Phytochrome and potassium uptake by mung bean hypocotyl sections

Uptake of potassium (K) and ⁸⁶rubidiumlabelled potassium (⁸⁶Rb) by sub-hypocotyl hook sections of Phaseolus aureus L. was inhibited by red light. The effect was reversible with far red light. Using short exposures of high irradiance the effect on ⁸⁶Rb-labelled K uptake was observed after 5 min. The response showed no specificity for a particular anion. Uptake of ⁸⁶Rb-labelled K by sections cut immediately below the cotyledons was enhanced by red light after 10 min incubation and was also far red reversible. These results are interpreted as a rapid phytochrome-induced change in membrane properties resulting in modified K uptake.Abbreviations P Phytochrome - Pr red absorbing form of P - Pfr far red absorbing form of P - R red light - F far red light     

Sugar uptake into strawberry fruit is stimulated by abscisic acid and indoleacetic acid

Changes in sugar uptake into strawberry fruits with maturation and the hormonal effect on uptake mechanisms, though important to fruit development, are not known. Therefore, the kinetics of sugar uptake into strawberry ( Fragaria x ananassa Duch cv. Nyoho) fruit tissue and the effects of abscisic acid (ABA) and indoleacetic acid (LAA) on the mechanism of uptake were investigated at 25 and 35 days after pollination (DAP). Uptake of ¹⁴C-sugar was measured over the concentration range of 2 to 30 m M. Uptake kinetics showed a biphasic response to increasing external concentration of ¹⁴C-sugars, and indicated the presence of P -chlorormercuribenzenesulfonic acid (PCMBS)-sensitive and PCMBS-insensitive uptake. The K_m value for each sugar was in the range of 10 to 20 m M. Stage of development had no effect on K_m. but V_max for glucose decreased with maturation. Further, sucrose was not taken up through a PC-MBS-sensitive transport at 35 DAP. ABA, especially 10 μ M , at 25 DAP stimulated uptake of all sugars, mostly through enhanced PCMBS-insensitive uptake but not PC-MBS-sensitive uptake. In contrast to ABA, stimulation of sugar uptake by IAA was most effective at 1 μ M . The PCMBS-insensitive uptake of each sugar was also stimulated by IAA. Further, the PCMBS-sensitive uptake of glucose was enhanced. The developmental change of PCMBS-sensitive sugar uptake and the effect of ABA and IAA on uptake mechanism in this study are considered to be important in influencing the development and enlargement of fruits.     

Evidence on the mechanism of enhanced sucrose uptake at low cell turgor in leaf discs of Phaseolus coccinius

Plants often tolerate water deficits by lowering the osmotic potential of their cell sap. This may be achieved by accumulation of solutes which results in the maintenance of a positive turgor potential. In this study, the effect of water deficit on sugar uptake was investigated in leaf discs of Phaseolus coccinius L. (cv. Scarlet). Evidence is presented that cell turgor affects the kinetics of sugar transport at the membrane level. Uptake kinetics of sucrose, glucose and 3-O-methyl glucose by tissues equilibrated in solutions of relatively high (200–400 mOsm) osmotic concentration consisted of a sat-urable and a linear component. Low external osmotic concentration i.e., high cellular turgor inhibited the saturating component of sucrose uptake, resulting in a linear uptake profile. However, high cell turgor had no effect on glucose or 3-O-methyl glucose uptake kinetics. The effect of turgor versus osmotic component of water potential was differentiated by comparing responses to non-penetrating (manmtol) or polyethylene glycol, (3350) and penetrating (ethylene glycal) osmotica. Changes in sucrose uptake rates and kinetics were due to changes in cellular turgor and not osmotic potential. Furthermore, at low cellular turgor, a net increase in sucrose uptake occurred as a consequence of enhanced influx rates and not as a result of reduced efflux rates. The data are consistent with previous findings that sugar uptake rates are enhanced under low turgor. We present first evidence indicating that the mechanism by which higher rates of sucrose uptake are maintained underwater deficit conditions is by the activation of the saturable transport system. This mechanism supports previous suggestions that changes in cell turgor are sensed and manifested at the membrane level.     

In Vitro Sugar Transport in Zea mays L. Kernels : I. Characteristics of Sugar Absorption and Metabolism by Developing Maize Endosperm

Short-term transport studies were conducted using excised whole Zea mays kernels incubated in buffered solutions containing radiolabeled sugars. Following incubation, endosperms were removed and rates of net ¹⁴C-sugar uptake were determined. Endogenous sugar gradients of the kernel were estimated by measuring sugar concentrations in cell sap collected from the pedicel and endosperm. A sugar concentration gradient from the pedicel to the endosperm was found. Uptake rates of ¹⁴C-labeled glucose, fructose, and sucrose were linear over the concentration range of 2 to 200 millimolar. At sugar concentrations greater than 50 millimolar, hexose uptake exceeded sucrose uptake. Metabolic inhibitor studies using carbonylcyanide-m-chlorophenylhydrazone, sodium cyanide, and dinitrophenol and estimates of Q₁₀ suggest that the transport of sugars into the developing maize endosperm is a passive process. Sucrose was hydrolyzed to glucose and fructose during uptake and in the endosperm was either reconverted to sucrose or incorporated into insoluble matter. These data suggest that the conversion of sucrose to glucose and fructose may play a role in sugar absorption by endosperm. Our data do not indicate that sugars are absorbed actively. Sugar uptake by the endosperm may be regulated by the capacity for sugar utilization (i.e. starch synthesis).     

The uptake and adsorption of diquat and paraquat by tomato, sugar beet and cocksfoot

Experiments on leaves of tomato, sugar beet and cocksfoot show that uptake of diquat and paraquat, although rapid in the light, is increased by darkness and therefore takes place through the cuticle and not through stomata. Darkness for as little as 4 hr. increased uptake almost twofold. Diquat and paraquat are rapidly and strongly adsorbed both to leaf tissue and to extraneous matter on the leaf surface. Uptake in the field is so rapid that rain immediately after treatment has little adverse effect.     

Energy Supply and Light-enhanced Chloride Uptake in Wheat Laminae

The light-supported component of ³⁶Cl uptake from 5 mM K³⁶Cl by green laminae, either chopped or vacuum-infiltrated, of Triticum aestivum L. seedlings has been determined by subtraction of dark uptake values from light uptake values and the energy sources for the uptake elucidated on the basis of the effects of 3-(3,4-dichlorophenyl)-1,1 dimethylurea (DCMU), carbonyl cyanide p-trifluoromethoxy-phenylhydrazone (FCCP), antimycin A₁ (AA), and N₂ on light and dark uptake. The light-supported Cl⁻¹ uptake is shown to be partially inhibited by DCMU or AA but unaffected or stimulated by FCCP or N₂. There is no additive effect on inhibition caused by DCMU + N₂ or FCCP + AA but there is an added inhibition caused by DCMU + AA, DCMU + FCCP, and by FCCP or AA in anaerobic conditions. The effect of these inhibitors on photosynthetic gas exchange of chopped tissue has also been determined. On the basis of the results it is concluded that the uptake of Cl⁻, supported in the dark by oxidative phosphorylation, is enhanced by light and may be supported by cyclic and non-cyclic electron-flow energy. Uptake is not obligatorily linked to any one energetic pathway and there may be switching from one source to another.     

Phloem Loading in Coleus blumei in the Absence of Carrier-Mediated Uptake of Export Sugar from the Apoplast

Phloem loading in Coleus blumei Benth. leaves cannot be explained by carrier-mediated transport of export sugar from the apoplast into the sieve element-companion cell complex, the mechanism by which sucrose is thought to load in other species that have been studied in detail. Uptake profiles of the export sugars sucrose, raffinose, and stachyose into leaf discs were composed of two components, one saturable and the other not. Saturable (carrier-mediated) uptake of all three sugars was almost completely eliminated by the inhibitor p-chloromercuribenzenesulfonic acid (PCMBS). However, when PCMBS was introduced by transpiration into mature leaves it did not prevent accumulation of ¹⁴C-photosynthate in minor veins or translocation of labeled photosynthate from green to nonchlorophyllous regions of the leaf following exposure to ¹⁴CO₂. The efficacy of introducing inhibitor solutions in the transpiration stream was proven by observing saffranin O and calcofluor white movement in the minor veins and leaf apoplast. PCMBS introduced by transpiration completely inhibited phloem loading in tobacco leaves. Phloem loading in C. blumei was also studied in plasmolysis experiments. The carbohydrate content of leaves was lowered by keeping plants in the dark and then increased by exposing them to light. The solute level of intermediary cells increased in the light (phloem loading) in both PCMBS-treated and control tissues. A mechanism of symplastic phloem loading is proposed for species that translocate the raffinose series of oligosaccharides.     

Interactions of light, temperature and inorganic nitrogen in controlling planktonic nitrogen utilisation in the Tagus estuary

The effect of light, temperature and ammonium on inorganic nitrogen uptake by phytoplankton was investigated from June 1994 through December 1995 at three sites in the Tagus estuary (Portugal), during high tide of neap tides. Ammonium concentrations higher than 10 M reduced nitrate uptake down to 24% but never prevented it. Below this threshold concentration, nitrate uptake was neither inhibited nor changed. Uptake of both nitrate and ammonium as a function of light intensity exhibited a saturation response. Uptake reduction occurred in the near bottom phytoplankton populations, particularly for nitrate. The ammonium uptake system was less limited by light than the nitrate uptake system, indicating the importance of ammonium as a nitrogen source for the phytoplankton which is likely to experience high changes in light in the well-mixed water column of this estuarine environment. Ammonium uptake was exponentially related to temperature in the upper estuary whereas in the mid and lower estuary this relationship was linear. The effect of temperature on nitrate uptake was linear but far less marked than for ammonium uptake.     

Uptake and Release of Abscisic Acid by Isolated Photoautotrophic Mesophyll Cells, Depending on pH Gradients

Uptake and release of abscisic acid (AbA) by isolated mesophyll cells of Papaver somniferum is characterized by the following observations: (a) Uptake rate is a linear function of the external AbA concentration in the range from 10⁻⁶ to 5 × 10⁻⁵ molar, and decreases with increasing pH. At any pH, uptake rate is linearly related to the concentration of undissociated abscisic acid, calculated from the pK = 4.7 according to the Henderson-Hasselbalch equation. At low external pH (5.0), AbA accumulation in the cells is about 10-fold. (b) Uptake of AbA is completely inhibited by salts such as KNO₂ or sodium acetate, which decrease the pH gradient between medium and cells. KCN or m-chlorocarbonylcyanide phenylhydrazone inhibits AbA uptake only after longer incubation periods (20-40 minutes). (c) Uptake rate as well as equilibrium concentration is significantly higher in light than in darkness. (d) At low external pH, release of AbA from preloaded cells is strongly stimulated by KNO₂. It is concluded that AbA is distributed between leaf cells and free space according to pH gradients, with the undissociated abscisic acid being the main penetrating species. Uptake and release occur via diffusion, without participation of a carrier.     

Red light and auxin effects on rubidium uptake by oat coleoptile and pea epicotyl segments

Apical segments of etiolated oat (Avena sativa L. cv. Victory) coleoptiles showed enhanced uptake of [⁸⁶Rb⁺] when tested 30 minutes after a 5-minute red irradiation. The response was partly reversible by far red light. Uptake was sensitive to carbonyl cyanide m-chlorophenyl hydrazone, but not to isotonic mannitol. Indoleacetic acid (10⁻⁷ molar) caused a very pronounced and rapid stimulation of uptake. Basal coleoptile segments also exhibited a red light-enhanced uptake, but not an effect of red light on changes in the pH of the medium. The [⁸⁶Rb⁺] uptake of third internode segments from etiolated peas (Pisum sativum L. cv. Alaska) was not affected by either red light or auxin. This tissue also showed no red light effect on acidification of the medium. It is concluded that alteration of [⁸⁶Rb⁺] flux is not a general feature of phytochrome action.     

Regulation of the glucose phosphotransferase system in Brochothrix thermosphacta by membrane energization.

Uptake of 2-deoxyglucose, alpha-methylglucopyranoside, and glucose into intact cells of Brochothrix thermosphacta (formerly Microbacterium thermosphactum, ATCC 11509) was stimulated by KCN or CCCP. The glucose analogs were recovered almost totally as the sugar phosphates. Membrane vesicles were isolated from protoplasts and shown to be right side out by freeze fracturing and by using ATPase as a marker for the cytoplasmic membrane surface. Uptake of glucose into vesicles was dependent on the presence of phosphoenolpyruvate. NADH oxidation, K+ -diffusion gradients, and externally directed lactate gradients (pH greater than 7 initially) were used to generate transmembrane potentials across membrane vesicles. Above a threshold value of about -50 mV, uptake of glucose into membrane vesicles was reduced. Likewise, the maximum uptake of glucose and its two analogs into cells occurred when the protonmotive force was less than about -50 mV.     

Sugar uptake by maize endosperm suspension cultures

Maize (Zea mays L.) endosperm suspension cultures are a useful model system for studying biochemical and physiological events in developing maize endosperm. In this report, sugar uptake by the cultures is characterized. Uptake of ¹⁴C-labeled fructose and l-glucose was linear with time, while the rate of uptake of radioactivity from sucrose increased over a 120 min period. Both saturable and linear components of uptake were observed for fructose, glucose, sucrose, 1′-deoxy-1′-fluorosucrose, and maltose. Uptake of mannitol, sorbitol, and l-glucose took place at lower rates and was linear with concentration. Rates of incorporation of radioactivity from fructose and glucose exceeded that of sucrose at all concentrations tested. Kinetics of 1′-deoxy-1′-fluorosucrose uptake indicated that ¹⁴C from sucrose can be taken up by a saturable carrier of intact sucrose as well as by invertase hydrolysis and subsequent uptake of hexoses. Cell wall invertase was demonstrated histochemically. Further study of fructose uptake at a concentration at which the saturable component predominated revealed sensitivity to metabolic inhibitors, respiratory uncouplers, the nonpermeant sulfhydryl reagent p-chloromercuribenzenesulfonic acid, and nigericin. Uptake was not affected by valinomycin plus K⁺ and was stimulated by fusicoccin. Fructose and glucose uptake was not pH-sensitive below pH 7.0, whereas uptake of radioactivity from sucrose and 1′-deoxy-1′-fluorosucrose declined as the pH was increased above 5.0. Fructose uptake was not completely inhibited by glucose and vice versa, suggesting the presence of specific carriers. These results indicate that maize endosperm suspension cultures (a) absorb fructose via a typical, energy-requiring, carrier-mediated proton cotransport system; (b) possess saturable carriers for glucose and sucrose; and (c) also absorb sucrose via hexose uptake after sucrose hydrolysis by extracellular invertase.     

Taenia crassiceps: absorption of hexoses and partial characterization of Na + -dependent glucose absorption by larvae

The uptake of ¹⁴C-fructose by T. crassiceps larvae was linear with respect to concentration. Uptake of 0.05 mM¹⁴C-fructose was not inhibited by 5.0 mM unlabeled fructose, tagatose, or sorbose. Fructose appears to enter larvae by diffusion only. The uptake of radioglucose and radiogalactose was not linear with respect to concentration at low substrate concentrations; at high substrate concentrations, the uptake of both hexoses was linear with respect to concentration. Inhibitor studies indicated that both glucose and galactose enter larvae by a combination of diffusion and a mediated process, and that these hexoses are mutually competitive inhibitors of one another. The uptake of glucose and galactose was also inhibited by α-and β-methyl glucoside, fucose, and phlorizin, but not by several amino acids, certain sugar analogs, nor ouabain. Glucose transport is Na⁺ sensitive; K⁺ was demonstrated to be a competitive inhibitor of Na⁺ activation of glucose uptake. After a 90-min incubation in 5 mM unlabeled glucose, larvae accumulated glucose against an apparent concentration difference. Although larvae appear freely permeable to ouabain, this compound had no apparent effect on glucose accumulation. The results of this study are compared with previous studies on Hymenolepis diminuta, Calliobothrium verticillatum, Hydatigera (Taenia) taeniaeformis, and mammalian systems.     

Uptake of cyclic guanosine-3',5'-monophosphate and its metabolism in 3T6 cells

Uptake of 3H-cGMP by cultured murine 3T6 cells was studied. The cells were shown to contain radioactivity 1 minute after its addition, with the level of radioactivity increasing during a 3 hour incubation period. By this time, the intracellular non-metabolized cGMP corresponded to 1-5% of the whole intracellular radioactivity. In the presence of theophylline the uptake of 3H-cGMP by cells was seen decreasing, however, the portion of non-metabolized cGMP reached 45-50% of the whole intracellular radioactivity. Thus, the presence of theophylline made it possible to maintain the high level of intracellular cGMP. It is concluded that the incubation of cell cultures in the medium with cGMP may be useful for achieving an elevating intracellular cGMP concentration and for studying the biological effect of cyclic nucleotide.     

Overproduction of lysine by mutant strains of Escherichia coli with defective lysine transport systems

Mutants selected on the basis of their resistance to S-(β-aminoethyl)cysteine and overproduction of lysine were found to be defective in the lysine transport system. The overproduction of lysine was not due to mutation affecting either of the two regulatory enzymes aspartokinase and dihydrodipicolinic acid synthetase. Uptake of labeled lysine by the lysine-specific transport system was reduced to a negligible level, while uptake by the lysine, ornithine, arginine system was also affected. A hypothesis regarding the nature of these mutations and their effects on the regulation of lysine biosynthesis is discussed.     

Transport of Glycerate across the Envelope Membrane of Isolated Spinach Chloroplasts

Uptake of d, l-glycerate into the chloroplast stroma has been studied using the technique of silicone oil filtering centrifugation. Glycerate uptake was 3 to 5 times higher in the light than in darkness, the stimulation by light being abolished by the proton ionophore carbonyl cyanide p-trifluoromethoxyphenyl hydrazone. The pH optimum for uptake was 7.0 at 2°C and 8.5 at 20°C, but at all pH values the rate of uptake was higher at 20°C than at 2°C. Uptake was concentration dependent, saturating above 8 millimolar glycerate. At 2°C, the K_m was 0.3 millimolar and the V_max was 13 micromoles per milligram of chlorophyll per hour. At 20°C initial rates of glycerate uptake were higher than 40 micromoles per milligram of chlorophyll per hour.     

Characterization of the Permeability of Excised Apple Tissue 2

Sorbitol uptake from a bathing solution into the compartmentedspace and into the diffusible or apparent free space of excisedparenchyma tissue from apple fruit (Pyrus malus L. cv. GoldenDelicious) was investigated. Uptake into the two cell compartmentswas measured after washing of ^l4C-loaded tissue for 1 h withan osmoticum-free bathing solution. Compartmental analysis showedthat this treatment released sorbitol taken up into the cytoplasmof the cell, which was considered to be part of the apparentfree space. Uptake of sorbitol into the apparent free space was dependenton the osmotic concentration of the incubation medium. Usingmannitol up to 200 mM, uptake decreased by 60%, and increasedagain above 600 mM mannitol, the external concentration whereturgor was eliminated. Uptake in the compartmented space wasabout 3 times lower and was hardly affected by the externalosmotic concentration. PCMBS inhibited sorbitol transport intothe apparent free space by 25% at 100 mM mannitol, but at 600mM the inhibitor had no effect. The results indicate that sorbitoltransport across the plasma membrane is possibly facilitatedby a turgor-sensitive carrier. Uptake of ^l4C-sorbitol into thefreely diffusible space of tissue discs also increased by 200%after storage of unripe fruit for 70 d. This increase in agedtissue did not occur when uptake was measured at 4C or in thepresence of 200 mM PEG. Enhanced uptake was concomitant withan increased release of endogenous sugars from aged tissue. It would appear that the effect of a hypotonic bathing solutionon the permeability of excised apple tissue is related to structuralchanges, such as stretching of the plasma membrane. This effect,which becomes more marked as unripe fruit ages, is probablybrought about by turgor-driven relaxation of the tissue. Itmay increase non-specific leakage of sugars but could also bea factor affecting carrier-mediated transport of sorbitol atthe plasma membrane. Key words: Apple, sugar transport, sorbitol, plasma membrane, apoplast     

Putrescine uptake in saintpaulia petals

Putrescine uptake and the kinetics of this uptake were studied in petals of Saintpaulia ionantha Wendl. Uptake experiments of [³H] or [¹⁴C] putrescine were done on single petals at room temperature at various pH values. The results show that putrescine uptake occurs against a concentration gradient at low external putrescine concentration (0.5-100 micromolar) and follows a concentration gradient at higher external putrescine concentrations (100 micromolar to 100 millimolar). 2,4-Dinitrophenol and carbonylcyanide-m-chlorophenylhydrazone, two uncouplers, had no effect on putrescine uptake. Uptake rates were constant for 2 hours, reaching a maximum after 3 to 4 hours. Putrescine uptake depended markedly on the external pH and two maxima were observed: at low external concentrations of putrescine, the optimum was at pH 5 to 5.5; at higher concentrations the optimum was at pH 8.     

Effects of fusicoccin and abscisic acid on glucose uptake into isolated beetroot protoplasts

Uptake of glucose, 3-O-methylglucose and sucrose into beetroot protoplasts is considerably stimulated by 10^–6M fusicoccin. This effect is decreased in the presence of 10mM Na⁺ or K⁺, 2 mM Mg²⁺ or Ca²⁺. Whereas fusicoccin causes no change in the pH-optimum of the sugar uptake (pH 5.0), the apparent K_m of this uptake which obeys a biphasic kinetics is decreased by the action of fusicoccin. In the protoplast suspension, fusicoccin induces an acidification which is suppressed by uncoupling agents. Correspondingly, uncouplers as well as vanadate and diethylstilbestrol markedly inhibit the effect of fusicoccin on sugar uptake. The present data support the view that glucose uptake into beetroot protoplasts depend on the proton-pumping activity of the plasmalemma-ATPase. cis–Abscisic acid diminishes significantly the fusicoccin-enhanced glucose uptake. By using a radioimmunoassay, the internal abscisic acid content of the protoplast was estimated to be in the range of 10^–6 M. Protoplasts isolated from bundle tissue contain twice as much abscisic acid as those derived from storage parenchyma. Because protoplasts from the bundle tissue were shown to take up sugars much faster than those from the storage cells, the observed effect of abscisic acid might reflect an involvement of this hormone in the regulation of carbohydrate partitioning in the beet.Abbreviations ABA cis–abscisic acid - bundle protoplast protoplasts isolated from the conducting tissue of beetroots - DES diethylstilbestrol - FC fusicoccin - 3-OMG 3-O-methylglucopyranose - PCMBS p–chloromercuribenzenesulfonic acid - storage protoplasts protoplasts isolated from storage parenchyma