Effects of Growth and Assay Temperatures on Unidirectional K+ Fluxes in Roots of Rye (Secale cereale)

The effects of growth and assay temperature on unidirectionalK⁺ fluxes in excised roots of rye (Secale cereale cv. Rheidol)were studied using ⁸⁶Rb⁺ as a tracer. Both K⁺ influx to thevacuole, estimated as K⁺ uptake between 3 and 12 h after transferof unlabelled roots to radioactive solution, and movement ofK⁺ to the xylem were determined directly. Other fluxes weredetermined on excised roots of plants, which had been labelledwith ⁸⁶Rb⁺ since germination, by conventional triple exponentialefflux analysis. When assayed at 20°C, roots of plants previously grown at20°C(WG roots) had lower rates of net K⁺ uptake than rootsof low temperature-acclimated plants, grown with a temperaturediferential between roots (87°C) and shoots (20°C) eithersince germination (DG roots) or for 3 d prior to experiments(DT roots). This resulted from a greater unidirectional K⁺ effluxacross the plasma membrane and a reduced K⁺ flux to the xylemin WG roots, compared to DG or DT roots, rather than a decreasein unidirectional K⁺ influx or a decrease in the net K⁺ fluxto the vacuole. Indeed, although WG roots had lower rates ofK⁺ influx and K⁺ efflux across the tonoplast at 20°C thanDG or DT roots, roots of plants from all growth temperaturetreatments showed an equivalent net K⁺ flux to the vacuole. Although all unidirectional K⁺ fluxes in roots from plants grownunder all temperature regimes were reduced by lowering the temperatureof the root, these fluxes were differentially affected in rootsof plants from contrasting growth temperature treatments. Rapidcooling to 8°C of WG roots resulted in a lower rate of K+influx and a transient increase in K⁺ efflux across both theplasma membrane and tonoplast, compared to DG and DT roots.Furthermore, since the K⁺ flux to the xylem was lower in WGroots, the net K⁺ uptake at 8°C into WG roots was considerablyreduced compared to DG and DT roots. These results suggest thatlow temperature-acclimation of K⁺ fluxes in rye roots may involvea reduction in the temperature sensitivity of K⁺ influx anda curtailment of K⁺ efflux across both the plasma membrane andtonoplast at low temperatures. Key words: K⁺influx, K⁺ efflux, low temperature, potassium, rye (Secale cereale cv. Rheidol)     

Potassium Transport in Enteromorpha intestinalis (L.) Link: MEASUREMENT OF INTRACELLULAR K+, EXCHANGE FLUXES AND THERMODYNAMIC ANALYSIS

Potassium transport has been studied in the marine euryhalinealga, Enteromorpha intestimlis cultured in seawater and in low-salinitymedium (Artificial Cape Banks Spring Water, ACBSW; 25·5mol m^–3 Cl^–, 20·4 mol m^–3 Na⁺, 0·5mol m^–3 K⁺). K⁺ fluxes were measured using ⁴²K⁺ and ⁸⁶Rb⁺although ⁸⁶Rb⁺ does not act as an efficient K⁺ analogue in thisplant. ⁴²K⁺ experiments on seawater plants typically exhibiteda single protoplasmic exchange phase whereas ⁸⁶Rb⁺ exhibitedtwo exchange phases. Compartmental analysis of ⁸⁶Rb⁺ effluxexperiments on seawater-grown Enteromorpha plants were usedto deduce the intracellular partition of K⁺ between the cytoplasm(279±38 mMolal) and vacuole (405±68 mMolal). Theplasmalemma K⁺ flux in plants in seawater was greater in thelight than in the dark (563±108 nmol m^–2 s^–1versus 389±66·7 nmol m^–2 s^–1). Inlow-salinity plants, separate cytoplasmic and vacuolar exchangephases were apparent. Analysis of ⁴²K⁺ efflux experiments onlow-salinity plants yielded a cytoplasmic K⁺ of 222±38mMolal and a vacuolar K⁺ of 82±11 mMolal. The plasmalemmaand tonoplast flux was 23±4·5 nmol m^–2 s^–1. The Nernst equation showed that, although K⁺ was close to electrochemicalequilibrium, active accumulation of K⁺ across the plasmalemmaoccurred in plants in seawater and ACBSW both in the light anddark. K⁺ was also actively transported inwards across the tonoplastin low-salinity plants. The electrochemical potential for K⁺across the plasmalemma ranged from 2·41±0·60kJ mol^–1 in plants grown in seawater in the light to 5·79±0·87kJ mol^–1 for plants in ACBSW in the light. Although K⁺is close to electrochemical equilibrium, the flux of K⁺ in plantsin both seawater and ACBSW media is high, hence the power consumptionof K⁺ transport is high. The permeability of K⁺ (P_K⁺) was significantlyhigher in the light than in the dark in plants in seawater (about7·0 versus 2·5 nm s^–1) but in plants inlow-salinity (ACBSW) medium the permeability was independentof light (about 12 nm s^–1). The energy requirements ofactive K⁺ transport by ATP-dependent pumps is discussed. Key words: Enteromorpha, Potassium transport, Ionic relations, Saltwater, Low salinity, Thermodynamics     

A Comparison between the Uptake of Potassium by Plants from Solutions of Constant Potassium Concentration and during Depletion

A comparison was made between two methods of measuring the relationshipbetween the external [K⁺] and the flux of K⁺ into whole plantsof Lolium perenne and Raphanus sativus. The values of flux obtainedfrom solutions of 1.2 µM K⁺ held constant around the rootswere three and six times greater for Lolium and Raphanus respectivelythan the values obtained at the same concentration in a depletionexperiment in which the solutions, initially 100 µM K⁺,were depleted to below 1.2 µM K⁺ by plant uptake. In thedepletion experiment with Lolium, the flux was higher into plantsgrown at low [K⁺] than into plants grown at 100 µM eventhough [K⁺] within the plant was about the same for all groupsof plants. It is suggested that Lolium grown at low [K⁺] hasan efficient mechanism for K⁺ uptake which continues to operatefor some time after the plants have been transferred to a higherconcentration. With both species, K_m was 15–20 µMin the depletion experiment and below 1 µM when concentrationswere held constant.     

The regulation of K+ influx into roots of rye (Secale cereale L.) seedlings by negative feedback via the K+ flux from shoot to root in the phloem

The uptake of K⁺ by plant roots is matched to the demand forK⁺ for growth. The growing shoot must communicate its K⁺ requirementto the root. It has been suggested that this might be effectedby varying the amount of K⁺ retranslocated from the shoot tothe root via the phloem. It is predicted that less K⁺ is returnedto the roots in K⁺-deficient plants and that this promotes compensatoryK⁺ uptake from the external medium. These experiments addressthis hypothesis. Rye (Secale cereale L.) was grown hydroponically in completenutrient solutions containing either 100 aM or 400 µMK⁺. Plant development, shoot fresh weight (FW) and plant drymatter accumulation did not differ between seedlings grown atthese K⁺ concentrations. However, root FW was lower in seedlingsgrown in solutions containing 100 µM K⁺, which resultedin a greater shoot/root FW ratio. Seedlings from both treatmentshad similar shoot K⁺ concentrations, but the root K⁺ concentrationof seedlings grown In solutions containing 100 µM K⁺ wasless than their counterparts grown at 400 µM K⁺. When assayed at the same K⁺ concentration, unidirectional K⁺(⁸⁶Rb⁺) influx into 14-d-old seedlings grown with 100 µMK⁺ in the nutrient solution was greater than that into seedlingsgrown with 400 µM K⁺ in the nutrient solution, indicatingan increased K⁺ influx capacity in the former. Furthermore,K⁺ (⁸⁶Rb⁺) influx into seedlings grown and assayed at 100 µMK⁺ was greater than that into seedlings grown and assayed at400 µM K⁺. Since net K⁺ uptake was lower in the seedlingsgrown at 100 µM K⁺, this indicates a greater unidirectionalK⁺ efflux from roots in solutions containing 100 µM K⁺. An empirical model, based on the immobility of calcium in thephloem, was used to describe quantitatively K⁺ fluxes in seedlings14 d after sowing. As primary data, the composition of xylemsap and the accumulation of elements in root and shoot tissueswere determined. Xylem sap was collected either as root-pressureexudate or from excised roots immersed in nutrient solutionand subjected to a pneumatic pressure of 0.4 MPa. The K:Ca ratioin these saps differed, and led to contrasting conclusions concerningthe effect of K⁺ nutrition on the recirculation of K⁺. Basedon the K:Ca ratio in the sap obtained following the applicationof pneumatic pressure, which is thought to resemble that ofintact transpiring plants, it was calculated that the K⁺ fluxfrom the shoot to the root was higher in seedlings maintainedin solutions containing higher K⁺ concentrations. This suggeststhat a negative feedback mechanism based on K⁺ recirculationfrom the shoot to the root via the phloem could be a primarysignal decreasing K⁺ influx. Key words: K⁺ influx, K⁺ recirculation, regulation, root, rye, Secale cereale L     

Potassium Transport in Enteromorpha intestinalis (L.) Link: II. EFFECTS OF MEDIUM COMPOSITION AND METABOLIC INHIBITORS

In springwater (25.5 mol m^–3 Cl^–, 20.4 mol m^–3Na⁺, 0.14 mol m^–3 K⁺) Enteromorpha intestinalis couldnot survive for more than a few weeks unless provided with 0.5mol m^–3 K⁺ in the medium or alternatively exposed to seawaterfor 1 day per week. Maintenance of a cytoplasmic K⁺ level ofabout 200 mol m^–3 is critical for the maintenance of normalmetabolic activity. Net gains of intracellular K⁺ occurred whenthe plants were transferred from low-salinity to seawater; converselylarge net losses occurred when plants were transferred fromseawater to springwater. These two processes were not simplythe reverse of one another; net gain of K⁺ involved a largeincrease in the tracer flux both into and out of the cell butnet loss of K⁺ virtually halted the tracer flux into the cell.Any injury incurred by rapid salinity changes was short-lived;plants were rapidly able to adjust intracellular [K₁.K⁺). K⁺(orto some extent Rb⁺) was found to be necessary in the effluxmedium for ⁴²K⁺ exchange to occur. The osmotic concentrationof the medium was also important but extracellular Na⁺ and Cl^–concentrationswere not critical. K⁺ influx and efflux in both springwaterand seawater were largely independent of light and were sensitivein varying degrees to a range of common metabolic inhibitorsand uncouplers. The results are best explained by the presenceof an active K⁺ influx, generated by an ATP-dependent K⁺ pumpat the plasmalemma. Key words: Enteromorpha, Potassium transport, Salinity changes, Uncouplers, Inhibitors     

Ionic Relations of Garden Orache, A triplex hortensis L.: Growth and Ion Distribution at Moderate Salinity and the Function of Bladder Hairs

The growth of garden orache, A triplex hortensis was studiedunder conditions of mild NaCl or Na₂SO₄ salinity. Growth, drymatter production and leaf size were substantially stimulatedat 10 mM and 50 mM Na⁺ salts. Increased growth, however, appearedto be due to a K⁺-sparing effect of Na⁺ rather than to salinityper se. The distribution of K⁺ and Na⁺ in the plant revealeda remarkable preference for K+ in the roots and the hypocotyl.In the shoot the K/Na ratio decreased strongly with leaf age.However, the inverse changes in K⁺ and Na⁺ content with leafage were dependent on the presence of bladder hairs, which removedalmost all of the Na⁺ from the young leaf lamina. Measurementsof net fluxes of K⁺ and Na⁺ into roots and shoots of growingAtriplex plants showed a higher K/Na selectivity of the netion flux to the root compared to the shoot. With increasingsalinity the selectivity ratio S_{K, Na}^* of net ion fluxes tothe roots and to the shoots was increased. The data suggestthat recirculation of K⁺ from leaves to roots is an importantlink in establishing the K/Na selectivity in A. hortensis plants.The importance of K⁺ recirculation and phloem transport forsalt tolerance is discussed. Key words: Atriplex hortensis, Salinity, Potassium, Sodium, K⁺ retranslocation, Bladder hairs, Growth stimulation     

Effects of univalent cations on the formation of nitrate reductase and nitrite reductase in rice seedlings

An investigation was made to determine the effects of univalentcations as activators on the formation of nitrate reductaseand nitrite reductase in rice seedlings. K⁺ functioned moreeffectively as a univalent cation activator than did other univalentcations examined. Substitution of Rb⁺ for K⁺ resulted in stimulationof nitrate reductase formation at about half the rate obtainedwith K⁺. There was no effect on nitrite reductase formation.Na⁺ could be partially substituted for K⁺ in the formation ofboth enzymes. NH₄⁺ slightly inhibited formation of the enzymes.In the absence of univalent cations, enzyme formation proceededat a slower rate during the initial 15-hr period, but thereafterproceeded at a higher rate. This delayed formation was not observedin the presence of K⁺. Results from inhibitor experiments suggestthat K⁺ stimulates the formation of nitrate reductase and nitritereductase. In conclusion, when nitrate nitrogen is supplied to rice plantsutilization of the nitrogen may be accelerated by increasedformation of enzymes involved in nitrate assimilation in thepresence of K⁺. (Received February 21, 1969; )     

Abnormal Stomatal Behaviour and Ion Imbalance in Capsicum scabrous diminutive

An attempt was made to explain the abnormal behaviour of stomatain Capsicum scabrous diminutive, a wilty pepper mutant. Stomatalmovement in the pepper plant was found to be associated withchanges in the ion content of the guard cells. These changeswere smaller in the mutant than in the normal plants. In addition,total ion content was higher in the mutant under both controland NaCl treatments. Na⁺ substituted K⁺ in its function in stomatalmovement under high salinity. This phenomenon was more pronouncedin the mutant plants. Analysis of whole root systems and leavesof plants grown on solutions of high NaCl or KCl concentrationconfirmed that the regulation of K⁺ and Na⁺ uptake mechanismswas not functioning properly in the mutant. Evidence was presentedthat the difference in K⁺ staining between mutant and normalepidermal cells is an artefact resulting from the differencein leaf anatomy.     

The Potassium Relations of Lemna minor L.: II. THE MECHANISM OF POTASSIUM UPTAKE

The experiments described in this paper concern the nature ofthe K⁺-influx mechanism in Lemna minor L. It is establishedthat K⁺ influx is ‘site restricted’ and that theaffinity of the system involved is in close agreement with thatof the well-documented System I mechanisms that are found inmany plants (Epstein, 1972). Experiments with ATP and CCCP suggestthat K⁺ influx is an active process, that the influx machineryresides at the plasmalemma and that this machinery containsan ATPase activity. Membrane vesicles isolated from the plantsabsorb K⁺(Rb⁺) with an affinity comparable to that of the systeminvolved in physiological K⁺ influx.     

Identification of a K+ Channel from Potato Leaves by Functional Expression in Xenopus Oocytes

Poly(A)⁺ mRNA was isolated from leaves of potato plants (Solatiumtuberosum L. cv. Desiree) according to standard protocols. Thispoly(A)⁺ mRNA was injected via glass microcapillaries into oocytesthat were surgically removed from the African clawed toad Xenopuslaevis. As a control, oocytes were either injected with H₂0or remained untreated. Three days after injection the oocyteswere analyzed by two electrode voltage clamping. Current voltageanalysis revealed that a K⁺ channel from potato was functionallyexpressed in injected oocytes. The identity of this K⁺ channelwas confirmed by its substrate specificity and a shift in thereversal potential. In particular, when the outside K⁺ concentrationwas increased the reversal potential of poly(A)⁺ injected oocytesshifted to more positive values. Furthermore, K⁺ outward currentsdeclined when the outside K⁺ concentration was raised from 0.1to 100 mM. Inward currents increased with an elevation of theK⁺ concentration. Several Pharmaceuticals were tested for theirpotential to block this K⁺ channel. As a result, the channelwas completely blocked by BaCl₂. A three state reaction kineticmodel was used to simulate the currents through the K⁺ transportprotein as function of the extracellular K⁺ concentration. Inparticular, the simulation revealed current voltage relationsthat exactly matched the measured ones. Saturation of currentvoltage curves emerged from the simulation as a consequenceof high extracellular potassium concentration. (Received November 7, 1997; Accepted March 21, 1998)     

Interrelationships between Potassium, Calcium, and Magnesium Nutrition of Zea mays L.

The uptake of potassium, calcium, and magnesium ions by maizeand the interrelationships among the cations have been investigatedat 48 K: Ca: Mg ratios in culture solutions. Calcium was foundto stimulate K⁺ and Mg⁺⁺ uptake at certain cation ratios butinhibit it at others. Potassium did the same for Ca⁺⁺ uptake,and Mg⁺⁺ for Ca⁺⁺ and K⁺. The uptake of Mg⁺⁺ was generally enhancedby K⁺. The sum of the cations in the plants expressed in meqwas fairly constant for treatments of the same K⁺ concentrationat the low to moderate levels of K⁺, but at considerably higher(> 24 meq l^–1) K⁺ levels the constancy was not dependenton K⁺ concentration. Potassium depressed, but Mg⁺⁺ stimulatedphosphorus accumulation. Calcium stimulated phosphate absorptionat certain cation ratios but had no effect at others. The plantyield increased with increasing K⁺ up to 24 meq l^–1 ofK⁺ after which the yield tended to fall with further increasein K⁺. The yield was also increased by Ca⁺⁺. Magnesium increasedthe yield at certain cation ratios and either depressed it orwas without effect at others.     

Effect of Potassium on Photosynthate Unloading from Seed Coats of Phaseolus vulgaris L. Specificity and Membrane Location

Rates of ¹⁴C-photosynthate unloading from excised seed-coathalves of Phaseolus vulgaris L. plants were stimulated by externalKCI concentrations in excess of 10 mM with an optimal responseat 100–150 mM KCI. The cellular pattern of ¹⁴C-photosynthatemetabolism was not altered by KCI but the treatment preferentiallystimulated the release of sucrose from the seed-coats. Photosynthateunloading was insensitive to Cl^– and was stimulated bya range of membrane-permeable cations (Na⁺, Mg²⁺ and tetraphenylphosphoniumion) in addition to K⁺. The K⁺ ionophore, valinomycin, abolishedthe K⁺ stimulation of ¹⁴C-photosynthate unloading. A switchto a wash solution containing K⁺ elicited a rapid burst of ¹⁴C-photosynthateunloading; the rate constant for the final phase of ¹⁴C-efflux(probably across the tonoplast) was unaffected by K⁺. The KCItreatment did not change the passive permeability of eitherthe plasmalemma or tonoplast. While sucrose influx across theplasmalemma was insensitive to K⁺, sucrose transfer to the vacuolewas slowed. The results obtained support the postulate thatK⁺ (and other membrane permeable cations) preferentially stimulatesucrose efflux across the plasmalemma of the unloading cellsby serving to carry positive charge in the opposite direction. Phaseolus vulgaris, bean, photosynthate unloading, potassium stimulation, seed-coat     

Changes in Outward K+ Currents in Response to Two Types of Sweeteners in Sweet Taste Transduction of Gerbil Taste Cells

Using the whole cell patch clamp technique, we measured changesin outward K⁺ currents of gerbil taste cells in response todifferent kinds of sweeteners. Outward K⁺ currents of the tastecell induced by depolarizing pulses were suppressed by sweetstimuli such as 10 mM Na-saccharin. The membrane-permeable analogof cAMP, cpt-cAMP, also decreased outward K⁺ currents. On theother hand, the K+ currents were enhanced by amino acid sweetenerssuch as 10 mM D-tryptophan. The outward K⁺ current was enhancedby external application of Ca²⁺-transporting ionophore, 5 µMionomycin, and intracellular application of 5 µM inositol-1,4,5-trisphosphate(IP₃). The outward K⁺ currents were no longer suppressed by10 mM Na-saccharin containing 20 µM gurmarin, but werestill enhanced by 10 mM D-tryptophan containing 20 µMgurmarin. These results suggest that sweet taste transductionfor one group of sweeteners such as Na-saccharin in gerbilsis concerned with an increase of the intracellular cAMP level,and that the transduction for the other group of sweetenerssuch as D-tryptophan is concerned with an increase of the intracellularIP₃ level which releases Ca²⁺ from the internal stores. Chem.Senses 22: 163–169, 1997.     

Ion Imbalance in Capsicum annum scarbrous diminutive a Wilty Mutant of pepper: II.RUBIDIUM FLUX

Root tips of the wilty pepper mutant scarbrous diminutive accumulateless rubidium than those of the normal genotype. This phenomenonwas evident in root tips excised from plants maintained for2 d in CaSO₄ solution (low salt plants), especially in the lowerexternal concentration range (0.1– 1.0 mM) of RbCl. Theefflux rate of Rb⁺ from mutant root tips was twice as high asin normal root tips. These results indicate that the ability of the mutant rootsto absorb and accumulate Rb⁺ and K⁺ is impaired. This defectcould be a consequence of either an impaired Na⁺/K⁺ carriersystem, or increased leakiness of mutant membranes, or both. The fact that the normal roots can accumulate Rb⁺ much fasterthan mutant roots supports the first alternative, i.e. thatthe high affinity carrier system was impaired in the mutantroots. However, the higher efflux rate of Rb⁺ from the mutantroots suggests that membrane leakiness was also affected.     

K+Nutrition and Na+Toxicity: The Basis of Cellular K+/Na+Ratios

The capacity of plants to maintain a high cytosolic K⁺/Na⁺ratiois likely to be one of the key determinants of plant salt tolerance.Important progress has been made in recent years regarding theidentification and characterization of genes and transportersthat contribute to the cytosolic K⁺/Na⁺ratio. For K⁺uptake,K⁺efflux and K⁺translocation to the shoot, genes have been isolatedthat encode K⁺uptake and K⁺release ion channels and K⁺carriersthat are coupled to either a H⁺or Na⁺gradient. Although thepicture is less clear for the movement of Na⁺, one pathway,in the form of non-selective ion channels, is likely to playa role in Na⁺uptake, whereas Na⁺efflux and compartmentationare likely to be mediated by H⁺-coupled antiport. In addition,several proteins have been characterized that play prominentroles in the regulation of K⁺and/or Na⁺fluxes. In this BotanicalBriefing we will discuss the functions and interactions of thesegenes and transporters in the broader context of K⁺nutritionand Na⁺toxicity. Copyright 1999 Annals of Botany Company Salinty, K⁺/N⁺ratio, transporter, membrane.     

Na+, K+ and Cl- in Xylem Sap Flowing to Shoots of NaCl-Treated Barley

Munns, R. 1985. Na⁺, K⁺ and Cl^– in xylem sap flowing toshoots of NaCl-treated barley.—J. exp. Bot. 36: 1032–1042. Na⁺, Cl^– and K⁺ concentrations were measured in xylemsap obtained by applying pressure to the roots of decapitatedbarley plants grown at external [NaCl] of 0, 25, 50, 100, 150and 200 mol m^–3. For any given NaCl treatment, ion concentrationsin the xylem sap were hyperbolically related to the flux ofwater. Ion concentrations in sap collected at very low volumefluxes (without applied pressure) were 5–10 times higherthan in sap collected at moderate fluxes (under pressure). Fora given moderate volume flux, Na+ concentration in the xylemsap, [Na⁺]_x, was only 4.0 mol m^–3 at external [NaCl] of25–150 mol m^–3, and increased to 7.0 mol m^–3at 200 mol m^–3. [Cl-]x showed a similar pattern. Thisshows there would be little difference in the rate of uptaketo the shoot of plants at 25–150 mol m^–3 externalNaCl and indicates little change even at 200 mol m^-3 NaCl becausetranspiration rates would be much lower. Thus the reduced growthof the shoot of plants at high NaCl concentrations is not dueto higher uptake rates of Na⁺ or Cl^–. The fluxes of Na⁺, Cl^– and K^– increased non-linearlywith increasing volume flux indicating little movement of saltin the apoplast. The flux of K⁺ increased even when [K⁺]_x wasgreater than external [K⁺], indicating that membrane transportprocesses modify the K⁺ concentration in the transpiration streamas it flows through the root system. Key words: -Xylem sap, Na⁺, K⁺, Cl^– fluxes, salinity, barley     

Salt Tolerance in the HalophyteHalosarcia pergranulatasubsp.pergranulata

Halosarcia pergranulata(P. G. Wilson) subsp.pergranulatais amember of the Salicornioideae and is native to Australia. Salttolerance inH. pergranulatasubsp.pergranulatawas assessed bygrowing plants for 83 d at seven NaCl concentrations from 10to 800 mol m^-3. Shoot biomass was greatest for plants grownat 10 to 200 mol m^-3NaCl, while at salinities of 300 mol m^-3orhigher it was inhibited. There was little increase in succulencein response to NaCl, and it even declined at the highest salinities.The K⁺[ratio]Na⁺molar ratio in succulent shoot tissues decreasedfrom 0.30[ratio]1 in plants grown at 10 mol m^-3NaCl to 0.02[ratio]1in plants at 600 mol m^-3, due to a three-fold increase in tissueNa⁺concentration and a five-fold decline in tissue K⁺. The osmoticpotential of sap (     

Relationship Between the Development and Growth of Rye (Secale cereale L.) and the Potassium Concentration in Solution

The development and growth of rye (Secale cereale L. cv. Rheidol)was studied in seedlings grown hydroponically in complete nutrientsolutions containing between 10 and 600 µM K⁺. The phyllochron(defined as the interval between the appearance of successiveleaves) was used as a developmental timescale to compare plants.The pattern of both shoot and root development was strictlyordered on a phyllochron basis and was unaffected by solutionK⁺ concentration, with the exception that tillers in plantsgrown at the lowest K⁺ concentrations were occasionally eithernot initiated or aborted at an early stage of development. However,both the rate of leaf appearance on the main stem and successivetillers and the rate of tiller appearance were slower in plantsgrown at lower K⁺ concentrations. The rate of leaf appearanceon the main stem was reduced to below 90% of its maximal valueat solution concentrations below about 50 µM K⁺. Plantrelative growth rate (RGR) was also reduced by lowering theK⁺ concentration of the nutrient solution and fell to below90% of its maximal value at solution concentrations below about200 µM K⁺. There was a complex relationship between tissueK⁺ concentration and the K⁺ concentration of the nutrient solution,which differed between leaves and root. Leaf K⁺ concentrationincreased steadily from about 50 µmol g^-1 f. wt to about200 µmol g^-1 f. wt as solution K⁺ concentration was increasedfrom 10 to 400 µM. In contrast, root K⁺ concentrationexhibited a sigmoidal dependence on solution K⁺ concentration,maintaining a minimal value of approximately 20 µmol g^-1f. wt at concentration below 100 µM K⁺, then increasingprogressively to about 120 µmol g^-1 f. wt at a solutionconcentration of 600 µM K⁺. The 'critical' leaf K⁺ concentration,i.e. the concentration at which either plant RGR or plant developmentwas reduced 90% of its maximal value, was 86 µmol g^-1f. wt for plant RGR and 150 µmol g^-1 f. wt for plant development.The 'critical' root K⁺ concentration was 24 µmol g^-1 f.wt K⁺ for both RGR and development. A decline in tissue K⁺ concentrationbelow these thresholds reduced plant growth considerably. RootK⁺ concentration was a sensitive indicator of the K⁺ statusof the plant with respect to potential growth since plant growthdeclined abruptly as root K⁺ concentration approached its 'critical'value, whereas plant growth showed a less defined relationshipwith shoot K⁺ concentration.Copyright 1993, 1999 Academic Press Critical K⁺ concentration, development, potassium, relative growth rate (RGR), rye, Secale cereale L. cv. Rheidol     

Purification and some properties of phosphomannomutase from corms of Amorphophallus konjac C. Koch

Phosphomannomutase [Glazer et al.: Biochim. Biophys. Acta 33:522–625 (1959)] was purified 1700-fold in a 39% yieldfrom cell-free extract of konjak (Amorphophallus konjac C. Koch)corms. The molecular weight of the enzyme as determined by gelfiltration was about 62,000. The enzyme required both Mg²+ and-D-glucose-l,6-bisphosphate for activity, although Mg²+ waspartially replaceable by either Co²+ or Ni²+. An apparent equilibriumconstant, K_eq=(mannose-6-phosphate) (mannose-1-phosphate), wasdetermined to be 8.5. Activity was maximal at pH 6.5 to 7.0.Activation energy was 11.1 kcal/mole. The enzyme was the moststable at pH 7.5. The addition of substrate or cofactor markedlyincreased enzyme stability toward heat denaturation. The enzymewas more labile to heat than phosphoglucomutase from konjakcorms. Treatment with various metal ions in Tris buffer inhibited theenzyme. Cu²+ and Zn²+ were the most potent inhibitors amongthe metal ions tested, while Co²+ and Ni²+ were weak. When theenzyme was treated with metal ions in the presence of histidinebuffer, Cu²+ and Zn²+ showed no inhibitory effect on the enzyme,whereas Be²+ inhibited it to an extent similar to that in Trisbuffer. Plots of 1/v versus l/(mannose-l-phosphate) at different fixedconcentrations of glucose-1,6-bisphosphate and 1/v versus 1/(glucose-1,6-bisphosphate)at different fixed concentrations of mannose-1-phosphate wereseries of converging lines. Mannose-1-phosphate at high concentrationswas found to inhibit the enzyme competitively with respect toglucose-l,6-bisphosphate. Apparent K_m and K₁ values for mannose-1-phosphatewere calculated to be 0.2 mM and 1.2 mM, respectively. The K_mvalue for glucose-1,6-bisphosphate was 1.8 µM. ¹This paper constitutes part 5 of a series of studies on konjakmannan biosynthesis. (Received May 24, 1976; )     

Biosynthesis of folic acid compounds in plants VI. The occurrence and properties of the dihydrofolate-synthesizing enzyme in pea seedlings

An enzyme, which catalyzes the formation of dihydrofolate fromdihydropteroic acid and L-glutamic acid, was found in pea seedlings.The enzyme was purified approximately 25-fold from the crudeextracts of pea seedlings, and its some properties were investigated.Optimum pH for the enzyme activity was found to be 8.8. Pteroicand tetrahydropteroic acids were not active as substrate. Theenzymatic reaction required as cofactors ATP, divalent (Mg²⁺or Mn²⁺) and univalent (K⁺, NH₄⁺ or Rb⁺) cations. The productwas characterized as dihydrofolic acid by bioautography. MICHAELIS constants for L-glutamic acid, ATP, dihydropteroicacid and Mg²⁺ were 7.0x10^–4, 9.0x10^–5, 3.5x10^–6and 1.2x10^–3 M, respectively. The MICHAELIS constant forMn²⁺ was 3.0x10^–4. The enzyme was inhibited by PCMB orsilver nitrate and, to some extent, by L-aspartic acid. Inhibitionby PCMB was completely reversed by addition of 2-mercaptoethanol.Enzyme activity was distributed widely among plants. The importanceof magnesium and potassium ions for enzyme catalysis is discussed. ¹For the previous paper, Part V, see Reference (30). (Received March 28, 1970; )