Determination of Unidirectional Sodium Fluxes in Roots of Intact Sunflower Seedlings

The method of compartmental analysis was applied to study sodiumfluxes in roots of intact seedlings of Helianthus annuus L.By measuring sodium uptake and transport to the shoots of theseedlings in parallel experiments, transport of tracer sodiumto shoots and net accumulation of Na⁺ in the roots during theflux measurements was accounted for. The steady-state sodiumfluxes in the intact sunflower roots were similar in size tothose in excised roots but in general they were somewhat higher.This indicates more metabolic activity in the intact tissues.Using whole plants it is possible to study the response of ionfluxes in roots to ecophysiological stimuli received by theshoots, and in the present experiments the effect of continuouslight versus long-day growth conditions was investigated. Potassium,when continually present, depressed all fluxes and the cytoplasmiccontent of sodium but tended to increase the vacuolar sodiumcontent, in particular when this was related to the cytoplasmiccontent. When added to sodium-loaded roots, potassium stimulatedthe plasmalemma sodium efflux but slightly, suggesting a lowefficiency of K⁺-Na⁺ exchange across the plasmalemma in intactas well as excised sunflower roots. Subsequently, however, potassiuminduced a transient decrease in the ²²Na efflux that was followedby oscillations in tracer efflux. These changes were attributedto potassium-induced transfer of sodium to vacuoles. Moreover,the oscillations seem to indicate the operation of negativefeedback control of sodium fluxes.     

Artificially Induced Water and Sulfate Transport through Sunflower Roots

A new experimental method is used to determine simultaneously the quantity and composition of the sap exuded by a detopped root system at the same time that a pressure deficit of desired magnitude can be applied to the stem stump. The technique was used in a study of the transport of radioactive sulfate through the roots of young sunflower plants placed on complete nutrient solutions labelled with ³⁵S. The complications by the time factor on the composition and rate of the sap stream in experiments of this type were observed and discussed. The time of detopping the roots was very critical as the conditions of sulfate transport were greatly changed some time after the excision. A rectilinear connection existed between the rate of sulfate transport in the sap and the water flow at sap flow velocities comparable with transpiration rates. When the transport of water was very slow, the rate of sulfate transport became constant and independent of the water stream. It was suggested that diffusion or water flow could act as motive force for the ion transport in some non-metabolic phase of transfer in the roots. The addition of 2,4-DNP to the test solution severely interfered with the water and sulfate transport conditions in the roots.     

The Effect of Cycloheximide on Uptake and Transport of lons by Sunflower Roots

The effect of cycloheximide on uptake and transport of saltsby sunflower roots was investigated. Treatment with cycloheximideresulted in a reduction of uptake and transport of K⁺ and NO₃^–to the xylem. Cycloheximide stimulated O₂ uptake and appearedto act as an uncoupler of oxidative phosphorylation. The implicationsof these results regarding the use of cycloheximide as a meansof distinguishing between uptake and transport components ofion movement to the xylem are discussed.     

Regulation of Rubidium Uptake in Sunflower Roots

Uptake of Rb⁺ was investigated in 12-day-old intact plants of sunflower (Helianthus annum L. var. californicus) which had been cultivated or pretreated in nutrient solutions with various K⁺ concentrations. The relationship between Rb⁺ influx and K⁺ concentration of the roots indicated regulation of Rb⁺ uptake by allosteric inhibition of the uptake mechanism. A constant passive influx occurred contemporaneously with the active uptake as shown by experiments at 0°C or with 2,4-dinitrophenol. The allosteric regulation of ion carrier activity occurred after a time lag of up to 1 h after the change of external solution. In experiments involving Rb⁺ treatments of K⁺-deficient plants, the synthesis of carriers for transport of Rb⁺ could be demonstrated. A model including allosteric regulation of Rb⁺ uptake in roots is discussed.     

Ion Uptake Efficiency of Sunflower Roots

The term ion uptake efficiency is used for the rate of uptake of a particular ion from nutrient solutions holding a standard concentration of that ion (0.5 mM sulphate, 1.5 mM phosphate or 2.0 mM rubidium). The uptake efficiency for rubidium and phosphate in roots of intact sunflower plants depended on the salt status of the plants and on the concentration of the ion under investigation in pretreatment solutions. The effect of pretreatment was a rapid process causing differences of more than 300% in ion uptake efficiency within 1 h, depending on the composition of the pretreatment solution. At concentrations above 0.1 mM the rate of uptake of rubidium in the root was higher than the net potassium uptake necessary for adequate growth. The rate of sulphate uptake was related to potassium uptake but not to phosphate uptake. It is suggested that ion uptake of the roots is regulated by internal factors as well as by direct interactions between the medium and the absorbing surfaces.     

Sodium and Potassium Compartmentation and Transport across the Roots of Intact Spergularia marina

The Na⁺ and K⁺ transport characteristics of Spergularia marina (L.) Griseb. were considered in order to compare the systems by which these two physiologically different cations are managed during initial acquisition and subsequent partitioning in midvegetative plants. Uptake of ²²Na⁺ and ⁴²K⁺ and redistribution of labels in pulse-chase studies were compared under steady state growth conditions or with the concentration of one of the ions elevated. At high external concentrations, the initial ⁴²K⁺ accumulation and transport to the shoot was associated with a small, rapidly exchanging, cellular compartment similar to that previously indicated for Na⁺ (D Lazof, JM Cheeseman 1986 Plant Physiol 81: 742-747). At 1 mol m⁻³, K⁺ was conducted to the shoot through a root compartment, the specific activity of which rose much more slowly than the rapidly exchanging compartment. After a lag of approximately 5 minutes, ⁴²K⁺ translocation approached a constant rate with a half-time of 14 minutes compared to 5 minutes for ²²Na⁺ or for ⁴²K⁺ at higher external levels. At all external levels, prolonged translocation of ⁴²K⁺ was measured when a 10 minute pulse was followed by an unlabeled chase, again suggesting a conducting compartment distinct from that for Na⁺. It is suggested that the K⁺ conducting compartment, possibly the `bulk cytoplasm,' is associated with the active K⁺ transport system generally found in higher plants.     

Nonosmotic Effects of Polyethylene Glycols upon Sodium Transport and Sodium-Potassium Selectivity by Rice Roots

Addition of polyethylene glycol (PEG) as an osmotic agent (at −230 kilopascals) dramatically lessened the toxicity of NaCl (at 50 moles per cubic meter) to rice (Oryza sativa L.) seedlings. This was explained by a reduction in the uptake of NaCl. This reduction was much greater than could be accounted for by the lowered transpiration rate resulting from the solute potential changes due to the PEG.

Low concentrations of PEG (−33 kilopascals and less) had no effect upon transpiration rate but reduced sodium uptake (from 10-50 moles per cubic meter NaCl) by up to 80%. PEG (at −33 kilopascals) also reduced chloride uptake but had no effect upon the uptake of potassium from low (0.5-2.0 moles per cubic meter) external concentrations. However, the increased uptake of potassium occurring between 2 and 10 moles per cubic meter external concentration was abolished by PEG. Similar concentrations of mannitol had no effect upon sodium uptake in rice. PEG, in similar conditions, had much less effect upon sodium uptake by the more salt-resistant species, barley.

²²Na studies showed that PEG reduced the transport of sodium from root to shoot, but had a long half time for maximal effect (several days).

¹⁴C-labeled PEG was shown to bind to microsomal membranes isolated from rice roots; it is suggested that this is due to multipoint attachment of the complex ions of PEG which exist in aqueous solutions. It is argued that this reduces passive membrane permeability, which accounts for the large effect of PEG on sodium influx in rice and the different effects on sodium influx and (carrier-dependent) potassium influx.

     

Extrusion of Sodium Ions by Barley Roots: III. The Effect of High Salinity on Long-distance Sodium Ion Transport

The quantity of sodium transported to the shoots of intact barleyplants was stimulated by 0·5 mM ouabain when the sodiumchloride level of the bathing medium was below 100 mM. At sodiumchloride concentrations of 100 mM or more this ouabain-stimulatedsodium transport was not observed. Equiosmotic mannitol, equimolarpotassium chloride or equivalent calcium chloride solutionsdid-not affect the ouabain-stimulated sodium transport froma basic medium containing 10 mM sodium chloride. It is suggestedthat under the present experimental conditions the increasedsodium uptake by the root cells at sodium chloride concentrationsof 100 mM or more masks the extrusion process.     

Properties of (Sodium Potassium)-Stimulated ATPases in English Ryegrass Roots and Implications in Ion Transport

Maximum ATPase activities in the cell wall fraction of English ryegrass (Lolium perenne L.) roots were stimulated by foru discrete millimole ratios of (Na⁺+ K⁺); 40:0, 35:5, 5:35, and 0:40. The optimal pH for stimlation was found to be 6.5. Contrary to data in the literature, Mg²⁺ inhibited all stimulatory ratios of (Na⁺+ K⁺) when plants were cultured on an adequate nutrient solution. When grown on a dilute solution, Mg²⁺ enhanced (Na⁺+ K⁺)-stimulated ATPase activity in this membrane preparation. The single optimal combined concentration of (Na⁺+ K⁺) for all stimulatory ratios was 40 MM. The ratios of (Na⁺+ K⁺) which stimulated ATPase activity in the cell wall fraction varied with position along the root axis such that all rarely existed simultaneously nor did any exist in the terminal millimetre of the root. Both cell wall and microsomal fractions showed stimulation by (Na⁺+ K⁺) at all the above ratios indicating the possible presence of plasma membrane fragments in both fractions. Only the 35:5 ratio was stimulations were found in the supernatant. Implications of ion-stimulated ATPase involvement in ion transport were drawn from the appearance of ATPase activity at a 40:0 ratio of (Na⁺+ K⁺) and the disappearance of stimulations at 35:5, 5:35, and 0:40 ratios when plants were moved from a strong (35 mM total concentration) to a dilute (0.75 mM) nutrient solution.     

Water Transport in Isolated Maize Roots

A simple model has been devised which predicts the concentration,C^x_s, of salt (e.g. KCl) in the exudate from isolated roots asa function of the salt concentration, C⁰_s, in the medium. Thechief assumption, made in deriving the relationship betweenC^x_s and C⁰₅, is that the exudation of water, J, from the rootsconsists of two components (one being osmotic, Ø, inorigin and the other, Ø⁰, flowing in the absence of anosmotic gradient). The exudation of salt, J_s, calculated asJ C^x_s, was found to be dependent on C⁰_s. Our investigationson maize roots were concerned with estimations of L_p and Ø^vand determinations of C^x_s as a function of C⁰_s. Satisfactoryagreement between prediction and experiment was found in thesepreliminary studies. It is considered that water movement inisolated roots can be explained by a simple osmotic model withthe additional possibility that a relatively small non-osmoticwater flow occurs.     

Rapid Effects of Abscisic Acid on Ion Uptake in Sunflower Roots

Short-term effects of ABA, ABA + kinetin and kinetin on ion (⁸⁶Rb-potassium and phosphate) and water uptake in sunflower plants (Helianthus annuus var. californicus) were examined with a continuous-recording technique. Ion uptake in the roots and transport to the shoots were also investigated by conventional tracer uptake experiments and by sap bleeding experiments with excised roots. After addition of 5 × 10^?6-4 × 10^?5M ABA to the root medium there was an immediate decrease (30–70%) in the rate of ion uptake which lasted 30–70 min. The rate of water uptake was not significantly affected as measured with this method. Ion transport to the shoots and to the bleeding sap of excised roots was decreased by ABA. ABA-induced inhibition of ion uptake was abolished by the presence of kinetin, and uptake was slightly stimulated by 2 × 10^?5M kinetin alone. We suggest that concentration gradients of ABA or rapid changes in the ABA-kinetin balance in the roots affect ion uptake and transport.     

Extrusion of Sodium Ions by Barley Roots: II. Localization of the Extrusion Mechanism and its Relation to Long-distance Sodium Ion Transport

An active sodium effux hasbeen demonstrated in excised barleyroots. Isolated cortical and stelar tissues from the same materialdo not show any evidence of such an efflux mechanism. When intactbarley plantswere allowed to absorb ²²Na active sodium extrusionwas not observed, although the percentage of ²²Na transportedto the shoot was stimulated 20 to 40 percent by ouabain at 5x 10^–4 M. Absorption and long-distance transport of phosphatewas unaffected by ouabain at this concentration. Interpretationof these data lead to the conclusion.that the sodium-effluxpump in barley roots is located at or near the endodermis externalto the Casparian band. This pump thus provides a mechanism wherebysodium can be partially excluded from the shoots of barley plants.     

The Micronutrient Boron Causes the Development of Adventitious Roots in Sunflower Cuttings

Three-day-old light-grown sunflower seedlings were de-rootedand incubated in nutrient solutions that either contained orlacked boric acid (B). In the absence of B, in the majorityof the seedlings, no adventitious roots were formed. The micronutrientB caused the development of numerous roots in the lower partof the hypocotyl. The effect of B occurred without the supplyof any phytohormones. A dose-response curve of B-induced rootingyielded an optimum concentration of 0.1 mM boric acid. Histologicalstudies revealed that cell divisions occurred in the controlbut no root primordia developed. In cuttings that were incubatedin B (0.1 m M) root primordia were observed that rapidly developedinto well-differentiated adventitious roots. Sunflower cuttingsthat were planted with their cut end in vermiculite that wasmoistened with nutrient solutions without B degenerated afterseveral weeks. In the presence of B the cuttings formed numerousadventitious roots that entirely replaced the tap root systemof intact seedlings. The rooted cuttings developed into sturdyadult sunflower plants. Our results are discussed with respectto the possible role of B in the evolution of vascular fromprevascular plants.Copyright 1999 Annals of Botany Company Adventitious roots, boron, cuttings, organogenesis, sunflower seedlings.     

The Effect of Salinity on the Membrane Potential of Sunflower Roots

The effect of NaCl and Na₂SO₄ on the electrical potential difference(PD) across the membrane in the outer cortical cells of theroot of sunflower (Helianthus annuus) has been investigated.The effect on the PD of a range of concentrations of these saltsgiven as a shock treatment was compared with the long-term effectwhere plants were grown in a range of salinities for 2 weeks. Considerable effects on the PD were observed. In low concentrationsof salt (approximately 1 mM) the PD was enhanced whilst in highconcentrations there was a progressive reduction in the PD.Na2SO4 had a relatively greater effect on the PD than NaCl. Using a transient recorder it was possible to measure the PDacross the plasmalemma and the tonoplast separately. The PDacross the plasmalemma was found to be markedly affected bysalinity. The tonoplast PD was relatively small compared tothat across the plasmalemma and appeared to be little affectedby the salts.     

A Labile-Bound Component of Phosphate in the Free Space of Sunflower Plant Roots

An experimental method is described by which it is possible to follow continuously the uptake and release of certain radioactive isotopes and the uptake of water by the roots of young plants. The method has been used in a study of the ion fractions related to the initial uptake of phosphate in the roots of sunflower plants. Two non-bound fractions were identified: one which was leachable in distilled water and another which was released from the roots only by a bathing medium containing inactive phosphate. Experiments using conventional analysis technique corroborated the results obtained by the recording technique.     

Evidence for Symplasmic Ion Transport in Maize Roots

Excised maize roots, placed in saturated water vapour to limitthe external ionic supply, continued to produce exudates attheir basal ends for at least 24 h. The mean rate of fluid exudationfrom roots in water vapour was about 28 per cent of the correspondingrate in ‘control’ roote placed in a solution containing0.1 mil CaCl₂ and 1 mM KC1. Moreover, the net fluxes (mean ±S.E.)of potassium and calcium ions into the exudate were reducedfrom (35.8±3.2) x 10^– and (4.37±0.39) xlO^–9 mole cm^–2 h^– for roots in solution to(10.9±0.6) x 10^–9 and (l.00±0.06)x 10^–9molecm^–2 h^–1 respectively for roots in vapour. It isconsidered that the observation of a prolonged exudation ofwater and ions from the roots placed in water vapour demonstratesthe existence of an alternative ionic supply within the roottissue itself and that this parallel route of ion transportto the exudate constitutes the cortical symplasmic pathway. Pre-treatment of the excised roots with 0.8 M mannitol beforeexudation studies in water vapour and solution led to a significantreduction in the rates of fluid and ion exudation which hadbeen observed in untreated roots under similar conditions. Itis concluded that the plas-molysis, induced by mannitol, disruptedthe symplasmic connections between root cells and that thisperturbation significantly reduced the operation of the symplasmicmode of ion transport into the exudate.     

Transport of Sodium in Intact Peanut Seedling

Peanut (Arachis hypogea L.) seedlings readily transported Nato the shoot. The amount of Na transported was linearly relatedto the absorption period (which ranged from 1 to 8 h) and alsoto the external Na level which varied from 0.05 to 1 mM.     

Oxidative Stress Induced in Sunflower Seedling Roots by Aqueous Dry Olive-Mill Residues

The contamination of soils with dry olive-mill residue can represent a serious problem as being an environmental stressor in plants. It has been demonstrated that inoculation of aqueous extract of olive oil-mill residue (ADOR) with saprobe fungi removes some phenolic compounds. In this paper we studied the effect of ADOR uninoculated or inoculated with saprobe fungi in sunflower seedling roots. The germination and root growth, O₂·^- generation, superoxide dismutase (SOD) and extracellular peroxidases (EC-POXs) activities, and the content of some metabolites involved in the tolerance of stress were tested. The roots germinated in ADOR uninoculated show a decrease in meristem size, resulting in a reduction of the root length and fresh weight, and in the number of layers forming the cortex, but did not alter the dry weight, protein and soluble amino acid content. ADOR caused the decreases in O₂·^- generation and EC-POX′s activities and protein oxidation, but enhanced SOD activity, lipid peroxidation and proline content. Fluorescence imaging showed that ADOR induced O₂·^- and H₂O₂ accumulation in the roots. The increase in SOD and the decrease in EC-POX′s activities might be involved in the enhancement of H₂O₂ content and lipid peroxidation. Control roots treated with ADOR for 10 min show an oxidative burst. Roots germinated in ADOR inoculated with saprobe fungi partially recovered normal levels of ROS, morphological characteristics and antioxidant activities. These results suggested that treatment with ADOR caused a phytotoxic effect during germination inducing an oxidative stress. The inoculation of ADOR with saprobe fungi limited the stress.     

Potassium Uptake and Transport in Roots of Ricinus communis

The uptake, accumulation, and transport of potassium by excisedroots of Ricinus communis were investigated. It was found that: 1. The transport of potassium to the vessels was closely dependenton the supply of the ion in the medium. 2. The flux of potassium to the vessels was only a fractionof the total potassium taken up and 99 per cent, of that absorbedwas accumulated in the root tissues. Once retained in this waypotassium appeared to become unavailable for transport to thevessels. 3. There was a fixed relationship between the potassium concentrationsof the medium and the exudate when potassium was supplied aspotassium nitrate in the medium at a range of concentrations.This relationship was altered by the presence of other ionsin the medium.