Cloning and functional analysis of the K+ transporter, PhaHAK2, from salt-sensitive and salt-tolerant reed plants

We isolated PhaHAK2 cDNAs from salt-tolerant and salt-sensitive reed plants. PhaHAK2 belongs to group II by phylogenetic analysis, and was predicted to be a high-affinity plasma membrane K⁺ transporter. Yeast transformed with the PhaHAK2-u from salt-sensitive reed plants (Phragmites australis) had a decreased ability to take up K⁺ in the presence of NaCl and showed a higher Na⁺ permeability than yeast transformed with PhaHAK2-n or PhaHAK2-e from two salt-tolerant reed plants. These results suggest a possibility that the continuous K⁺ uptake by PhaHAK2 and maintenance of high K⁺/Na⁺ ratio under salt stress condition is one of the causes of the salt-tolerance in reed plants.     

Salt tolerance inNitellopsis obtusa

Summary The mechanism of salt tolerance was studied using isolated internodal cells of the charophyteNitellopsis obtusa grown in fresh water. When 100 mM NaCl was added to artificial pond water (0.1 mM each of NaCl, KC1, CaCl₂), no cell survived for more than one day. Within the first 30 minutes, membrane potential (E_m) depolarized and membrane resistance (R_m) decreased markedly. Simultaneously, cytoplasmic Na⁺ increased and K⁺ decreased greatly. At steady state the increase in Na⁺ content was roughly equal to the decrease in K⁺ content. The Cl^– content of the cytoplasm did not change. These results suggest that Na⁺ enters the cytoplasm by exchange with cytoplasmic K⁺. Both the entry of Na⁺ and the exit of K⁺ are assumed to be passive and the latter being caused by membrane depolarization. Vacuolar K⁺, Na⁺, and Cl^– remained virtually constant, suggesting that rapid influx of Na⁺ from the cytoplasm did not occur.In 100 mM NaCl containing 10 mM CaCl₂, membrane depolarization, membrane resistance decrease and changes in cytoplasmic [Na⁺] and [K⁺] did not occur, and cells survived for many days. When cells treated with 100 mM NaCl were transferred within 1 hour to 100 mM NaCl containing 10 mM CaCl₂, Em decreased, Rm increased, cytoplasmic Na⁺ and K⁺ returned to their initial levels, and cells survived. Two possible mechanisms for the role of Ca²⁺ in salt tolerance inNitellopsis are discussed; one a reduction in plasmalemma permeability to Na⁺ and the other a stimulation of active Na⁺-extrusion.     

Influence of abscisic acid on unidirectional fluxes and intracellular compartmentation of K+ and Na+ in excised barley root segments

Using compartmental analysis, unidirectional fluxes of K⁺ and Na⁺ and their intracellular compartmentation in excised barley (Hordeum distichon L. cv. Kocher-perle) root segments have been measured during a steady state in the presence or absence of ABA. Almost all flux rates were altered in the presence of external ABA, in particular the xylem transport R’ and the plasmalemma influx Ø_oc (see below) were strongly inhibited in the steady state. At the same time the presence of ABA induced a strong increase in the vacuolar K⁺ and Na⁺ content Q_v and a decrease in the cytoplasmic one (Q_c). Since the fluxes of an ion and its vacuolar or, in particular, cytoplasmic concentrations are interrelated, the ratios of fluxes originating from the cytoplasm and the cytoplasmic ion content were taken into account. On this basis ABA had the following effects: a) the secretion of K⁺ or Na⁺ to the xylem vessels was drastically inhibited; b) the plasmalemma K⁺ or Na⁺ efflux Ø_co was moderately stimulated and c) the tonoplast influx Ø_cv of Na⁺ was stimulated, while the tonoplast influx of K⁺ appeared to be unchanged (the decrease in Ø_cv being due to the decreased cytoplasmic K⁺ content). By a similar argument, also the apparent inhibition of the plasmalemma influx Ø_oc of K⁺ and Na⁺ in the steady state merely is an indirect effect of ABA. It only reflects the strong ABA-induced decrease in the xylem transport, that governs the magnitude of Ø_oc in the steady state. The results are discussed with reference to possible regulatory functions of ABA. In this respect it is suggested that – in particular under conditions of stress – ABA might regulate cellular metabolic processes by changing the cytoplasmic K⁺ level.     

NaCl胁迫对不同耐盐性玉米自交系萌动种子和幼苗离子稳态的影响

盐胁迫影响植物组织的离子分布,不同品种间存在差异。以玉米耐盐自交系81162和8723及盐敏感自交系P138为材料,研究了不同浓度(0、60、140、220 mmol/L)Na Cl胁迫下萌动期种子和幼苗的不同部位中Na+、K+、Ca2+含量以及K+/Na+和Ca2+/Na+比值的变化,旨在探讨不同自交系耐盐性差异的原因。结果表明,在萌动种子中,3个玉米自交系中的Na+积累量表现为种皮胚胚乳,K+累积表现为胚种皮胚乳;幼苗中,Na+积累表现为根茎叶。随着Na Cl浓度的增加,3个玉米自交系萌动种子和幼苗中的Na+含量逐渐升高,但是萌动种子中耐盐自交系81162和8723的Na+增加幅度小于盐敏感自交系P138,Na+含量小于盐敏感自交系P138;幼苗中耐盐自交系81162和8723的Na+增加幅度大于盐敏感自交系P138,幼苗根中Na+含量大于盐敏感自交系P138;茎叶中的Na+含量小于盐敏感自交系P138。随着Na Cl浓度的增加,萌动种子和幼苗中的K+和Ca2+含量逐渐降低。K+离子在耐盐自交系81162和8723萌动种子和幼苗中的降低幅度小于盐敏感自交系P138;Ca2+离子在耐盐自交系81162和8723幼苗中的降低幅度小于盐敏感自交系P138;而在萌动种子中3个自交系Ca2+的流失差异不大。耐盐自交系81162和8723萌动种子和幼苗中K+含量都大于盐敏感自交系P138。耐盐自交系81162和8723的萌动种子和幼苗根中Ca2+含量都大于盐敏感自交系P138;幼苗叶片中则小于盐敏感自交系P138。萌动种子和幼苗中K+/Na+和Ca2+/Na+均随着Na Cl浓度的升高而降低,K+/Na+比值表现为耐盐自交系81162和8723大于盐敏感自交系P138。耐盐自交系81162和8723通过调节离子平衡维持萌动种子和幼苗中较高的K+/Na+比值从而提高耐盐性。     

Physiological responses of the mangrove Rhizophora mangle grown in the absence and presence of NaCl

Abstract. Propagules of the mangrove, Rhizophora mangle L., were precultivated for 9 months in a greenhouse. The young plants were transferred into unaerated nutrient solutions without and with 200 mol m ³ NaCl and subsequently their growth, their water relations and the photosynthetic properties of their leaves were studied. Growth of the salttreated plants was significantly increased, while the control plants gradually died off after finishing the experiments. The shoot water potential and the stomatal resistance of the leaves were lowered while the chlorophyll contents and the chlorophyll a/b ratio in the leaves of salt-treated plants were increased by NaCl, the net result being an enhanced rate of CO₂ assimilation. The leaves of both sets of plants showed diurnal fluctuations in malic acid concentration which were more pronounced in the leaves of salt treated plants which, additionally, were more succulent. However, the plants showed no net CO₂ fixation at night, indicating that Rhizophora mangle is a CAM-cycling plant. After 200 d of cultivation without or with NaCl, the Na⁺, Cl and K⁺ concentrations in tissues and vacuoles were measured. Energy-dispersive X-ray microprobe analyses on root vacuoles of control plants reveal Na⁺ preference, on those of salt treated plants a strong K⁺ preference. Vacuolar K⁺ concentrations are neither affected by NaCl nor do they vary across the root radius. High vacuolar Na⁺ and Cl concentrations are found in the hypodermis followed by a stepwise decrease towards the inner root cortex cells. Ion concentrations of the photosynthetically active leaf tissues seem to be regulated by (1) radial filtration across the root cortex: (2) ion exchange of the xlem parenchyma cells: and (3) sequestration of Na⁺ and Cl in the hypodermal water storage tissue of the leaves.     

Sodium and potassium fluxes and compartmentation in roots of atriplex and oat

K⁺ and Na⁺ fluxes and ion content have been studied in roots of Atriplex nummularia Lindl. and Avena sativa L. cv Goodfield grown in 3 millimolar K⁺ with or without 3 or 50 millimolar NaCl. Compartmental analysis was carried out with entire root systems under steady-state conditions.

Increasing ambient Na⁺ concentrations from 0 to 50 millimolar altered K⁺, in Atriplex, as follows: slightly decreased the cytoplasmic content (Q_c), the vacuolar content (Q_v), and the plasma membrane influx and efflux. Xylem transport for K⁺ decreased by 63% in Atriplex. For oat roots, similar increases in Na⁺ altered K⁺ parameters as follows: plasma membrane influx and efflux decreased by about 80%. Q_c decreased by 65%, and xylem transport decreased by 91%. No change, however, was observed in Q_v for K⁺. Increasing ambient Na⁺ resulted in higher (3 to 5-fold) Na⁺ fluxes across the plasma membrane and in Q_c of both species. In Atriplex, Na⁺ fluxes across the tonoplast and Q_v increased as external Na⁺ was increased. In oat, however, no significant change was observed in Na⁺ flux across the tonoplast or in Q_v as external Na⁺ was increased. In oat roots, Na⁺ reduced K⁺ uptake markedly; in Atriplex, this was not as pronounced. However, even at high Na⁺ levels, the influx transport system at the plasma membrane of both species preferred K⁺ over Na⁺.

Based upon the Ussing-Teorell equation, it was concluded that active inward transport of K⁺ occurred across the plasma membrane, and passive movement of K⁺ occurred across the tonoplast in both species. Na⁺, in oat roots, was actively pumped out of the cytoplasm to the exterior, whereas, in Atriplex, Na⁺ was passively distributed between the free space, cytoplasm, and vacuole.

     

Ouabain switches Na+ transport to Na+/Na+ equivalent exchange in normal and apoptotic lymphoid cells

A theory of change of the ionic fluxes in the lymphoid cells in their transition from normal to apoptosis we have developed previously is applied to the analysis of Na⁺/Na⁺ exchange fluxes in human lymphoid cells U937 exposed to ouabain. We solve a system of equations describing changes in the intracellular concentrations of Na⁺, K⁺ and Cl^?, membrane potential and cell volume. It is shown that the Na⁺ influx (I _Na/Na) and output flux through the Na⁺/Na⁺ tract increased 4 times in 8 h after disconnecting Na⁺/K⁺-ATPase for normal cell U937. These fluxes increased 2.6 times for apoptotic cells. The value of I _Na/Na after 8 h off pump by ouabain is 97% of the total Na⁺ input for both cell types. It is concluded that ouabain not only inhibits the Na⁺/K⁺-ATPase, but also increases Na⁺ exchange fluxes through the Na⁺/Na⁺ tract, thereby switching sodium transport across the membrane of lymphoid cells to Na⁺/Na⁺ equivalent exchange.     

The uptake and extrusion of salts by the halotolerant bacterium,Ba1

Ion fluxes were studied in the cells of the moderately halophilic, halotolerant bacterium, Ba₁. K⁺ and Rb⁺ ions rapidly penetrated the cell membrane in contrast to Na⁺ and Li⁺ ions which were found to be nearly nonpenetrant. Moreover, Na⁺ ions at millimolar concentrations inhibited the passive penetration of K⁺ and Rb⁺ into the cytosol. Under energized conditions a powerful pump mechanism became functional causing extrusion of the penetrated K or Rb salt, as revealed by light-scattering changes as well as by tracer methods. From the pattern of action of agents which are known to interfere with respiration or energy transformation it was concluded that this pump is activated by the electrochemcial proton gradient without the mediation of ATP. In the range of pH values between 7.0 and 8.0 the rate of respiration and that of the extrusion of the salt declined gradually and in a near-parallel manner. The presence of Na⁺ ions at low concentrations prevented this drop in the two activities. Some evidence is brought in favor of the view that the pump activity is controlling respiratory rate rather than vice versa. It is suggested that the pump responsible for the extrusion of the K salt may be involved in the regulation of the intracellular salt concentration and, as a corollary, in the mechanism of adaptation of Ba₁ to variation in the salinity of the environment.     

Responses of salt-tolerant and intolerant wheat genotypes to sodium chloride: Photosynthesis,antioxidants activities,and yield

Physiological responses of two wheat (Triticum aestivum L.) genotypes (salt-tolerant DK961 and salt-sensitive JN17) to increased salt concentrations (50, 100, 150 mM NaCl: NaCl⁵⁰, NaCl¹⁰⁰, NaCl¹⁵⁰) were studied. Photosynthetic capacity, irradiance response curves, contents of soluble sugars, proteins, and chlorophyll (Chl), K⁺/Na⁺ ratio, and activities of antioxidant enzymes (superoxide dismutase, peroxidase, and catalase) in flag leaves were measured on 7 d after anthesis. In control (NaCl⁰) plants, non-significant (p>0.05) differences were found in gas exchange and saturation irradiance (SI) between salt-tolerant (ST) and salt-sensitive (SS) wheat genotypes. However, we found higher soluble sugar and protein contents, K⁺/Na⁺ ratio, and antioxidant enzyme activities, but lower Chl content and yield in ST wheat. Salinity stresses remarkably increased soluble sugar and protein contents and the antioxidant activities, but decreased K⁺/Na⁺ ratio, Chl contents, SI, photosynthetic capacities, and yield, the extent being considerably larger in JN17 than DK961. Although the soluble sugar and protein contents and the antioxidant activities of JN17 elevated more evidently under salt stresses, those variables never reached the high levels of DK961. The antioxidant enzyme activities of SS wheat increased in NaCl⁵⁰ and NaCl¹⁰⁰, but decreased rapidly when the NaCl concentration reached 150 mM. Thus the ST wheat could maintain higher grain yield than the SS one by remaining higher osmoregulation and antioxidative abilities, which led to higher photosynthetic capacity. Hence the ST wheat could harmonize the relationship between CO₂ assimilation (source) and the grain yield (sink) under the experimental conditions.     

Comparative salt tolerance analysis between Arabidopsis thaliana and Thellungiella halophila, with special emphasis on K(+)/Na(+) selectivity and proline accumulation

The eco-physiology of salt tolerance, with an emphasis on K⁺ nutrition and proline accumulation, was investigated in the halophyte Thellungiella halophila and in both wild type and eskimo-1 mutant of the glycophyte Arabidopsis thaliana, which differ in their proline accumulation capacity. Plants cultivated in inert sand were challenged for 3 weeks with up to 500 mM NaCl. Low salinity significantly decreased A. thaliana growth, whereas growth restriction was significant only at salt concentrations equal to or exceeding 300 mM NaCl in T. halophila. Na⁺ content generally increased with the amount of salt added in the culture medium in both species, but T. halophila showed an ability to control Na⁺ accumulation in shoots. The analysis of the relationship between water and Na⁺ contents suggested an apoplastic sodium accumulation in both species; this trait was more pronounced in A. thaliana than in T. halophila. The better NaCl tolerance in the latter was associated with a better K⁺ supply, resulting in higher K⁺/Na⁺ ratios. It was also noteworthy that, despite highly accumulating proline, the A. thaliana eskimo-1 mutant was the most salt-sensitive species. Taken together, our findings indicate that salt tolerance may be partly linked to the plants’ ability to control Na⁺ influx and to ensure appropriate K⁺ nutrition, but is not linked to proline accumulation.     

Energetic basis of development of salt-tolerant transport in a moderately halophilic bacterium,Vibrio costicola

Active transport of -aminoisobutyric acid (AIB) in Vibrio costicola utilizes a system with affinity for glycine, alanine and, to some extent, methionine. AIB transport was more tolerant of high salt concentrations (3–4 M NaCl) in cells grown in the presence of 1.0 M NaCl than in those grown in the presence of 0.5 M NaCl. The former cells could also maintain much higher ATP contents than the latter in high salt concentrations.Transport kinetic studies performed with bacteria grown in 1.0 M NaCl revealed three effects of the Na⁺ ion: the first effect is to increase the apparent affinity (K _t) of the transport system for AIB at Na⁺ concentrations <0.2 M, the second to increase the maximum velocity (V _max) of transport (Na⁺ concentrations between 0.2 and 1.0 M), and the third to decrease the V _max without affectig K _t (Na⁺ concentrations >1.0 M). Cells grown in the presence of 0.5 M or 1.0 M NaCl had similar affinity for AIV. Thus, the differences in salt response of transport in these cells do not seem due to differences in AIB binding. Large, transport-inhibitory concentrations of NaCl resulted in efflux of AIB from cells preloaded in 0.5 M or 1.0 M NaCl, with most dramatic efflux occurring from the cells whose AIB transport was more salt-sensitive. Our results suggest that the degree to which high salt concentrations affect the transmembrane electrochemical energy source used for transport and ATP synthesis is an important determinant of salt tolerance.Abbreviations AIB -aminoisobutyric acid - pmf proton motive force     

Ionic and osmotic components of salt stress specifically modulate net ion fluxes from bean leaf mesophyll

Ionic mechanisms of salt stress perception were investigated by non‐invasive measurements of net H⁺, K⁺, Ca²⁺, Na⁺, and Cl^? fluxes from leaf mesophyll of broad bean (Vicia faba L.) plants using vibrating ion‐selective microelectrodes (the MIFE technique). Treatment with 90 m M NaCl led to a significant increase in the net K⁺ efflux and enhanced activity of the plasma membrane H⁺‐pump. Both these events were effectively prevented by high (10 m M ) Ca²⁺ concentrations in the bath. At the same time, no significant difference in the net Na⁺ flux has been found between low‐ and high‐calcium treatments. It is likely that plasma membrane K⁺ and H⁺ transporters, but not the VIC channels, play the key role in the amelioration of negative salt effects by Ca²⁺ in the bean mesophyll. Experiments with isotonic mannitol application showed that cell ionic responses to hyperosmotic treatment are highly stress‐specific. The most striking difference in response was shown by K⁺ fluxes, which varied from an increased net K⁺ efflux (NaCl treatment) to a net K⁺ influx (mannitol treatment). It is concluded that different ionic mechanisms are involved in the perception of the ‘ionic’ and ‘osmotic’ components of salt stress.     

Bumetanide-sensitive Ion Fluxes in Vascular Smooth Muscle Cells: Lack of Functional Na+, K+, 2 Cl− Cotransport

To examine the involvement of Na⁺,K⁺,2Cl⁻ cotransport in monovalent ion fluxes in vascular smooth muscle cells (VSMC), we compared the effect of bumetanide on ⁸⁶Rb, ³⁶Cl and ²²Na uptake by quiescent cultures of VSMC from rat aorta. Under basal conditions, the values of bumetanide-sensitive (BS) inward and outward ⁸⁶Rb fluxes were not different. Bumetanide decreased basal ⁸⁶Rb uptake by 70–75% with a K _i of ∼0.2–0.3 μm. At concentrations ranging up to 1 μm, bumetanide did not affect ³⁶Cl influx and reduced it by 20–30% in the range from 3 to 100 μm. In contrast to ⁸⁶Rb and ³⁶Cl influx, bumetanide did not inhibit ²²Na uptake by VSMC. BS ⁸⁶Rb uptake was completely abolished in Na⁺- or Cl⁻-free media. In contrast to ⁸⁶Rb, basal BS ³⁶Cl influx was not affected by Na⁺ _o and K⁺ _o . Hyperosmotic and isosmotic shrinkage of VSMC increased ⁸⁶Rb and ³⁶Cl influx to the same extent. Shrinkage-induced increments of ⁸⁶Rb and ³⁶Cl uptake were completely abolished by bumetanide with a K _i or ∼0.3 μm. Shrinkage did not induce BS ⁸⁶Rb and ³⁶Cl influx in (Na⁺ or Cl⁻)- and (Na⁺ or K⁺)-depleted media, respectively. In the presence of an inhibitor of Na⁺/H⁺ exchange (EIPA), neither hyperosmotic nor isosmotic shrinkage activated ²²Na influx. Bumetanide (1 μm) did not modify basal VSMC volume and intracellular content of sodium, potassium and chloride but abolished the regulatory volume increase in isosmotically-shrunken VSMC. These data demonstrate the absence of the functional Na⁺,K⁺,2Cl⁻ cotransporter in VSMC and suggest that in these cells basal and shrinkage-induced BS K⁺ influx is mediated by (Na⁺ _o + Cl⁻ _o )-dependent K⁺/K⁺ exchange and Na⁺ _o -dependent K⁺,Cl⁻ cotransport, respectively. Received: 30 January 1996/Revised: 20 May 1996     

Effects of salinity on uptake and distribution of Na+, Cl- and K+ in two wheat cultivars

Plants of two wheat (Triticum aestivum L.) cultivars differing in salt tolerance were grown in sand with nutrient solutions. 35-d-old plants were subjected to 5 levels of salinity created by adding NaCl, CaCl₂ and Na₂SO₄. Growth reduction caused by salinity was accompanied by increased Na⁺ and Cl^- concentrations, Na⁺/K⁺ ratio, and decreased concentration of K⁺. The salt tolerant cv. Kharchia 65 showed better ionic regulation. Salinity up to 15.7 dS m^-1 induced increased uptake of Na⁺ and Cl^- but higher levels of salinity were not accompanied by further increase in uptake of these ions. Observed increases in Na⁺ and Cl^- concentrations at higher salinities seemed to be the consequence of reduction in growth. Uptake of K⁺ was decreased; more in salt sensitive cultivar. This was also accompanied by differences in its distribution.     

Irreversible zinc ion inhibition of (Na+ -K+)-adenosinetriphosphatase, Na+ -phosphorylation, and K+-p-nitrophenylphosphatase of Electrophorus electricus electroplax.

Zinc ion in micromolar concentrations is an irreversible inhibitor of Electrophorus electricus electroplax microsomal (Na⁺-K⁺)-ATPase. The rate of inhibition is dependent on [ZnCl₂] and the extent of inhibition varies with the ratio of ZnCl₂ to microsomal protein. The same kinetics are observed for inhibition of K⁺ -p-nitrophenylphosphatase and steady-state levels of Na⁺ -dependent enzyme phosphorylation. The observations suggest that a Zn²⁺ -sensitive conformational restraint is important to both kinase and phosphatase activities. The fact that inhibition is irreversible has implications for models seeking to relate zinc effects in tissue to inhibition of (Na⁺-K⁺)-ATPase.     

Ion Secretion and Isotonic Transport in Frog Skin Glands

The aim of this study was to clarify the mechanism of isotonic fluid transport in frog skin glands. Stationary ion secretion by the glands was studied by measuring unidirectional fluxes of ²⁴Na⁺, ⁴²K⁺, and carrier-free ¹³⁴Cs⁺ in paired frog skins bathed on both sides with Ringer's solution, and with 10⁻⁵ m noradrenaline on the inside and 10⁻⁴ m amiloride on the outside. At transepithelial thermodynamic equilibrium conditions, the ¹³⁴Cs⁺ flux ratio, J ^out _Cs/J ⁱⁿ _Cs, varied in seven pairs of preparations from 6 to 36. Since carrier-free ¹³⁴Cs⁺ entering the cells is irreversibly trapped in the cellular compartment (Ussing & Lind, 1996), the transepithelial net flux of ¹³⁴Cs⁺ indicates that a paracellular flow of water is dragging ¹³⁴Cs⁺ in the direction from the serosal- to outside solution. From the measured flux ratios it was calculated that the force driving the secretory flux of Cs⁺ varied from 30 to 61 mV among preparations. In the same experiments unidirectional Na⁺ fluxes were measured as well, and it was found that also Na⁺ was subjected to secretion. The ratio of unidirectional Na⁺ fluxes, however, was significantly smaller than would be predicted if the two ions were both flowing along the paracellular route dragged by the flow of water. This result indicates that Na⁺ and Cs⁺ do not take the same pathway through the glands. The flux ratio of unidirectional K⁺ fluxes indicated active secretion of K⁺. The time it takes for steady-state K⁺ fluxes to be established was significantly longer than that of the simultaneously measured Cs⁺ fluxes. These results allow the conclusion that — in addition to being transported between cells — K⁺ is submitted to active transport along a cellular pathway.Based on the recirculation theory, we propose a new model which accounts for stationary Na⁺, K⁺, Cl⁻ and water secretion under thermodynamic equilibrium conditions. The new features of the model, as compared to the classical Silva-model for the shark-rectal gland, are: (i) the sodium pumps in the activated gland transport Na⁺ into the lateral intercellular space only. (ii) A barrier at the level of the basement membrane prevents the major fraction of Na⁺ entering the lateral space from returning to the serosal bath. Thus, Na⁺ is secreted into the outside bath. It has to be assumed then that the Na⁺ permeability of the basement membrane barrier (P ^BM _Na) is smaller than the Na⁺ permeability of the junctional membrane (P ^JM _Na), i.e., P ^JM _Na/P ^BM _Na > 1. The secretory paracellular flow of water further requires that the Na⁺ reflection coefficients (σ_Na) of the two barriers are governed by the conditions, σ^BM _Na > 0, and σ^BM _Na > σ^JM _Na. (iii) Na⁺ channels are located in the apical membrane of the activated gland cells, so that a fraction of the Na⁺ outflux appearing downstream the lateral intercellular space is recirculated by the gland cells. Based on measured unidirectional fluxes, a set of equations is developed from which we estimate the ion fluxes flowing through major pathways during stationary secretion. It is shown that 80% of the sodium ions flowing downstream the lateral intercellular space is recycled by the gland cells. Our calculations also indicate that under the conditions prevailing in the present experiments 1.8 ATP molecule would be hydrolyzed for every Na⁺ secreted to the outside bath. Received: 30 January 1996/Revised: 12 March 1996     

Salt-tolerance in plants. II. In vitro translation of m-RNAs from salt-tolerant and salt-sensitive plants on wheat germ ribosomes. Responses to ions and compatible organic solutes

Abstract Messenger RNA from salt-sensitive and salt-tolerant plants Triticum aestivum. Beta vulgaris, Pisum sativum, Chenopodium album and Atriplex nummularia was translated in vitro in a wheatgerm translation system. The optimal monovalent and divalent ion concentrations for translation were independent of the salt tolerance of the plants from which the m-RNAs were derived. Translation was optimal in 100 120 mol m⁻³ potassium acetate and 1.5–2.0 mol m⁻³ Mg²⁺. Substitution of Na⁺ for K⁺, or of Cl⁻ for acetate, was inhibitory. The pattern of polypeptides synthesized from cytoplasmic m-RNAs of salt-sensitive and salt-tolerant plants remained constant in all the conditions examined. The effects of adding the ‘compatible' organic solutes glycine-betaine and mannitol were examined in the wheat-germ system primed with RNA from the leaves of Triticum aestivum or Beta vulgaris. The rate of translation, the optimum ionic concentrations and the distribution of polypeptide products were maintained in organic solute concentrations of up to 500 mol m⁻³. Proline above 300 mol m⁻³ and surcose above 100 mol m⁻³ did inhibit translation. The results indicate that translation in plants is unlikely in cytoplasmic K⁺ concentrations exceeding 180 mol m⁻³, but would proceed in the presence of up to 500 mol m⁻³ mannitol or glyinebetaine, or of up to 300 mol m⁻³ proline.     

短期NaCl胁迫对不同小麦品种幼苗K+吸收和Na+、K+积累的影响

为研究盐胁迫下小麦幼苗生长及Na⁺、K⁺的吸收和积累规律,以中国春、洲元9369和长武134等3种耐盐性不同小麦品种为材料,采用非损伤微测技术检测盐胁迫2 d后的根系K⁺离子流变化,并对植株体内的Na⁺、K⁺含量进行测定。结果表明:短期(2d)盐胁迫对小麦生长有抑制作用,且对根系的抑制大于地上部,耐盐品种下降幅度小于盐敏感品种。盐胁迫下,小麦根际的 K⁺大量外流,盐敏感品种中国春K⁺流速显著高于耐盐品种长武134,最高可达15倍。小麦幼苗地上部分和根系均表现为Na⁺积累增加,K⁺积累减少,Na⁺/K⁺比随盐浓度增加而上升。中国春限Na⁺能力显著低于长武134,Na⁺/K⁺则显著高于长武134。综上所述,盐胁迫下造成小麦组织器官中Na⁺/K⁺比上升的主要原因是根系K⁺大量外流和Na⁺的过量积累,耐盐性不同的小麦品种间差异显著,并认为根系对K⁺的保有能力可能是作物耐盐性评价的一个重要指标。     

Sodium and potassium interactions with Na+-ATPase of inside-out membrane vesicles from high-K+ and low-K+ sheep red cells

Na⁺-ATPase of high-K⁺ and low-K⁺ sheep red cells was examined with respect to the sidedness of Na⁺ and K⁺ effects, using inside-out membrane vesicles and very low ATP concentrations (?2 μM). With varying amounts of Na⁺ in the medium, i.e., at the cytoplasmic surface, Na_cyt⁺, the activation curves show that high-K⁺ Na⁺-ATPase has a higher affinity for Na_cyt⁺ compared to low-K⁺. The apparent affinity for Na_cyt⁺ is also increased by increasing the ATP concentrations in high-K⁺ but not low-K⁺. With Na_cyt⁺ present, Na⁺-ATPase is stimulated by intravesicular Na⁺, i.e., Na⁺ at the originally external surface, Na_ext⁺, to a greater extent in low-K⁺ than high-K⁺. Intravesicular K⁺ (K_ext⁺) activates Na⁺-ATPase in high-K⁺ but not in low-K⁺ vesicles and extravesicular K⁺ (K_cyt⁺) inhibits low-K⁺ but not high-K⁺ Na⁺-ATPase. Thus, the genetic difference between high-K⁺ and low-K⁺ is expressed as differences in apparent affinities for both Na⁺ and K⁺ and these differences are evident at both cytoplasmic and external membrane surfaces.     

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.