Effect of light intensity on proton extrusion by roots of intact maize plants

Shading of maize plants ( Zea mays L. cv. Blizzard) reduced net H⁺ extrusion by roots and increased K⁺ release, whereas there was no significant effect on anion efflux in deionized water. With lower light intensity the concentrations of carbohydrates in the roots decreased, but ATP levels and energy charge remained unchanged. Also, shading raised the tissue pH of roots and made the cytoplasmic pH of root cells drop. There was a significant influence of light intensity on H⁺ uptake by roots from an acidified test solution and CCCP (carbonylcyanide-3-chlorophenylhydrazone)-in-duced H⁺ uptake was modified by shading.
It is concluded that low light intensity does not limit active H⁺ release by plasmalemma ATPase activity in the root cells, but that a reduced carbohydrate supply brings about a change in biochemical reactions which alter the membrane permeability for protons. An increased passive reflux of H⁺ into the cells rather than a reduced H⁺ ATPase activity explains the decrease of net H⁺ release by roots of intact maize plants under low light intensity.     

Ion transport and osmotic adjustment in Escherichia coli in response to ionic and non-ionic osmotica

Bacteria respond to osmotic stress by a substantial increase in the intracellular osmolality, adjusting their cell turgor for altered growth conditions. Using Escherichia coli as a model organism we demonstrate here that bacterial responses to hyperosmotic stress specifically depend on the nature of osmoticum used. We show that increasing acute hyperosmotic NaCl stress above ∼1.0 Os kg⁻¹ causes a dose-dependent K⁺ leak from the cell, resulting in a substantial decrease in cytosolic K⁺ content and a concurrent accumulation of Na⁺ in the cell. At the same time, isotonic sucrose or mannitol treatment (non-ionic osmotica) results in a gradual increase of the net K⁺ uptake. Ion flux data are consistent with growth experiments showing that bacterial growth is impaired by NaCl at the concentration resulting in a switch from net K⁺ uptake to efflux. Microarray experiments reveal that about 40% of upregulated genes shared no similarity in their responses to NaCl and sucrose treatment, further suggesting specificity of osmotic adjustment in E. coli to ionic and non-ionic osmotica. The observed differences are explained by the specificity of the stress-induced changes in the membrane potential of bacterial cells highlighting the importance of voltage-gated K⁺ transporters for bacterial adaptation to hyperosmotic stress.     

Passive uptake of K+(86Rb+) in sunflower roots at low external K+ concentrations

Passive fluxes of K⁺ (⁸⁶Rb) into roots of sunflower ( Helianthus annuus L. cv. Uniflorus) were determined at low K⁺ concentration (0.1 and 1.0 mM K⁺) in the ambient solution. Metabolic uptake of K⁺ was inhibited by 10⁻⁴M 2,4-dinitrophenol (DNP). K⁺ (⁸⁶Rb) fluxes were studied both continuously and by time differentiation of uptake. In high K⁺ roots passive uptake was directly proportional to the K⁺ concentration of the uptake solution, indicating free diffusion. This assumption was supported by the fact that passive Rb⁺ uptake was not affected by high K⁺ concentrations. In low K⁺ roots the passive uptake of K⁺ was higher than in high K⁺ roots. The increase was possibly due to carrier-mediated K⁺ transport. As K⁺ effluxes were quantitatively similar to influxes, it is suggested that passive K⁺ fluxes represent exchange diffusion without relation to net K⁺ transport.     

ENERGETICS OF AMINO ACID TRANSPORT INTO BRAIN SLICES: EFFECTS OF K+ DEPLETION AND Rb+ OR Cs+ SUBSTITUTION ON AMINO ACID UPTAKE

Abstract— Mouse brain slices were depleted of K⁺ by three 10-min incubations-in oxygenated HEPES-buffered medium lacking glucose and K⁺. Addition of K⁺ or Rb⁺ (or Cs⁺, to a smaller degree) with glucose, or with succinate, malate, and pyruvate (SMP) before incubation at 37°C with ¹⁴C-amino acids restored active low-affinity transport of d -Glu, α-aminoisobutyrate (AIB), GABA, Gly, His, Val, Leu, Lys, and Orn. Ouabain at 1–2μ m with Rb⁺ was more inhibitory with SMP than with glucose, suggesting that the glycoside may affect specific energy coupling to transport. Valinomycin, in contrast, showed no specificity of inhibition of amino acid uptake with glucose or SMP and K⁺ or Rb⁺. Cs⁺ partially restored amino acid uptake, but Li⁺ was less effective than Cs ⁺. NaF at 10 m m with SMP + Rb⁺, or SMP + K⁺ did not inhibit amino acid uptake. Therefore, it was possible to dissociate glycolysis and Na⁺, K ⁺ -ATPase activity from amino acid transport. The ion replacements for K ⁺ that supported active amino acid transport indicate that the specificity of ions in possible ionic gradients for transport energetics should be reexamined.     

Influence of exogenously supplied potassium and sodium salts on the abscisic acid-induced proline accumulation in barley leaf segments

A stimulation of the abscisic acid (ABA)-induced increase in proline was observed in leaf segments of barley ( Hordeum vulgare L. cv. Georgie) if K⁺ or Na⁺ were supplied in the external medium as salts of monovalent anions such as NO₃, Br, Cr and I, but not when sulphate or phosphate were used. To a lesser extent, the effect was evident also with RbCl, but it did not occur when chlorides of Li⁺. Cs⁺, NH₄⁺, Mg:⁺ and Ca²⁺ were used. Both KC1 and NaCl in the concentration range 2–100 m M influence the ABA-dependent proline accumulation to the same extent; the increase induced was about 100% at 10 m M , and reached a maximum between 60 and 100 m M. The effect is not due to the osmotic activity of the salts and does not seem to depend on changes in K⁺ and Na⁺ levels within the leaf tissue, but it is somehow linked to their external concentration. The existence of a specific interaction between ABA and K⁺ or Na⁺, possibly at the cell membrane level, is proposed.     

Long-term effects of abscisic acid on K+ transport in young wheat plants of different K+ status

The effects of abscisic acid (ABA) on growth, uptake and translocation of potassium ions, K⁺,Mg²⁺-ATPase activity and transpiration were investigated in young wheat ( Triticum aestivum L. cv. Martonvásári-8) plants grown at different K⁺ supplies. Long-term treatment with ABA (10 μ M ) reduced growth in high-K⁺ plants, but had less effect under low-K⁺ conditions. K⁺(⁸⁶Rb) uptake was inhibited by about 70 and 40% in low- and high-K⁺ plants, respectively. The stimulation by K⁺ of the Mg²⁺-ATPase activity in the root microsomal fraction was lost with ABA treatment. It is suggested that the inhibitory effect of ABA on K⁺ uptake may be related to this effects on the K⁺,Mg²⁺-ATPase. Translocation of K⁺ to the shoot was inhibited in low-K⁺ plants only, and it was not affected in high-K⁺ plants. In parallel to this, ABA treatment reduced transpiration by about 50% in low-K⁺ plants, whereas a much smaller effect was seen in high-K⁺ plants. These observations suggest that the regulation by ABA of the stomatal movements is strongly counteracted by high-K⁺ status.     

Long distance transport of potassium in cereals during grain filling in intact plants

Translocation of labeled potassium (K⁺) from the root to the ear and its distribution within the culm during 4, 8 and 12 h of uptake was studied in intact wheat plants ( Triticum aestivum L. cv. Kolibri) 3 and 5 weeks after anthesis at 0.5 and 5.0 m M K⁺ concentration in the uptake solution. Uptake of labeled K⁺ into the shoot was proportional to the K⁺ concentration applied. After 4 h of uptake about 2% and after 12 h about 7% of labeled K⁺ applied to the roots were taken up into the shoot at both K⁺ concentrations. After 12 h of uptake only 6% of the total label in the culm had reached the ear, while about 40% of the label was found in the upper three internodes. In spite of an increasing concentration of labeled K⁺ during 12 h in the uppermost internode (peduncle), translocation of K⁺ into the rachis was low. The low and uniform K⁺ content found generally in grain dry weight seems therefore to be due to a controlled K⁺ supply to the ear.     

Effect of ABA, fusicoccin and thiourea on germination and K+ and glucose uptake in chick-pea seeds at different temperatures

Germination and growth of seeds of Cicer arietinum L. are retarded or inhibited either by temperatures above 30°C or by the presence of abscisic acid (ABA). Both treatments affect the uptake of K⁺ and glucose. ABA may affect the properties of the cell membrane or the kinetics of the enzymes bound to it. Fusicoccin (FC) and thiourea (TU) are to a greater or lesser extent able to counteract the inhibition produced by ABA and high temperatures. Both FC and TU increase the uptake of K⁺ by the embryonic axis. FC seems to act directly on H⁺ extrusion, but the mode of action of TU appears to be different. The results seem to indicate the importance of the activation of the ion transport process for the growth of the embryonic axis during early germination, and suggest a control of the growth of the embryo by part of the cotyledons. Physiological inhibitors such as ABA might be implied in the control mechanism.     

Changes in potassium fluxes in cells of carrot storage tissue related to turgor pressure

The effect of increasing external osmotic pressure on potassium fluxes in aged and fresh-cut discs of Daucus carota L. storage tissue was investigated. An increase of the external osmotic pressure by 5 bars of mannitol solution increased the rate of K⁺ net uptake of aged discs to 180% of their control rate. At 3°C and in 0.1 m M azide, in which a net efflux of potassium was observed, the mannitol treatment caused a reduction in the net efflux. In fresh-cut discs, in which the capability of net influx was rather low and a substantial net release of potassium was noted, the increase in the external osmotic pressure by mannitol caused a 70% inhibition in the net efflux. This effect was also observed at 3°C.
Measurements of separate fluxes confirmed the assumption that the mannitol treatment brought about two distinct effects on K⁺ fluxes: (a) raised the metabolically-dependent influx and (b) lowered the membrane permeability-dependent efflux. When a permeating solute (ethylene glycol) was used instead of mannitol, no effect on K⁺ flux was detectable. Reasons are given for relating the observed changes in K⁺ fluxes to the reduction in turgor pressure of the cells.     

Growth, contents of K+ and kinetics of K+(86Rb) uptake in barley cultured at different low supply rates of potassium

Plants of barley ( Hordeum vulgare L. cv. Salve) were grown with 6.5–35% relative increase of K⁺ supply per day (RKR) using a special computer-controlled culture unit. After a few days on the culture solution the plants adapted their relative growth rate (RGR) to the rate of nutrient supply. The roots of the plants remained in a low salt status irrespective of the rate of nutrient supply, whereas the concentration of K⁺ in shoots increased with RKR. Both V_max and K_m for K⁺(⁸⁶Rb) influx increased with RKR. It is concluded that with a continuous and stable K⁺ stress, the K⁺ uptake system is adjusted to provide an effective K⁺ uptake at each given RKR. Allosteric regulation of K⁺ influx does not occur and efflux of K⁺ is very small.     

Potassium transport in wheat seedlings grown with different potassium supplies.

The K⁺(⁸⁶Rb) uptake into the roots and the translocation to the shoots of 11-day-old intact wheat seedlings ( Triticum aestivum L. cv. Martonvásári 8) were investigated using plants grown with different K⁺ supplies. The effects of environmental conditions (darkness, humidity) and of metabolic and transport inhibitors (oligomycin, disalicylidene-propanediamine, 2,4-dinitriphenol, diethylstilbestrol, colchicine) were also studied. Plants with K content of about 0.2 mmol/g dry weight in the root and 0.5 mmol/g dry weight in the shoot (low K status) showed high K⁺ uptake into the roots and high translocation rates to the shoots. Both transport processes were very low in plants with K content of more than 1.5 and 2.2 mmol/g dry weight in the root and shoot, respectively (high K status).
Darkness and a relative humidity of the air of 100% did not influence K⁺ uptake by roots, but did inhibit upward translocation and water transport. Inhibition of photosynthesis and treatments with diethylstilbestrol (10⁻⁵ mol/dm³), as well as with colchicine resulted in inhibition of translocation in plants of low K status, but these inhibitors had little effect on K⁺ uptake by the roots. Oligomycin, 2,4-dinitrophenol and diethylstilbestrol (10⁻⁴ mol/dm³), however, inhibited K⁺ uptake by the roots. In general, K⁺ transport processes were almost unchanged in plants of high K status. It is concluded that only plants of low K status operating with active K⁺ transport mechanisms are responsive to environmental factors. In high K⁺ plants the transport processes are passive and are uncoupled from the metabolic energy flow.     

Net Glutamate Release from Astrocytes Is Not Induced by Extracellular Potassium Concentrations Attainable in Brain

Abstract: Elevated extracellular potassium concentration ([K⁺]_e) has been shown to induce reversal of glial Na⁺-dependent glutamate uptake in whole-cell patch clamp preparations. It is uncertain, however, whether elevated [K⁺]_e similarly induces a net glutamate efflux from intact cells with a physiological intracellular milieu. To answer this question, astrocyte cultures prepared from rat and mouse cortices were incubated in medium with elevated [K⁺]_e (by equimolar substitution of K⁺ for Na⁺), and glutamate accumulation was measured by HPLC. With [K⁺]_e elevations to 60 m M , medium glutamate concentrations did not increase during incubation periods of 5–120 min. By contrast, 45 min of combined inhibition of glycolytic and oxidative ATP production increased medium glutamate concentrations 50–100-fold. Similar results were obtained in both rat and mouse cultures. Studies were also performed using astrocytes loaded with the nonmetabolized glutamate tracer d -aspartate, and parallel results were obtained; no increase in medium d -aspartate content resulted from [K⁺]_e elevation up to 90 m M , whereas a large increase occurred during inhibition of energy metabolism. These results suggest that a net efflux of glutamate from intact astrocytes is not induced by any [K⁺]_e attainable in brain.     

Synergistic activation of plasma membrane H+– ATPase in Arabidopsis thaliana cells by turgor decrease and by fusicoccin

The regulation of the H⁺-ATPase of plasma membrane is a crucial point in the integration of transport processes at this membrane. In this work the regulation of H⁺-ATPase activity induced by changes in turgor pressure was investigated and compared with the stimulating effect of fusicoccin (FC). The exposure of cultured cells of Arabidopsis thaliana L. (ecotype Landsberg 310–14-2) to media containing mannitol (0. 15 or 0. 3 M ) or polyethylene glycol 6000 (PEG) (15. 6% or 22% w/v) resulted in a decrease in the turgor pressure of the cells and in a strong stimulation of H⁺ extrusion in the incubation medium. The osmotica-induced H⁺ extrusion was (1) inhibited by the inhibitor of plasma membrane H⁺-ATPase, erythrosin B (EB), (2) dependent on the external K⁺ concentration, (3) associated with a net K⁺ influx, and (4) lead to an increase of cellular malate content. These results show that the reduction of external osmotic potential stimulates the activity of plasma membrane H⁺-ATPase
The effect of mannitol was only partially inhibited by treatments with cycloheximide (CH) and cordycepin, which block protein and mRNA synthesis, respectively. All the effects of osmotica were qualitatively and quantitatively similar to those induced by 5 μ M FC. However, when FC and mannitol (or PEG) were fed together, their effects on H⁺ extrusion appeared synergistic, irrespective of whether FC was present at suboptimal or optimal concentrations. This behaviour suggests that the modes of action of FC and of the osmotica on H⁺-ATPase activity differ at least in some step(s)     

Kinetics of efflux of K+ from cultured rose cells treated with ultraviolet light

Irradiation of cultured rose ( Rosa damascena Mill. cv. Gloire de Guilan) cells with ultraviolet light caused of loss of K⁺, which occurred with sigmoid kinetics. The kinetics of loss of K⁺ were not changed when the extracellular concentration of K⁺ was held constant during the period of efflux. Furthermore, the rate of loss of K⁺ was approximately the same even though the K⁺ concentration in the medium was increased from 0.1 to 10 m M . The kinetics of uptake of the lipophilic methyltriphenylphosphonium cation, an indicator of the plasma membrane potential, were linear throughout the period of K⁺ efflux, suggesting that the starting and stopping of K⁺ efflux do not reflect a passive response to changes in the membrane potential of the cells. The results are interpreted in terms of activation and inactivation of an efflux channel or pump for K⁺.     

Outward-rectifying K+ channel activities regulate cell elongation and cell division of tobacco BY-2 cells

Potassium ions (K⁺) are required for plant growth and development, including cell division and cell elongation/expansion, which are mediated by the K⁺ transport system. In this study, we investigated the role of K⁺ in cell division using tobacco BY-2 protoplast cultures. Gene expression analysis revealed induction of the Shaker -like outward K⁺ channel gene, NTORK1 , under cell-division conditions, whereas the inward K⁺ channel genes NKT1 and NtKC1 were induced under both cell-elongation and cell-division conditions. Repression of NTORK1 gene expression by expression of its antisense construct repressed cell division but accelerated cell elongation even under conditions promoting cell division. A decrease in the K⁺ content of cells and cellular osmotic pressure in dividing cells suggested that an increase in cell osmotic pressure by K⁺ uptake is not required for cell division. In contrast, K⁺ depletion, which reduced cell-division activity, decreased cytoplasmic pH as monitored using a fluorescent pH indicator, SNARF-1. Application of K⁺ or the cytoplasmic alkalizing reagent (NH₄)₂SO₄ increased cytoplasmic pH and suppressed the reduction in cell-division activity. These results suggest that the K⁺ taken up into cells is used to regulate cytoplasmic pH during cell division.     

Long distance transport of potassium in cereals during grain filling in detached ears

Ears of wheat plants ( Triticum aestivum L. cv. Kolibri), which were given different and uniform K⁺-nutrition in two experiments, were cut at 2, 4 and 6 weeks after anthesis at 15 cm below the ear. These detached ears were fed 30 m M (experiment 1) or 15, 30, 60 or 90 m M ⁸⁶Rb-K₂ malate (experiment 2) and 146 m M [¹⁴C]-sucrose. After a pulse period of 6 and 4 h, respectively, the ears were transferred to identical non-labeled solutions for additional 0, 4, 8 or 20 h.
About 50% of the K⁺ and sucrose supplied was absorbed by detached ears. This rate declined with plant age and decreasing transpiration. Within the 6 and 4 h uptake period less than 7% of the absorbed K⁺, but 20% of the sucrose taken up were incorporated into the grain. During the chase period labeled K⁺ in the grain increased to 15% and ¹⁴C even to 50% of total tracer uptake. Incorporation of labeled K⁺ into the grain was not affected by the previous K⁺ nutrition of the plant and was proportional to the K⁺ concentration in the uptake solution. Transition of K⁺ from xylem into phloem during its acropetal transport is assumed. No evidence was found that the grain itself could control its uptake of K⁺.     

Effect of potassium status on K+ (Rb)+ uptake and transport in sunflower roots

Young sunflower plants ( Helianthus annuus L. cv. Halcón), grown in nutrient solution at two K⁺ levels (0.25 and 2.5 m M ) were used to study the effect of K⁺ content in the root on uptake and transport of K⁺ to the exuding stream of decapitated plants. Roots of plants grown in low K⁺ gave higher exudation flux, higher K⁺ concentration in exudate and higher K⁺ flux than high K⁺ roots. After 6 h of uptake the K⁺ flux in low K⁺ roots was about three times that in high K⁺ roots. When the roots were kept in a nutrient solution in which Rb⁺ replaced K⁺, low K⁺ roots exuded much more Rb⁺ than K⁺ after the first 2 h, whereas high K⁺ roots exuded about similar amounts of K⁺ and Rb⁺. In intact plants grown at three different K⁺ levels (0.1, 1.0 and 10.0 m M ), there was an inverse relationship between the K⁺ level in the nutrient solution and the Rb⁺ accumulated in the roots or transported to the shoot. The results suggest that the transport of ions from xylem parenchyma to stele apoplast may be controlled by ions coming down from the shoot in sieve tubes.     

Effect of fusicoccin and gibberellic acid on primary dormant Avena fatua caryopses

Fusicoccin induced germination in dormant and partially afterripened dormant caryopses of Avena fatua L. The rate of caryopsis germination was slower and final percentage germination lower in the highly dormant inbred line M73 at a given concentration of fusicoccin than in the dormant caryopses of line AN265. Gibberellic acid was more effective than fusicoccin in breaking dormancy in both lines. Promotion of germination of dormant caryopses by fusicoccin was inhibited by a 6-day pretreatment with (2-chloroethyl)trimethylammonium chloride.
The basal rate of proton efflux from embryos isolated from dormant and fully afterripened line AN265 caryopses was similar. Addition of fusicoccin increased the rate of proton efflux from the isolated embryos of dormant and afterripened caryopses by nearly 400%. Gibberellic acid had no effect on the rate of proton extrusion. The uptake of ⁸⁶Rb⁺ in dormant and afterripened A. fatua embryos was similar after a 2 h uptake period. The addition of fusicoccin to the medium doubled the uptake of ⁸⁶Rb⁴ by dormant and afterripened embryos. Gibberelleic acid had no effect on the uptake of ⁸⁶Rb⁺ by isolated embryos from either dormant or afterripened caryopses. The experimental results indicate that gibberellic acid is more versatile in its action than fusicoccin, and gibberellic acid may facilitate dormant A. fatua caryopsis germination by stimulating mechanisms other than the direct H⁺ efflux and K⁺ uptake at the membrane level.