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1.
M. de Agazio  R. Federico  S. Grego 《Planta》1989,177(3):388-392
The inhibition of K+ uptake through the plasma membrane resulting from injury caused by cutting, or from application of polyamines (PAs), has been investigated in root segments of maize (Zea mays L.) and pea (Pisum sativum L.). It was found, for both treatments, that K+ uptake recovered if the segments were washed for 2 h. The K+ uptake inhibited by cutting and that inhibited by spermidine treatment were stimulated to the same extent by fusicoccin. In addition, there was a correlation between the extent of the recovery of K+ uptake caused by washing and the distribution, along the root axis, of both PAs and the activities of enzymes responsible for PA degradation. In apical segments of maize, where the PA content and the activity of the degradative enzyme polyamine oxidase (EC 1.5.3.3) were higher than in the more distal segments, the recovery of K+ uptake caused by washing was also higher. On the other hand, the opposite trend was observed in root segments of pea, where the PA content and the activity of the degradative enzyme diamine oxidase (EC 1.4.3.6) were higher in distal segments in which K+ uptake was greatly stimulated by washing. The effect of the amine-oxidase inhibitor, aminoguanidine, indicates that the degradation products of PAs are involved in the mechanism of inhibition of K+ uptake by PAs. The data also seem to indicate that PAs and their degradation products are responsible for the inhibition of K+ uptake occurring as a result of injury sustained by cutting roots into segments.Abbreviations DAO diamine oxidase - FC fusicoccin - PA polyamine - PAO polyamine oxidase - PUT putrescine - SPD spermidine  相似文献   

2.
It has recently been reported that plasmalemma electron transport may be involved in the generation of H+ gradients and the uptake of ions into root tissue. We report here on the influence of extracellular NADH and ferricyanide on K+ (86Rb+) influx, K+ (86Rb+) efflux, net apparent H+ efflux, and O2 consumption in 2-centimeter corn (Zea mays [A632 × Oh43]) root segments and intact corn roots. In freshly excised root segments, NADH had no effect on O2 consumption and K+ uptake. However, after the root segments were given a 4-hour wash in aerated salt solution, NADH elicited a moderate stimulation in O2 consumption but caused a dramatic inhibition of K+ influx. Moreover, net apparent H+ efflux was significantly inhibited following NADH exposure in 4-hour washed root segments.

Exogenous ferricyanide inhibited K+ influx in a similar fashion to that caused by NADH, but caused a moderate stimulation of net H+ efflux. Additionally, both reagents substantially altered K+ efflux at both the plasmalemma and tonoplast.

These complex results do not lend themselves to straightforward interpretation and are in contradiction with previously published results. They suggest that the interaction between cell surface redox reactions and membrane transport are more complex than previously considered. Indeed, more than one electron transport system may operate in the plasmalemma to influence, or regulate, a number of transport functions and other cellular processes. The results presented here suggest that plasmalemma redox reactions may be involved in the regulation of ion uptake and the `wound response' exhibited by corn roots.

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3.
Abstract Fusicoccin (FC)-stimulated K+ (86Rb) uptake and proton extrusion of maize (Zea mays) root apical segments were inhibited by pretreatment of 4-day-old seedlings with the herbicide Chlorsulfuron. In the range of Chlorsulfuron concentrations 0.01-10 mmol m?3, the percentage of inhibition was 15% at 0.01 mmol m?3 and progressively increased with Chlorsulfuron concentration up to 60% at 10 mmol m?3. At the maximum concentration tested (10 mmol m?3), the inhibition was evident after 1.5 h of pre-treatment. The binding of FC to microsomal fractions of root segments from Chlorsulfuron-pretreated seedlings was inhibited by 30%. It is suggested that Chlorsulfuron causes an alteration at the plasmalemma level involving the FC binding sites. The ineffectiveness of Chlorsulfuron in inhibiting FC-stimulaled K+ uptake when administered to excised segments, while inhibiting the enzyme acetolactate synthase, pointed out by Ray (1984) as the site of action of Chlorsulfuron in pea plants, suggests that the observed inhibition of K+ uptake and H+ extrusion is not induced by Chlorsulfuron inhibition of this enzyme. An alternative site of action of Chlorsulfuron is hypothesized in maize plants.  相似文献   

4.
The effect of a toxin extract of Helminthosporium maydis, race T on K+ (86Rb) uptake by excised root segments of normal (N) and Texas cytoplasmic male-sterile (T) versions of corn inbred W64A was investigated. The uptake of K+ was inhibited in both N and T roots by the toxin. This was true for both basal (freshly excised) and augmented (pretreated with aeration) K+ uptake. Augmented uptake was more toxin-sensitive than basal uptake (irrespective of cytoplasm type), and the augmented uptake in T roots was seven to eight times more toxin-sensitive than in N roots.  相似文献   

5.
A detailed examination was conducted on the linear, or first-order kinetic component for K+(86Rb+) influx into root segments of both low- and high-salt grown corn seedlings (Zea mays [A632 × Oh 43]). In tissue from both low- and high-salt grown roots, replacement of Cl in the uptake solution by either SO42−, H2PO4, or NO3 caused a significant (50-60%) and specific inhibition of the linear component of K+ influx. The anion transport inhibitor, 4,4′-diisothiocyano-2,2′-disulfonic acid, was found to abolish saturable Cl influx in corn roots while causing a significant (50-60%) and specific inhibition of the linear K+ uptake system; this inhibition was identical to that observed when Cl was replaced by other anions in the K+ uptake solution. Additionally, the quaternary ammonium cation, tetraethylammonium, which has been shown to block K+ channels in nerve axons, also caused a dramatic (70%) and specific inhibition of the linear component of K+ influx, but this was obtained only in high-salt roots. The reasons for this difference are discussed with respect to the differing abilities of low- and high-salt roots to absorb tetraethylammonium.

Our present results indicate that the linear component of K+ influx may occur by a passive process involving transmembrane K+ channels. Fluxes through these K+ channels may be partly coupled to a saturating Cl influx mechanism.

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6.
Segments of oat (Avena sativa L.) roots which had been exposed to 1 millimolar CdSO4 in quarter-strength Hoagland No. 1 solution exhibited decreased respiratory rates, ATP levels, membrane-bound ATPase activity, and reduced K+ fluxes. Respiration and ATP levels were decreased after a 2-hour treatment with 1 millimolar CdSO4 to 65 and 75%, respectively, of control rates. A membrane-bound, Mg2+-dependent, K+-stimulated acid ATPase was rapidly inhibited to 12% of control activity in the presence of 1 millimolar CdSO4. Potassium uptake into root segments was inhibited to 80% of control values after 30 minutes in the presence of CdSO4. A 2-hour pretreatment of root segments with CdSO4 inhibited K+ uptake to 15% of control values. Cytoplasmic K+ efflux was inhibited with 1 millimolar CdSO4.

The rates and the degree of Cd2+ inhibition of the parameters listed above suggest that one of the first sites of Cd2+ action is the plasmalemma K+ carrier (ATPase) in oat roots.

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7.
The relative transport capabilities of the cells of the root periphery and cortex were investigated using a variety of experimental techniques. Brief (30 seconds to 1 minute) exposures with the penetrating sulfhydryl reagent, N-ethyl maleimide (NEM), and the impermeant reagent, p-chloromercuribenzene sulfonic acid (PCMBS), dramatically reduced 86Rb+ (0.2 millimolar RbCl) uptake into 2 centimeter corn (Zea mays [A632 × (C3640 × Oh43)]) root segments. Autoradiographic localization studies with [3H]NEM and [203Hg]PCMBS demonstrated that, during short term exposures with either reagent, sulfhydryl binding occurred almost exclusively in the cells of the root periphery.

Corn root cortical protoplasts were isolated, and exhibited significant K+(86Rb+) influx. The kinetics for K+ uptake were studied; the influx isotherms were smooth, nonsaturating curves that approached linearity at higher K+(Rb+) concentrations (above 1 millimolar K+). These kinetics were identical in shape to the complex kinetics previously observed for K+ uptake in corn roots (Kochian, Lucas 1982 Plant Physiol 70: 1723-1731), and could be resolved into a saturable and a first order kinetic component.

The existence of a hypodermal apoplastic barrier was investigated. The apoplastic, cell wall binding dye, Calcofluor White M2R, appeared to be excluded from the cortex by the hypodermis. However, experiments with damaged roots indicated that this result may be an artifact resulting from the binding of dye to the epidermal cell walls. Furthermore, [203Hg] PCMBS autoradiography demonstrated that the hypodermis was not a barrier to apoplastic movement of PCMBS.

These results suggest that although cortical cells possess the capacity to absorb ions, K+ influx at low concentrations is limited to the root periphery. Cortical cell uptake appears to be repressed under these conditions. At higher concentrations, cortical cells may function to absorb K+. Such a model may involve regulation of cortical cell ion transport capacity.

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8.
Abscisic acid (ABA) was found to increase the accumulation of 36Cl, total Cl, 22Na and total Na+ in roots of intact bean seedlings. After an initial promotion. ABA inhibited longdistance transport of these ions from the root to the shoot. However, it consistently inhibited both uptake and transport of 42K and total K+ in intact bean seedlings. A promotion of net 36Cl influx (ψoc) and its accumulation in the root (Q*v) concomitant decrease in transport index (long-distance transport as percentage of total influx) showed that ABA stimulates -36Cl transport at the tonoplast. It inhibited H4 extrusion and net 86Rb influx which agrees with a cation exchange theory K+/Rb+ transport.  相似文献   

9.
Abstract Dissociation of active H+ extrusion (?ΔH+) from K+ uptake in pea and maize root segments was attempted by substituting K+ in the incubation medium with lipophilic cations assumed to enter the cell by passive, non-specific, permeation through the lipid component of the plasmalemma. Among the compounds tested, tributylbenzylammonium significantly stimulated ?ΔH+ in the absence of other monovalent cations in the medium. This effect was much more evident when the experiment was carried out in the presence of fusicoccin, which strongly stimulates proton extrusion and monovalent cation uptake, and hyperpolarizes the trans-membrane electric potential in these materials. Also the lipophilic cations tetraphenylphosphonium, dimethyldibenzylammonium and hexylguanidine markedly stimulated FC-promoted ?ΔH+. Octylguanidine at a low concentration induced an early stimulation followed by a strong inhibition of ?ΔH+. A complete lack of additivity was observed between the effects of lipophilic cations and that of K+ on H+ extrusion. Lipophilic cations severely inhibited K+ uptake. These data are interpreted as supporting the view of an electric, rather than a chemical, (namely, involving the same carrier system) nature of the coupling of active H+ extrusion with K+ influx.  相似文献   

10.
The action of exogenous polyamines (putrescine, spermidine, and spermine) on `washing' and fusicoccin-stimulated K+ uptake and H+ extrusion through the plasmamembrane in maize (Zea mays L., hybrid line Plenus S 516) root apical segments was studied. The results showed that polyamines inhibit the washing-stimulated K+ influx and H+ extrusion without interfering with K+ uptake and H+ extrusion stimulated by fusicoccin. Spermidine appeared to be the most effective in inhibiting K+ uptake and H+ extrusion while putrescine showed a smaller inhibiting action with respect to the others. The analysis of kinetic constants indicated that the polyamines behave as competitive inhibitors with respect to K+.  相似文献   

11.
We report here on the putative coupling between a high affinity K+ uptake system which operates at low external K+ concentrations (Km = 10-20 micromolar), and H+ efflux in roots of intact, low-salt-grown maize plants. An experimental approach combining electrophysiological measurements, quantification of unidirectional K+(86Rb+) influx, and the simultaneous measurement of net K+ and H+ fluxes associated with individual cells at the root surface with K+- and H+-selective microelectrodes was utilized. A microelectrode system described previously (IA Newman, LV Kochian, MA Grusak, and WJ Lucas [1987] Plant Physiol 84: 1177-1184) was used to quantify net ion fluxes from the measurement of electrochemical potential gradients for K+ and H+ ions within the unstirred layer at the root surface. No evidence for coupling between K+ uptake and H+ efflux could be found based on: (a) extremely variable K+:H+ flux stoichiometries, with K+ uptake often well in excess of H+ efflux; (b) dramatic time-dependent variability in H+ extrusion when both fluxes were measured at a particular location along the root over time; and (c) a lack of pH sensitivity by the high affinity K+ uptake system (to changes in external pH) when net K+ uptake, unidirectional K+(86Rb+) influx, and K+-induced depolarizations of the membrane potential were determined in uptake solutions buffered at pH values from pH 4 to 8. Based on the results presented here, we propose that high affinity active K+ absorption into maize root cells is not mediated by a K+/H+ exchange mechanism. Instead, it is either due to the operation of a K+-H+ cotransport system, as has been hypothesized for Neurospora, or based on the striking lack of sensitivity to changes in extracellular pH, uptake could be mediated by a K+-ATPase as reported for Escherichia coli and Saccharomyces.  相似文献   

12.
We have investigated the effects of hyperpolarization and depolarization, and the presence of K+ and/or Ca2+, on 22Na+ influx into corn (Zea mays L.) root segments. In freshly excised root tissue which is injured, Na+ influx is unaffected by hyperpolarization with fusicoccin, or depolarization with uncoupler (protonophore), or by addition of K+. However, added Ca2+ suppresses Na+ influx by 60%. In washed tissue which has recovered, Na+ influx is doubled over that of freshly excised tissue, and the influx is increased by fusicoccin and suppressed by uncoupler. This energy-linked component of Na+ influx is completely eliminated by low concentrations of K+, leaving the same level and kind of Na+ influx seen in freshly excised roots. The K+-sensitive energy linkage appears to be by the carrier for active K+ influx. Calcium is equally inhibitory to Na+ influx in washed as in fresh tissue. Other divalent cations are only slightly less effective. Net Na+ uptake was about 25% of 22Na+ influx, but proportionately the response to K+ and Ca2+ was about the same.

The constancy of K+-insensitive Na+ influx under conditions known to hyperpolarize and depolarize suggests that if Na+ transport is by means of a voltage-sensitive channel, the rise or fall of channel resistance must be proportional to the rise or fall in potential difference. The alternative is a passive electroneutral exchange of 22Na+ for endogenous Na+. The data suggest that an inwardly directed Na+ current is largely offset by an efflux current, giving both a small net uptake and isotopic exchange.

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13.
The adenine nucleotide content of the 2-centimeter segments excised from tray-grown corn (Zea mays L., WF9 × Mo17) roots declines for the first hour after excision. Concomitant with the loss of adenine nucleotides is a decline in respiration and a leakage of K+. With continued washing, these parameters partially or completely recover and increased phosphate influx develops. Increasing the wound effect by cutting 0.5-centimeter segments gives a more rapid and pronounced degradation of adenine nucleotides and slower recovery. Conversely, the mild injury caused by submerging intact roots induces less degradation and produces greater net adenine nucleotide synthesis during recovery; adding auxin to the washing medium produces a similar result. With all treatments, there is stabilization of energy charge at about 0.85.  相似文献   

14.
High Na+ concentrations may disrupt K+ and Ca2+ transport and interfere with growth of many plant species, cotton (Gossypium hirsutum L.) included. Elevated Ca2+ levels often counteract these consequences of salinity. The effect of supplemental Ca2+ on influx of Ca2+, K+, and Na+ in roots of intact, salt-stressed cotton seedlings was therefore investigated. Eight-day-old seedlings were exposed to treatments ranging from 0 to 250 millimolar NaCl in the presence of nutrient solutions containing 0.4 or 10 millimolar Ca2+. Sodium influx increased proportionally to increasing salinity. At high external Ca2+, Na+ influx was less than at low Ca2+. Calcium influx was complex and exhibited two different responses to salinity. At low salt concentrations, influx decreased curvilinearly with increasing salt concentration. At 150 to 250 millimolar NaCl, 45Ca2+ influx increased in proportion to salt concentrations, especially with high Ca2+. Potassium influx declined significantly with increasing salinity, but was unaffected by external Ca2+. The rate of K+ uptake was dependent upon root weight, although influx was normalized for root weight. We conclude that the protection of root growth from salt stress by supplemental Ca2+ is related to improved Ca-status and maintenance of K+/Na+ selectivity.  相似文献   

15.
Yan H  Li K  Ding H  Liao C  Li X  Yuan L  Li C 《Journal of plant physiology》2011,168(10):1067-1075
The primary objective of this study was to better understand how root morphological alteration stimulates N uptake in maize plants after root growth restriction, by investigating the changes in length and number of lateral roots, 15NO3 influx, the expression level of the low-affinity Nitrate transporter ZmNrt1.1, and proteomic composition of primary roots. Maize seedlings were hydroponically cultured with three different types of root systems: an intact root system, embryonic roots only, or primary roots only. In spite of sufficient N supply, root growth restriction stimulated compensatory growth of remaining roots, as indicated by the increased lateral root number and root density. On the other hand, there was no significant difference in 15NO3 influx between control and primary root plants; neither in ZmNrt1.1 expression levels in primary roots of different treatments. Our data suggested that increased N uptake by maize seedlings experiencing root growth restriction is attributed to root morphological adaptation, rather than explained by the variation in N uptake activity. Eight proteins were differentially accumulated in embryonic and primary root plants compared to control plants. These differentially accumulated proteins were closely related to signal transduction and increased root growth.  相似文献   

16.
We report here on an experimental system that utilizes ion-selective microelectrodes to measure the electrochemical potential gradients for H+ and K+ ions within the unstirred layer near the root surface of both intact 4-day-old corn seedlings and corn root segments. Analysis of the steady state H+ and K+ electrochemical potential gradients provided a simultaneous measure of the fluxes crossing a localized region of the root surface. Net K+ influx values obtained by this method were compared with unidirectional K+ (86Rb+) influx kinetic data; at any particular K+ concentration, similar values were obtained by either technique. The ionspecific microelectrode system was then used to investigate the association between net H+ efflux and net K+ influx. Although the computed H+:K+ stoichiometry is dependent upon the choice of diffusion coefficients, the values obtained were extremely variable, and net K+ influx rarely appeared to be charge-balanced by H+ efflux. In contrast to earlier studies, we found the cortical membrane potential to be highly K+ sensitive within the micromolar K+ concentration range. Simultaneous measurements of membrane potential and K+ influx, as a function of K+ concentration, revealed similar Km values for the depolarization of the potential (Km 6-9 micromolar K+) and net K+ influx (Km 4-7 micromolar K+). These data suggest that K+ may enter corn roots via a K+-H+ cotransport system rather than a K+/H+ antiporter.  相似文献   

17.
The relationships of concentration gradients to electropotential gradients resulting from passive diffusion processes, after equilibration, are described by the Nernst equation. The primary criterion for the hypothesis that any given ion is actively transported is to establish that it is not diffusing passively. A test was made of how closely the Nernst equation describes the electrochemical equilibrium in seedling tissues. Segments of roots and epicotyl internodes of pea (Pisum sativum var. Alaska) and of roots and coleoptiles of oat (Avena sativa var. Victory) seedlings were immersed and shaken in defined nutrient solutions containing eight major nutrients (K+, Na+, Ca2+, Mg2+, Cl, NO3, H2PO4 and SO42−) at 1-fold and 10-fold concentrations. The tissue content of each ion was assayed at 0, 8, 24, and 48 hours. A near-equilibrium condition was approached by roots for most ions; however, the segments of shoot tissue generally continued to show a net accumulation of some ions, mainly K+ and NO3. Only K+ approached a reasonable fit to the Nernst equation and this was true for the 1-fold concentration but not the 10-fold. The data suggest that for Na+, Mg2+, and Ca2+ the electrochemical gradient is from the external solution to the cell interior; thus passive diffusion should be in an inward direction. Consequently, some mechanism must exist in plant tissue either to exclude these cations or to extrude them (e.g., by an active efflux pump). For each of the anions the electrochemical gradient is from the tissue to the solution; thus an active influx pump for anions seems required. Root segments approach ionic equilibrium with the solution concentration in which the seedlings were grown. Segments of shoot tissue, however, are far removed from such equilibration. Thus in the intact seedling the extracellular (wall space) fluid must be very different from that of the nutrient solution bathing the segments; it would appear that the root is the site of regulation of ion uptake in the intact plant although other correlative mechanisms may be involved.  相似文献   

18.
Diethard Köhler 《Planta》1968,84(2):158-165
Summary Five to 6 day old dark-adapted dwarf and tall pea seedlings grown in water culture were illuminated for ten minutes with red light and/or ten minutes with far-red light, and 90 to 170 minutes later their roots were immersed in a 0.2 mM K+ solution containing labeled 86Rb+. After two hours uptake the fresh-weights and radioactivities of the shoot organs were determined. It was found that red light inhibits K+uptake into internodes and promotes uptake into the plumula. The red-light effect on K+transport precedes the red-light induced growth inhibition of internodes and growth promotion of leaves and is abolished by far-red light given immediately after red. The red-light effect on K+transport is independent of the concentration of K+ given to the roots in the range between 0.2 to 125 mM.  相似文献   

19.
Vale FR  Jackson WA  Volk RJ 《Plant physiology》1987,84(4):1416-1420
Potassium influx into roots of dark-grown decapitated maize seedling (Zea mays L., cv Pioneer 3369A) was examined in presence and absence of ambient ammonium and at various root potassium concentrations. Six-day old seedlings which were dependent on the endosperm reserves for their energy source were exposed to KCl (labeled with 86Rb) ranging from 5 to 200 micromolar. At both low (13 micromoles per gram fresh weight) and high (100 micromoles per gram fresh weight) root potassium concentration, isotherms indicated two potassium influx systems, one approaching saturation at 50 to 100 micromolar potassium and an additional one tentatively considered to be linear. A mixed-type inhibition by ammonium for the low-concentration saturable system was indicated by a concomitant decrease in Vmax and increase in Km. High root potassium concentration decreased Vmax but had little effect on Km of this system. The rate constant for the second quasilinear system was decreased by ambient ammonium and by high root potassium status. Transfer of high potassium roots to potassium-free solutions resulted in an increase in influx within 2 hours; by 24 hours influx significantly exceeded that of roots not previously exposed to potassium. In roots of both low and high root potassium concentrations, potassium influx was restricted progressively as ambient ammonium increased to about 100 micromolar, but there was little further inhibition as ammonium concentrations increased beyond that to 500 micromolar. The data imply that potassium influx has two components, one subject to inhibition by ambient ammonium and one relatively resistant.  相似文献   

20.
Allosteric and Non-Allosteric Regulation of Rubidium Influx in Barley Roots   总被引:1,自引:0,他引:1  
Uptake of Rb+ was investigated in 6–8-day-old intact barley plants (Hordeum vulgare cv. Kristina), which had been cultivated or pretreated in nutrient solutions with various K+ concentrations. The relationship between Rb+ influx and the K+ concentration of roots appeared to be sigmoidal for plants grown in solutions containing K+, indicating regulation of Rb+ uptake by allosteric inhibition of the uptake mechanism. Pretreatment of the roots in K+-free solutions changed the pattern of uptake and caused the Rb+ influx to become linearly related to the chemical Rb+ potential of the uptake solution. Pretreatment in K+-free solutions probably abolishes the allosteric inhibition of a carrier system.  相似文献   

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