Modification of the culture medium to produce aluminum toxicity in cell suspensions of coffee (Coffea arabica L.)

Coffee (Coffea arabica) plants are usually grown in soils containing high levels of organic materials. Under these conditions, aluminum (Al) is toxic because of the acidic nature of the soils. Al is the most abundant metal found in the earth's crust and occurs in a number of different forms in soil. In acid soils, Al toxicity is a global problem that limits crop productivity. A major problem in obtaining cellular lines displaying Al tolerance in culture is the composition of the medium. In the experiments presented here, we modified the composition of the culture medium for a C. arabica cell line to produce Al toxicity. Murashige-Skoog media was used, complete (MS) and half ionic strength (MSHIS), at either pH 5.8 or 4.3. We found that MSHIS and pH 4.3 provided the optimal conditions to obtain Al toxicity as measured by the ability to grow in a range of Al concentrations (25-1,000 µM). The lethal dose (LD₅₀) under these conditions was 25 µM. The concentrations of free Al in the culture medium were corroborated by the fluorescent compound Morin. Al was found to enter the cell after 30 min, and the signal was then retained for up to 2 h.     

Aluminium,boron and molybdenum availability and uptake by rice in acid sulfate soils

Although Al toxicity is believed to be a problem in acid sulfate soils cropped to rice (Oryza, sativa L.), little is known about the behavior of other trace metals such as B and Mo in these soils. The objectives of this study were to measure the availability of Al, B, and Mo in these soils, to determine what governs the availability of these metals and to investigate the relationships between metal availability and uptake by rice. Metal availability and uptake by rice were evaluated in 134 flooded acid sulfate soils in the Central Plains region of Thailand and in a growth chamber study using 50 of the same soils. Soil and plant metal analyses were conducted at the panicle differentiation stage of growth in both studies and in the soil prior to transplanting in the growth chamber study. Metal activities were determined with GEOCHEM. The mineral phases believed to be governing Al³⁺ activities were jurbanite under low pH conditions and amorphous Al(OH)₃ at high pH. The Al chemistry is believed to be intimately linked to the redox-pH cycle, which is driven by the monsoonal climate. Mortality of rice associated with Al toxicity was observed under field and growth chamber conditions. Interference in P uptake and/or assimilation was believed to be the mechanism of Al toxicity. Activities of B(OH) ₄ ^– and B(OH) ₃ ⁰ were found to be highly correlated to pH and ionic strength, respectively, with the latter being the dominant B ion found in these soils. Activities of MoO ₄ ^2– were positively correlated to pH and appeared to be controlled by wulfenite. Leaf Mo contents were found to be positively correlated with MoO ₄ ^2– activity.     

Simulating the Mineral Environment of Aluminium Toxic Soils in Plant Cell Culture

The development of a medium for studying aluminium toxicityin plant cell cultures is described. To prevent the precipitationof Al added to the standard cell culture medium, it was necessaryto lower the phosphate concentration from 1250 mmol m^–3to10 mmol m^–3, and the pH from 5.8 to 4-0. Two additionalmodifications were the use of unchelated iron and a reductionin the calcium concentration from 3.0 mol m^–3 to 0.1 molm^–3. Since the gelling properties of agar are inhibitedat pH 4.0, cells were cultured on filter paper supported bypolyurethane foam sturated with liquid medium. The only limitationto the growth of plated Nicotiana plumbaginifolia Viv. cellson the modified medium was the reduced phosphate concentration.This was partly overcome by ‘preloading’ the cellswith phosphate prior to each experiment. In addition, the filterpaper with adhering cells was transferred to fresh medium everysecond day to replenish phosphate, and to re-establish the initialpH of4.0 (which otherwise drifts upward). With the modifiedmedium, Al toxicity was observed in plated N. plumbaginifoliacells at both 200 mmol m^–3 and 400 mmol m^–3 Al.There was no toxicity at these Al concentrations when the normalphosphate concentration or pH were restored to the modifiedmedium. Partial alleviation of Al toxicity occurred with restorationof the normal calcium concentration or chelated iron. Chelationof Al with citrate or EDTA also mitigated Al toxicity. In additonto Al toxicity, the modified medium should also prove usefulfor studying other metal toxicities in plant cell culture. Key words: Al toxicity, Cell culture, Nicotiana plumbaginifolia     

改良剂对酸性土壤上伴矿景天铝毒缓解作用及镉锌吸收性的影响

为探究不同改良剂对酸性土壤铝(Al)胁迫条件下镉(Cd)锌(Zn)超积累植物伴矿景天Sedum plumbizincicola生长以及镉和锌吸取修复效率的影响,分别添加不同种类改良剂(钙镁磷肥(CMP)、MgCO3、KH2PO4)和不同浓度CMP进行温室盆栽试验。结果表明,CMP能够一定程度上提高土壤pH值并降低土壤交换性Al的浓度,MgCO3能够显著提高土壤pH值和降低土壤交换性Al的浓度,KH2PO4能够降低土壤中交换性Al浓度但未改变土壤pH值。施用适量的CMP(9.39 mg/kg)能够提高伴矿景天生物量和Cd、Zn吸取修复效率,用量过高会抑制伴矿景天生长和Cd、Zn修复效率;施用MgCO3可增大伴矿景天生物量和Cd、Zn修复效率,施用KH2PO4反而抑制了伴矿景天生长。酸性土壤上施用适量的CMP和MgCO3能够缓解伴矿景天的铝毒作用,维持较高的重金属吸收效率。     

The effect of aluminium in Atlantic salmon (Salmo salar) with special emphasis on alkaline water

Atlantic salmon (Salmo salar) parr were exposed to aluminium under both steady state and non-steady state chemical conditions in alkaline water. Under alkaline (pH 9.5) steady state conditions, approximately 350 microg Al l(-1) (predominantly aluminate, Al(OH)(4)(-)) had no acute toxic effect on the salmon. The fish, however, showed a physiological response after 3 weeks of exposure ( approximately 300% increase in blood glucose concentration, about 30% increase in blood haematocrit, and about 15% decrease in plasma Cl(-) concentration). No increase in toxicity was evident under non-steady state conditions, i.e. lowering Al solubility as pH was lowered from 9.5 to 7.5. The results indicate that the toxicity of the aluminate ion (Al(OH)(4)(-)) is low, and particularly lower than the corresponding toxicity of cationic Al hydroxides. The effects observed in fish exposed to Al-rich water at pH 9.5 were counteracted as Al solubility was decreased by lowering pH to 7.5. This is contrary to previous observations where Al solubility has been lowered by increasing pH from 5.0 to 6.5.     

Soil pH Effects on Uptake of Cd and Zn by Thlaspi caerulescens

For phytoextraction to be successful and viable in environmental remediation, strategies that can optimize plant uptake must be identified. Thlaspi caerulescens is an important hyperaccumulator of Cd and Zn, whether adjusting soil pH is an efficient way to enhance metal uptake by T. caerulescens must by clarified. This study used two soils differing in levels of Cd and Zn, which were adjusted to six different pH levels. Thlaspi caerulescens tissue metal concentrations and 0.1 M Sr(NO₃)₂ extractable soil metal concentrations were measured. The soluble metal form of both Cd and Zn was greatly increased with decreasing pH. Lowering pH significantly influenced plant metal uptake. For the high metal soil, highest plant biomass was at the lowest soil pH (4.74). The highest shoot metal concentration was at the second lowest pH (5.27). For low metal soil, due to low pH induced Al and Mn toxicity, both plant growth and metal uptake was greatest at intermediate pH levels. The extraordinary Cd phytoextraction ability of T. caerulescens was further demonstrated in this experiment. In the optimum pH treatments, Thlaspi caerulescens extracted 40% and 36% of total Cd in the low and high metal soils, respectively, with just one planting. Overall, decreasing pH is an effective strategy to enhance phytoextraction. But different soils had various responses to acidification treatment and a different optimum pH may exist. This pH should be identified to avoid unnecessarily extreme acidification of soils.     

Fluorescence characterization of the interaction of Al3+ and Pd2+ with Suwannee River fulvic acid in the absence and presence of the herbicide 2,4-dichlorophenoxyacetic acid

In an effort to understand the role of environmental metal ions in the interaction of charged pesticides with humic substances, a fluorescence study of the interaction of the widely-used herbicide 2,4-dichlorophenoxyacetic acid (DCPAA) with Al(3+) and Pd(2+) and Suwannee River fulvic acid (SRFA) was undertaken. Initial fluorescence experiments on binary solutions clearly indicated that both Al(3+) and Pd(2+) strongly interact with both SRFA and DCPAA when alone in solution with the metal ion. Titrations of SRFA with Al(3+) at pH values of 4.0, 3.0 and 2.0 revealed decreased degrees of fluorescence emission enhancement (at lambda(emission, max)=424 nm) with decreasing pH, consistent with the expected loss of rigidity in the SRFA-Al(3+) complexes formed as pH is lowered. In contrast, titrations of SRFA with Pd(2+) at all of these pH values resulted in significant fluorescence quenching. Al(3+) additions to solutions of DCPAA at pH values above the pK(a) (2.64) of DCPAA resulted primarily in significant changes in the wavelength of maximum emission (without significant quenching or enhancement of emission intensity), while Pd(2+) additions to DCPAA solutions resulted primarily in very significant fluorescence quenching. The DCPAA fluorescence results strongly support the formation of an Al(3+)-DCPAA complex at pH values above the pK(a) of DCPAA. The fluorescence results obtained for solutions of Pd(2+) and DCPAA are best explained by a collisional quenching mechanism, that is, energy transfer from excited DCPAA molecules to Pd(2+) following the collision of these two species in solution. Excitation-emission matrix plots obtained on ternary solutions (at environmentally-relevant pH 4.0) containing SRFA, DCPAA and metal ions (i.e., either Al(3+) or Pd(2+)) provides evidence (especially for systems containing Al(3+)) for the existence of ternary complexes between fulvic acid species, the herbicide DCPAA and metal ion, suggesting (at least at pH 4.0, where the predominant DCPAA species is negatively-charged) that metal ions may function to "bridge" negatively-charged fulvic acids to negatively-charged pesticides.     

Aluminum Inhibition of the Inositol 1,4,5-Trisphosphate Signal Transduction Pathway in Wheat Roots: A Role in Aluminum Toxicity?

In crop plants, aluminum (Al) rhizotoxicity is a major problem worldwide; however, the cause of Al toxicity remains elusive. The effects of Al on the inositol 1,4,5-trisphosphate (Ins[1,4,5]P3)-mediated signal transduction pathway were investigated in wheat roots. Exogenously applied Al (50 [mu]M) rapidly inhibited root growth (<2 hr) but did not affect general root metabolism. An Ins(1,4,5)P3 transient was generated in root tips, either before or after exposure to Al for 1 hr, by treating the roots with H2O2 (10 mM). Background (unstimulated) levels of Ins(1,4,5)P3 were similar in both Al-treated and Al-untreated root apices. However, H2O2-stimulated levels of Ins(1,4,5)P3 in root apices showed a significant (>50%) reduction after Al exposure in comparison with untreated controls, indicating that Al may be interfering with the phosphoinositide signaling pathway. When phospholipase C (PLC) was assayed directly in the presence of Al or other metal cations in microsomal membranes, AlCl3 and Al-citrate specifically inhibited PLC action in a dose-dependent manner and at physiologically relevant Al levels. Al exposure had no effect on inositol trisphosphate dephosphorylation or on a range of enzymes isolated from wheat roots, suggesting that Al exposure may specifically target PLC. Possible mechanisms of PLC inhibition by Al and the role of Ins(1,4,5)P3 in Al toxicity and growth are discussed. This study provides compelling evidence that the phytotoxic metal cation Al has an intracellular target site that may be integrally involved in root growth.     

Nitrate and ammonium uptake and solution pH changes for Al-tolerant and Al-sensitive sorghum (Sorghum bicolor) genotypes grown with and without aluminium

Plant tolerance to Al toxicity has been associated with differential nitrate and ammonium uptake and solution pH changes. Sorghum [Sorghum bicolor (L.) Moench] genotypes with tolerance (SC283) and sensitivity (ICA-Nataima) to Al toxicity were grown with different nitrate/ammonium ratios (39:1, 9:1, and 3:1) at 0 and 300 μM Al to determine genotypic differences in nitrate and ammonium uptake, changes in nutrient solution pH, and relationships of these traits to Al toxicity tolerance in the genotypes. ICA-Nataima had greater reductions in nitrate and ammonium uptake than SC283 when plants were grown with Al, but SC283 had higher nitrate and ICA-Nataima had higher ammonium uptake when plants were grown without Al. Differences in nitrate and ammonium uptake were associated with changes in solution pH; pH decreased as long as ammonium was in solution and increased when ammonium was depleted from solution. Greater changes in solution pH occurred when plants were grown with 39:1 compared to 9:1 and 3:1 nitrate/ammonium ratios. Solution pH values were lower when plants were grown with than without Al. The genotypes maintained their relative differences in Al toxicity tolerance when plants were grown separately or together in the same container with Al and with different nitrate/ammonium ratios.     

Protective mechanism of sodium molybdate against the acute toxicity of cadmium in rats. II. Prevention of cytoplasmic acidification

In order to clarify the protective mechanism of sodium molybdate against the acute toxicity of cadmium chloride in rat, the effect of in vivo sodium molybdate pretreatment on the cytotoxic action of cadmium in isolated hepatocytes was studied. The cytosolic pH of hepatocytes isolated from untreated rats immediately decreased with incubation in either neutral Hank's balanced salt solution (HBS), pH 7.4, containing 5 µM cadmium chloride minimum or acidic HBS (pH 7.1, 6.8, 6.5, and 6.2). The presence of 5 µM cadmium in HBS adjusted to pH 7.1 aggravated cytosalic acidification induced by the acidic medium alone. Cell viability of hepatocytes incubated in HBS at pH 6.2 was significantly reduced as compared to that of control cells in HBS at pH 7.4, but the presence of cadmium in the acidic HBS had no aggravating action against such a toxic action of the acidic medium although cellular uptake of the metal in the medium increased, as compared to that in HBS at pH 7.4. Molybdenum pretreatment alleviated cytoplasmic acidification induced by the treatment with HBS at pH 7.4 or 7.1 containing cadmium or by extracellular acid load wothout cadmium. This pretreatment also prevented the loss of cell viability induced by the treatment with HBS at pH 6.2 but could not attenuate that when cadmium was present in the medium.These facts suggest that molybdenum pretreatment alleviated the acute toxicity of cadmium in rat by preventing cytoplasmic acidification caused by the harmful metal.     

A rapid assay for aluminium phytotoxicity at submicromolar concentrations

Investigations of Al phytotoxicity, including the identification of the Al species responsible for toxicity, require a rapid assay procedure employing very low concentrations of Al and a chemically simple rooting medium. Root elongation in newly germinated red clover ( Trifolium pratense L. cv. Kenland) was inhibited by submicromolar concentrations of Al. Ca²⁺ at concentrations of at least 0.2 m M was essential for optimal elongation in control seedlings. Ca²⁺ also relieved Al toxicity with the net effect that maximum reduction of elongation by 1 μ M Al was achieved at 0.2 m M Ca²⁺. Elongation in control seedlings was at least 90% of maximum from pH 4.5 to 5.7. Increases in pH relieved Al toxicity so that maximum sensitivity to 1 μ M Al occurred at pH 4.7. As a consequence of these experiments and other considerations we chose for our basic assay a medium composed of 0.2 m M CaSO₄ adjusted to pH 4.5 with H₂SO₄, variously supplemented with Al₂(SO₄)₃.
Day-old seedlings were incubated in this aerated medium in the dark at 23°C for one day. No additions of other solutes increased the sensitivity of the assay, but amelioration of Al toxicity was effected by Mg²⁺, F^-, phosphate and citrate. Increases in ionic strength per se had comparatively little effect on the toxic effects of Al. Two barley cultivars ( Hordeum vulgare L. cv. Dayton and Kearney) and two wheat cultivars ( Triticum aestivum L. cv. Hart and Thorne) known to differ in sensitivity to Al were reliably separated at submicromolar Al concentrations by the assay procedure, which was slightly modified. Suggestions for the improvement of the assay and for applications to future research are offered.     

Effect of pH and phosphate on soluble soil aluminium and on growth and composition of kikuyu grass

Summary Kikuyu (Pennisetum clandestinium Hochst) grew relatively poorly on the Wollongbar krasnozem at soil pH values below 4.36. At these low pH values dry matter yields were increased by raising the pH or by application of high rates of phosphate. Both treatments decreased the concentration of soluble soil-Al on which the concentration of Al in tops was linearly dependent (r=0.95). The inverse relationship found between plant growth and Al concentration, when present in excess of ∼1.5 μg/g soil and ∼90 μg/g tops, is suggestive of Al toxicity. However, at Al concentrations causing severe yield reductions, the Ca concentration in kikuyu tops was approaching deficiency levels. The Al-Ca antagonism was further demonstrated by the reduction in Ca-uptake caused by increased concentrations of soluble soil-Al under constant conditions of exchangeable Ca and of pH. The yield-reducing effects of Al toxicity per se and Al-induced Ca deficiency are therefore confounded.     

RESPONSE OF CHLORELLA PYRENOIDOSA TO ALUMINUM AND LOW pH12

Chlorella pyrenoidosa, a green alga which has no measurable Ca requirement, tolerated much higher Al concentrations in solution than higher plants which require considerable Ca. This alga also gave significant positive yield responses to Al concentrations between 1.5 and 12 ppm (added at pH 4.6). The positive Al response was not attributable to V, Cr, Ni, Co, W, or Ti contaminants in the Al salt. A strain of C. pyrenoidosa having even greater Al tolerance was isolated, by subjecting the original Strain I (Fitzgerald) culture to increasing Al stress. This strain, I-Al, grew in stagnant cultures containing 48 ppm Al at an initial pH of 4.2. Its yield also was not significantly decreased by 48 ppm Al in aerated cultures when both inoculum and solution pH were 4.6. Under the same conditions the original Strain I organism was injured by 3 and 6 ppm Al and was killed by 12 ppm. Algal strains which differ in Al tolerance may be useful in (1) studies on the mechanism of Al toxicity and mineral nutrition in general; and (2) in raising the pH, precipitating Al, and thereby detoxifying Al-containing acid mine drainage water and commercial wastes.     

Aluminium resistance requires resistance to acid stress: a case study with spinach that exudes oxalate rapidly when exposed to Al stress

Spinach is a vegetable with a high oxalate concentration in its tissues. Oxalate efflux from spinach (Spinacia oleracea L. cv. Quanneng) roots was rapidly stimulated (within 30 min) by aluminium (Al) treatment. The efflux was constant within 6 h, but increased with increasing Al concentration. The efflux was confined to the root tip (0-5 mm), which showed a 5-fold greater efflux than the root zone distal to the tip (5-10 mm). Oxalate efflux could not be triggered by treatment with the trivalent cation lanthanum or by phosphorus deficiency, indicating that the efflux was specific to the Al treatment. All this evidence suggested that spinach possesses Al-resistance mechanisms. However, spinach was found to be as sensitive to Al toxicity as the Al-sensitive wheat line ES8, which had no Al-dependent organic acids efflux. The Al accumulated in the apical 5 mm of the roots of spinach which was also similar to that in the Al-sensitive wheat after 24 h treatment with 50 microM AlCl(3), indicating a non-exclusion mechanism. In addition, root elongation in spinach was significantly inhibited at pH 4.5, compared with that at pH 6.5. Based on this evidence, it is concluded that the sensitivity to acid stress in spinach could mask the potential role for oxalate to protect the plant roots from Al toxicity.     

The amelioration of aluminium toxicity by silicon in higher plants: Solution chemistry or an in planta mechanism?

Aluminium (Al) toxicity is a very important factor limiting the growth of plants on acidic soils. Recently, a number of workers have shown that, under certain conditions, silicon (Si) can ameliorate the toxic effects of A1 in hydroponic culture. The mechanism of the amelioration is unclear, but three suggestions have been put forward: Si‐induced increase in solution pH during the preparation of hydroponic solutions; reduced availability of Al due to the formation of hydroxyaluminosilicate (HAS) species in those solutions during plant growth; or in planta detoxification. It is now known that it is possible to make up Al and Si solutions in an order in which pH is lowered prior to Al addition; in these cases amelioration has still been observed. Amelioration has also been noted in experiments where HAS formation is minimal. These observations would suggest that, at least under some circumstances, there is an in planta component to the amelioration phenomenon. Several microanalytical investigations have noted codeposition of Al and Si in root cell walls. We propose a model in which root cell walls are the main internal sites of aluminosilicate (AS) and/or HAS formation and of Al detoxification. Factors promoting AS/HAS formation in this compartment include: high apoplastic pH; the presence of organic substances (e.g. malate); and the presence of suitable local concentrations of reactive forms of Al and Si, on or within the surfaces of the wall matrix. All these are likely to be important in the amelioration of Al toxicity.     

Toxicity of Al to Desulfovibrio desulfuricans

The toxicity of Al to Desulfovibrio desulfuricans G20 was assessed over a period of 8 weeks in a modified lactate C medium buffered at four initial pHs (5.0, 6.5, 7.2, and 8.3) and treated with five levels of added Al (0, 0.01, 0.1, 1.0, and 10 mM). At pH 5, cell population densities decreased significantly and any effect of Al was negligible compared to that of the pH. At pHs 6.5 and 7.2, the cell population densities increased by 30-fold during the first few days and then remained stable for soluble-Al concentrations of <5 x 10(-5) M. In treatments having total-Al concentrations of > or =1 mM, soluble-Al concentrations exceeded 5 x 10(-5) M and limited cell population growth substantially and proportionally. At pH 8.3, soluble-Al concentrations were below the 5 x 10(-5) M toxicity threshold and cell population density increases of 20- to 40-fold were observed. An apparent cell population response to added Al at pH 8.3 was attributed to the presence of large, spirilloidal bacteria (accounting for as much as 80% of the cells at the 10 mM added Al level). Calculations of soluble-Al speciation for the pH 6.5 and 7.2 treatments that showed Al toxicity suggested the possible presence of the Al(13)O(4)(OH)(24)(H(2)O)(12)(7+) "tridecamer" cation and an inverse correlation of the tridecamer concentration and the cell population density. Analysis by (27)Al nuclear magnetic resonance spectroscopy, however, yielded no evidence of this species in freshly prepared samples or those taken 800 days after inoculation. Exclusion of the tridecamer species from the aqueous speciation calculations at pHs 6.5 and 7.2 yielded inverse correlations of the neutral Al(OH)(3) and anionic Al(OH)(4)(-) monomeric species with cell population density, suggesting that one or both of these ions bear primary responsibility for the toxicity observed.     

Comparative hydrolysis and sorption of Al and La onto plant cell wall material and pectic materials

Pectin, which is an important component of plant cell walls, strongly binds Al and this may play a role in expression of Al toxicity. Sorption of aluminium (Al) and lanthanum (La) from aqueous solutions onto pectic acid, Ca-pectate and plant cell wall material was pH dependent. For Al at pH 3, sorption was less than the available sorption sites (i.e., the cation exchange capacity) on all three sorbents, whereas at pH 4, sorption of Al was in excess of available sorption capacity. By contrast, sorption of the trivalent Al analogue La corresponded to the available sorption capacity on all three sorbents at pH 5. This indicates, therefore, that Al hydrolyses at ≥ pH 4, and hydrolysis increases with Al concentration in solution. Further, it is proposed that the sorption of Al to pectin leads to deprotonation of the galacturonic acid (GalA) residues. Sorption of Al to pectin limits hydrolysis of Al, thereby shifting the pH of hydrolysis to a higher value. Hydrolysis of Al results in its sorption in excess of the stoichiometric equivalent (assuming the free Al³⁺ ion), leading to oversaturation of the pectin with Al. Staining of the metal-pectate complexes with the metachromatic dye eosin showed that with increasing Al saturation (but not La saturation), the complex developed a positive net charge, due to formation of some positively charged Al-complexes. The development of a positive charge on the Al-pectate complex may have major effects on cellular transmembrane potential and nutrient acquisition by plant roots. This is the first report of Al binding in excess of binding sites and development of a net positive charge on Al-pectate.     

Aluminium accumulation in root cell walls coincides with inhibition of root growth but not with inhibition of magnesium uptake in Norway spruce

High levels of aluminium in the soil solution of forest soils cause stress to forest trees. Within the soil profile, pH and aluminium concentration in the soil solution vary considerably with soil depth. pH strongly influences the speciation of A1 in solution, and is a factor when considering toxicity of A1 to roots. Norway spruce ( Picea abies [L.] Karst.) seedlings were grown for 7 weeks in nutrient solutions at pH 3.2, 4.0 or 5.0 containing 0, 100 or 400 µ M A1. At the end of this period, seedling growth, the cation exchange capacity of the roots and the amount of exchangeable Ca and Mg in roots were determined. A1 concentrations in whole roots, root segments, and in needles were measured. Using X‐ray microanalysis, the concentrations of Al, Ca, Mg and P were determined in cortical cell walls. We wanted to test the hypotheses that (1) the amount of Al bound to cation exchange sites can be used as a marker for Al toxicity and (2) the Mg concentration of needles is controlled by the amount of Mg bound to cation exchange sites. Low pH reduced the inhibition of Al on root growth and shoot length. Both low pH and Al lowered the concentration of Ca and Mg in needles. Al concentrations in the roots decreased as the pH decreased. In the roots, Al displaced Mg and Ca from binding sites at the root cortical cell walls. A comparison of the effects of Al at the different pH values on root growth and Mg concentration in the needles, suggests that, at pH 5.0, an Al fraction in the apoplast inhibits root growth, but does not affect Mg uptake. This fraction of Al is not available for transport to the shoots. In contrast, Mg uptake is strongly affected by Al at pH 3.2, although only very low levels of Al were detected in the roots. Thus, Al accumulation in the apoplast is a positive marker for Al effects on root growth, but not Mg uptake. The Mg concentration of needles is not controlled by the amount of Mg bound to cation exchange sites.     

Silicon amelioration of aluminium toxicity and cell death in suspension cultures of Norway spruce (Picea abies (L.) Karst.)

A role for silicon (Si) in the amelioration of aluminium (Al) toxicity in gymnosperms is suggested by their codeposition in planta, including within needles. This study was designed to investigate Al/Si interactions at the cellular level using suspension cultures of Norway spruce. Toxic effects of Al were dependent on duration of Al exposure, concentration of Al, and pH. Toxicity was reduced when Si was present, and the effect was enhanced at pH 5.0 compared to pH 4.2. Study of the ultrastructure of Al-treated cells indicated that changes in cell wall thickening, degree of vacuolation, and the degeneration of mitochondria, Golgi bodies, ER and nucleus preceded cell death, and significant amelioration was noted when Si was also present. When the fluorescent dye Morin was employed to localise free Al, cells treated with Al and Si in combination showed less fluorescence than the cells treated with Al alone. Intensity of fluorescence depended on the concentration of Al, duration of treatment and pH. Notably, presence of Si reduced the concentration of free Al in the cell wall in parallel with amelioration of Al toxicity. We therefore propose that formation of aluminosilicate complexes in the wall and apoplasm provide a significant barrier to Al penetration and cell damage in Norway spruce.     

COMPARATIVE pH DEPENDENT METAL INHIBITION OF NUTRIENT UPTAKE BY SCENEDESMUS QUADRICAUDA(CHLOROPHYCEAE)1,2

Cadmium and copper inhibition of nutrient uptake by the green alga Scenedesmus quadricauda is highly pH dependent in an inorganic medium; both metals are less toxic at low pH. The alga was grown in chemostats with both N and P approaching limiting levels; it was then possible to study metal toxicity to the nitrate, ammonium, and phosphate uptake systems of algae in an identical physiological state. When the logarithm of the Cd concentration causing 25% inhibition of nitrate, ammonium, and phosphate uptake was regressed against pH almost perfect linear relationships were obtained. This was also true at the 50% inhibition level, except for a smaller than predicted increase in Cd toxicity to ammonium uptake at pH 8, which may be due to the beginning of Cd precipitation at this pH. Cu²⁺ toxicity was linearly related to pH for ammonium and phosphate uptake and although, its toxicity for nitrate uptake also increased with pH, the increase was not perfectly linear. The toxicity of total Cu showed no linear relationship to pH. Cd²⁺ and Cu²⁺ toxicity increased by up to four orders of magnitude from pH 5 to 8. Competition between free metal and hydrogen ions for uptake sites on the cell surface is suggested as a mechanism increasing the toxicity of free metal, ions as the hydrogen ion content decreases (i.e. at higher pH).