Conostegia xalapensis (Melastomataceae): an aluminum accumulator plant

In acidic soils, an excess of Al³⁺ is toxic to most plants. The Melastomataceae family includes Al‐accumulator genera that tolerate high Al³⁺ by accumulating it in their tissues. Conostegia xalapensis is a common shrub in Mexico and Central America colonizing mainly disturbed areas. Here, we determined whether C. xalapensis is an Al accumulator, and whether it has internal tolerance mechanisms to Al. Soil samples collected from two pastures in the state of Veracruz, Mexico, had low pH and high Al³⁺ concentrations along with low Ca²⁺ levels. Leaves of C. xalapensis from pastures showed up to 19 000 mg Al kg^?1 DW (dry weight). In laboratory experiments, 8‐month‐old seedlings treated with 0.5 and 1.0 mM AlCl₃ for 24 days showed higher number of lateral roots and biomass. Pyrocatechol violet and hematoxylin staining evidenced that Al localized in epidermis and mesophyll cells in leaves and in epidermis and vascular pith in roots. Scanning electron microscope‐energy dispersive X‐ray microanalysis of Al‐treated leaves corroborated that Al is in abaxial and adaxial epidermis and in mesophyll cells (31.2%) in 1.0 mM Al‐treatment. Roots of Al‐treated plants had glutathione reductase (EC 1.6.4.2) and superoxide dismutase (EC 1.15.1.1) activity higher, and low levels of O and H ₂O₂. C. xalapensis is an Al‐accumulator plant that can grow in acidic soils with higher Al³⁺ concentrations, and can be considered as an indicator species for soils with potential Al toxicity.     

Effects of litter incorporation and nitrogen fertilization on the contents of extractable aluminium in the rhizosphere soil of tea plant (Camallia sinensis (L.) O. Kuntze)

The effects of litter incorporation and nitrogen application on the properties of rhizosphere and bulk soils of tea plants (Camellia sinensis (L.) O. Kuntze) were examined in a pot experiment. Total of 8 treatments included four levels of tea litter additions at 0, 4.9, 9.8, and 24.5 g kg^–1 in combination with two N levels (154.6 mg kg^–1 and without). After 18 months of growth the rhizosphere soil was collected by removing the soil adhering to plant roots and other soil was referred to as bulk soil. The dry matter productions of tea plants were significantly increased by N fertilization and litter incorporation. The effect of litter was time-depending and significantly decreased the content of exchangeable Al (Al_ex, by 1 mol L^–1 KCl) and Al saturation at 9 months after litter incorporation whereas soil pH was not affected, although the litter contained high Al content. After 18 months, the contents of extractable Al by dilute CaCl₂, CuCl₂ + KCl, NH₄OAC, ammonium oxalate and sodium citrate (Al_CaCl2, Al_Cu/KCl, Al_NH4OAC, Al_Oxal, and Al_Cit respectively) and Al_ex, were not affected by litter application, except that of Al_CaCl2 in the rhizosphere soil which was decreased following litter additions. Nitrogen fertilization with NH₄ ⁺ (urea and (NH₄)₂SO₄) significantly reduced soil pH, the contents of exchangeable Ca, K, Mg and base saturation while raised extractable Al levels (Al_CaCl2, Al_Cu/KCl, Al_NH4OAC, and Al_ex). In the rhizosphere soils exchangeable K accumulated in all treatments while exchangeable Ca and Mg depleted in treatments without litter application. The depletions of Ca and Mg were no longer observed following litter incorporation. This change of distribution gradients in rhizosphere was possibly due to the increase of nutrient supplies from litter decomposition and/or preferable root growth in soil microsites rich in organic matter. Lower pH and higher extractable Al (Al_CaCl2, Al_ex, and Al_NH4OAC) in the rhizosphere soils, regardless of N and litter treatments, were distinct and consistent in all treatments. Such enrichments of extractable Al in the rhizosphere soil might be of importance for tea plants capable of taking up large amounts of Al.     

Models for relating pH measurements in water and calcium chloride for a wide range of pH,soil types and depths

Soil pH is commonly measured in water (pHw) or 0.01 M CaCl₂ (pH_Ca). The need to convert between these methods has led to the publication of linear, quadratic and cubic polynomial relationships for limited suites of soils. Concerns over the applicability of such relationships when mapping a wide range of soils and pH led to the establishment of a database of pH_W and pH_Ca values on each of 7894 samples from soil survey and field experimental sites in Queensland. The relationship between pH_W and pH_Ca across all soils was investigated and preliminary results examining the effect of soil depth and soil type on the relationship are presented.For all soils and depths, a linear regression accounted for 93.2% of the variation but did not predict pH_Ca well at very high or low pH_W values. The inclusion of second and third powers of pH_W accounted for significantly more of the variation (R²=0.94) in pH_Ca and the resultant curve matched the data better at high and low pH.Analysis of surface, sub-surface and subsoil groupings did not reveal any appreciable differences in the relationship between pH_W and pH_Ca attributable to depth. In contrast, differences in the relationship were evident between soil types. Generally, the mildly leached soils had linear relationships, while the weathered soils were distinctly curvilinear at low pH.     

Phosphate and calcium uptake in beech (Fagus sylvatica) in the presence of aluminium and natural fulvic acids

In the present study we examine the effects of Al on the uptake of Ca²⁺ and H₂PO^-₄ in beech (Fagus sylvatica L.) grown in inorganic nutrient solutions and nutrient solutions supplied with natural fulvic acids (FA). All the solutions used were chemically well characterized. The uptake of Al by roots of intact plants exposed to solutions containing 0, 0.15 or 0.3 mM AlCl₃ for 24 h, was significantly less if FA (300 mg l⁻¹) were also present in the solutions. The Ca²⁺(⁴⁵Ca²⁺) uptake was less affected by Al in solutions supplied with FA than in solutions without FA. There was a strong negative correlation between the Al and Ca²⁺ uptake (r²=0.98). When the Al and Ca²⁺ (⁴⁵Ca²⁺) uptake were plotted as a function of the Al³⁺ activity (or concentration of inorganic mononuclear Al), almost the same response curves were obtained for the -FA and +FA treatments. We conclude that FA-complexed Al was not available for root uptake and therefore could not affect the Ca²⁺ uptake. The competitive effect of Al on the Ca²⁺ uptake was also shown in a 5-week cultivation experiment, where the Ca concentration in shoots decreased at an AlCl₃ concentration of 0.3 mM. The effect of Al on H₂PO⁻₄ uptake was more complex. The P content in roots and shoots was not significantly affected, compared with the control, by cultivation for 5 weeks in a solution supplied with 0.3 mM AlCl₃, despite a reduction of the H₂PO⁻₄ concentration in the nutrient solution to about one-tenth. At this concentration Al obviously had a positive effect on H₂PO⁻₄ uptake. The presence of FA decreased ³²P-phosphate uptake by more than 60% during 24 h, and the addition of 0.15 or 0.3 mM AlCl₃ to these solutions did not alter the uptake of ³²P-phosphate.     

Plant cation-anion balance as affected by the ionic composition of the growing medium

Plants that remove an excess of cations over anions may cause soil acidification. The acidification potential of plants has been evaluated using solution culture techniques, but the influence of ionic composition of the medium on the plant cation-anion balance remains unclear. Our objective was to determine how electrolyte concentration and salt type affect the cation- anion balance of two test plants [barley (Hordeum vulgare L.) and kochia (Kochia scoparia L. Schrad.)]. Seedlings were grown in sand culture and irrigated with nutrient solution (Hoagland’s solution), which was adjusted to a range of electrolyte concentrations (target electrical conductivity of 7.5, 17.5 and 27.5 dS m⁻¹) using either chloride or sulphate salts. Increase in electrolyte concentration reduced yield of kochia, a salt-tolerant plant, by up to 38%. Total cation (Ca + Mg + K + Na) equivalents in kochia exceeded those of anions (Cl + S + P + NO₃) by 250 to 280 cmol_c kg⁻¹ of dry matter. Electrolyte concentration had no effect on the cation-anion balance of kochia, but excess cation values were significantly greater in the sulphate than in the chloride system. Kochia had a large content of water-soluble oxalate (194 to 226 cmol_c kg⁻¹), which was linearly related to the excess cation content. Growth of barley was severely restricted at the intermediate and high electrolyte concentrations. Cations exceeded anions by 21 to 59 cmol_c kg⁻¹ of barley dry matter. Excess cation content was greater in the sulphate than in the chloride medium, but electrolyte concentration did not have a consistent effect on the cation-anion balance. The small amounts of oxalate found in barley (0.9 to 2.6 cmol_c kg⁻¹) were insufficient to balance the cation excess.     

Calcium supply effects on wheat cultivars differing in salt resistance with special reference to leaf cytosol ion homeostasis

Salinity causes changes in cytosolic Ca²⁺, [Ca²⁺]_cyt, Na⁺, [Na⁺]_cyt and pH, pH_cyt, which induce specific reactions and signals. Reactions causing a rebalancing of the physiological homeostasis of the cytosol could result in plant resistance and growth. Two wheat cultivars, Triticum aestivum, Seds1 and Vinjett, were grown in nutrient solution for 7 days under moderate salinity (0 and 50 mM NaCl) with and without extra addition of 5 mM CaSO₄ to investigate the seedling‐ion homeostasis under salinity. In the leaf protoplasts [Ca²⁺]_cyt, [Na⁺]_cyt and pH_cyt were detected using acetoxymethyl esters of the ion‐specific dyes, Fura 2, SBFI and BCECF, respectively, and fluorescence microscopy. In addition, both cultivars were grown for 3 weeks at 0, 50 and 125 mM NaCl with, or without, extra addition of 5 mM CaSO₄ to detect overall Na⁺ and Ca²⁺ concentrations in leaves and salinity effects on dry weights. In both cultivars, salinity decreased [Ca²⁺]_cyt, while at extra Ca²⁺ supplied, [Ca²⁺]_cyt increased. The [Ca²⁺]_cyt increase was accompanied by increase in the overall Ca²⁺ concentrations in leaves and decrease in the overall Na⁺ concentration. Moreover, irrespective of Ca²⁺ treatment under salinity, the cultivars reacted in different ways; [Na⁺]_cyt significantly increased only in cv. Vinjett, while pH_cyt increased only in cv. Seds1. Even at rather high total Na⁺ concentrations, the cytosolic concentrations were kept low in both cultivars. It is discussed whether the increase of [Ca²⁺]_cyt and pH_cyt can contribute to salt tolerance and if the cytosolic changes are due to changes in overall Ca²⁺ and Na⁺ concentrations.     

Interactions of intracellular pH and intracellular calcium in primary cultures of rabbit corneal epithelial cells

Summary Homeostasis of intracellular calcium ([Ca⁺⁺]_i) and pH (pH_i) is important in the cell's ability to respond to growth factors, to initiate differentiation and proliferation, and to maintain normal metabolic pathways. Because of the importance of these ions to cellular functions, we investigated the effects of changes of [Ca⁺⁺]_i and pH_i on each other in primary cultures of rabbit corneal epithelial cells. Digitized fluorescence imaging was used to measure [Ca⁺⁺]_i with fura-2 and pH_i with 2′,7′-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF). Resting pH_i in these cells was 7.37±0.05 (n=20 cells) and resting [Ca⁺⁺]_i was 129±10 nM (n=35 cells) using a nominally bicarbonate-free Krebs Ringer HEPES buffer (KRHB), pH 7.4. On exposure to 20 mM NH₄Cl, which rapidly alkalinized cells by 0.45 pH units, an increase in [Ca⁺⁺]_i to 215±14 nM occurred. Pretreatment of the cells with 100 μM verapamil or exposure to 1 mM ethylene bis-(oxyethylenenitrilo)-tetraacetic acid (EGTA) without extracellular calcium before addition of 20 mM NH₄Cl did not abolish the calcium increase, suggesting that the source of the calcium transient was from intracellular calcium stores. On removal of NH₄Cl or addition of 20 mM sodium lactate, there were minimal changes in calcium even though pH_i decreased. Treatment of CE cells with the calcium ionophores, ionomycin and 4-bromo A23187, increased [Ca⁺⁺]_i, but produced a biphasic change in pH_i. Initially, there was an acidification of the cytosol, and then an alkalinization of 0.10 to 0.11 pH units above initial values. When [Ca⁺⁺]_i was decreased by treating the cells with 5 mM EGTA and 20 μM ionomycin, pH_i decreased by 0.35±0.02 units. We conclude that an increase in pH_i leads to an increase in [Ca⁺⁺]_i in rabbit corneal epithelial cells; however, a decrease in pH_i leads to minor changes in [Ca⁺⁺]_i. The ability of CE cells to maintain proper calcium homeostasis when pH_i is decreased may represent an adaptive mechanism to maintain physiological calcium levels during periods of acidification, which occur during prolonged eye closure.     

Leaf litter ash alkalinity and neutralisation of soil acidity

Soil acidification is a major factor limiting the sustainability of agricultural production systems throughout the world. Liming may not always be economically possible and therefore alternative methods or complementary methods of amelioration are required. Leaf litter collected from several tree species was examined for ash alkalinity (as an estimate of organic anion content) and ability to ameliorate an acid soil. Ash alkalinity measured by titration of the ash and excess cation values obtained by calculation as the difference between cation and anion content were correlated. Values obtained by the latter method ranged from 247 cmol _c kg^-1 for Melia azedarach (white cedar) to 36 cmol _c kg^-1 for Eucalyptus globoidea (white stringybark). There was a significant linear correlation between ash alkalinity and the Ca concentration in the litter. When added to an acid soil (pH 4.04 measured in 0.01 M calcium chloride) and incubated for 8 weeks, leaf litter raised the pH. Species differed markedly with Melia azedarach having the greatest effect. The increase in pH was proportional to the quantity of ash alkalinity (organic anions) added, expressed as calcium carbonate equivalents. Aluminium levels on the exchange complex were lowered by treatment with leaf litter through direct precipitation of a solid phase and again Melia azedarach litter was most effective. There was also indirect evidence of organo-Al complexes affecting the concentration of monomeric Al in soil treated with litter from Liquidambar styraciflua (liquidambar), Quercus robur (English oak) and Pinus radiata (radiata pine).     

Signaling Pathways in the Biphasic Effect of ANG II on Na<Superscript>+</Superscript>/H<Superscript>+</Superscript> Exchanger in T84 Cells

The effect of ANG II on pH_i, [Ca²⁺]_i and cell volume was investigated in T84 cells, a cell line originated from colon epithelium, using the probes BCECF-AM, Fluo 4-AM and acridine orange, respectively. The recovery rate of pH_i via the Na⁺/H⁺ exchanger was examined in the first 2 min following the acidification of pH_i with a NH₄Cl pulse. In the control situation, the pH_i recovery rate was 0.118 ± 0.001 (n = 52) pH units/min and ANG II (10⁻¹² M or 10⁻⁹ M) increased this value (by 106% or 32%, respectively) but ANG II (10⁻⁷ M) decreased it to 47%. The control [Ca²⁺]_i was 99 ± 4 (n = 45) nM and ANG II increased this value in a dose-dependent manner. The ANG II effects on cell volume were minor and late and should not interfere in the measurements of pH_i recovery and [Ca²⁺]_i. To document the signaling pathways in the hormonal effects we used: Staurosporine (a PKC inhibitor), W13 (a calcium-dependent calmodulin antagonist), H89 (a PKA inhibitor) or Econazole (an inhibitor of cytochrome P450 epoxygenase). Our results indicate that the biphasic effect of ANG II on Na⁺/H⁺ exchanger is a cAMP-independent mechanism and is the result of: 1) stimulation of the exchanger by PKC signaling pathway activation (at 10⁻¹² – 10⁻⁷ M ANG II) and by increases of [Ca²⁺]_i in the lower range (at 10⁻¹² M ANG II) and 2) inhibition of the exchanger at high [Ca²⁺]_i levels (at 10⁻⁹ – 10⁻⁷ M ANG II) through cytochrome P450 epoxygenase-dependent metabolites of the arachidonic acid signaling pathway.     

Short-term regulation of cytosolic Ca2+, cytosolic pH and vacuolar pH under NaCl stress in the charophyte alga Nitellopsis obtusa

Isolated characean internodal cells of Nitellopsis obtusa can be stored in artificial pond water for many days, but they cannot survive in 100mol m^?3 NaCl solution unless more than several mol m^?3 Ca²⁺ is added. Short-term effects of NaCl stress on the cytosolic concentration of Ca²⁺ ([Ca²⁺]_c), cytosolic pH (pH_c) and vacuolar pH (pH_v) were studied in relation to the external concentration of Ca²⁺ ([Ca²⁺]_e). Changes in [Ca²⁺]_c were measured with light emission from a Ca²⁺-sensitive photoprotein, semisynthetic fch-aequorin which had been injected into the cytosol. Both pH_c and pH_v were measured with double-barrelled pH-sensitive microelectrodes. When internodal cells were treated with 100 mol m^?3 NaCl (0–1 mol m^?3 NaCl (0.1 mol m^?3 [Ca²⁺]_e), [Ca²⁺]_c increased and then recovered to the original level within 60 min. The time course of the transient change in [Ca²⁺]_c was not influenced by the level of [Ca²⁺]_c (0.1 and 10 mol m^?3). In some cases, the transient increase in [Ca²⁺]_c was induced only by increasing external osmotic pressure with sorbitol. In response to treatment with 100 mol m^?3 NaCl (0.1 mol m^?3 [Ca²⁺]_c), pH_c decreased by 0.1–0.2 units after 10min but recovered after 30–60 min, while pH_v increased by 0.4–0.5 units after 2–50 min and tended to recover after 60 min. The initial changes in both pH_c and pH_v were suppressed when [Ca²⁺]_e was raised from 0.1 to 10mol m^?3. These results show that the charophyte alga Nitellopsis can regulate [Ca²⁺]_c, pH_c and pH_v under NaCl stress in the short term and that the protective effect of Ca²⁺ on salinity stress is apparently unrelated to perturbation of Ca²⁺ and pH homeostasis.     

Effects of aluminum chloride on the kinetics of rat cortex synaptosomal ATP diphosphohydrolase 9EC 3.6.1.5)

Aluminum chloride (AlCl₃), a neurotoxic compound, inhibited ATP diphosphohydrolase activity of synaptosomes obtained from cerebral cortex of adult rats. The metal ion significantly inhibited ATPase and ADPase activities of the enzyme at all concentrations tested in vitro (0.01, 0.05, 0.5, 5 and 10 mM) in the presence of 1.5 mM calcium. When tested in the absence of Ca²⁺, and with increasing amounts of Al³⁺, enzyme activity remained below basal levels, suggesting that the trivalent cation Al³⁺ is not a substitute for the divalent cation Ca²⁺ in ATP-Ca²⁺ and ADP-Ca²⁺ complexes. The Al³⁺ inhibition was competitive with respect to Ca²⁺. The enzyme inhibition was reversed by the addition of deferoxamine (DFO). NaF significantly inhibited ATP diphosphohydrolase activity, and this inhibition was reversed by the addition of Ca²⁺ to the medium. Such inhibition was not potentiated by AlF₄, which is an inhibitor of cation-transport ATPases.     

Response of soil chemistry to forest dieback after bark beetle infestation

We evaluated changes in the chemistry of the uppermost soil horizons in an unmanaged spruce forest (National Park Bohemian Forest, Czech Republic) for 3 years after dieback caused by a bark beetle infestation, and compared these changes with a similar undisturbed forest area. The soils below the disturbed forest received 2–6 times more elements via litter fall compared to the unaffected plot. The subsequent decomposition of litter and reduced nutrient uptake by trees resulted in a steep increase in soil concentrations of soluble N (NH₄-N, organic-bound N) and P forms in the disturbed plot. The average concentrations of NH₄-N and soluble reactive P increased from 0.8 to 4.4 mmol kg^?1 and from 0.04 to 0.9 mmol kg^?1, respectively, in the uppermost soil horizon. Decomposition of litter at the disturbed plot elevated soil concentrations of Ca²⁺, Mg²⁺ and K⁺, which replaced Al³⁺ and H⁺ ions from the soil sorption complex. Consequently, soil concentrations of exchangeable base cations increased from 120 to 200 meq kg^?1, while exchangeable Al³⁺ and H⁺ decreased 66 and 50 %, respectively, and soil base saturation increased from 40 to 70 %. The Al³⁺ liberation did not elevate concentrations of ionic Al in the soil solution, because most of the liberated Al³⁺ was rapidly complexed by dissolved organic carbon (DOC) and transformed to DOC–Al complexes. The chemical parameters investigated at the unaffected plot remained stable during the study.     

Parallel control of the inward-rectifier K+ channel by cytosolic free Ca2+ and pH inVicia guard cells

The influence of cytosolic pH (pH_i) in controlling K⁺-channel activity and its interaction with cytosolic-free Ca²⁺ concentration ([Ca²⁺]_i) was examined in stomatal guard cells ofVicia faba L. Intact guard cells were impaled with multibarrelled microelectrodes and K⁺-channel currents were recorded under voltage clamp while pH_i or [Ca²⁺]_i was monitored concurrently by fluorescence ratio photometry using the fluorescent dyes 2,7-bis (2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF) and Fura-2. In 10 mM external K⁺ concentration, current through inward-rectifying K⁺ channels (I_K,in) was evoked on stepping the membrane from a holding potential of –100 mV to voltages from –120 to –250 mV. Challenge with 0.3-30 mM Na+-butyrate and Na⁺-acetate outside imposed acid loads, lowering pH_i from a mean resting value of 7.64 ± 0.03 (n = 25) to values from 7.5 to 6.7. The effect on pH_i was independent of the weak acid used, and indicated a H⁺-buffering capacity which rose from 90 mM H⁺/pH unit near 7.5 to 160 mM H⁺/pH unit near pH_i 7.0. With acid-going pH_i, (I_K,in) was promoted in scalar fashion, the current increasing in magnitude with the acid load, but without significant effect on the current relaxation kinetics at voltages negative of –150 mV or the voltage-dependence for channel gating. Washout of the weak acid was followed by transient rise in pH_i lasting 3–5 min and was accompanied by a reduction in (I_K,in) before recovery of the initial resting pH_i and current amplitude. The pH_i-sensitivity of the current was consistent with a single, titratable site for H⁺ binding with a pK_a near 6.3. Acid pH_i loads also affected current through the outward-rectifying K⁺ channels (I_K,out) in a manner antiparallel to (I_K,in) The effect on I_K, out was also scalar, but showed an apparent pK_a of 7.4 and was best accommodated by a cooperative binding of two H⁺. Parallel measurements showed that Na⁺-butyrate loads were generally without significant effect on [Ca²⁺]_i, except when pH_i was reduced to 7.0 and below. Extreme acid loads evoked reversible increases in [Ca²⁺]_i in roughly half the cells measured, although the effect was generally delayed with respect to the time course of pH_i changes and K⁺-channel responses. The action on [Ca²⁺]_i coincided with a greater variability in (I_K,in) stimulation evident at pH_i values around 7.0 and below, and with negative displacements in the voltage-dependence of (I_K,in) gating. These results distinguish the actions of pH_i and [Ca²⁺]_i in modulating (I_K,in) they delimit the effect of pH_i to changes in current amplitude without influence on the voltage-dependence of channel gating; and they support a role for pH_i as a second messenger capable of acting in parallel with, but independent of [Ca²⁺]_i in controlling the K⁺ channels.Abbreviations BCECF 2,7-bis (2-carboxyethyl)-5(6)-carboxy fluorescein - [Ca²⁺]_i cytosolic free Ca²⁺ concentration - g_K ensemble (steady-state) K⁺-channel conductance - I_K,out, I_K,in outward-, inward-rectifying K⁺ channel (current) - IN current-voltage (relation) - Mes 2-(N-morpholinolethanesulfonic acid - pH_i cytosolic pH - V membrane potential     

Is the Ca:Al ratio superior to pH,Ca or Al concentrations of soils in accounting for the distribution of plants in deciduous forest?

The distributions of vascular plants in south Swedish deciduous forests were related to exchangeable (exc) and soil solution concentrations of H⁺ (pH), Ca, Al and the Ca:Al ratios within these fractions. Topsoils (0–5 cm) of 172 sites with a pH_KCl of 3.2–3.9 (corresponding to 3.7–4.4 in soil solution) were used. In the soil solution both total Al_t and quickly reacting Al_r were determined. Exchangeable concentrations were generally well related to plant distributions, the highest correlation coefficients usually being given by pH_KCl>Ca_exc>Al_exc.>(Ca:Al)_exc. The (Ca:Al)_exc ratio was clearly inferior. Out of the soil solution variables studied, Ca concentration, followed by pH, was best correlated with plant distributions, Al_t, Al_r, and the Ca:Al ratios having similar and lower coefficients. It is concluded that the use of Ca:Al ratios as a general measure of Al toxicity in controlling plant distributions is rather problematic. It seems difficult to apply evidence for Ca-Al interactions from solution culture experiments to field conditions when measured as exchangeable or soil solution concentrations of the soil.     

Cytosolic acidification leads to Ca mobilization from intracellular stores in single and populational parietal cells and platelets

Regulatory relationship and gain control between cytosolic free Ca²⁺ concentration (Ca_i) and cytosolic pH (pH_i) were evaluated by two different cell types, gastric parietal cells, and blood platelets. Studies were carried out in both single cells and populations of cells, using Ca²⁺-indicative probe fura-2 (1-(2-(5′-carboxyoxazol-2′-yl)-6-aminobenzofuran-5-oxy)-2-(2′-amino-5′-methylphenoxy) ethane-N,N,N′,N′-tetraacetic acid) and pH-indicative probe BCECF (2′,7′-bis(carboxyethyl) carboxyfluorescein). Stimulation of single and populational parietal cells and platelets with gastrin and thrombin, respectively, resulted in an increase in Ca_i. In both populational cell types, an initial change in pH_i during agonist stimulation occurred almost simultaneously with the mobilization of Ca²⁺; an initial transient decrease in pH_i was followed by a slower increase in pH_i above the prestimulation level. When populational platelets were preloaded with the Ca²⁺ chelator BAPTA (1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′tetraacetic acid), the thrombin-induced initial large increase in Ca_i was apparently inhibited, whereas the pH_i decrease induced by thrombin was not altered. This suggests that the initial Ca_i change is not a prerequisite for the pH_i change. The effect of pH_i on Ca_i was examined next. In both single and populational cell types, application of the K⁺-H⁺ ionophore nigericin, which induced a transient decrease in pH_i, led to the release of Ca²⁺ from intracellular stores. In single parietal cells double-labeled with fura-2 and BCECF, a temporal decrease in pH_i preceded the rise in Ca_i after stimulation with nigericin. A decrease in pH_i, and an increase in Ca_i occurred at 1.5 and 4 s, respectively. In single parietal cells, replacement of medium Na⁺ with N-methyl- -glucamine (NMG⁺), which also induced a decrease in pH_i, resulted in repetitive Ca²⁺ spike oscillations. The source of Ca²⁺ utilized for the Ca²⁺ oscillation that was induced by NMG⁺ originated from the agonist-sensitive pool. Thus, several maneuvers, which were capable of decreasing pH_i, led to an increase in Ca_i. Cytosolic acidification may be a part of the trigger for Ca²⁺ mobilization from intracellular stores in both parietal cells and platelets.     

Aluminium effects on transmembrane potential in cells of fibrous roots of sugar beet

The effect of aluminium (Al) on the electrical transmembrane potential of epidermal and outer cortical root cells of intact seedlings of sugar beet (Beta vulgaris L. cv. Monohill) was studied. The potential difference to the surrounding medium was recorded with microelectrodes inserted into the vacuoles (PD_v) and cytoplasm (PD_c) of intact roots. Both long-term effects of AlCl₃ (100, μM present during cultivation) and immediate effects of AlCl₃ (10, 50, or 100 μM present in the assay medium), were measured. The effect of Al was measured at pH 4.0, 5.0 and 6.5 in order to obtain information on the toxicity of different Al forms existing at different pH values. Low pH and/or the presence of AlCl₃ during cultivation caused large depolarizations of the PD_v. Since the immediate effect of 2,4-dinitrophenol (DNP) on the resting potential of cells from Al-cultivated plants was negligible, it is likely that Al affects the metabolic component of the transmembrane potential. Aluminium also had an immediate effect on the PD in root cells of plants cultivated without Al. Addition of 10 or 50 μM Al to the assay medium caused hyperpolarization of PD_v in the presence of 0.5 mM Ca²⁺ at all pH values studied, depolarization of PD_c at pH 6.5, and hyperpolarization of PD_c at lower pH. At 1 mM Ca²⁺, or in the presence of K⁺ (≥ 2 mM), however, the same Al concentrations had little effect on PD_c. The strongest depolarizing effects of 10 or 50 μM Al in short-term treatments were obtained at pH 6.5, and were probably due to the soluble species Al(OH)₃, which is more frequent at pH 6.5 than at a lower pH. Addition of 50 μM Al caused alkalinization of the root medium at pH 6.5, but not at pH 4.0. Therefore, it is possible that Al at pH 6.5 is bound to, or translocated across, the membrane without the accompanying hydroxide ions. It is likely that most of the Al is bound to the root cells, since removal of Al from the buffer surrounding the roots did not cause the changed PD values to return to the original values. Aluminium also interacts with effects of Ca²⁺ and K⁺ on the membrane potential, since changes in PD, induced by changes in concentrations of Ca²⁺ and K⁺ are different in the absence and presence of Al.     

The effects of low concentrations of aluminium on the growth and uptake of nitrate-N by white clover

Summary The effects of aluminium (Al³⁺) at concentrations of 0, 25, 50 and 100 μM on the growth of white clover, dependent upon N supplied as NO ₃ ⁻ , were examined in flowing solution culture. Plants were established with a normal nutrient supply for 7 weeks and then grown with carefully controlled pH (at 4.5) and P concentrations, and with 0, 25, 50 or 100 μM Al³⁺ for a further three weeks. There were rapid visual effects (i.e. symptoms of P deficiency and reduction in root extension) and the dry weights of shoots and roots were reduced at 50 and 100 μM. Less than 10% of Al absorbed from solution was transported to the shoots. The uptake of P, and its transport between roots and shoots, were reduced in plants grown with Al. The uptake of NO ₃ ⁻ stopped immediately after the introduction of 50 or 100 μM Al, and was significantly reduced at 25 μM after three weeks. During a second phase of the experiment, plants previously grown at 0, 25, 50 and 100 μM Al, were grown for a further 2 weeks either with NO ₃ ⁻ (with and without 50 μM Al³⁺) or without NO ₃ ⁻ but with inoculation by Rhizobia (and with or without 50 μM Al³⁺). The effects of the previous treatments with Al on N uptake were small during the second phase, but uptake by all plants was restricted when Al was present. Inoculation did not result in nodulation in the second phase when Al³⁺ was present in the solution, but Al already in the plant from the first phase did not prevent nodulation in the absence of Al during the second phase.     

Intracellular pH,intracellular free Ca,and junctional cell-cell coupling

Summary Intracellular pH (pH _i ) and intracellular Ca²⁺ ([Ca²⁺] _i ) were determined inChironomus salivary gland cells under various conditions of induced uncoupling. pH _i was measured with aThomas-type microelectrode, changes in [Ca²⁺] _i and their spatial distribution inside the cell were determined with the aid of intracellularly injected aequorin and an image intensifier-TV system, and cell-to-cell coupling was measured electrically. Treatments with NaCN (5mm), DNP (1.2mm), or ionophore A23187 (2m) caused fall in junctional conductance (uncoupling) that was correlated with [Ca²⁺] _i elevation, as was shown before (Rose & Loewenstein, 1976,J. Membrane Biol. 28:87) but not with changes in pH _i : during the uncoupling induced by CN, the pH _i (normally 7.5) decreased at most by 0.2 units; during the uncoupling induced by the ionophore, pH _i fell by 0.13 or rose by 0.3; and in any one of these three agents' uncouplings, the onset of uncoupling and recovery of coupling were out of phase with the changes in pH _i . Intracellular injection of Ca-citrate or Ca-EGTA solutions buffered to pH 7.2 or 7.5 produced uncoupling with little or no pH _i change when their free [Ca²⁺] _i was >10^–5 m. On the other hand, such a solution at pH 4, buffered to [Ca²⁺]<10^–6 m, lowered pH _i to 6.8 but produced no uncoupling. Thus, a decrease in pH _i is not necessary for uncoupling in any of these conditions. In fact, uncoupling ensued also during increase in pH _i : exposure to NH₄HCO₃ or withdrawal of propionate following exposure to a propionate-containing medium caused pH _i to rise to 8.74, accompanied by [Ca²⁺] _i elevation and uncoupling at pH _i >7.8.Cell acidification itself can cause elevation of [Ca²⁺] _i : injection (iontophoresis) of H⁺ invariably caused [Ca²⁺] _i elevation and uncoupling. These effects were produced also by an application of H⁺-transporting ionophore Nigericin at extracellular pH 6.5 which caused pH _i to fall to 6.8. Exposure to 100% CO₂ produced a fall in pH _i , associated in 10 out of 25 cases with [Ca²⁺] _i elevation and, invariably, with uncoupling. The absence of a demonstrable [Ca²⁺] _i elevation in a proportion of these trials is attributable to depression in Ca²⁺-measuring sensitivity; inin vivo tests, detection sensitivity for [Ca²⁺] _i by aequorin was found to be depressed by the CO₂ treatment. Upon CO₂ washout, pH _i and coupling recovered, but onset of recoupling set in at pH _i as low as 6.32–6.88, generally lower than at the pH _i at which uncoupling had set in. Exposure to 5% CO₂ lowered pH _i on the average by 0.3 and depressed coupling (in initially poorly coupled cells). After CO₂-washout, pH _i and coupling recovered. During the recovery phase [Ca²⁺] _i was elevated, an elevation associated with renewed uncoupling or decrease in rate of recoupling. The results are discussed in connection with possible regulatory mechanisms of junctional permeability.     

Effects of External pH Variations on Brain Presynaptic Sodium and Calcium Channels; Repercussion on the Evoked Release of Amino Acid Neurotransmitters

The effects of external pH (pH _out) variations on the Na⁺ and on the Ca²⁺ dependent fractions of the evoked amino acid neurotransmitter release were separately investigated, using GABA as a model transmitter. In [³H]GABA loaded mouse brain synaptosomes, the external acidification (pH _out6.0) markedly decreased the Na⁺ dependent fraction of [³H]GABA release evoked by veratridine (10 M) in the absence of external Ca²⁺, as well as the Ca²⁺ dependent fraction of [³H]GABA release evoked by high (20 mM) K⁺ in the absence of external Na⁺. The depolarization-induced elevation of [Na _i ] (monitored in synaptosomes loaded with the Na⁺ indicator dye, SBFI) and the depolarization-induced elevation of [Ca _i ] (monitored in synaptosomes loaded with the Ca²⁺ indicator dye fura-2) were also markedly decreased at pH _out 6. On the contrary, the external alkalinization (pH _out 8) facilitated all the above responses. A slight increase of the baseline release of the [³H]GABA was observed when pH _out was changed from 7.4 to 8. This effect was only observed in the presence of Ca²⁺. pH _out changes from 7.4 to 6 or to 7 did not modify the baseline release of the transmitter. All the effects of pH _out variations on [³H]GABA release were independent on the presence of HCO-₃. It is concluded that external H⁺ regulate amino acid neurotransmitter release by their actions on presynaptic Na⁺ channels, as well as on presynaptic Ca²⁺ channels.     

Ca2+ responses to acetylcholine and adenosine triphosphate in the otocyst of chick embryo

The action of acetylcholine and adenosine triphosphate (ATP) on cytoplasmic Ca²⁺ concentration ([Ca²⁺]_i) was studied in the otocyst epithelium of embryonic day 3 chicks with Ca²⁺-sensitive fluorescence measurements. Increases in [Ca²⁺]_i were evoked by the bath application of acetylcholine (1 μM or higher). The rise in [Ca²⁺]_i was due to the release of Ca²⁺ from intracellular Ca²⁺ stores, since the Ca²⁺ response occurred even in a Ca²⁺-free medium. The Ca²⁺ response to acetylcholine was mediated by muscarinic receptors. Atropine of 1 μM abolisehd the response to 10 μM acetylcholine; muscarine and carbamylcholine (100 μM each) evoked Ca²⁺ rises. Increases in [Ca²⁺]_i were also evoked by the bath application of ATP (10 μM or higher). The Ca²⁺ rise by ATP was evoked even in a Ca²⁺-free medium. Adenosine (500 μM) did not cause any Ca²⁺ response. Suramin and reactive blue 2 (200 μM each) completely blocked the Ca²⁺ response to 500μM ATP. Uridine triphosphate (500 μM) caused comparable Ca²⁺ responses with those to 500 μM ATP. These results suggested the involvement of P_2U purinoceptors. The potentiation of Ca²⁺ rise was observed when acetylcholine and ATP were co-applied at submaximal concentrations (10 μM and 100 μM, respectively). We conclude that undifferentiated cells in the otocyst epithelium have CaCa²⁺ mobilizing systems activated by acetylcholine and ATP. © 1995 John Wiley & Sons, Inc.