Effects of sulphate and pH on the plant-availability of phosphate adsorbed on goethite

The adsorption of phosphate on metal (hydr)oxides may be influenced by the pH and by the adsorption of other ions. In this study, the influence of sulphate and pH on phosphate adsorption on goethite and the availability to plants of adsorbed phosphate was examined. Maize plants were grown on suspensions of goethite with adsorbed phosphate, containing the same total amount of phosphate and either 0.11 mM or 2.01 mM sulphate at pH 3.7, 4.6 or 5.5. The uptake of phosphorus by the plants increased with the larger sulphate concentration and decreasing pH. Mean P uptake in the treatment with 2.01 mM sulphate and pH 3.7 was 55 µmol plant^-1, whereas in the treatment with 0.11 mM sulphate and pH 5.5 it was 2 µmol plant^-1. Batch adsorption experiments using³² P and speciation modelling of ion adsorption showed that in the presence of sulphate, the phosphate concentration in solution strongly increased with decreasing pH, due to competitive adsorption between sulphate and phosphate on goethite. Modelled phosphate concentrations in solution in the uptake experiment were all below 0.6 µM and correlated well with the observed P uptake. This correlation indicates that the strong influence of the sulphate concentration and pH on the plant-availability of adsorbed phosphate results from the competition between sulphate and phosphate for adsorption on goethite.     

Die pH-Abhängigkeit der Aufnahme von H2PO 4 - , SO 4 = , Na+ und K+ und ihre gegenseitige Beeinflussung bei Ankistrodesmus braunii

Summary Ion uptake was studied using ³²P, ³⁵S, ²²Na and ⁴²K as tracers in synchronized cells of Ankistrodesmus, which were slightly starved with respect to the ions to be investigated. In the light and in the dark, phosphate uptake is maximal between pH 5.5 and 6.5. Whereas Na⁺ in comparison to K⁺ enhances phosphate uptake in the light (8 to 9-fold) and in the dark, Ca⁺⁺ exerts only a slightly stimulatory effect. The stimulation of phosphate binding by Na⁺ occurs rapidly, even after less than 5 sec of incubation, and also in the presence of an equimolar concentration of K⁺.The pH-dependence of Na⁺-uptake in the light and in the dark is comparable to a dissociation curve: Na⁺-uptake increases with decreasing extracellular H⁺-concentration and is inversely proportional to phosphate uptake in the absence of Na⁺. The light:dark ratio of Na⁺-uptake at pH 8 amounts to 7:1. Mere adsorption of Na⁺ is similarly dependent on the pH. K⁺ strongly competes with Na⁺-uptake, even at pH 8. K⁺-uptake proceeds in a quite different manner from Na⁺-uptake and has an optimum at pH 7.Sulfate is taken up linearly in a biphasic process as a function of time; the pH-optimum lies between pH 7.5 and 8. K⁺ but not Na⁺ slightly enhances sulfate uptake.The Na⁺-enhancement of phosphate uptake can be related neither to a sodium-potassium exchange pump nor to a photosynthesis-dependent ion-exchange reaction.The results suggest that the uptake of phosphate, Na⁺ and K⁺, and the influence of alkali cations on phosphate uptake, but not sulfate uptake, are strongly dependent on fixed charges of the plasmalemma or even of the cell wall. These fixed charges may even prevent an active ion uptake.     

Der Einfluß von Natrium- und Kaliumionen auf die Phosphataufnahme bei Ankistrodesmus braunii

Summary The uptake of phosphate as influenced by sodium and potassium ions was investigated in the light and in the dark. It was found to be a function of the external phosphate concentration. At a low concentration (up to 10^–5 mol/l) in the presence of Na⁺ phosphate is quickly absorbed and hence phosphate is the limiting factor for further labelling. In the presence of K⁺ phosphate uptake is constant over a long period.The enhancement of phosphate uptake by Na⁺ is also found when the external concentration of P is raised up to 10^–4 mol/l. Then the gross uptake proceeds over six hours, with the greatest Na⁺-dependent increase occurring in the label of the TCA-insoluble phosphate fraction (P_u).The phosphate uptake is strongly dependent on the pH of the reaction mixture. In the presence of Na⁺ it is highest between pH 5.6 and 7. As the uptake in the presence of K⁺ parallels the dissociation curve of the dihydrogen form H₂PO ₄ ^– , the Na⁺-enhancement is optimal in the alkaline pH range (pH 8).On the basis of a comparison between the pH-dependence of phosphate uptake and the dependence of the uptake on the external phosphate concentration analysed by a method of enzyme kinetics, it is suggested that Ankistrodesmus metabolically transports H₂PO ₄ ^– but not HPO ₄ ⁼ . Moreover, it is concluded from the absence of light stimulation and the weak inhibition of the uptake by DCMU or CCCP in the presence of K⁺ that at low P-concentrations the diffusion is limiting the uptake. Only at higher concentrations is an active phosphate uptake measured.Furthermore it is concluded that the observed Na⁺-stimulation of the ³²P-labelling of the TCA-soluble and insoluble compounds inside the cell is indirect and depends only on the action of Na⁺ and K⁺ ions at the first transport site in the plasmalemma.     

Uptake by Yeast: Interaction of Rb+, Na+ and Phosphate

It has been shown that addition of phosphate to phosphate deficient yeast gives rise to an immediate increase in the rate of Na⁺ uptake and an immediate decrease in the rate of Rb⁺ uptake. In addition, phosphate uptake is enhanced specifically by Na ions presumably by a process with a very high affinity for phosphate with a K_m of about 2 × 10⁻⁶M at pH 7.2, whereas the K_m for phosphate uptake of the Na⁺ independent process amounts to 1.3 × 10⁻⁴M.     

Characterization of phosphate absorption in maize root cortex segments

Uptake of phosphate ions by 1 mm segments of isolated maize root cortex layers was studied. Cortex segments (from roots of 8 days old maize plants) absorb phosphate ions from 1 mM KH₂PO₄ in 0.2 mM CaSCO₄ at the average rate of 34.3 ±3.2 μg P_i g^?1 (fr. m.) h^?1,i.e. 0.35± 0.02 μmol P_i g^?1 (fr. m.) h^?1. Phosphate uptake considerably increases after a certain period of “augmentation”,i.e. washing in aerated 0.2 mM CaSO₄. This increase is completely blocked by the presence of 10 μg ml^?1 cycloheximide. The relation of uptake rate to phosphate concentration in the medium was shown to have 3 phases in the concentration range of 0.02 - 40 mM. Transition points were found between 0.8–1 mM and 10–20 mM. Following K_m and V_max values were found: K_m[mM] : 0.37 - 3.82 - 27.67 V_max[μg P_i g^?1 (fr. m.) h^?1] : 3.33 - 39.40 - 66.67 We have found no sharp pH optimum for phosphate uptake. It proceeds at almost constant rate till pH 6.0 and then the uptake rate drops with increasing pH. At low phosphate concentrations (1 mM) the lowest uptake rate was found at 5 and 13 °C, while the uptake is higher at 5 °C than at 13 °C at phosphate concentrations higher than 1 mM. At these concentrations uptake rate at 35 °C is lower than at 25 °C. Phosphate uptake considerably decreased in anaerobic conditions. DNP and iodoacetate (0.1 mM) completely blocked phosphate uptake from 1 mM KH₂PO₄, while uptake from 5 and 10 mM KH₂PO₄ was left unaffected by these substances. The inhibitors of active - SH groups NEM and PCMB inhibited phosphate uptake: 10^?3 M NEM by 81.6%, 10⁴ M NEM by 42% and 10^?4 M PCMB by 42%.     

Proton/Phosphate Stoichiometry in Uptake of Inorganic Phosphate by Cultured Cells of Catharanthus roseus (L.) G. Don

Upon absorption of phosphate, cultured cells of Catharanthus roseus (L.) G. Don caused a rapid alkalinization of the medium in which they were suspended. The alkalinization continued until the added phosphate was completely exhausted from the medium, at which time the pH of the medium started to drop sharply toward the original pH value. Phosphate exposure caused the pH of the medium to increase from pH 3.5 to values as high as 5.8, while the rate of phosphate uptake was constant throughout (10-17 micromoles per hour per gram fresh weight). This indicates that no apparent pH optimum exists for the phosphate uptake by the cultured cells. The amount of protons cotransported with phosphate was calculated from the observed pH change up to the maximum alkalinization and the titration curve of the cell suspension. Proton/phosphate transport stoichiometry ranged from less than unity to 4 according to the amount of phosphate applied. At low phosphate doses, the stoichiometries were close to 4, while at high phosphate doses, smaller stoichiometries were observed. This suggests that, at high phosphate doses, activation of the proton pump is induced by the longer lasting proton influx acidifying the cytoplasm. The increased H⁺ efflux due to the proton pump could partially compensate protons taken up via the proton-phosphate cotransport system. Thus, the H⁺/H₂PO₄⁻ stoichiometry of the cotransport is most likely to be 4.     

Phosphate transport in fertilized sea urchin eggs. I. Kinetic aspects

Properties of the fully developed phosphate transport system in the fertilized egg of the sea urchin, Strongylocentrotus purpuratus, were investigated. The rates of phosphate transport at concentrations of external phosphate of 1 to 44 μM, both in the absence and in the presence of 100 μM arsenate, exhibit typical saturation kinetics. At sea water concentrations of 2 μM phosphate, the rate of uptake is about 2 × 10^?9 μm/egg/minute at 15°C. Arsenate is a competitive inhibitor of phosphate transport, fully and immediately reversible in its effects, yielding K_i values ranging from 10.5 to 14.1 × 10^?6 M in comparison to the corresponding apparent K_M (Michaelis-Menten) constants for phosphate of 5.6 to 7.5 × 10^?6 M (pH 8.0, 15°C). The rate of arsenate uptake in a phosphate deficient medium amounts to 2.8 to 2.9 × 10^?10 μm arsenate/egg/minute at an arsenate concentration of 2.9 to 10.2 μM arsenate (HAsO₄^??), which is 9.5 and 5.6% of the rate of phosphate uptake at corresponding phosphate concentrations. Arsenate has essentially the same developmental effects at initial concentrations of 5–10 μM and 100 μM arsenate, namely no observable effects for exposure periods of 7.5 hours, although longer periods result in blockage of development at the early blastula stage. Outward flux of phosphate ions cannot be demonstrated by washing prelabelled eggs with sea water containing low or high concentrations of phosphate, even when phosphorylation has been blocked by exposing the eggs to a metabolic inhibitor. Phosphate uptake rates measured in the pH range from 5.0 to 10.0 reveal a sharp optimum at pH 8.8–8.9. Reference to the apparent pK' values of the phosphoric acid system indicate that the entering species is the HPO₄^?? ion. The effects on rates of phosphate uptake of exposure to sea water at pH values between 7 and 10 for 30 minute periods are fully reversible, but at lower pH values, reversal is delayed, and is only partial. Sodium molybdate (0.01 M), sodium pyrophosphate (1.5 × 10^?4 M), and adenosine triphosphate (1–5 × 10^?4 M) for exposure periods ranging from 40 to 180 minutes did not significantly affect phosphate uptake. Omission of Ca⁺⁺ ion from artificial sea water is without effect on phosphate uptake but the absence of both Ca⁺⁺ and Mg⁺⁺ results in profound and irreversible depression of both phosphate uptake and development. The data of this and the following paper are consistent with the conclusion that the transport of phosphate involves a surface located carrier. The apparent secondary and tertiary ionization constants of phosphoric acid in sea water (ionic strength = 0.6885) were measured, resulting in a value for pK′₂ = 6.14 and for pK′₃ = 10.99, at 15°C and phosphate at infinite dilution.     

Beziehungen der Aufnahme von Nitrat,Nitrit und Phosphat zur photosynthetischen Reduktion von Nitrat und Nitrit und zum ATP-Spiegel bei Ankistrodesmus braunii

Summary The pH-dependence of NO ₃ ^- and NO ₂ ^- uptake is different from that of phosphate uptake, but similar to that of sulfate uptake, with optima between pH 7.4 and 8.2 and smaller peaks at higher H⁺-concentration.Since the ATP-level is not affected by addition of ions and since phosphate uptake is not depressed by NO ₃ ^- , the inhibition of phosphate uptake by K⁺ reported in former papers cannot be explained by competition for the available energy(ATP) at the site of uptake.NO ₃ ^- uptake is strongly dependent on the activity of the NO ₃ ^- reducting system, as can be seen from the inhibition of NO ₃ ^- uptake in light by N₂ compared with that in air. Furthermore, the pH-dependences of NO ₃ ^- and NO ₂ ^- uptake correspond to the pH-optima known for the reductases.Phosphate uptake is enhanced by NO ₃ ^- and NO ₂ ^- in N₂. Since the enhancement of phosphate uptake is sensitive to DCMU and since this DCMU-sensitive phosphate uptake is accompanied by O₂-evolution, it is probably due to an NO ₃ ^- -stimulated noncyclic photophosphorylation which enhances the ATP-turnover and hence the incorporation of phosphate into organic compounds.

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Abkürzungen TCE Trichloressigsäure - P Orthophosphat - P₀ TCE-lösliche organische Phosphatverbindungen - P_u TCE-unlösliche Phosphatverbindungen - GP Gesamtphosphat     

Mesophyll Resistances to SO(2) Fluxes into Leaves

Uptake of label from solutions containing ³⁵SO₂, H³⁵SO₃⁻ and ³⁵SO₃²⁻ into mesophyll protoplasts, vacuoles, and chloroplasts isolated from young barley leaves was measured at different pH values. Uptake was fast at low pH, when the concentration of SO₂ was high, and low at high pH, when the concentration of SO₂ was low. When the resistance (R) of plasmalemma, tonoplast, and chloroplast envelope to the penetration of SO₂ was calculated from rates of uptake of label, comparable values were obtained for the different biomembranes at low pH values. R was close to 8000 seconds per meter and permeability coefficients were close to 1.25 × 10⁻⁴ meters per second. Under these conditions R may describe resistance to SO₂ diffusion across a lipid bilayer. At higher pH values, R decreased. As R was calculated on the assumption that SO₂ is the only penetrating molecular species, the data suggest that carrier-mediated anion transport contributes to the uptake of sulfur at physiological pH values thereby decreasing apparent R_SO2. The contribution of anion transport appeared to be smaller for transfer across the plasmalemma than for transfer across the tonoplast. It was large for transfer across the chloroplast envelope. The phosphate translocator of the chloroplast envelope catalyzed uptake of SO₃²⁻ into chloroplasts at neutral pH. Uptake was decreased in the presence of high levels of phosphate or sulfate and by pyridoxal phosphate. SO₂ transfer into cells leads to the intracellular liberation of one or two protons, depending on pH and oxidizing conditions. When the divalent sulfite anion is exchanged across the chloroplast envelope, bisulfite formation results in proton uptake in the chloroplast stroma, whereas SO₂ uptake into chloroplasts lowers the stroma pH.     

Adaptation to phosphate deprivation in osteoblast-like cells

The rat osteosarcoma cell line UMR-106–01 has an osteoblast-like phenotype. When grown in monolyer culture these cells transport inroganic phosphate and L-alanine via Na⁺-dependent transport systems. Exposure of these cells to a low phosphate medium for 4 h produced a 60–70 per cent increase in Na⁺-dependent phosphate uptake compared to control cells maintained in medium with a normal phosphate concentration. In contrast, Na⁺-dependent alanine uptake and Na⁺-independent phosphate uptake were not changed during phosphate deprivation. The increased phosphate uptake was due, in part, to an increased V_max and was blocked completely by pretreatment with cycloheximide (70 μM). In these cells recovery of intracellular pH after acidification with NH₄Cl is due primarily to the Na⁺/H⁺ exchange system. The rate of this recovery process, monitored with a pH sensitive indicator (BCECF), was decreased by more than 50 per cent in phosphate-deprived cells compared to controls indicating that Na⁺/H⁺ exchange was inhibited during phosphate deprivation.     

Transport du sulfate à travers la double membrane limitante, ou enveloppe, des chloroplastes d'épinard

Evidence is presented for low rates of carriermediated uptake of sulphate, thiosulphate and sulphite into the stroma of the C3 plant Spinacia oleracea. Uptake of sulphate in the dark was followed using two techniques (1) uptake of sulphate [³⁵S] as determined by silicon oil centrifugal filtration and (2) uptake as indicated by inhibition of CO₂-dependant O₂ evolution rates after addition of sulphate.Sulphate, thiosulphate and sulphite were transported across the envelope leading to an accumulation in the chloroplasts. Sulphate transport had saturation kinetics of the Michaelis-Menten type (Vmax : 25 μmoles . mg⁻¹ chl . h⁻¹ at 22°C ; Km : 2.5 mM). The rate of transport for sulphate was not influenced either by illumination or pH change in the external medium. Phosphate was a competitive inhibitor of sulphate uptake by chloroplasts (Ki : 0.7 mM, fig. 1). The rate of transport for phosphate appeared to be much higher than for sulphate. When the chloroplasts were pre-loaded with labelled sulphate, radioactivity was rapidly released after addition of phosphate into the external medium. Consequently, the transport of sulphate occurs by a strict counter-exchange : for each molecule of sulphate entering the chloroplast, one molecule of phosphate leaves the stroma, and vice-versa.The uptake of sulphate by isolated intact chloroplasts exchanging for internal free phosphate induced a lower rate of photophosphorylation, which in turn inhibited CO₂-dependent O₂ evolution.The presence, on the inner membrane of the chloroplast envelope, of a specific sulphate carrier, distinct from the phosphate translocator, is discussed.     

Mechanisms of phosphate uptake into brush-border membrane vesicles from goat jejunum

This study concerns the uptake of inorganic phosphate into brush-border membrane vesicles prepared from jejunal tissues of either control or Ca-and/or P-depleted goats. The brush-border membrane vesicles showed a time-dependent accumulation of inorganic phosphate with a typical overshoot phenomenon in the presence of an inwardly directed Na⁺ gradient. The Na⁺-dependent inorganic phosphate uptake was completely inhibited by application of 5 mmol·l^-1 sodium arsenate. Half-maximal stimulation of inorganic phosphate uptake into brush-border membrane vesicles was found with Na⁺ concentrations in the order of 5 mmol·l^-1. Inorganic phosphate accumulation was not affected by a K⁺ diffusion potential (inside negative), suggesting an electroneutral transport process. Stoichiometry suggested an interaction of two or more Na ions with one inorganic phosphate ion at pH 7.4. Na⁺-dependent inorganic phosphate uptake into jejunal brush-border membrane vesicles from normal goats as a function of inorganic phosphate concentration showed typical Michaelis-Menten kinetic with V _max=0.42±0.08 nmol·mg^-1 protein per 15 s^-1 and K _m=0.03±0.01 mmol·l^-1 (n=4, x ±SEM). Long-term P depletion had no effect on these kinetic parameters. Increased plasma calcitriol concentrations in Ca-depleted goats, however, were associated with significant increases of V _max by 35–80%, irrespective of the level of P intake. In the presence of an inwardly directed Na⁺ gradient inorganic phosphate uptake was significantly stimulated by almost 60% when the external pH was decreased to 5.4 (pH_out/pH_in=5.4/7.4). The proton gradient had no effect on inorganic phosphate uptake in absence of Na⁺. In summary, in goats Na⁺ and calcitriol-dependent mechanisms are involved in inorganic phosphate transport into jejunal brush-border membrane vesicles which can be stimulated by protons.Abbreviations AP activity of alkaline phosphatase - BBMV brush-border membrane vesicles - EGTA ethyleneglycol-triacetic acid - n _app apparent Hill coefficient - P _i inorganic phosphate - PTH parathyroid hormone     

Examination of the substrate stoichiometry of the intestinal Na+/phosphate cotransporter

Summary The substrate stoichiometry of the intestinal Na⁺/phosphate cotransporter was examined using two measures of Na⁺-dependent phosphate uptake: initial rates of uptake with [³²P] phosphate and phosphate-induced membrane depolarization using the potential-sensitive dye diSC₃(5). Isotopic phosphate measures electrogenic and electroneutral Na⁺-dependent phosphate uptake, while phosphate-induced membrane depolarization measures electrogenic phosphate uptake. Using these measures of Na-dependent phosphate uptake, three parameters were compared: substrate affinity; phenylglyoxal sensitivity and labeling; and inhibiton by mono- and di-fluorophosphates. Na⁺/phosphate cotransport was found to have similar Na⁺ activations (apparentK _0.5's of 28 and 25mm), apparentK _m 's for phosphate (100 and 410 m), andK _0.5's for inhibition by phenylglyoxal (70 and 90 m) using isotopic phosphate, uptake and membrane depolarization, respectively. Only difluorophosphate inhibited Na⁺-dependent phosphate uptake below 1mm at pH 7.4.Difluorophosphate also protected a 130-kDa polypeptide from FITC-PG labeling in the presence of Na⁺ with apparentK _0.5 for phosphate of 200 m; similar to the apparentK _m for phosphate uptake, andK _0.5 for phosphate protection against FITC-PG inhibition of Na⁺-dependent phosphate uptake and FITC-PG labeling of the 130-kDa polypeptide. These results indicate that the intestinal Na⁺/phosphate cotransporter is electrogenic at pH 7.4, that H₂PO ₄ ^– is the transport-competent species, and that the 130-kDa polypeptide is an excellent candidate for the intestinal Na⁺/phosphate cotransporter.     

Factors influencing phosphate exchange across the sediment-water interface of eutrophic reservoirs

The results of a survey of the sediment chemistry of 7 East Anglian reservoirs are presented as part of a regional study on the assessment and control of eutrophication. The influence of water quality (dissolved oxygen, pH, temperature) on phosphate (PO₄) adsorption by sediment from hypertrophic Ardleigh Reservoir is also examined. Extractable phosphate-P (extr.-P) varied between 92 and 383 mg kg^–1 dry matter. Extractable P varied between 5.3 and 16.6% of the total phosphate-P (Tot. P) content and increased with the concentration of dissolved reactive phosphate-P (DRP) in the overlying water column. Organically complexed iron (organic Fe) was the determinand which correlated most closely with phosphate adsorption capacity, PAC (r = 0.8). Organic Fe was also related inversely to Extr. P. The rate and extent of PO₄ adsorption by Ardleigh Reservoir sediment increased with the initial concentration of DRP and adsorption equilibria were reached after 24 h. The equilibrium DRP concentration, [DRP], was 0.7 mg P 1^–1 under aerobic conditions indicative of a high potential for PO₄ exchange. The rate and extent of PO₄ adsorption was greater at 7 °C than at 22 °C PO₄ adsorption increased markedly with dissolved oxygen status. Ardleigh sediment exhibited a marked buffering capacity to a change in pH; however, PO₄ adsorption was greatest at an equilibrium pH of 5.6 and decreased progressively either side of this pH value.Options for the artificial control of sediment PO₄ release are discussed in relation to the seasonal variation in sediment PO₄ exchange observed for Ardleigh Reservoir.     

Multiphasic Uptake of Sulfate by Barley Roots I. Effects of Analogues, Phosphate, and pH

For sulfate uptake by barley roots, competition studies reveal that uptake and phase transitions are caused by interaction of ions with separate sites on or in the plasmalemma. Uptake is competitively, and unequally, inhibited by sulfate analogues but not by other divalent anions. In contrast, divalent phosphate and di- and trivalent pyrophosphate are equally effective in causing transitions. Phosphate is taken up mainly or entirely as H₂PO₄^? by a similar but separate multiphasic mechanism. At pH 8, sulfate uptake is mediated by fewer phases than at low and intermediate pH.     

Seawater pH,and not inorganic nitrogen source,affects pH at the blade surface of Macrocystis pyrifera: implications for responses of the giant kelp to future oceanic conditions

Ocean acidification (OA), the ongoing decline in seawater pH, is predicted to have wide‐ranging effects on marine organisms and ecosystems. For seaweeds, the pH at the thallus surface, within the diffusion boundary layer (DBL), is one of the factors controlling their response to OA. Surface pH is controlled by both the pH of the bulk seawater and by the seaweeds' metabolism: photosynthesis and respiration increase and decrease pH within the DBL (pH_DBL), respectively. However, other metabolic processes, especially the uptake of inorganic nitrogen (N_i; NO₃^? and NH₄⁺) may also affect the pH_DBL. Using Macrocystis pyrifera, we hypothesized that (1) NO₃^? uptake will increase the pH_DBL, whereas NH₄⁺ uptake will decrease it, (2) if NO₃^? is cotransported with H⁺, increases in pH_DBL would be greater under an OA treatment (pH = 7.65) than under an ambient treatment (pH = 8.00), and (3) decreases in pH_DBL will be smaller at pH 7.65 than at pH 8.00, as higher external [H⁺] might affect the strength of the diffusion gradient. Overall, N_i source did not affect the pH_DBL. However, increases in pH_DBL were greater at pH 7.65 than at pH 8.00. CO₂ uptake was higher at pH 7.65 than at pH 8.00, whereas HCO₃^? uptake was unaffected by pH. Photosynthesis and respiration control pH_DBL rather than N_i uptake. We suggest that under future OA, Macrocystis pyrifera will metabolically modify its surface microenvironment such that the physiological processes of photosynthesis and N_i uptake will not be affected by a reduced pH.     

The labyrinth of nutrient cycles and buffers in wetlands: results based on research in the Camargue (southern France)

Wetlands, especially in the Mediterranean area, are subject to severe eutrophication. This may upset the equilibrium between phytoplankton production in undesirable quantities and a quantitatively desirable macrophyte production. In order to manage this equilibrium, a quantitative knowledge of nutrient input and fluxes is essential and the role of sediments in these processes must be understood. This knowledge can be useful even for agriculture, e.g. rice cultivation, where optimal utilization of fertilizers can lead to an economic benefit.In this article different aspects of nutrient cycles are discussed in view of approaching a sufficiently precise quantification. The nutrient input balance of the Camargue was therefore measured which showed that the input of nutrients with the irrigation water, taken from the river Rhone, roughly equals the quantity of fertilizers added.Phytoplankton growth can be approached reasonably with the Monod model, although there are still many practical problems, such as the influence of the pH on P uptake and the problem of measuring P uptake in the field. The situation is worse for macrophyte growth; quantitative data are scarce and studies have often been carried out with unrealistic nutrient concentrations or without addressing the influence of the sediment. This influence can also include negative factors, such as high concentrations of Fe²⁺, H₂S or FeS, but cannot yet be quantified.The nitrogen cycle in wetlands is dominated by denitrification. Most wetlands have sediments with high concentrations of organic matter, therefore with a large reducing capacity. Besides this process, we have shown that denitrification can also be controlled by FeS. In the Camargue sediments this denitrification is mediated by bacteria from the sulfur cycle; this appeared to be the major pathway. It was shown that a stoicheiometric relation exists between nitrate reduced and sulphate produced. The influence of the temperature was quantified and appeared to be stronger at high organic matter concentrations than at lower ones. Denitrification with FeS means that the bacteria use nitrate also for their N demands, while this is not necessarily the case during denitrification with organic matter.Mineralization of macrophytes is a much slower process than that of phytoplankton, probably because of their high C/N ratio. We could, however, not confirm the general assumption that the addition of nitrogen stimulates this mineralization. On the contrary, we found that two amino acids both with a C/N ratio of 6 had different mineralization rates. The amino acid composition of dead macrophytes and the C/N ratio may be of equal importance.Unlike nitrogen, phosphate is always strongly adsorbed onto sediments. The two mechanisms of the adsorption of inorganic phosphate onto sediments, i.e. the adsorption onto Fe(OOH) and the precipitation of apatite, have been quantified. The adsorption of phosphate onto Fe(OOH) can be satisfactory described with the Freundlich adsorption isotherm: P_ads = A^* (o-P)^B. The adsorption coefficient A depends on the pH of the system and the Ca²⁺ concentration of the overlying water and can be quantified preliminarily by A = a.10(^–0.416*pH).(2.86 – (1.86.e^–Ca2+)). B can be approached by 0.333, which means the cube root of the phosphate concentration. The second mechanism is the solubility of apatite. We found a solubility product of 10^–50 for hard waters. The two mechanisms are combined in solubility diagrams which describe equilibrium situations for specific lakes.The conversion of Fe(OOH) to FeS has a strong influence on phosphate adsorption, although the partial reduction of Fe(OOH) P by H₂S does not release significant quantities of phosphate. Even after complete conversion to FeS only a small part of the bound phosphate was released.Besides the two inorganic phosphate compounds, we established the existence of two organic pools, one soluble after extraction with strong acid (ASOP), the other one with strong alkali. The first pool is probably humic bound phosphate, while the larger part of the second pool was phytate. The ASOP was remineralized during the desiccation of a Camargue marsh; this drying up oxidized FeS, thus improving the phosphate adsorption and decreasing the denitrification capacity. It can, therefore, be an important tool for management. The phytate was strongly adsorbed onto Fe(OOH), which explains the non-bioavailability towards bacteria.The fact that the sediment phosphate concentration can be approached by multiplying the relevant sediment adsorption constant with % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGak0Jf9crFfpeea0xh9v8qiW7rqqrFfpeea0xe9Lq-Jc9% vqaqpepm0xbba9pwe9Q8fs0-yqaqpepae9pg0FirpepeKkFr0xfr-x% fr-xb9adbaqaaeGaciGaaiaabeqaamaabaabaaGcbaWaaOqaaeaaca% WGVbGaeyOeI0IaamiuaaWcbaGaaG4maaaaaaa!3B8D!\[\sqrt[3]{{o - P}}\] concentration has the consequence that much larger quantities of phosphate accumulate in the sediments than in the overlying water. This means that even if the phosphate input is stopped, the eutrophication will only be reversed very slowly, and not at all, if the shallow waters in wetlands have no through flow — as is often the case in many marshes in Mediterranean wetlands.Abbreviations used o-P = dissolved ortho phosphate (or its concentration) - N_part, P_part = particulate N or P - Tot-N_inorg = Total inorganic nitrogen (= NH₃ + NO ₂ ^– + NO ₃ ^– ) This paper, giving an overview of the research in the sediments of the Camargue, was read during the symposium Nutrient Cycles — A Joy Forever, on the occasion of my retirement, 19th of May 1993 at the I.H.E. in Delft (Netherlands).     

A new alkalitolerant <Emphasis Type="Italic">Yarrowia lipolytica</Emphasis> yeast strain is a promising model for dissecting properties and regulation of Na<Superscript>+</Superscript>-dependent phosphate transport systems

A newly isolated osmo-, salt-, and alkalitolerant Yarrowia lipolytica yeast strain is distinguished from other yeast species by its capacity to grow vigorously at alkaline pH values (9.7), which makes it a promising model organism for studying Na⁺-dependent phosphate transport systems in yeasts. Phosphate uptake by Y. lipolytica cells grown at pH 9.7 was mediated by several kinetically discrete Na⁺-dependent systems specifically activated by Na⁺. One of these, a low-affinity transporter, operated at high concentrations of extracellular phosphate. The other two, high-affinity systems, maximally active in phosphate-starved cells, were repressed or derepressed depending on the prevailing extracellular phosphate concentration and pH value. The contribution of Na⁺/P_i-cotransport systems to the total cellular phosphate uptake progressively increased with increasing pH, reaching its maximum at pH 9.Translated from Biokhimiya, Vol. 69, No. 11, 2004, pp. 1607–1615.Original Russian Text Copyright © 2004 by Zvyagilskaya, Persson.     

Citrate transport in corn mitochondria

Citrate uptake by corn mitochondria (Zea mays L. B73 × Mol9) was investigated by osmotic swelling and [¹⁴C]citrate accumulation. Uptake driven by passive influx, ammonium gradients, and respiration was followed. There was no requirement for phosphate and/or malate to secure citrate uptake, although under some conditions these additives were promotive. Inhibition of the phosphate and dicarboxylate carriers did not eliminate citrate uptake. Citrate_in/malate_out exchange occurs, but at a rate too slow to account for observed citrate uptake, and depletion of endogenous malate only reduced citrate uptake by 38%. It was concluded that citrate can be rapidly accumulated by a mechanism other than by exchange for dicarboxylates. The effect of uncoupler on respiration-driven [¹⁴C]citrate accumulation, and studies of passive swelling using ionophores and uncouplers indicated that the major avenue of citrate uptake is by H⁺/citrate co-transport with a pH optimum near 4.5. The in vivo role of this mechanism is not yet understood.     

Extra- and Intracellular pH and Membrane Potential Changes Induced by K, Cl, H(2)PO(4), and NO(3) Uptake and Fusicoccin in Root Hairs of Limnobium stoloniferum

Short-term ion uptake into roots of Limnobium stoloniferum was followed extracellularly with ion selective macroelectrodes. Cytosolic or vacuolar pH, together with the electrical membrane potential, was recorded with microelectrodes both located in the same young root hair. At the onset of chloride, phosphate, and nitrate uptake the membrane potential transiently decreased by 50 to 100 millivolts. During Cl⁻ and H₂PO₄⁻ uptake cytosolic pH decreased by 0.2 to 0.3 pH units. Nitrate induced cytosolic alkalinization by 0.19 pH units, indicating rapid reduction. The extracellular medium alkalinized when anion uptake exceeded K⁺ uptake. During fusicoccin-dependent plasmalemma hyperpolarization, extracellular and cytosolic pH remained rather constant. Upon K⁺ absorption, FC intensified extracellular acidification and intracellular alkalinization (from 0.31 to 0.4 pH units). In the presence of Cl⁻ FC induced intracellular acidification. Since H⁺ fluxes per se do not change the pH, recorded pH changes only result from fluxes of the stronger ions. The extra- and intracellular pH changes, together with membrane depolarization, exclude mechanisms as K⁺/A⁻ symport or HCO₃⁻/A⁻ antiport for anion uptake. Though not suitable to reveal the actual H⁺/A⁻ stoichiometry, the results are consistent with an H⁺/A⁻ cotransport mechanism.