Association of 14-3-3 proteins with the C-terminal autoinhibitory domain of the plant plasma-membrane H+-ATPase generates a fusicoccin-binding complex

The plant plasma-membrane H⁺-ATPase (EC 3.6.1.35) contains a C-terminal autoinhibitory domain whose displacement from the catalytic site is caused by treatment of intact plant tissue with the phytotoxin fusicoccin (FC). The FC-induced activation of the H⁺-ATPase was proposed to involve a direct interaction of 14-3-3 proteins with the H⁺-ATPase. By analysing plasma membranes derived from leaves of Commelina communis L., direct biochemical evidence has now been obtained for a complex between the C-terminus of the H⁺-ATPase and a 14-3-3 dimer. Stabilization of this complex was achieved by FC treatment in vivo or in vitro. Furthermore, the C-terminal domain of the H⁺-ATPase in association with a 14-3-3 dimer is essential for the creation of a functional FC-binding complex. Received: 1 August 1998 / Accepted: 15 September 1998     

Calcium-dependent phosphorylation regulates the plasma-membrane H+-ATPase activity of maize (Zea mays L.) roots

Phosphorylation/dephosphorylation of the plasma-membrane H⁺-ATPase (EC 3.6.1.35) could act as a regulatory mechanism to control its activity. In this work, a plasmalemma-enriched fraction from maize roots and a partially purified H⁺-ATPase were used to investigate the effects of Ca²⁺ and calmodulin on the H⁺-ATPase activity and on its phosphorylation status. Both the hydrolytic and the proton-pumping activities were reduced approximately 50% by micromolar Ca²⁺ concentrations while calmodulin did not show any effect either alone or in the presence of Ca²⁺. The lack of effect of calmodulin antagonists indicated that calmodulin was not involved in this response. The addition of staurosporine, a kinase inhibitor, abolished the inhibitory effect of Ca²⁺. Phosphorylation of plasma membrane and partially purified H⁺-ATPase showed the same behavior. In the presence of Ca²⁺ a polypeptide of 100 kDa was phosphorylated. This polypeptide cross-reacted with antibodies raised against the H⁺-ATPase of maize roots. The autoradiogram of the immunodetected protein clearly showed that this polypeptide, which corresponds to the H⁺-ATPase, was phosphorylated. Additional clear evidence comes from the immunoprecipitation experiments: the data obtained show that the H⁺-ATPase activity is indeed influenced by its state of phosphorylation. Received: 19 October 1998 / Accepted: 23 February 1999     

Transient and persistent tetrodotoxin-sensitive sodium currents in squid olfactory receptor neurons

Squid olfactory receptor neurons are primary bipolar sensory neurons capable of transducing water-born odorant signals into electrical impulses that are transmitted to the brain. In this study, we have identified and characterized the macroscopic properties of voltage-gated Na⁺ channels in olfactory receptor neurons from the squid Lolliguncula brevis. Using whole-cell voltage-clamp techniques, we found that the voltage-gated Na⁺ channels were tetrodotoxin sensitive and had current densities ranging from 5 to 169 pA pF⁻¹. Analyses of the voltage dependence and kinetics revealed interesting differences from voltage-gated Na⁺ channels in olfactory receptor neurons from other species; the voltage of half-inactivation was shifted to the right and the voltage of half-activation was shifted to the left such that a “window-current” occurred, where 10–18% of the Na⁺ channels activated and did not inactivate at potentials near action potential threshold. Our findings suggest that in squid olfactory neurons, a subset of voltage-gated Na⁺ channels may play a role in generating a pacemaker-type current for setting the tonic levels of electrical activity required for transmission of hyperpolarizing odor responses to the brain. Accepted: 1 October 1998     

Differential activation of H+-ATPase genes by an arbuscular mycorrhizal fungus in root cells of transgenic tobacco

In arbuscular mycorrhizas, H⁺-ATPase is active in the plant membrane around arbuscules but absent from plant mutants defective in arbuscule development (Gianinazzi-Pearson et al. 1995, Can J Bot 73: S526–S532). The proton-pumping H⁺-ATPase is encoded by a family of genes in plants. Immunocytochemical studies and promoter-gusA fusion assays were performed in transgenic tobacco (Nicotiana tabacum L.) to determine whether the periarbuscular enzyme activity results from de-novo activation of plant genes by an arbuscular mycorrhizal fungus. The H⁺-ATPase protein was localized in the plant membrane around arbuscule hyphae. The enzyme was absent from non-colonized cortical cells. Regulation of seven H⁺-ATPase genes (pma) was compared in non-mycorrhizal and mycorrhizal roots by histochemical detection of β-glucuronidase (GUS) activity. Two genes (pma2, pma4) were induced in arbuscule-containing cells of mycorrhizal roots but not in non-mycorrhizal cortical tissues or senescent mycorrhiza. It is concluded that de-novo H⁺-ATPase activity in the periarbuscular membrane results from selective induction of two H⁺-ATPase genes, which can have diverse roles in plant-fungal interactions at the symbiotic interface. Received: 23 October 1999 / Accepted: 7 February 2000     

The turnover of cell surface proteins of carrot protoplasts

The covalent modification of cell surface proteins with N-hydroxysuccinimide esters of biotin was used to develop a strategy for following the turnover of proteins on the surface of carrot (Daucus carota L.) protoplasts. A biotinylation/internalisation assay was established which enabled the turnover of cell surface proteins to be examined by biochemical and immunocytochemical techniques. The detection of biotinylated proteins after sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting indicated that a variety of proteins on the surface of the protoplasts were covalently modified. Immunolocalisation of biotinylated proteins in protoplasts directly after their derivatisation, demonstrated that the proteins were initially restricted to the cell surface. Incubation of biotinylated protoplasts at 25 °C for 1 h resulted in the detection of biotin-labelled proteins on the cell surface and intracellularly. A small proportion of these proteins was associated with coated pits, the Golgi apparatus and vacuolar compartments. Biochemical analysis of internalised proteins revealed that a polypeptide of approximate M_r 100 000 was internalised by the protoplasts. Immunolabelling of a biotinylated protein of M_r 100 000 by an antibody raised against an isoform of a tobacco plasma-membrane H⁺-ATPase, strongly suggests that the plasma-membrane H⁺-ATPase is internalised by carrot protoplasts. The implications of these results are discussed within the context of endocytosis in plants. Received: 13 July 1998 / Accepted: 11 November 1998     

Defensive Role of Quercetin Against Imbalances of Calcium,Sodium, and Potassium in Galactosemic Cataract

Galactosemic cataracts are characterized by electrolyte disturbances resulting in osmotic imbalance and loss of transparency. We have studied the defensive role of quercetin, a bioflavonoid, against the alterations of calcium (Ca²⁺), sodium (Na⁺), and potassium (K⁺) concentrations in galactose-induced cataract in a rodent model. The experimental study was conducted on weanling male Wistar rats with an average body weight of 34 ± 0.9 g. Different groups received normal stock AIN 93 diet (group A, n = 8), AIN 93 diet with 30% galactose (group B, n = 8), and AIN 93 diet with 30% galactose + quercetin at 400 mg/100 g diet (group C, n = 8). Aldose reductase activity and protein content and concentrations of Ca²⁺, Na+, and K⁺ were determined in normal and cataractous lenses. Treatment with quercetin resulted in a significant decrease in Na⁺ and Ca²⁺ and aldose reductase levels and an increase in K⁺ and protein levels in galactosemic cataractous lenses. These results imply that inclusion of quercetin contributes to lens transparency through the maintenance of characteristic osmotic ion equilibrium and protein levels of the lens.     

The effect of light on membrane potential, apoplastic pH and cell expansion in leaves of Pisum sativum L. var. Argenteum.

The connection between three light responses of green leaf cells-membrane potential (V_m), H⁺ net efflux and growth, was analyzed. Illumination of mesophyll cells in leaves from Argenteum peas caused two rapid responses: (i) a de- and repolarization of V_m and (ii) an alkalinization of the apoplast. The rapid responses were completely eliminated by the photosynthetic inhibitor 3-(3′,4′-dichlorophenyl)-1,1-dimethylurea (DCMU) but not affected by ortho-vanadate, an inhibitor of the plasma membrane (PM) H⁺-ATPase. The rapid changes were followed by a set of delayed responses: (i) a slow, gradual hyperpolarization of V_m, (ii) a gradual acidification of the mesophyll apoplast and (iii) an increased rate of elongation. These three light responses persisted under DCMU but were completely eliminated by vanadate. The data show that the delayed (in contrast to the rapid) responses were due to a stimulation of PM H⁺ pumps which occurred independently of non-cyclic photosynthetic electron transport and the “dark” processes depending on it. When the rapid responses were blocked by DCMU, light-induced acidification, hyperpolarization of the membrane potential and growth proceeded simultaneously. A shared (4-min) lag phase indicated slower signal processing in mesophyll than in epidermal cells where light stimulation of PM H⁺ pumps was rapid. Received: 3 September 1998 / Accepted: 15 October 1998     

High salt‐treatment‐induced Na+ extrusion and low salt‐treatment‐induced Na+ accumulation in suspension‐cultured cells of the mangrove plant,Bruguiera sexangula

A suspension‐cultured cell strain of the mangrove plant (Bruguiera sexangula) was established from a callus culture and maintained in an amino acid medium in the absence of NaCl. NaCl non‐adapted cells were transferred to media containing 0–200 mm NaCl. The initial growth rate decreased gradually with increasing salt concentrations. However, at up to 150 mm NaCl, cell number growth at the highest point was almost the same as that at lower salt concentrations. Cells even continued to grow in the presence of 200 mm NaCl. Cells incubated in a medium containing 50 mm NaCl for 3 weeks accumulated Na⁺, while those incubated in 150 mm NaCl for 2 d showed only a transient increase in Na⁺ and Cl^– concentrations. In the latter treatment, the intracellular concentration of Na⁺ returned to the original low level within 2 weeks. It took a longer time for Cl^– to return to its original level. As a result, the Na⁺ and Cl^– concentrations in cells cultured with 50 mm NaCl were much larger than those in cells cultured with 150 mm NaCl. The intracellular distribution of ions after transfer to the medium containing 150 mm NaCl was analysed by isolating the vacuoles. Treatment with amiloride, an inhibitor of the Na⁺/H⁺ antiporter, suppressed the recovery of Na⁺ to the original level in the cells. Treatment with 150 mm NaCl for 3 d stimulated the activities of both the vanadate‐dependent H⁺‐ATPase and the Na⁺/H⁺ antiporter in the plasma membrane fraction.     

Pinitol, a Compatible Solute in Mesembryanthemum crystallinum L.?

The irrigation of Mesembryanthemum crystallinum L. plants with400 mol m^–3 NaCl to induce crassulacean acid metabolism(CAM) was accompanied by the accumulation of pinitol. Pinitolconstituted 71% of the soluble carbohydrate fraction and 9.7%dry weight in the CAM form. Pinitol in the C₃ form did not exceed5% of the soluble carbohydrate fraction. Pinitol appeared metabolicallyinert: it was not readily degraded during 96 h of darkness inthe CAM form or during CAM deinduction. Preparations of CAMM. crystallinum protoplasts, vacuoles and chloroplasts showedpinitol to be chloroplastic at a concentration of about 230mol m^–3 and cytosolic at about 100 mol m^–3. No pinitolwas detected in vacuoles. CAM leaf extracts possessed a highermyo-inositol phosphate synthesising capacity than C₃ extracts,revealing greater activity in the CAM form of glucose-6-phosphatecycloaldolase, an enzyme in the pathway of pinitol synthesis. Although pinitol accumulation and CAM induction could not beseparated and appeared to be specific responses to water stress,there may not be a causal link between them. Pinitol may functionas a compatible solute in the cytosol and especially the chloroplaststo counteract the presence of high concentrations of Na⁺ andCl^– ions in the vacuole. The accumulation of pinitol,though apparently not directly related to CAM may, like CAM,be viewed as an aspect of the adaptation of the plant to a reductionin water availability. Key words: pinitol, Mesembryanthemum crystallinum L, CAM, compatible solute     

Auxin augments conductance of K+ inward rectifier in maize coleoptile protoplasts

Potassium is taken up by maize (Zea mays L.) coleoptile cells via a typical plant inward rectifier (K _ir ). Sufficient conductance of this channel is essential in order to maintain auxin-stimulated cell elongation. It was therefore investigated whether the activity of this channel is subject to direct or indirect control by this growth hormone. Patch-clamp measurements of whole coleoptile protoplasts revealed no appreciable effect of externally applied 10 μM or 100 μM α-naphthaleneacetic acid (NAA) on the activity of K _ir over test periods of ≥ 18 or ≥ 8 min, respectively. When, however, K _ir was recorded in the cell-attached configiuration and 10 μM NAA administered to the bath medium, the conductance of K _ir increased significantly in 13 out of 18 protoplasts over the control. This rise occurred at a fixed protoplast voltage after a lag period of less than 10 min and exhibited no voltage dependency. The absence of response to NAA of protoplasts in the whole-cell configuration indicates that auxin perception and channel control is linked via a soluble cytoplasmic factor and that this mediator is washed out or modified upon perfusion of the cytoplasm with pipette solution. To search for this expected diffusible factor the K _ir current was recorded before and after elevation of Ca²⁺ and H⁺ in the cytoplasm. In the whole-cell configuration the increase in Ca²⁺ from a nanomolar value to >1 μM by means of Ca²⁺-release from the caged precursor Na₂-DM-nitrophen left K _ir unaffected. The whole-cell K _ir conductance was also not affected upon addition of 10 mM Na⁺-acetate to the bath medium, an operation used to lower the cytoplasmic pH. This excludes a primary role for the known auxin-evoked rise in cytoplasmic Ca²⁺ and H⁺ in K _ir activity. We postulate that another, as yet unknown, mechanism mediates the auxin-evoked stimulation of the number of active K _ir channels in the plasma membrane. Received: 13 May 1998 / Accepted: 9 November 1998     

Involvement of 14-3-3 proteins in the osmotic regulation of H+-ATPase in plant plasma membranes

 Taking the binding of fusicoccin to plasma membranes as an indicator of complex formation between the 14-3-3 dimer and H⁺-ATPase, we assessed the effect of osmotic stress on the interaction of these proteins in suspension-cultured cells of sugar beet (Beta vulgaris L.). An increase in osmolarity of the cell incubation medium, accompanied by a decrease in turgor, was found to activate the H⁺ efflux 5-fold. The same increment was observed in the number of high-affinity fusicoccin-binding sites in isolated plasma membranes; the 14-3-3 content in the membranes increased 2- to 3-fold, while the H⁺-ATPase activity changed only slightly. The data obtained indicate that osmotic regulation of H⁺-ATPase in the plant plasma membrane is achieved via modulation of the coupling between H⁺ transport and ATP hydrolysis, and that such regulation involves 14-3-3 proteins. Received: 10 February 2000 / Accepted: 31 March 2000     

Establishment of stable NaCl-resistant rice plant lines from anther culture: distribution pattern of K+/Na+ in callus and plant cells

 Soil salinity markedly suppresses the growth of rice (Oryza sativa L.). We established rice anther culture to select for rice callus lines adapted to NaCl stress and regenerated plant progenies resistant to a NaCl stress of E.C. 16–18 mS. When exposed to NaCl, NaCl-adapted rice calli lost K⁺ and accumulated little Na⁺. Conversely, plant cells lost relatively little K⁺ and accumulated Na⁺. It is plausible that, NaCl-resistant mechanisms are different at callus and plant level. The stable NaCl-resistant lines produced have potential use in elucidating the molecular mechanisms behind NaCl resistance in rice and in rice breeding. Received: 27 February 1997 / Accepted: 4 April 1997     

Herbivore exposure alters ion fluxes and improves salt tolerance in a desert shrub

Plants have evolved complex mechanisms that allow them to withstand multiple environmental stresses, including biotic and abiotic stresses. Here, we investigated the interaction between herbivore exposure and salt stress of Ammopiptanthus nanus, a desert shrub. We found that jasmonic acid (JA) was involved in plant responses to both herbivore attack and salt stress, leading to an increased NaCl stress tolerance for herbivore-pretreated plants and increase in K⁺/Na⁺ ratio in roots. Further evidence revealed the mechanism by which herbivore improved plant NaCl tolerance. Herbivore pretreatment reduced K⁺ efflux and increased Na⁺ efflux in plants subjected to long-term, short-term, or transient NaCl stress. Moreover, herbivore pretreatment promoted H⁺ efflux by increasing plasma membrane H⁺-adenosine triphosphate (ATP)ase activity. This H⁺ efflux creates a transmembrane proton motive force that drives the Na⁺/H⁺ antiporter to expel excess Na⁺ into the external medium. In addition, high cytosolic Ca²⁺ was observed in the roots of herbivore-treated plants exposed to NaCl, and this effect may be regulated by H⁺-ATPase. Taken together, herbivore exposure enhance s A. nanus tolerance to salt stress by activating the JA-signalling pathway, increasing plasma membrane H ⁺ - ATPase activity, promoting cytosolic Ca²⁺ accumulation, and then restricting K⁺ leakage and reducing Na⁺ accumulation in the cytosol.     

Adaptation of the tonoplast V-type H+-ATPase of Mesembryanthemum crystallinum to salt stress, C3–CAM transition and plant age

Plants of the facultative halophyte and CAM species Mesembryanthemum crystallinum L. (Aizoaceae) were stressed for 8 d with 400 mol m⁻³ NaCl in the root medium. NaCl was then removed from the substratum, and the plants were watered again with NaCl-free solution. A second set of plants was maintained as controls. A small degree of CAM, as indicated by day-night changes in malate levels, was expressed during ageing of the plants. Salinity-stress-dependent CAM induction was reversible by the removal of salt, as indicated by similar Δ malate levels in previously salt-stressed plants and in non-stressed plants on day 19 of the experiment. Tonoplast vesicles were isolated from leaves during the time-course of stress application, stress removal and ageing. Parameters of the tonoplast H⁺-ATPase were correlated to the application of salinity, the expression of CAM and ageing. It was concluded, first, that a pronounced increase in the amount of tonoplast H⁺-ATPase is related to salinity per se and a smaller increase to ageing; secondly, that there is an increase in the specific activity of the enzyme related to ageing; thirdly, that the induction of two new polypeptides with molecular masses of 32 and 28 kDa is correlated in time with the expression of CAM, and, fourthly, that the two new polypeptides are part of the tonoplast H⁺-ATPase holoenzyme.     

Localization of pyrophosphatase in membranes of cauliflower inflorescence cells

Using a polyclonal antiserum specific for the tonoplastic H⁺-pyrophosphatase (tPPase), significant amounts of antigenic polypeptides of the correct molecular mass were detected in Western blots of plasma membrane isolated from cauliflower (Brassica oleracea L.) inflorescence by phase-partitioning and subsequent sucrose density centrifugation. Potassium iodide-stripped plasma membranes continued to give a strong positive signal, indicating that the PPase antigen detected was not a result of contamination through soluble PPase released during homogenisation. The same preparation contained negligible vacuolar (v)H⁺-ATPase activity and the A subunit of the vATPase could not be detected by immunoblotting. Plasma membrane fractions exhibited a proton-pumping activity with ATP as substrate, but such an activity was not measurable with pyrophosphate, although the hydrolysis of this substrate was recorded. By contrast, pyrophosphate supported proton pumping in tonoplast-containing fractions. Immunogold electron microscopy confirmed the presence of PPase at the plasma membrane as well as at the tonoplast, trans Golgi network, and multivesicular bodies. The density of immunogold label was higher at the plasma membrane than at the tonoplast, except for membrane fragments occurring in the lumen of the vacuoles which stained very conspicuously. Received: 29 June 1998 / Accepted: 9 November 1998     

Paxillus involutus mycorrhiza attenuate NaCl-stress responses in the salt-sensitive hybrid poplar Populus×canescens

In order to characterise the effect of ectomycorrhiza on Na⁺-responses of the salt-sensitive poplar hybrid Populus × canescens, growth and stress responses of Paxillus involutus (strain MAJ) were tested in liquid cultures in the presence of 20 to 500 mM NaCl, and the effects of mycorrhization on mineral nutrient accumulation and oxidative stress were characterised in mycorrhizal and non-mycorrhizal poplar seedlings exposed to 150 mM NaCl. Paxillus involutus was salt tolerant, showing biomass increases in media containing up to 500 mM NaCl after 4 weeks growth. Mycorrhizal mantle formation on poplar roots was not affected by 150 mM NaCl. Whole plant performance was positively affected by the fungus because total biomass was greater and leaves accumulated less Na⁺ than non-mycorrhizal plants. Energy dispersive X-ray microanalysis using transmission electron microscopy analysis of the influence of mycorrhization on the subcellular localisation of Na⁺ and Cl⁻ in roots showed that the hyphal mantle did not diminish salt accumulation in root cell walls, indicating that mycorrhization did not provide a physical barrier against excess salinity. In the absence of salt stress, mycorrhizal poplar roots contained higher Na⁺ and Cl⁻ concentrations than non-mycorrhizal poplar roots. Paxillus involutus hyphae produced H₂O₂ in the mantle but not in the Hartig net or in pure culture. Salt exposure resulted in H₂O₂ formation in cortical cells of both non-mycorrhizal and mycorrhizal poplar and stimulated peroxidase but not superoxide dismutase activities. This shows that mature ectomycorrhiza was unable to suppress salt-induced oxidative stress. Element analyses suggest that improved performance of mycorrhizal poplar under salt stress may result from diminished xylem loading of Na⁺ and increased supply with K⁺.     

Na<Superscript>+</Superscript>/H<Superscript>+</Superscript> exchange activity in the alkaliphile halotolerant cyanobacterium <Emphasis Type="Italic">Aphanothece halophytica</Emphasis>

The activity of Na⁺/H⁺ exchanger to remove toxic Na⁺ is important for growth of organisms under high salinity. In this study, the halotolerant cyanobacterium Aphanothece halophytica was shown to possess Na⁺/H⁺ exchange activity since exogenously added Na⁺ could dissipate a pre-formed pH gradient, and decrease extracellular pH. Kinetic analysis yielded apparent K _m (Na⁺) and V _max of 20.7 ± 3.1 mM and 3,333 ± 370 nmol H⁺ min⁻¹ mg⁻¹, respectively. For cells grown under salt-stress condition, the apparent K _m (Na⁺) and V _max was 18.3 ± 3.5 mM and 3,703 ± 350 nmol H⁺ min⁻¹ mg⁻¹, respectively. Three cations with decreasing efficiency namely Li⁺, Ca²⁺, and K⁺ were also able to dissipate pH gradient. Only marginal exchange activity was observed for Mg²⁺. The exchange activity was strongly inhibited by Na⁺-gradient dissipators, monensin, and sodium ionophore as well as by CCCP, a protonophore. A. halophytica showed high Na⁺/H⁺ exchange activity at neutral and alkaline pH up to pH 10. Cells grown at pH 7.6 under high salinity exhibited higher Na⁺/H⁺ exchange activity than those grown under low salinity during 15 days of growth suggesting a role of Na⁺/H⁺ exchanger for salt tolerance in A. halophytica. Cells grown at alkaline pH of 9.0 also exhibited a progressive increase of Na⁺/H⁺ exchange activity during 15 days of growth.     

Escherichia coli and Lactobacillus plantarum responses to osmotic stress

Escherichia coli and Lactobacillus plantarum were subjected to final water potentials of −5.6 MPa and −11.5 MPa with three solutes: glycerol, sorbitol and NaCl. The water potential decrease was realized either rapidly (osmotic shock) or slowly (20 min) and a difference in cell viability between these conditions was only observed when the solute was NaCl. The cell mortality during osmotic shocks induced by NaCl cannot be explained by a critical volume decrease or by the intensity of the water flow across the cell membrane. When the osmotic stress is realized with NaCl as the solute, in a medium in which osmoregulation cannot take place, the application of a slow decrease in water potential resulted in the significant maintenance of cell viability (about 70–90%) with regard to the corresponding viability observed after a sudden step change to same final water potential (14–40%). This viability difference can be explained by the existence of a critical internal free Na⁺ concentration. Received: 20 May 1998 / Received revision: 31 July 1998 / Accepted: 31 July 1998     

Spatial distributions and net deposition rates of mineral elements in the elongating wheat (Triticum aestivum L.) leaf under saline soil conditions

Wheat leaf growth is known to be spatially affected by salinity. The altered spatial distribution of leaf growth under saline conditions may be associated with spatial changes in tissue mineral elements. The objective of this study was to evaluate the spatial distributions of mineral elements and their net deposition rates in the elongating and mature zones of leaf 4 of the main stem of spring wheat (Triticum aestivum L. cv. Lona) during its linear growth phase under saline soil conditions. Plants were grown in an illitic-chloritic silty loam with 0 and 120 mM NaCl. Three days after emergence of leaf 4, sampling was begun at 3 and 13 h into the 16-h light period. Spatial distributions of fresh weight (FW), dry weight (DW), and Na⁺, K⁺, Cl⁻, NO⁻ ₃, Ca²⁺, Mg²⁺, total P, and total N in the elongating and mature tissues were determined on a millimeter scale. The patterns of spatial distribution of Na⁺, Cl⁻, K⁺, NO₃ ⁻, and Ca²⁺ in the growing leaves were affected by salinity, while those of Mg²⁺, total P, and total N were not. Sodium, K⁺, Cl⁻, Ca²⁺, Mg²⁺, and total N concentrations (mmol · kg⁻¹ FW) were consistently higher at 120 mM NaCl than at 0 mM NaCl along the leaf axis from the leaf base, whereas NO₃ ⁻ concentration was lower at 120 mM NaCl. Deposition rates of all nutrients were greatest in the elongation zone. The elongation zone was the strongest sink for mineral elements in the leaf tissues. Local net deposition rates of Na⁺, Cl⁻, Ca²⁺, and Mg²⁺ (mmol · kg⁻¹ FW · h⁻¹) in the most actively elongating zone were enhanced by 120 mM NaCl, whereas for NO₃ ⁻ this was depressed. The lower supply of NO⁻ ₃ to growing leaves may be responsible for the inhibition of growth under saline conditions. Higher tissue concentrations of Na⁺ and Cl⁻ may cause ion imbalance but probably did not result in ion toxicity in the growing leaves. Potassium, Ca²⁺, Mg²⁺, total P, and total N are less plausibly responsible for the reduction in leaf growth in this study. Higher tissue K⁺ and Ca²⁺ concentrations at 120 mM NaCl are probably due to the presence of high Ca²⁺ in the soil of this study. Received: 13 March 1997 / Accepted: 9 June 1997