Effect of gibberellic acid on K+ (Rb+) uptake and transport in sunflower roots

Four-week-old sunflower plants ( Helianthus annuus L. cv. Halcón), grown in different nutrient solutions, were used to study the effects of gibberellic acid (GA₃) on K⁺ (Rb⁺) uptake by roots or transport to the shoot. Gibberellic acid application to the nutrient solution did not affect the exudation process of excised roots. When GA₃ was sprayed on leaves 2 to 6 days before excising the roots, the rate of exudation and the K⁺ flux increased. When the exudation study was done keeping the roots in a nutrient solution in which Rb⁺ replaced K⁺, the GA₃ effects were evident also on Rb⁺ uptake and transport. In intact plants, GA₃ increased the Rb⁺ transported to the shoot but did not affect Rb⁺ accumulation in the root. It is suggested that these GA₃ effects can be explained if it is assumed that GA₃ acts on the transport of ions to the xylem vessels.     

Separation of metabolic and non-metabolic steps of Rb+ uptake in spring wheat roots with different K+ status

Uptake of Rb⁺ from a complete nutrient solution with 2.0 mM Rb⁺ was studied in roots of spring wheat seedlings ( Triticum aestivum L. cv. Svenno) with different K⁺ levels. The relationship between Rb⁺ uptake and concentration of K⁺ in the roots indicated a negative feedback mechanism operating through allosteric control. The Rb⁺ uptake process in root cells was divided into two steps: (1) binding of the ion in the free space, and (ii) transmembrane transport into the cytoplasm. Metabolic and non-metabolic components of uptake were separated by addition of the metabolic inhibitor 2,4-dinitrophenol (DNP) to the nutrient solution. It is suggested that metabolic Rb⁺ uptake requires energy in two uptake steps (for binding to the carrier entity in the free space and for transmembrane transport) or in one step only (for transmembrane transport), dependent on the K⁺ status of the roots. The change from metabolic to non-metabolic binding in the free space is accomplished by changing the conformational state of the carrier (slow/fast transitions). There may be a hysteretic effect on metabolic Rb⁺ uptake through a slow transition between carrier states. This is superimposed on the negative cooperativity, strengthening further cooperativity at intermediate K⁺ levels in the roots. Non-metabolic Rb⁺ uptake probably consists of two components, a carrier-mediated (facilitated diffusion) and a parallel diffusive component.     

Rubidium Uptake and Accumulation in Peripheral Myelinated Internodal Axons and Schwann Cells

Abstract: To study mechanisms of K⁺ transport in peripheral nerve, uptake of rubidium (Rb⁺), a K⁺ tracer, was characterized in rat tibial nerve myelinated axons and glia. Isolated nerve segments were perfused with zero-K⁺ Ringer's solutions containing Rb⁺ (1–20 m M ) and x-ray microanalysis was used to measure water content and concentrations of Rb, Na, K, and Cl in internodal axoplasm, mitochondria, and Schwann cell cytoplasm and myelin. Both axons and Schwann cells were capable of removing extracellular Rb⁺ (Rb⁺_o) and exchanging it for internal K⁺. Uptake into axoplasm, Schwann cytoplasm, and myelin was a saturable process over the 1–10 m M Rb⁺_o concentration range, although corresponding axoplasmic uptake rates were higher than respective glial velocities. Mitochondrial accumulation was a linear function of axoplasmic Rb⁺ concentrations, which suggests involvement of a nonenzymatic process. At 20 m M Rb⁺_o, a differential stimulatory response was observed; i.e., axoplasmic Rb⁺ uptake velocities increased more than fivefold relative to the 10 m M rate, and glial cytoplasmic uptake rose almost threefold. Finally, Rb⁺_o uptake rate into axons and glia was completely inhibited by ouabain (2–4 m M ) exposure or incubation at 4°C. These results suggest that Rb⁺ uptake into peripheral nerve internodal axons and Schwann cells is mediated by Na⁺,K⁺-ATPase activity and implicate the presence of axonal- and glial-specific Na⁺ pump isozymes.     

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.     

Effects of copper on active and passive Rb+ influx in roots of winter wheat

The effects of copper (CuCl₂) on active and passive Rb⁺(⁸⁶Rb⁺) influx in roots of winter wheat grown in water culture for 1 week were studied. External copper concentrations in the range of 10–500 μ M in the uptake nutrient solution reduced active Rb⁺ influx by 20–70%, while passive influx was unaffected (ca 10% of the Rb⁺ influx in the Cu-free solution). At external Rb⁺ concentrations of up to 1 m M , Cu exposure (50 μ M decreased V_max to less than half and increased K_m to twice the value of the control. Short Cu exposure reduced the K⁺ concentration in roots of low K⁺ status. Pretreatment for 5 min in 50 μ M CuCl₂ prior to uptake experiments reduced Rb⁺ influx by 26%. After 60 min pretreatment with Cu, the corresponding reduction was 63%. Cu in the cultivation solution impeded growth, especially of the roots. The Cu concentration in the roots increased linearly with external Cu concentration (0–100 μ M ) while Cu concentration in the shoots was relatively unchanged. The K⁺ concentration in both roots and shoots decreased significantly with increased Cu in the cultivation solutions. Possible effects of Cu on membranes and ion transport mechanisms are discussed.     

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.     

Nickel and rubidium uptake by whole oat plants in solution culture

Nickel and rubidium uptake by oat plants ( Avena sativa L. cv. Victory) were examined in relation to solution temperature, solution concentrations, metabolic inhibitors, anaerobic root conditions, transpiration and time. Over a 4-h period, uptake rates for both Ni²⁺ and Rb⁺ remained constant at 23°C. Decreasing temperatures to 2°C, 20 μ M concentrations of 2,4-dinitrophenol (DNP), or anaerobic root conditions decreased Ni²⁺ and Rb⁺ uptake rates by 97 to 86% in whole plants. Treatment of excised roots with 20 μ M DNP decreased Ni²⁺ uptake by 93%. Nickel and Rb⁺ uptake rates measured as a function of the external solution concentration followed a typical parabolic curve. K_m (0.012 m M ) and V_max [2.72 μmol (g dry weight)^-1 h^-1] values for Ni²⁺ were nearly 7 times lower than those for Rb⁺ [0.09 m M and 19.2 μmol (g dry weight)^-1 h^-1]. In all experiments, Ni²⁺ and Rb⁺ showed qualitatively similar uptake patterns, but Rb⁺ uptake was quantitatively more sensitive than Ni²⁺ to experimental manipulations.     

Potassium channels in barley: cloning, functional characterization and expression analyses in relation to leaf growth and development

It is not known how the uptake and retention of the key osmolyte K⁺ in cells are mediated in growing leaf tissue. In the present study on the growing leaf 3 of barley, we have cloned the full-length coding sequence of three genes which encode putative K⁺ channels ( HvAKT1 , HvAKT2 , HvKCO1 / HvTPK1 ), and of one gene which encodes a putative K⁺ transporter ( HvHAK4 ). The functionality of the gene products of HvAKT1 and HvAKT2 was tested through expression in Xenopus laevis oocytes. Both are inward-rectifying K⁺ channels which are inhibited by Cs⁺. Function of HvAKT1 in oocytes requires co-expression of a calcineurin-interacting protein kinase ( At CIPK23) and a calcineurin B-like protein (AtCBL9) from Arabidopsis , showing cross-species complementation of function. In planta , HvAKT1 is expressed primarily in roots, but is also expressed in leaf tissue. HvAKT2 is expressed particularly in leaf tissue, and HvHAK4 is expressed particularly in growing leaf tissue. Within leaves, HvAKT1 and HvAKT2 are expressed predominantly in mesophyll. Expression of genes changes little in response to low external K⁺ or salinity, despite major changes in K⁺ concentrations and osmolality of cells. Possible contributions of HvAKT1 , HvAKT2 , HvKCO1 and HvHAK4 to regulation of K⁺ relations of growing barley leaf cells are discussed.     

Effects of desiccation on the 77°K fluorescence properties of the liverwort Porella navicularis and the isolated lichen green alga Trebouxia pyriformis

The uptake of the auxin type herbicide 2,4-D into rice seedlings ( Oryza sativa L. cv. Dunghan Shali) and its effects on the K⁺, NH⁺₄ and NO₃ ion uptake and the K⁺ content were investigated at different pH values. A short incubation of the roots in 0.01 m M 2,4-D caused a marked ion uptake inhibition only at low pH. The non-auxin type herbicide benthiocarb did not produce such an inhibitory effect. Lowering of the pH in the external medium led to an increased 2,4-D uptake by the roots. These results can be explained by the increased H⁺ permeability of the membranes, allowing a more rapid entrance of 2,4-D into the root cells, thereby inhibiting the active ion uptake. Rice roots not subjected to 2,4-D treatment responded to H⁺ stress with an increased anomalous K⁺ uptake and a decreased K⁺ content. With reference to the effects of pH changes on the ion and 2,4-D uptake, possible transport mechanism of NH⁺₄ and 2,4-D are briefly discussed.     

Compartmental analysis of Rb+ efflux from roots of intact high-salt barley plants

The validity of compartmental analysis of Rb⁺ efflux from roots of intact high-salt barley plants ( Hordeum vulgare L. cv. Salve) was examined. ⁸⁶Rb⁺ was used as a tracer. Rb⁺ (1 m M ) was included together with 3 m M K⁺ in the growth medium, and steady-state conditions were assumed to prevail during the experiment. Three phases of efflux were revealed with half-times of 23 min, 109 min and 12 h, respectively; and the time span of the experiment had to be at least 20 h to make determination of the slow phase possible. We cannot state what compartments in the root the 3 different slopes represent. A comparison of slopes was made between the plots of In efflux vs time and In content vs time. In spite of correction for tracer transport from the roots to the shoot, the slopes for the slow phase did not agree unless up to 85% of the root content of Rb⁺ is assumed not to participate in efflux.     

K/Na selectivity at the plasmalemma of cortical root cells and preferential release of sodium to the xylem vessels in roots of Atriplex hortensis

Using excised roots of Atriplex hortensis L., cv. Gelbe Gartenmelde, the uptake, accumulation and xylem transport of K⁺ and Na⁺ have been measured. Influx as well as xylem transport proved to discriminate little between K⁺ and Na⁺, when considered in relation to the external solution. Both K⁺ and Na⁺ inhibited the uptake and xylem transport of each other to about the same degree. Measurements of intracel-lular Na⁺ fluxes by means of compartment analysis indicated that the low degree of K/Na discrimination during uptake was due to low influx selectivity. Moreover, K⁺/Na⁺ exchange at the plasmalemma was not very efficient in Atriplex roots. In order to establish the basis of the low K/Na discrimination in xylem transport, the rates of K⁺ and Na⁺ transport were related to the cytoplasmic K⁺ and Na⁺ concentrations to yield the selectivity ratio of transport, S(transport) = (φ_cx(K) × [Na⁺]_c)/(φ_cx(Na) × [K⁺]_c). Under all conditions this ratio was far below one indicating that Na⁺ was favoured during xylem release in excised roots of Atriplex at low external concentrations. The implications of this discrimination in favour of Na+ are discussed with respect to salt tolerance of A. hortensis .     

Effects of aluminium on growth and kinetics of K+(86Rb) uptake in two cultivars of wheat (Triticum aestivum) with different sensitivity to aluminium

Two cultivars of wheat (Triticum aestivum L. cvs Kadett and WW 20299) were grown for 9 days with 20% relative increase in nutrient supply per day at pH 4.1. Aluminium at 50 μ M retarded the growth of roots more than that of shoots in both cultivars, thus decreasing the root/shoot ratio. The inhibition was largest in WW 20299. With long term Al treatment (9 days), K_m for K⁺(⁸⁶Rb) influx increased five times in both cultivars and V_max decreased in WW 20299. Efflux of K⁺(⁸⁶Rb) was little affected. When the roots were treated with aluminium for two days, only relative growth rate of roots was retarded, while growth of shoots was unaffected and influx of K⁺(⁸⁶Rb) adjusted to the actual K⁺ demand of the plants. It is concluded that the effects of aluminium on K⁺ uptake in these wheat cultivars are not primary factors contributing to aluminium sensitivity. However, in soil with Al the demand for a comparatively high concentration of K⁺ to maintain an adequate K⁺ uptake rate, in combination with a slow growth rate of the roots, may secondarily lead to K⁺ deficiency in the plants.     

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.     

Influence of light and darkness and the experimental design on kinetics of K+ (86Rb) influx in roots of intact spring wheat

Seedlings of spring wheat ( Triticum aestivum L. cv. Svenno) were cultivated at 20°C in continuous light or darkness with the roots in nutrient solutions for six days. The plants were starved for K⁺ during different periods of time to produce plants with various K⁺ status. In one cultivation light-grown plants were pretreated in darkness, and vice versa, before the uptake experiment. In all experiments, roots were put in a complete nutrient medium containing 2.0 m M K⁺ radiolabelled with ⁸⁶Rb. The uptake time was varied (5, 60 or 120 min).
The K⁺ concentration in the roots, [K⁺]_root, increased during the course of the uptake experiments, especially in light and at initially low [K⁺]_root, At the same time K⁺ (⁸⁶Rb) influx in the roots decreased. The simoidal relationship obtained between K⁺ (⁸⁶Rb) influx and [K⁺]_root was affected by these changes, and Hill plots gave various Hill coefficients, n_H, depending on the duration of the uptake experiments. n_H from three apparently straight line segments of the same plot, in different [K⁺]_root - intervals, indicated a falling degree of interaction between the binding sites as [K⁺]_root increased. For the dark-grown plants negative cooperativity could not be demonstrated.     

Volume changes of Commelina communis guard cell protoplasts in response to K+, light and CO2

The effects of K⁺ concentration, light intensity and CO₂ levels on the volume of Commelina communis L. guard cell protoplasts were studied. Two degrees of swelling response were observed, both dependent on an external supply of K⁺, but not necessarily on the supply of a permeant anion. The presence of K⁺ itself, independent of light or CO₂ level, stimulated swelling at a relatively slow rate. When K⁺, light and low CO₂ conditions were supplied together, the swelling was relatively rapid and of high magnitude. The rapid swelling was specific for K⁺ and Rb⁺ giving a half maximal effect after 2 h at a KCl concentration of about 18 mmol m⁻³. The addition of CaCl₂ at 1 mol m⁻³ inhibited K⁺-dependent swelling under all conditions tested. The response to light and low CO₂ levels by Commelina guard cell protoplasts is thought to reflect a high degree of physiological integrity.     

The effect of xanthoxylin (2-oxy, 4-6-dimethoxyacetophenone) on potassium-dependent acid: extrusion in wheat and maize root segments

Abstract. Xanthoxylin is a cytotoxic and fungicidal compound with the characteristics of a typical phytoalexin. At a concentration of 0.3 mol m⁻³ it inhibits K⁺-dependent acid extrusion and K⁺ net uptake (or uptake of equivalent alkaline cations such as Rb⁺ and CS⁺) by up to about 80% and hyperpolarizes by about 20% the membrane electrical potential. Its inhibition of the acid extrusion does not depend on altered ion exchange involving the anions in the media, a reduction of the metabolic energy available, or detectable changes in the permeability of the cell membrane to H⁺ ions. The drop in K⁺ net uptake depends on a decrease in the influx of K⁺ into the cell. In functional terms, xanthoxylin is an inhibitor of the K⁺ permeation mechanism and does not appear to interact with the mechanisms creating the electrochemical energy gradient.     

Growth and accumulation of N, K+, Ca2+ and Mg2+ in barley exposed to various nutrient regimes and root/shoot temperatures

Six cultivars of spring barley ( Hordeum vulgare L. cvs Salve, Nümberg II, Bomi, Risø 1508, Mona and Sv 73 608) were grown in water culture for three weeks with various combinations of mineral supply and differential roots/shoot temperatures during the growth period. Most important for growth and accumulation of N, K⁺, Ca²⁺ and Mg²⁺ was the mineral supply, followed by the root temperature and the choice of cultivar. Treatments with low mineral supply or low root temperature induced a uniform reduction in growth and accumulation of the ions studied. The effects of low mineral supply and low root temperature on growth and N accumulation was additive, which indicates that these factors exert their influence independently of each other.
Roots grown at 10°C were smaller and Rb⁺(⁸⁶Rb) influx was higher than in roots grown at 20°C. It is suggested that the control of Rb⁺(⁸⁶Rb) influx is affected by the root temperature and the age of the plants. The higher ⁸⁶Rb⁺ (⁸⁶Rb) influx into the low temperature roots could not compensate for the smaller root size. However, the lower total mineral accumulation made up for the needs of the smaller plants and cannot explain the reduction in growth.     

An effect of Na+ on K+ uptake in intact seedlings of Zea mays

Models for the regulation of K⁺ uptake in higher plant roots have become more complex as studies have moved from the level of excised low-salt roots to that of intact plants grown under fully autotrophic conditions. In this paper we suggest that some of the differences between the conditions are qualitative, possibly requiring fundamental changes to the model, rather than simply quantitative.
The uptake of K⁺ by low-salt roots of Zea mays L. [(A619 x Oh 43) x A632], was independent of Na⁺ concentration over a wide range. However, independence of Na⁺ was not the case in plants grown on complete nutrient medium in the light: inclusion of Na⁺ in the uptake medium enhanced K⁺ uptake. In the presence of Na⁺, K⁺ uptake rates were similar in whole plants with high root K⁺ contents to rates in excised or intact, low-salt roots.