Control of Na+ and K+ transport in Spergularia marina. III. Relationship between ion uptake and growth at moderate salinity

In this paper, we continue our analysis of Na⁺ and K⁺ uptake by mid-vegetative Spergularia marina (L.) Griseb. plants growing on 0.2x sea water medium, with attention to the relationship of ion uptake and growth. In the first part of the paper, growth analysis techniques are used to compare relative growth rates (RGR) and relative accumulation rates (RAR) for Na⁺ and K⁺. Under constant growth conditions, a high correlation between RGR and RAR indicated that growth and accumulation of both ions were well balanced, resulting in Na⁺ and K⁺ concentrations within the plants which were stable after adjustement to the saline medium. The analysis confirmed the existence of a Na⁺ -related growth stimulation in S. marina and an associated increase in the efficiency of K⁺ utilization for growth. When plants were subjected to more rapid salinization and step changes in the light intensity of the growth chamber, RGR and RAR were again similar, even through the discontinuities in growth conditions, suggesting that growth and ion accumulation were co-regulated rather than simply correlated. The growth analysis data were then transformed to give net uptake rates for Na⁺ and K⁺ and the results were compared to those of isotope studies under similar growth conditions. In roots, the rates estimated by the two techniques differed substantially; net uptake rates reflected primarily growth, while isotope studies indicated a substantial ion exchange rate between mature cells and the growth medium. The rates of transport of either Na⁺ or K⁺ to the shoot were very similar using the two estimation techniques. As the rates measured with isotopes were taken from studies lasting at most a few hours, this suggested a very rapid turnover of the upwardly mobile Na⁺ and K⁺ pools in the roots.     

Control of Na+ and K+ transport in Spergularia manrina. II. Effects of plant size, tissue ion contents and root-shoot ratio at moderate salinity

In this paper we continue our analysis of Na⁺ and K⁺ uptake in vegetative Spergularia marina (L.) Griseb. plants growing on 0.2x sea water in solution culture. We consider the relationships between isotope uptake and plant size, root: shoot ratio (RSR) and total ion contents, both individually (with the linear effects of time removed) and in combinations through stepwise multiple linear regression. The results differ from those of other studies, representing in our case inherent variability in a homogeneous population under steady-state growth conditions. The results were broadly similar for ²²Na⁺ and ⁴²K⁺. Total uptake was significantly and negatively correlated with RSR and root weight (W_r), and positively with root K⁺ content (K_r). These 3 variables were mutually correlated, however, and this was reflected in the multiple analyses as a reduction or loss of significance of one or more of the measures. Transport to the shoot was very highly correlated with total uptake (r² > 0.99 for both isotopes), resulting in nearly identical regression results. In multiple regression analyses of root data alone, accumulation was related only to root contents, but in a manner inconsistent with the allosteric regulation hypothesis, the most significant correlation being positive with K_r. The results were nearly identical for the two isotopes. The results were not consistent with a single factor regulatory system involving only initial root plamalemma ion influx. The observed Na⁺-K⁺ and root-shoot balances seem to require at least involvement of symplast-to-medium and symplast-to-xylem transport steps. Though the biochemical and biophysical signalling and transduction steps are not known, a physiological working hypothesis is presented, in which the positive correlation of uptake with root contents is balanced by a negative feedback signal deriving from plant size and by the diluting effects of growth. Considered over the vegetative period, these would produce the observed stability of plant contents during growth. The negative interaction with RSR is postulated to manifest the integrating system required to deliver ions to the shoot at the required rates.     

Long-term effects of plant hormones on K+ levels and transport in young wheat plants of different K+ status

Long-term effects of 1-naphtaleneacetic acid (NAA), benzyladenine (BA), gibberellic acid (GA₃), abscisic acid (ABA) and ethylene on K⁺ levels, K⁺ uptake and translocation to the shoot were studied in young wheat plants (Triticum aesticum L. cv. Martonvásári-8) grown at different K⁺ supplies. Na⁺ levels and K⁺/Na⁺ selectivity were also investigated. Both in shoots and roots, NAA, BA and ABA decreased K⁺ and Na⁺ levels more effectively in high-K⁺ plants than in low-K⁺ plants. GA, and ethylene did not influence K⁺ and Na⁺ levels. K⁺/Na⁺ selectivity in roots of low-K⁺ plants was increased in favour of K⁺ by BA, NAA and to a lesser extent by ABA. In high-K⁺ plants only BA increased the K⁺/Na⁺ ratio, whereas the effects of the other hormones were the opposite (NAA) or less pronounced (ABA). K⁺(⁸⁶Rb) uptake was inhibited by NAA and BA in low-K⁺ plants but not in high-K⁺ plants. K⁺(⁸⁶Rb) uptake was inhibited throughout by 10 μM ABA. K⁺(⁸⁶Rb) translocation to the shoot was influenced by the hormones similarly to the uptake patterns, with the exception of ABA, which inhibited translocation in low-K⁺ plants but not in high-K⁺ plants. The results show that hormonal effects may quantitatively and qualitatively be modified by K⁺ levels in the plant and that internal K⁺ concentration may play a role in the mechanisms regulating the effects of NAA, BA and ABA but probably not in those of GA₃ or ethylene.     

Effects of interrupted K+ supply on growth and uptake of K+, Ca2+, Mg2+ and Na+ in spring wheat

Effects of interrupted K⁺ supply on different parameters of growth and mineral cation nutrition were evaluated for spring wheat (Triticum aestivum L. cv. Svenno). K⁺ (2.0 mM) was supplied to the plants during different periods in an otherwise complete nutrient solution. Shoot growth was reduced before root growth after interruption in K⁺ supply. Root structure was greatly affected by the length of the period in K⁺ -free nutrient solution. Root length was minimal, and root branching was maximal within a narrow range of K⁺ status of the roots. This range corresponded to cultivation for the last 1 to 3 days, of 11 in total, in K⁺ -free nutrient solution, or to continuous cultivation in solution containing 0.5 to 2 mM K⁺. In comparison, both higher and lower internal/external K⁺ concentrations had inhibitory effects on root branching. However, the differing root morphology probably had no significant influence on the magnitude of Ca²⁺, Mg²⁺ and Na⁺ uptake. Uptake of Ca²⁺ and especially Mg²⁺ significantly increased after K⁺ interruption, while Na⁺ uptake was constant in the roots and slowly increased in the shoots. The two divalent cations could replace K⁺ in the cells and maintain electroneutrality down to a certain minimal range of K⁺ concentrations. This range was significantly higher in the shoot [110 to 140 μmol (g fresh weight)^?1] than in the root [20 to 30 μmol (g fresh weight)^?1]. It is suggested that the critical K⁺ values are a measure of the minimal amount of K⁺ that must be present for physiological activity in the cells. At the critical levels, K⁺ (⁸⁶Rb) influx and Ca²⁺ and Mg²⁺ concentrations were maximal. Below the critical K⁺ values, growth was reduced, and Ca²⁺ and Mg²⁺ could no longer substitute for K⁺ for electrostatic balance. In a short-term experiment, the ability of Ca²⁺ to compete with K⁺ in maintaining electroneutrality in the cells was studied in wheat seedlings with different K⁺ status. The results indicate that K⁺, which was taken up actively and fastest at the external K⁺ concentration used (2.0 mM), partly determines the size of Ca²⁺ influx.     

Control of K+ (Rb+) influx and transport with changed internal K+ concentration and age in two varieties of barley

The influx of Rb⁺ into the roots of two barley varieties (Hordeum vulgare L. cv. Salve and cv. Ingrid) from a K⁺-free ⁸⁶Rb-labelled nutrient solution with 2.0 mM Rb⁺, was checked at intervals from day 6 to day 18. The control plants were continuously grown in complete nutrient solution containing 5.0 mM K⁺, while two other groups of plants were grown in K⁺-free nutrient solution starting on day 6 and between day 6 and day 9, respectively. The pattern of Rb⁺ influx was similar for both varieties, although their efficiencies in absorbing Rb⁺ were different. The relationship between Rb⁺ influx and K⁺ concentration of the root could be interpreted in terms of negative feedback through allosteric control of uptake across the plasmalemma of the root cells. Hill plots were bimodal, but in the opposite direction. The Hill coefficients, reflecting the minimum number of interacting allosteric binding sites for K⁺ (Rb⁺), were low (≤–3.0). It is discussed whether the threshold value, that is the breaking point in the Hill plot, is indicative of a changed efficiency of transporting units for K⁺ (Rb⁺) transport to the xylem. Moreover, feedback regulation might be involved in transport of K⁺ between root and shoot. The variation in K⁺ concentrations in the roots and shoots of control plants were cyclic but in phase opposition despite an exponential growth. The average K⁺ concentration varied only slightly with age.     

The Effect of Dichlorophenyldimethylurea on Light-Stimulated 22Na+, 42K+ and 86Rb+ Absorption in Different Tissues of Phaseolus vulgaris Leaves

The light-stimulated absorption of ⁸⁶Rb⁺ by Phaseolus vulgaris L. leaf slices was found to be sensitive to dichlorophenyldimethylurea in air as well as in nitrogen, whereas light-stimulated ²²Na⁺ absorption in nitrogen was not sensitive to this inhibitor. The absorption of ²²Na⁺ is not affected by light in air. The absorption of ⁴²K⁺ is enhanced by a dichlorophenyldimethylurea-insensitive light effect under anaerobic conditions and further increased by light in the absence of the inhibitor. Light-enhanced ⁴²K⁺ absorption in air was also inhibited by dichlorophenyldimethylurea. Previous work showed that light-stimulated ⁸⁶Rb⁺ and ⁴²K⁺ absorption by Phaseolus vulgaris leaf slices is restricted to the guard cells. The present results are discussed with reference to the effect of light on stomatal opening.     

Uptake by Yeast: Interaction of Rb+, Na+ and Phosphate

It has been shown that addition of phosphate to phosphate deficient yeast gives rise to an immediate increase in the rate of Na⁺ uptake and an immediate decrease in the rate of Rb⁺ uptake. In addition, phosphate uptake is enhanced specifically by Na ions presumably by a process with a very high affinity for phosphate with a K_m of about 2 × 10⁻⁶M at pH 7.2, whereas the K_m for phosphate uptake of the Na⁺ independent process amounts to 1.3 × 10⁻⁴M.     

Na+/H+ antiport activity in tonoplast vesicles isolated from sunflower roots induced by NaCl stress

Na⁺ transport across the tonoplast and its accumulation in the vacuoles is of crucial importance for plant adaptation to salinity. Mild and severe salt stress increased both ATP- and PP_i-dependent H⁺ transport in tonoplast vesicles from sunflower seedling roots, suggesting the possibility that a Na⁺/H⁺ antiport system could be operating in such vesicles under salt conditions (E. Ballesteros et al. 1996. Physiol. Plant. 97: 259–268). During a mild salt stress, Na⁺ was mainly accumulated in the roots. Under a more severe salt treatment, Na⁺ was equally distributed in shoots and roots. In contrast to what was observed with Na⁺, all the salt treatments reduced the shoot K⁺ content. Dissipation by Na⁺ of the H⁺ gradient generated by the tonoplast H⁺-ATPase, monitored as fluorescence quenching of acridine orange, was used to measure Na⁺/H⁺ exchange across tonoplast-enriched vesicles isolated by sucrose gradient centrifugation from sunflower (Helianthus annuus L.) roots treated for 3 days with different NaCl regimes. Salt treatments induced a Na⁺/H⁺ exchange activity, which displayed saturation kinetics for Na⁺ added to the assay medium. This activity was partially inhibited by 125 μM amiloride, a competitive inhibitor of Na⁺/H⁺ antiports. No Na⁺/H⁺ exchange was detected in vesicles from control roots. The activity was specific for Na⁺. since K⁺ added to the assay medium slightly dissipated H⁺ gradients and displayed non-saturating kinetics for all salt treatments. Apparent K_m for Na⁺/H⁺ exchange in tonoplast vesicles from 150 mM NaCl-treated roots was lower than that of 75 mM NaCl-treated roots, V_max remaining unchanged. The results suggest that the existence of a specific Na⁺/H⁺ exchange activity in tonoplast-enriched vesicle fractions, induced by salt stress, could represent an adaptative response in sunflower plants, moderately tolerant to salinity.     

Ammonia excretion in the seawater blue crab (Callinectes sapidus) occurs by diffusion, and not Na+/NH4+ exchange

Summary A series of experiments was conducted to investigate whether ammonia is excreted across the seawater-acclimated blue crab's gills as ionized NH ₄ ⁺ or as the free base, NH₃. The net excretion rate of ammonia was not changed by transfer of the crabs to reduced (150 mM) Na⁺ solutions, by transfer to Na⁺- and K⁺-free artificial sea water, or by the sodium transport inhibitor amiloride. Ammonia excretion, therefore, does not appear to be linked to Na⁺ uptake in these animals, and appears to take place by passive diffusion. Since ammonia could diffuse either as NH ₄ ⁺ or NH₃, we examined two other kinds of evidence. The trans-epithelial potential was measured in sea water and the various artificial media. In spite of a 10 mV more negative potential in Na⁺-, K⁺-free medium, the ammonia excretion was not reduced. Also, in alkalinized seawater in which the partial pressure gradient of NH₃ was reduced, but the concentration gradient of NH ₄ ⁺ increased, ammonia excretion was reduced by about 70%. These results are consistent with the conclusion that ammonia excretion takes place by diffusion of the free base, NH₃.Abbreviations SW sea water - ASW artificial sea water - t.e.p. transepithelial potential The University of Texas Marine Science Institute Contribution No. 461Supported by NSF Grant PCM77-24358     

Compartmental analysis of efflux of K+(86Rb+) and Rb+(86Rb+) from roots of intact plants of barley (Hordeum vulgare) of high and low K+ status, and after excision of the shoot

The classic compartment analysis of ion efflux from roots is often applied with the assumption that there is a system of 3 compartments in series. However, complex ion transport across the root tissues, as well as influences from the shoot, may complicate the picture. The present experiments were performed to study the immediate effects that excision of the shoot before the experiment exerts on the efflux of Rb⁺(⁸⁶Rb⁺) and of K⁺(⁸⁶Rb⁺) from 9-day-old roots of plants of barley (Hordeum vulgare L. cv. Salve). The efflux from high K⁺ and low K⁺ roots of intact and detopped plants were compared. After excision of the shoot of high K⁺ plants, a marked increase in efflux was observed after 2.5 h with a maximum at about 7 h. The increase in efflux was seen as a peak in plots of efflux versus time. Excision of the shoot from low K⁺ roots did not give rise to a consistent increase in efflux. Regular K⁺ ion efflux curves were observed from roots of intact plants of high or low K⁺ status. Furthermore, after a pulse treatment of 9-day-old roots of intact plants of high or low K⁺ status with a solution containing Rb⁺(⁸⁶Rb⁺), the Rb⁺(⁸⁶Rb⁺) transport to the shoots was not reduced during the following 3 h in unlabelled solution. It is suggested that both the peak appearing in the efflux plots and the maintained tracer transport to the shoots after transfer of the roots to an unlabelled solution indicate the existence of a K⁺/Rb⁺ transport system in the symplasm of the roots that has only a slow exchange with the bulk cytoplasm and vacuoles.     

Mechanism for Cd2+ inhibition of (K++ Mg2+)ATPase activity and K+ (86Rb+) uptake join roots of sugar beet (Beta vulgaris)

Steady state kinetics were used to examine the influence of Cd²⁺ both on K⁺ stimulation of a membrane-bound ATPase from sugar beet roots (Beta vulgaris L. cv. Monohill) and on K⁺(⁸⁶Rb⁺) uptake in intact or excised beet roots. The in vitro effect of Cd²⁺ was studied both on a 12000–25000 g root fraction of the (Na⁺+K⁺+Mg²⁺)ATPase and on the ATPase when further purified by an aqueous polymer two-phase system. The observed data can be summarized as follows: 1) Cd²⁺ at high concentrations (>100 μM) inhibits the MgATPase activity in a competitive way, probably by forming a complex with ATP. 2) Cd²⁺ at concentrations <100 μM inhibits the specific K⁺ activation at both high and low affinity sites for K⁺. The inhibition pattern appears to be the same in the two ATPase preparations of different purity. In the presence of the substrate MgATP, and at K⁺ <5 mM, the inhibition by Cd²⁺ with respect to K⁺ is uncompetitive. In the presence of MgATP and K⁺ >10 μM, the inhibition by Cd²⁺ is competitive. 3) At the low concentrations of K⁺, Cd²⁺ also inhibits the 2,4-dinitrophenol(DNP)-sensitive (metabolic) K⁺(⁸⁶Rb⁺) uptake uncompetitively both in excised roots and in roots of intact plants. 4) The DNP-insensitive (non metabolic) K⁺(⁸⁶Rb⁺) uptake is little influenced by Cd²⁺. As Cd²⁺ inhibits the metabolic uptake of K⁺(⁸⁶Rb⁺) and the K⁺ activation of the ATPase in the same way at low concentrations of K⁺, the same binding site is probably involved. Therefore, under field conditions, when the concentration of K⁺ is low, the presence of Cd²⁺ could be disadvantageous.     

Na+ accumulation in shoot is related to water transport in K+-starved sunflower plants but not in plants with a normal K+ status

Sunflower plants (Helianthus annuus L. cv Sun-Gro 380) grown in nutrient solutions with different K⁺ levels were used to study the effect of potassium status on water uptake, Na⁺ uptake and Na⁺ accumulation in the shoot. Changes in nutrient potassium levels induced evident differences in internal potassium content. When both low and normal-K⁺ plants were exposed to 22 °C and salinity conditions (25 or 50 mM NaCl) during a short time period (9 h), water uptake in low-K⁺ plants was greater than in normal-K⁺ plants. In addition, K⁺ starvation favoured the Na⁺ uptake and the Na⁺ accumulation both in the root and in the shoot. When the plants were exposed to heat stress by a sharp increase of the temperature to 32 °C during the same period of time, the stimulating effect of K⁺ starvation on the water uptake was even greater. The high temperature increased Na⁺ uptake in both types of plants, but the Na⁺ accumulation in the shoot was only favoured in low-K⁺ plants. The results suggest that Na⁺ accumulation in the shoot is more dependent on the water uptake in low-K⁺ plants than in normal-K⁺ plants, and this effect could explain the greatest susceptibility to the salinity in K⁺-starved plants under high transpiration conditions, which are typical in dry climates.     

The effect of sodium chloride on the Ca2+, K+ and Na+ concentrations of the seed coat and embryo of Acacia tortilis and A. coriacea

The uptake of Na⁺ and the loss of Ca²⁺ and K⁺ by seeds of Acacia tortilis (Forsk.) Hayne (salt tolerant) and A. coriacea DC. (salt sensitive) were determined after 24 h soaking in 250 mol m^-1,3 NaCl or in distilled water. Na⁺ uptake was higher by the seed coat than by the embryo of both species and higher by A. coriacea than by A. tortilis. The greater Na⁺ uptake by A. coriacea was associated with greater Ca and K⁺ leakage. The Na⁺ concentration of solution imbibed by the embryo of both species was lower than the Na⁺ concentration in the external solution, indicating an exclusion of Na⁺. When A. tortilis and A. coriacea seeds were treated with a series of NaCl concentration (0–400 mol m^-1,3), the exclusion mechanism was particularly clear with A. tortilis at lower concentrations (50 and 150 mol m^-1,3) of NaCl. In contrast, the seed coat of both species accumulated Na⁺. Thus the seed coat may play an important role in ion exchange. These results show that it is important to consider the seed coat and embryo separately rather than the whole seed when considering ion exchange in relation to salinity tolerance.     

Kinetic studies of a (Na++ K++ Mg2+)ATPase in sugar beet roots. III. A proposed model for the (Na++ K+) activation and its significance for field properties

A microsomal (Na⁺+ K⁺+ Mg²⁺)ATPase preparation from sugar beet roots was used. The activation by simultaneous addition of Na⁺ and K⁺ at different levels was examined in terms of steady state kinetics. The observed data can be summarized in the following way: 1. The apparent affinity between the enzyme and the substrate MgATP depends on the ratio between Na⁺ and K⁺. At low Na⁺ concentration (below 5 mM), the apparent K_m decreases with increasing concentrations of K⁺ (1–20 mM). At 5 mM Na⁺, the K⁺ level does not change the apparent K_m, while at Na⁺ levels above 10 mM, the apparent K_m between enzyme and substrate increases with increasing concentration of K⁺. 2. When the MgATP concentration is kept constant, homotropic cooperativity (concerning one type of ligand) and heterotropic cooperativity (concerning different types of ligands) exist in the activation by Na⁺ and K⁺. The Na⁺ binding is cooperative with different K_m values and Hill coefficients (n) in the presence of low and high concentration of K⁺. At low Na⁺ level (< 5 mM). a negative cooperativity exists for Na⁺ (n_Na < 1) which is more pronounced in the presence of high [K⁺]. When the concentration of Na⁺ is raised the negative cooperativity disappears and turns into a positive one (n_Na > 1). Only K⁺ binding in the presence of low [Na⁺] shows cooperativity with a Hill coefficient that reflects changes from negative to positive homotropic cooperativity with increasing concentrations of K⁺ (n_K < 1 → n_K > 1). In the presence of [Na⁺] > 10 mM, the changes in n_k are insignificant. 3. A model is proposed in which one or two different K sites and one or two Na sites control the catalytic activity, with multiple interactions between Na⁺, K⁺ and MgATP. 4. In the presence of Na⁺ (< 10 mM), K⁺ is probably bound to two K sites, one of which translocates K⁺ through the membrane by an antiport Na⁺/K⁺ mechanism. This could be connected with an elevated K⁺ uptake in the presence of Na⁺ and could therefore explain some field properties of sugar beets.     

Involvement of cation channels and NH4+-sensitive K+ transporters in Na+ uptake by cowpea roots under salinity

Na⁺ accumulation was investigated in the roots of 11-d-old cowpea [Vigna unguiculata (L.) Walp.] plants. The relative contribution of different membrane transporters on Na⁺ uptake was estimated by applying Ca²⁺, K⁺, NH₄ ⁺, and pharmacological inhibitors. Na⁺ accumulation into the root symplast was decreased by half in the presence of 1 mM Ca²⁺ and it was almost abolished by 100 mM K⁺. The inhibitory effect of external NH⁴⁺ on Na⁺ accumulation was more pronounced in the roots of NH₄ ⁺-free growing plants. Na⁺ accumulation was reduced about 73 % by 0.1 mM flufenamate and it was almost blocked by 2 mM quinine. In addition, 20 mM tetraethylammonium and 1.0 mM Cs⁺ decreased Na⁺ accumulation by 28 and 30 %, respectively. These results evidenced that low-affinity Na⁺ uptake by cowpea roots depends on Ca²⁺-sensitive and Ca²⁺-insensitive pathways. The Ca²⁺-sensitive pathway is probably mediated by nonselective cation channels and the Ca²⁺-insensitive one may involve K⁺ channels and to a lesser extent NH₄ ⁺-sensitive K⁺ transporters.     

Kinetic Studies of a (Na++ K++ Mg2+) ATPase in Sugar Beet Roots

Kinetic studies of a dithiothreitol treated membrane ATPase fraction from sugar beet roots led to the following conclusions: 1) In the presence of MgATP, Na⁺ and K⁺ stimulate the ATPase activity in different ways following simple Michaelis-Menten kinetics. Thus separate sites for Na⁺ and K⁺ are suggested. 2) In the absence of K⁺, Na⁺ acts as an uncompetitive modifier raising the apparent K_m and V_max for MgATP. 3) In the absence of Na⁺, K⁺ activates non-competitively with respect to MgATP. Thus K⁺ increases V_max but does not affect the apparent affinity constant. 4) K⁺ and Na⁺ double the rate constants. 5) In the presence of Na⁺ or K⁺, Mg²⁺ in excess acts as a weak inhibitor to Na⁺ and/or K⁺ activity. 6) The temperature-activity dependence in the 5–40°C interval shows biphasic Arrhenius plots with the transition point between 15–18°C. The activation energy is lowered at temperatures > 18°C.     

Regulation of the uptake and distribution of Na+ in shoots of rice (Oryza sativa) variety Pokkali: role of Ca2+ in salt tolerance response

Soil salinity is a major factor affecting crop productivity worldwide. This study explores mechanisms that contribute to salt tolerance in rice (Oryza sativa L.). Hydroponically grown, 2-week-old salt tolerant and sensitive indica rice varieties, Pokkali and Jaya, respectively, were exposed to a 48-h stress period with NaCl (0–250 mM). When exposed to 200 mM NaCl, micromolar levels of external Ca²⁺ elevated survival of both varieties. The Ca²⁺ levels required were lower for Pokkali than for Jaya, but resulted in significantly higher survival. Estimates of Na⁺ and K⁺ in root and shoot compartments were made by flame photometry, while X-ray microanalysis was used to localize Na⁺ in the extracellular matrix of the shoot. Transpirational bypass flow was estimated using the apoplastic tracer, 8-hydroxypyrene-1,3,6-trisulphonic acid, trisodium salt. Our data demonstrate a Ca²⁺-dependent reduction in Na⁺ transport to shoots, which correlated with a decline in bypass flow and of Na⁺ in the transpirational stream. In addition, the Na⁺ that enters the shoot is partitioned among several distinct compartments. Survival is inversely correlated with Na⁺ levels in the shoot apoplastic fluid, which surrounds the cell and influences cytosolic composition. Pokkali maintained lower Na⁺ in its apoplast compared with the salt sensitive Jaya at the same total shoot Na⁺. Na⁺ in the apoplast appears to be regulated by sequestration into intracellular compartments. This sink supplements the primary response of reducing Na⁺ influx into the shoot and effectively buffers the apoplastic fluid in Pokkali. All of these mechanisms are operational in Jaya as well but are deployed less effectively.     

Differential effects of Na+, Mg2+, K+, Ca2+ and osmotic stress on the wild type and the NaCl-tolerant mutants stl1 and stl2 of Ceratopteris richardii

In order to identify physiological components that contribute to salinity tolerance, we compared the effects of Na⁺, Mg²⁺ and K⁺ salts (NaCl, Na₂SO₄, MgCl₂, MgSO₄, KCl and K₂SO₄), Ca₂₊ (CaSO₄), mannitol and melibiose on the wild type and the single-gene NaCl-tolerant mutants stl1 and stl2 of Ceratopteris richardii. Compared with gametophytic growth of the wild type, stl2 showed a low level of tolerance that was restricted to Na⁺ salts and osmotic stress. stl2 exhibited high tolerance to both Na⁺ and Mg²⁺ salts, as well as to osmotic stress. In response to short-term exposure (3 d) to NaCl, accumulation of K⁺ and Na⁺ was similar in the wild type and stl1. In contrast, stl2 accumulated higher levels of K⁺ and lower levels of Na⁺. Ca²⁺ supplementation (1.0 mol m^?3) ameliorated growth inhibition by Na⁺ and Mg²⁺ stress in wild type and stll, but not in stl2. In addition, under Na⁺ stress (175 mol m^?3) wild-type, stll and stl2 gametopbytes maintained higher tissue levels of K⁺ and lower levels of Na⁺ when supplemented with Ca²⁺ (1.0 mol m^?3). stl2 gametophytes were extremely sensitive to K⁺ supplementation. Growth of stl2 was greater than or equal to that of the wild type at trace concentrations of K⁺ but decreased substantially with increasing K⁺ concentration. Supplementation with K⁺ from 0 to 1.85 mol m^?3 alleviated some of the inhibition by 75 mol m^?3 NaCl in the wild type and in stl1. In stl2, growth at 75 mol m^?3 NaCl was similar at 0 and 1.85 mol m^?3 K⁺ supplementation. Although K⁺ supplementation above 1.85 mol m^?3 did not alleviate inhibition of growth by Na⁺ in any genotype, stl2 maintained greater relative tolerance to NaCl at all K⁺ concentrations tested.     

Properties of Mg2+-Stimulated and (Na++K+)-Activated Adenosine-5'-Triphosphatase from Sugar Beet Cotyledons

Sugar beet leaf homogenate contains Mg²⁺-stimulated ATPase activity with the highest specific activity in the 25,000–30,000 ×g-fraction. This fraction also has (Na⁺+ K⁺)-activated ATPase activity. Both activities have two pH optima, one stable at pH 7.9 and one variable at lower pH. When optimal conditions of Na⁺ and K⁺ were tested with 64 combinations of these ions, at least two mountains of activity were revealed. The (Na⁺+ K⁺)-ATPase had a high specificity for ATP. It had lost about 50% of its original activity after 56 days of storage at ?85°C. The activity drop was most pronounced at high ionic concentrations in the test medium. The (Na⁺+ K⁺)-ATPase shows four peaks of activity when tested at constant ionic strength. The idea is put forward that the four peaks reflect two ATPases, one in the tonoplast and one in the plasmalemma, which undergo conformational changes in relation to the ionic milieu.     

Sodium transport measured in plasma membrane vesicles isolated from wheat genotypes with differing K+/Na+ discrimination traits

Right-side-out plasma membrane vesicles were isolated from wheat roots using an aqueous polymer two-phase system. The purity and orientation of the vesicles were confirmed by marker enzyme analysis. Membrane potential (Ψ)-dependent ²²Na⁺ influx and sodium/proton (Na⁺/ H⁺) antiport-mediated efflux across the plasma membrane were studied using these vesicles. Membrane potentials were imposed on the vesicles using either K⁺ gradients in the presence of valinomycin or H⁺ gradients. The ΔΨ was quantified by the uptake of the lipophilic cation tetraphenylphosphonium. Uptake of Na⁺ into the vesicles was stimulated by a negative ΔΨ and had a K_m for extrav-esicular Na⁺ of 34.8 ± 5.9 mol m³. The ΔΨ-dependent uptake of Na⁺ was similar in vesicles from roots of hexaploid (cv. Troy) and tetraploid (cv. Langdon) wheat differing in a K⁺/Na⁺ discrimination trait, and was also unaffected by growth in 50 mol m^?3 NaCl. Inhibition of ΔΨ-dependent Na⁺ uptake by Ca²⁺ was greater in the hexaploid than in the tetraploid. Sodium/proton antiport was measured as Na⁺-dependent, amiloride-inhibited pH gradient formation in the vesicles. Acidification of the vesicle interior was measured by the uptake of ¹⁴C-methylamine. The Na⁺/H⁺ antiport had a K_m, for intravesicular Na⁺ of between 13 and 19 mol m^?3. In the hexaploid, Na⁺/H⁺ antiport activity was greater when roots were grown in the presence of 50 mol m^?3NaCl, and was also greater than the activity in salt-grown tetraploid wheat roots. Antiport activity was not increased in a Langdon 4D chromosome substitution line which carries a trait for K⁺/Na⁺ discrimination. It is concluded that neither of the transport processes measured is responsible for the Na⁺/K⁺ discrimination trait located on the 4D chromosome of wheat.