Involvement of epidermal cells in stomata movement: The effect of ABA

ABA affected K⁺ and solute transport between guard cells and epidermal cells as indicated by K⁺ staining and plasmolysis. ABA enhanced K⁺ (⁸⁶Rb) uptake into epidermal cells. To find out whether the ABA enhanced accumulation of K⁺ (⁸⁶Rb) in epidermal cells is active, uptake in the presence of exogenous ATP was studied. These studies hinted that K⁺ (⁸⁶Rb) uptake by epidermal cells is a passive process, while its release is an active one. This was verified by applying iodoacetate, which interferes with energy supply. The epidermal cells thus seem to play a role in stomatal movement.     

Sodium Transport from Blood to Brain: Inhibition by Furosemide and Amiloride

Abstract: Brain sodium uptake in vivo was studied using a modified intracarotid bolus injection technique in which the uptake of ²²Na ⁺ was compared with that of the relatively impermeable molecule, [³H]l-glucose. At a Na ⁺ concentration of 1.4 m M , Na ⁺ uptake was 1.74 ± 0.07 times greater than l -glucose uptake. This decreased to 1.34 ± 0.04 at 140 m M Na ⁺, indicating saturable Na ⁺ uptake. Relative Na ⁺ extraction was not affected by pH but was inhibited by amiloride ( K _i= 3 ± 10⁻⁷ M ) and by 1 m M furosemide. The effects of these two inhibitors were additive. Brain uptake of ⁸⁶Rb ⁺, a K ⁺ analogue, was measured to study interaction of K ⁺ with Na ⁺ transport systems. Relative ⁸⁶Rb ⁺ extraction was also inhibited by amiloride; however, it was not inhibited by furosemide. The results suggest the presence of two distinct transport systems that allow Na ⁺ to cross the luminal membrane of the brain capillary endothelial cell. These transport systems could play an important role in the movement of Na ⁺ from blood to brain.     

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.     

Membrane and ion transport properties in cereals under acidic and alkaline stress

The effects of pH on the growth and the K⁺ (⁸⁶Rb) uptake and K⁺ content of excised rice ( Oryza sativa L. cv. Dunghan Shali) and wheat ( Triticum aestivum L. cv. GK Szeged) roots were investigated. Rice roots responded to H⁺ stress with an increased K⁺(⁸⁶Rb) influx and a decreased K⁺ content, suggesting an increased exchange between the cytoplasmic K⁺ pool and the external medium. Under the same experimental conditions wheat did not show any anomalous K⁺(⁸⁶Rb) influx. Growth of both rice and wheat was relatively insensitive to pH between 4 to 10.     

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.     

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.     

Comparison of the effects of auxin and fusicoccin on phosphate absorption by aged potato tuber discs

Auxin (IAA, 5 × 10⁻⁵ M ) partially prevents the increase in the rate of phosphate uptake during ageing of potato tuber discs ( Solatium tuberosum L. cv. Bintje), whereas fusicoccin (FC, 10⁻⁵ M) stimulates it. After the development of enhanced phosphate transport capacity, the response to fusicoccin is greater than with fresh discs. Complementary experiments on K⁺ (⁸⁶Rb) absorption show that FC also slightly enhances the rate of K⁺ uptake, while IAA has no much effect. It is suggested that IAA acts specifically on the development of a mechanism which occurs during the ageing period, while FC action may be more directly linked to the system of phosphate transport itself.     

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.     

Effects of nitrate on influx, efflux and translocation of potassium in young sunflower plants

Influx, efflux and translocation of K⁺(⁸⁶Rb) were studied in the roots of sunflower seedlings ( Helianthus annuus L. cv. Uniflorus) treated with 0–4.0 m M NO₃⁻ during a 9 day growth period or a 24 h pretreatment period. Roots treated with high levels of NO₃⁻ absorbed and translocated more K⁺(⁸⁶Rb) than seedlings treated with low levels of NO₃⁻. The content of K⁺ in the shoots was, however, higher in seedlings treated with low levels of NO₃⁻, indicating a low rate of retranslocation of K⁺ in those plants. K⁺(⁸⁶Rb) efflux was highest into the low-NO₃⁻ solutions. All effects on K⁺(⁸⁶Rb)-fluxes were more obvious in high-K plants than in low-K plants. The results are discussed in relation to the Dijkshoorn-Ben Zioni hypothesis for K⁺+ NO₃⁻-uptake and translocation in plants.     

Effect of the crown on uptake and transport in embryonic shoots of Picea abies

Embryonic shoots of Picea abies (L.) Karst, isolated from 10-year-old trees, were excised either with or without the crown. Various short-term uptake experiments (3, 6 and 24 h) and one long-term uptake experiment (4 weeks) were performed with these shoots to obtain information about the physiological role of the crown as translocation barrier for different substances. Transport through the embryonic shoots was followed in both acropetal and basipetal directions using radiolabelled substances supplied in an agarified Schenk and Hildebrandt medium. The medium was labelled with [¹⁴C]-IAA and/or [³²P]-phosphate, or with [³⁵S]-sulphate and ⁸⁶Rb (as a tracer for K⁺). The experiments were conducted in light at 20°C, with the exception of one of the short-term experiments, which was carried out at 5°C to evaluate the connection between transport and metabolism. The main observation is that the crown in its collenchymatous stage of development acts as a selective barrier both acropetally and basipetally for transport of substances such as [¹⁴C]-IAA and [³²P]-phosphate or their metabolized forms. This could explain why the embryonic shoot when cultured plus or minus its crown shows different growth and developmental patterns in vitro.     

Effect of boron on stomatal opening in epidermal strips of Commelina communis

Boron enhances stomalal opening in epidermal strips of Commelina communis L. under conditions conductive to opening, i.e . light and KCI. This effect is obtained when boron is given together or before KCI application. Since boron increases ⁸⁶Rb net uptake in both illuminated epidermal strips and illuminated isolated guard cells, it is suggested that boron acts either by reducing potassium leakage or by increasing its uptake, thus leading to increased osmotic potential and, as a result, to stomatal opening.     

Heterosis and ion transport in hexaploid varieties of rye-wheat (triticale) compared to the parental species

Important winter and spring varieties of hexaploid rye-wheat (triticale cvs. 6048 and 5004) were selected for study of heterotic effects on growth and ion transport in the hybrids compared to the parental species rye ( Secale cereale L. cvs. MT 77 and Sv 6970) and wheat (Triticum aestivum L. cvs. Starke II and Sonett). After 3 days germination, seedlings were grown 11 days in water culture on a complete nutrient solution diluted to 1, 25 and 50%. Intracellular influx and transport to shoots of K⁺, Ca²⁺, sulphur and phosphate were determined by using radioactive tracers (⁸⁶Rb (for K⁺), ⁴⁵Ca, ³⁵S and ³²P). Varietal differences in the parameters studied were generally small compared to differences between species. The heterotic effect on growth of rye-wheat was mainly localized to the shoots at high ionic strengths (25% and 50%). There were no heterotic effects on ion influx or transport to the shoots. Ion influx and transport characteristics in rye-wheat appear to be inherited mainly from wheat. Growth of all species on 1% medium was severely reduced. At the low ionic strength ion influx was inhibited similarly for all species, except influx of K⁺ (⁸⁶Rb) which was higher in rye-wheat and wheat than in rye. Ion influx and transport in rye-wheat and wheat and in rye differed especially for 25% and 50% media. Rye had the highest ion influx and transport and the highest shoot/root fresh weight ratio at the high ionic strengths. To feed a comparatively large shoot, rye may compensate for a relatively small root system by efficient ion transport mechanisms.     

Transport of G AB A, β-Alanine and Glutamate into Perikarya of Postnatal Rat Cerebellum

Abstract: Cells dissociated from the postnatally developing rat cerebellum retain their high-affinity carrier-mediated transport systems for [³H]GABA ( K _t=1.9 μM, V = 1.8 pmol/10⁶ cells/min) and [³H]glutamate ( K _t= 10 μM, V = 7.9 pmol/10⁶ cells/min). Using a unit gravity sedimentation technique it was demonstrated that [3H]GABA was taken principally into fractions that were enriched in inhibitory neurons (Purkinje, stellate and basket cells). [³H]β-alanine (which is taken up specifically by the glial GABA transport system) and [³H]glutamate were concentrated by glial-enriched fractions. However [³H]glutamate uptake was minimal in fractions enriched in precursors of granule cells, which may utilise this amino acid as their neurotransmitter. These results are discussed in relation to reports of high-affinity [³H]glutamate uptake by glia. The role of glutamate transport in glutamatergic cells is also considered. The data suggest that high-affinity glutamate transport is a property of glial cells but not granule neurons.     

Potassium and Taurine Release Are Highly Correlated with Regulatory Volume Decrease in Neonatal Primary Rat Astrocyte Cultures

Abstract: Neonatal rat primary astrocyte cultures were swollen by exposure to hypotonic buffer. Using an electrical impedance method for determination of cell volume coupled with on-line measurements of efflux of radioactive ions or amino acids, we have investigated the role of K⁺ (using ⁸⁶Rb), taurine, and d -aspartate (an analogue of glutamate) in regulatory volume decrease (RVD). Addition of 1 m M quinine, 10 µ M nimodipine, 100 µ M BAPTA-AM, 10 µ M trifluoperazine, or a calcium-free buffer significantly ( p < 0.0001) inhibited RVD. This was accompanied by inhibition of ⁸⁶Rb release but an increase in d -[³H]-aspartate release, which was proportional to the degree to which RVD was inhibited. These results support a regulatory role for calcium in RVD and show that inhibition of calcium entry from the extracellular fluid, intracellular calcium sequestration, inhibition of calcium-activated K⁺ channels, and inhibition of calmodulin all inhibit RVD. Because d -[³H]aspartate efflux profiles increase as RVD is inhibited, it is unlikely that d -aspartate release is a main determinant of RVD. In contrast, [³H]taurine release was increased by 1 m M quinine and inhibited by 10 µ M trifluoperazine. The net release of K⁺ and taurine is highly correlated with the degree of RVD, implicating a regulatory role for both K⁺ and taurine release in RVD.     

Comparison of DABA and GABA Transport into Plasma Membrane Vesicles Derived from Synaptosomes

Abstract: Transport of GABA by a high-affinity transport system ( K _m≃ 10⁻⁵ M) is thought to terminate the action of this postulated neurotransmitter. 2,4-Diaminobutyric acid (DABA), a structural analogue, is taken up by neuronal elements and inhibits GABA uptake. Localization of [³H]DABA by auto-radiography has been used to identify neurons with the GABA high-affinity transport system. After reconstitution of lysed synaptosomal fractions in potassium salts, transfer of these membrane vesicles to sodium salts produces sodium and potassium ion gradients which drive [³H]GABA and [³H]DABA transport. For each, transport requires external sodium, is abolished by ionophores that dissipate the Na⁺ gradient, and is enhanced by conditions which make the intravesicular electromotive force more negative. Some characteristics of the transport of these substances, however, differ. For example, external chloride is required for GABA, but not DABA, transport. Internal potassium is required for DABA, but not GABA, transport. DABA is a competitive inhibitor ( K _i≃ 0.6 MM) of GABA transport into membrane vesicle and synaptosomes. GABA, however, is a feeble inhibitor of DABA uptake into the membrane vesicles. These differences suggest that the two substances are transported by different mechanisms and possibly by different carriers. In addition to these experiments, using enzymatic-fluorometric techniques, it was shown that the artificially imposed ion gradients drive net chemical transport of GABA into the vesicles.     

Early Signals in Serum-Induced Increases in Ouabain-Sensitive Na+-K+ Pump Activity and in Glucose Transport in Rat Skeletal Muscle Are Amiloride-Sensitive

Abstract: The acute effects of serum on sodium-potassium (Na⁺-K⁺) pump activity and glucose uptake in cultured rat skeletal muscle were studied. Addition of serum to myo-tubes in phosphate-buffered saline caused Na⁺-K⁺ pump activity (as measured by changes in the ouabain-sensitive component of both membrane potential and ⁸⁶Rb uptake) to increase, with peak effects obtained after 30 min. The effect was blocked completely by treatment with amiloride, but not by tetrodotoxin, which blocks voltage-dependent Na⁺ channels. On transfer of myotubes to Na⁺-free, choline buffer, resting Na⁺-K⁺ pump activity decreased to about 10% of that in phosphate-buffered saline. Addition of regular serum, but not Na⁺-free serum, caused Na⁺-K⁺ pump activity to increase slightly. Similar results were obtained with serum on glucose uptake, the peak effect being reached within 15 min. Stimulation of glucose uptake by serum was partially reduced by amiloride and was not altered by tetrodotoxin. Removal of external Na⁺ also eliminated serum effects on glucose uptake. The results demonstrate that there are similar signals involving Na⁺-H⁺ exchange for serum-induced increases in Na⁺-K⁺ pump activity and glucose transport. The lack of complete blockade of serum-induced elevation of glucose transport suggests an additional, as yet undefined, intracellular signal for stimulation of this transport system.     

Net uptake and partitioning of nitrogen and potassium in cultivars of barley during ageing

Experiments were carried out with barley cultivars ( Hordeum vulgare L.) grown in both pot- and water-culture. Net uptake of NO3₋ and K⁺ in the roots was followed in two barley cultivars grown on water-culture for 85 days. After an initial period of low net uptake of both ions, uptake increased until a maximum was reached after 30 to 45 days. Thereafter, net uptake of NO₃⁻ and K⁺ steadily decreased. In the pot experiments, effects of different mineral supply on day 4 to 18 upon the development of five barley cultivars of various earliness were investigated. The effect of earliness on fresh weight production was largest when mineral supply on day 4 to 18 was limited. The influence of limited mineral supply on day 4 to 18 on K-economy was independent of earliness of the cultivars. The maximal N-content was reached at the same time as maximal fresh and dry weight in fairly late cultivars; in early cultivars maximum N-level was reached later than maximum fresh and dry weight. Overall, maximal N-content was higher in the fairly late cultivars than in the early cultivars. The highest rate of ¹⁵N-transport was attained later in two of three fairly late cultivars than in early cultivars. Partitioning of dry weight, N and K in the shoots changed during ageing, ears being an important sink. Varietal differences in partitioning depended on the earliness of the cultivars. The largest fraction of recently supplied ¹⁵N, supplied as nitrate, and K⁺ (⁸⁶Rb) were found in the stems. In the oldest plants of the early cultivars the transport to the ears of these isotopes was gradually impaired, reflecting the decreasing function of the long distance transport system.     

Free space uptake and influx of Ni2+ in excised barley roots

The effects of switches between high and low nutrient supplies on growth and mineral nutrition of winter wheat ( Triticum aestivum L. cv. Martonvásári-8) were followed in four main developmental phases: tillering, shooting, heading and grain filling. Growth of the shoots was significantly affected by switches. Under low nutrient supply the life cycle was shortened. Root growth was only slightly affected by switches, but an early high nutrient supply followed by low nutrient supply gave an impetus for root development. In general, the growth data indicate that the nutrient status of the plants is determined by the nutrient level supplied during shooting. A high level of nutrients during shooting leads also to high vegetative growth, whereas the best grain yield was obtained by a high dose of nutrients during tillering followed by low nutrient conditions during the shooting stage and later. K⁺(⁸⁶Rb) influx in the roots decreased with age. The potential for K⁺ (⁸⁶Rb) influx was low in plants of high-salt status, but it became high in response to switching to low supply at shooting, whereas later switches had no influence on this function in high-salt plants. The highest K⁺(⁸⁶Rb) influx was found in plants starting with high nutrient supply followed by low-salt conditions; this plant group was outstanding also with respect to its high grain yield.     

Actual escape area and lateral escape from the xylem of the alkali ions Na+, K+, Rb+ and Cs+ in tomato

Approximation of the total escape area of the xylem in an inbred line of tomato (Ly-copersicon escutentum Mill. cv. Tiny Tim) with help of the frequency distribution of xylem vessel radii provides the possibility to calculate realistic escape constant values from uptake experiments of several elements into tomato stem segments. Comparison of the lateral escape rates of ²⁴Na⁺, ⁴²K⁺, ⁸⁶Rb⁺ and ¹³⁴Cs⁺ indicate that Na⁺ escape is rate-limited by its uptake into a rather constant number of surrounding cells, regardless of changes in the total escape area of the xylem vessels. The escape of K⁺, Rb⁺ and Cs⁺ seems to be proportional to the surface area of the xylem vessels and their escape is apparently controlled by their transport across the cell walls of the transport channels. The calculated small values for the escape rate constants (apparent permeability of the xylem cell walls, ca 2–3 · 10−⁹ m s−⁷) are probably due to the presence of lignin in the xylem cell walls, the discrimination between ions as a result of differing affinities and selectivities and the presence of other solutes in the applied solution.     

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.