurred, but to alesser extent than in the meristematic cells. Gradients werefound of decreasing ratio of Na/K from the epidermis to thestele. Measurements of chemical content of the roots and shoots ofthese plants showed that the ratio of Na/K was higher in theshoot than in the cortical cell vacuoles and higher again thanin the stele or meristematic cytoplasm. It is suggested thattransport of ions to the shoot of Atripiex spongiosa involvesselective exclusion of Na from the xylem parenchyma into thexylem, and that this may be general to other halophytes. Key words: Micro-analysis, X-ray, Cells, Atripiex spongiosa     

Intracellular Kinetics of Tracer Chloride and Bromide in Nitella translucens

The kinetics of transfer of ³⁶CI and ₈₂Br to the vacuole ofNitella translucens have been compared with the kinetics oftransfer of the flowing cytoplasm in half-cell labelling experiments.About 40 per cent of the activity entering the cell reachesthe flowing layers, much more than could be accounted for asthe fast cytoplasmic phase seen in vacuolar kinetics. Also thespecific activity in the flowing cytoplasm is proportional totime over the first 20 min of uptake, and is essentially constantduring a 10-min wash period. Hence the major fraction of tracerin the flowing cytoplasm is in the slow phase of the cytoplasm.In spite of this there is ready movement of tracer from theflowing cytoplasm to the stationary cytoplasmic layer in theinactive end. It is concluded that the bulk (watery) phase ofthe cytoplasm is to be identified as the slow phase, and thatthe fast phase (if it exists as a real cytoplasmic compartment)can be only a very small membrane-bound phase within the bulkcytoplasm. It is shown that in the presence of 10 mM Mn²⁺, whichmakes the cells inexcitable, there is little or no fast componentof vacuolar transfer. The alternative explanation of the fastcomponent, as an artefact arising from transfer of tracer associatedwith action potentials on cutting open the cell, may providea more satisfactory explanation of the kinetics. The kineticcharacteristics of the slow phase and the lack of discriminationbetween chloride and bromide in the face of the link betweenvacuolar transfer and influx are still held to suggest ion transferto the vacuole by the creation and discharge of salt-filledvesicles, rather than by processes of single ion transfer atpre-formed tonoplast.     

Salt Tolerance in the Triticeae: K/Na Discrimination in Barley

Concentrations of ions were measured in the youngest fully-expandedleaves of Triticum aestivum, T. durum, Hordeum vulgare, H. spontaneum,Secale cereale, and Aegilops squarrosa accessions grown in hydroponicculture in the presence of salt (NaCl+CaCl₂). Triticum aestivum,Secale cereale, and Ae. squarrosa had the low leaf Na and highleaf K concentrations typical of plants which contain the enhancedK/Na discrimination character originally found in Ae. squarrosa.T. durum and the Hordeum species did not have this character.The better growth of H. vulgare than of T. durum with similarsalt concentrations in the youngest fully-expanded leaves maybe a result of better compartmentation of Na, Cl, and K betweendifferent tisssues or between different compartments withincells. The enhanced K/Na discrimination character was expressedin disomic addition lines of H. vulgare chromosomes in Triticumaestivum. The H. vulgare variety Herta and its slender mutantboth had similar leaf cation concentrations, although they differedin growth rate when grown at 60 mol m^–3 NaCl. H. vulgareand T. durum seedlings grown in the absence of monovalent cationsaccumulated more ²²Na in their shoots than seedlings of otherspecies when incubated in 1.0 mol m^–3 NaCl labelled with²²Na. Key words: Salt, ion transport, I genome, barley, wheat     

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; )     

The Mineral Ion Relations of Mangroves I. Root Cell Compartments in a Salt Excluder and a Salt Secreter Species at Low Salinities

The efflux kinetics of ²²Na and ³⁶Cl from 48 h pre-loaded Rhizophoramangle and Avicennia germinans mangrove seedling roots grownin low salinity conditions have been documented. Estimates ofthe percent isotope distributed throughout the whole plant,the uptake rates, the amounts calculated to be present in theroot cell compartments, and the rate constants and half-timesof efflux were made. Little to no ²²Na or ³⁶Cl was noted asbeing distributed to leaf tissue in the red mangrove, Rhizophoramangle, substantiating its characterization as a non-secreting,salt excluding species. Significantly greater net uptake ratesfor Na and Cl were observed for the black mangrove, Avicenniagerminans, a non-excluding, salt-secreting species. The typicalintracellular compartmentation found in higher plant root tissueswas noted to be present in these mangroves and the efflux kineticsclearly indicated that Na and Cl were handled differentiallyby these compartments and their membranes. Rhizophora manglerevealed greater apparent contents of Na than Cl in both thevacuolar and cytoplasmic phases (5.4 and 7.7 fold greater, respectively).The inverse was found for Avicennia germinans, with the apparentvacuolar compartment amount for Cl being 4.6 times greater thanthe Na amount and the cytoplasmic Cl amount was 1.5 times morethan Na contents. The rate constants of efflux showed a patternof significantly greater Na than Cl efflux from each respectivered mangrove compartment, while the black mangroves had a patternof greater efflux rates of Cl than Na in each compartment. Thehalf-times of efflux for Na from the black mangrove cytoplasmiccompartment were 3.5 times longer, while vacuolar half-timesof efflux were near unity for Na and Cl. The results are discussedin terms of Scholander's model of salt secreting and salt excludingspecies. (Received March 26, 1984; Accepted June 26, 1984)     

Salt Tolerance in the Triticeae: Leymus sabulosus

Elymus dahuhcus, Leymus giganteus, L. angustus, L. sabulosusand, to a lesser extent, L. triticoides, were found to tolerate200 mol m^–3 NaCl in solution culture. Elymus dahuricusdiffered from the Leymus species in its ion-uptake characteristics,showing a greater uptake of Cl and Na and a greater loss ofK from the shoots. In a more detailed experiment on Leymus sabulosusit was found that transpiration rates altered rapidly in responseto changes in external salinity whereas the accumulation ofNa and Cl in the leaves exhibited a lag of several days. Insalt stressed L. sabulosus Cl partially replaced the high levelsof nitrate found in the leaves of control plants. Glycinebetainelevels increased in the leaves from 8.0 mol m^–3 plantsap in the controls to 28 mol m^–3 plant sap at 250 molm^–3 NaCl. Key words: Salt stress, Transpiration, Solute accumulation, Leymus     

Airway surface fluid volume and Cl content in cystic fibrosis and normal bronchial xenografts

We describe theuse of an in vivo human bronchial xenograft model of cystic fibrosis(CF) and non-CF airways to investigate pathophysiological alterationsin airway surface fluid (ASF) volume (V_s) and Cl content.V_s was calculated based on thedilution of an impermeable marker,[³H]inulin, duringharvesting of ASF from xenografts with an isosmotic Cl-free solution.These calculations demonstrated thatV_s in CF xenographs (28 ± 3.0 µl/cm²;n = 17) was significantly less thanthat of non-CF xenografts (35 ± 2.4 µl/cm²;n = 30). The Cl concentration of ASF([Cl]_s) wasdetermined using a solid-state AgCl electrode and adjusted for dilutionduring harvesting using the impermeable[³H]inulin marker.Cumulative results demonstrate small but significant elevations(P < 0.045) in[Cl]_s in CF (125 ± 4 mM; n = 27) compared with non-CF(114 ± 4 mM; n = 48) xenografts.To investigate potential mechanisms by which CF airways may facilitatea higher level of fluid absorption yet retain slightly elevated levelsof Cl, we sought to evaluate the capacity of CF and non-CF airways toabsorb both ²²Na and³⁶Cl. Two consistent findings wereevident from these studies. First, in both CF and non-CF xenografts,²²Na and³⁶Cl were always absorbed in anequal molar ratio. Second, CF xenografts hyperabsorbed (~1.5-foldhigher) both ²²Na and³⁶Cl compared with non-CFxenografts. These results substantiate previously documented findingsof elevated Na absorption in CF airways and also suggest that theslightly elevated[Cl]_s found in thisstudy of CF xenograft epithelia does not occur through a mechanism ofdecreased apical permeability to Cl.     

Ion Composition of the Chara Internode

Ion compositions of the cytoplasm and the vacuole of Chara australiswere analyzed according to Kishimoto and Tazawa (1964) and Kiyosawa(1979a). The ions in the cytoplasm and the vacuole analyzedwere K⁺, Na⁺, Ca²⁺, Mg²⁺, Cl^–, NO₃^– and H₂PO₄^–.Assuming that the volume of the cytoplasm V_p is 10% of thatof the whole cell V, the concentrations of K⁺, Na⁺, Ca²⁺, Mg²⁺,Cl^–, NO₃^– and H₂PO₄^– in the cytoplasm averaged70, 15, 13, 4.6, 31, 2.2 and 16 mM, respectively. If the volumeof the cytoplasm was assumed to be 5% of that of the whole cell,their averaged concentrations were 139, 31, 25, 9.2, 62, 4.4and 33 mM, respectively. The averaged ion compositions of thecell sap were K⁺, 111; Na⁺, 47; Ca²⁺, 4.4; Mg²⁺, 8.9; Cl^–,91; NO^–₃, 3.3 and H₂PO₄^–, 6.0 mM. These values,taking the concentrations and the charges of the protein (Kiyosawa1979b) and amino acids (Sakano and Tazawa 1984) into accountand assuming the presence of some uni- or oligovalent anionsand/or small nonelectrolyte molecules, could explain fairlywell both the electroneutrality and the osmotic pressure ofthe cell, except when V_p/V = 5%. (Received May 18, 1987; Accepted September 29, 1987)     

Ionic Interactions of Petiole and Lamina During the Life of a Leaf of Castor Bean (Ricinus communis L.) Under Moderately Saline Conditions

Ion (K⁺, Na⁺, Mg²⁺, Ca²⁺ and Cl) flows and partitioning in thepetiole and lamina of leaf 6 of castor bean {Ricinus communisL.) plants growing in the presence of a mean of 71 mol m^–3NaCl were described by an empirical modelling technique. Thiscombined data on changes in ion contents of petiole and lamina,ion: carbon molar ratios of phloem bleeding sap and pressure-inducedxylem exudates of the leaf with previously described informationon the economies of C and N in identical leaf material. Datawere expressed as daily exchanges of ions in xylem and phloem,or depicted as models of ion balance and transport activityof petiole and lamina during four consecutive phases of leaflife. The early import phase was characterized by high intakeof K and Mg through phloem, and of Ca mainly through xylem,but only limited intake of Na and Cl. The next phase up to fullleaf expansion showed similar relative differences in xylemintake between ions and the onset of rapid phloem export fromthe lamina of K and Mg, some export of Na and Cl but scarcelyany of Ca. The next mature phase, marked by maximal photosynthesisand transpiration by the leaf, showed high xylem intake of allions in xylem. This was more than matched by phloem export ofMg and K, but by only fractional re-export of Na and Cl andagain very limited cycling through the leaf of Ca. The finalpre-senescence phase exhibited similar behaviour, but with generallygreater contribution to phloem transport from mobilization ofion reserves of the lamina. The petiole retained particularlylarge amounts of Na and Cl in its early growth, thereby protectingthe lamina from excessive entry of salt, but these petiolarpools, together with those or other nutrient ions, were laterpartially mobilized to the lamina via the xylem stream. Datawere discussed in relation to the relatively high salt toleranceexhibited by the species. Key words: Ricinus communis, xylem and phloem transport, ion balance, K+ economy, Na+ exclusion, NaCl-stress, salt tolerance, leaf development