Effects of aluminium on canola roots

There is little information on the effects of aluminium (Al) on canola (Brassica napus var. napus L.), which is a commercially important crop species in many parts of the world. In this report, we describe the effects of Al on roots of canola seedlings grown hydroponically in a nutrient solution at pH 4.5. The morphological and ultrastructural changes that accompanied these growth effects were examined. Additions to the nutrient solution of Al at concentrations below 40 μM stimulated root growth of canola seedlings, increasing both the size and number of central cap cells. The stimulation of root growth did not appear to be due to the alleviation of a proton toxicity at the root surface. At concentrations of Al above 60 μM, root growth was strongly inhibited, with cellular damage being observed primarily in peripheral root cap cells. This revised version was published online in June 2006 with corrections to the Cover Date.     

The extremely high Al resistance of Penicillium janthineleum F-13 is not caused by internal or external sequestration of Al

Penicillium janthinellum F-13 has been isolated in previous work as a fungus tolerating the presence of high concentrations of Al (as high as 100 mM AlCl₃). Here its growth rate and yield in three acidic (pH 3.0) media of different composition with varying concentrations of Al are reported. The presence of Al did not affect these parameters, except that the growth yield was somewhat lower in GM (a glucose/peptone/yeast extract-containing medium) with the highest concentration tested (100 mM AlCl₃). The amount of Al found in the mycelium was so low that it cannot lead to a significant decrease in the medium for the higher Al concentrations applied. Although citric acid was excreted at growth on GM, and the presence of Al even promoted this, the concentration of this was far too low to diminish (by chelation) the high Al concentrations in the medium to a non-toxic level, i.e. the level (of approx. 1 mM) that is tolerated by low-resistance fungi. At growth on SLBM (a peptone/yeast extract/soil extract-containing medium), a rise in pH occurred. The same was found for SM (a glucose/mineral salts-containing medium), although in this case the picture was more complicated because the initial rise in pH was followed by a lowering due to the excretion of oxalic acid. Although both phenomena can diminish Al toxicity (by decreasing the external concentration of monomeric Al, regarded to be the toxic species), again the decrease is far too low to attain a non-toxic level when high Al concentrations are applied. Therefore, although in principal the metabolic phenomena observed for P. janthinellum F-13 at growth on different media can diminish Al toxicity, the tolerance of this organism for high external Al concentrations must be caused by another mechanism.     

Physiological responses of lupin roots to high pH

High pH seems to be a major constraint limiting the production of narrow-leafed lupin (Lupinus angustifolius L.) on alkaline soils. Whereas there has been much interest in soil acidity, relatively little is known about the effect of high pH on the growth of roots of higher plants.Elongation of roots of L. angustifolius was particularly sensitive to pH6.0 compared with other species. The effect of high pH in decreasing root elongation in L. angustifolius occurred within one hour. It was via an effect on cell elongation and not cell division and the effect was readily reversible. The mechanisms of the adverse effect of high pH are unknown. The permeability ratio of K⁺ to Na⁺ in the plasma-membrane of the root cortical cells was similar in solutions of both low pH and high pH. Reduced cell growth at high pH was not associated with an inefficiency of proton extrusion to the bulk solution by roots of this species. Nevertheless, increasing buffer concentration in the external solution decreased root elongation more in L. angustifolius than in Lupinus pilosus and Pisum sativum.     

Effect of exchangeable aluminium on the reduction of potato scab

The severity of the incidence of the fungal disease, potato scab, varies with different soil groups at the same soil pH. At a soil pH of 5.3, potato scab is easily controlled in soils of western Hokkaido (soil group A) by simply decreasing soil pH, but in soils from eastern Hokkaido (soil group B) it is not so easily controlled. The difference appears to be due to higher levels and exchangeable aluminium in Group A.Addition of sufficient aluminium or ferrous sulfate to a group B soil decreased the incidence of potato scab in a field experiment. Higher levels of aluminium sulfate depressed crop production. It is concluded that aluminium ions control the incidence of potato scab in acid soils. It is suggested that, in soils with low exchange acidity Y1, potato scab can be controlled by adding sufficient aluminium to increase their exchange acidity Y1 to above 7–8.     

Effects of aluminium and mineral nutrition on growth and chemical composition of hydroponically grown seedlings of five different forest tree species

Forest die-back and impaired tree vitality have frequently been ascribed to Al-toxicity and Al-induced nutritional disorders due to increased acidification of forest soils. Therefore, in this experiment effects of Al were studied on growth and nutrient uptake with seedlings of five different forest tree species. During growth in culture solutions with and without Al all five species proved to be very Al-tolerant, despite high accumulation of Al in roots. In the coniferous evergreens Douglas-fir and Scots pine shoot as well as root Al concentrations were significantly higher than in the deciduous broad-leaved species oak and birch. Larch showed intermediate Al levels. In none of the five species did Al reduce nutrient concentrations or the Ca/Al ratio to values below the critical level. Besides differences in Al accumulation, coniferous and broad-leaved species also differed with respect to uptake and assimilation of nitrogen. Due to extra NH ₄ ⁺ uptake, oak and birch showed a much higher N uptake and higher NH ₄ ⁺ preference than the coniferous species. Especially with oak this high NH ₄ ⁺ preference in combination with a low specific root surface area resulted in a high root proton efflux density. In comparison to both broad-leaved trees and Scots pine the NO ₃ ⁻ reduction capacity of larch and Douglas-fir was extremely low. This may have important consequences for both species if grown in NO ₃ ⁻ -rich soils.     

Control of nutrient solutions for studies at high pH

Little is known about factors effecting plant growth at high pH, with research often limited by the inability to separate nutritional deficiencies and HCO ₃ ⁻ toxicity from the direct limitations imposed under high pH conditions. Various methods of controlling dilute nutrient solutions for studies at high pH were investigated. For short-term studies, it was found that a solution without Cu, Fe, Mn and Zn and aerated with CO₂ depleted air, greatly reduced nutrient precipitation at high pH, thus eliminating nutritional differences between treatments. Manual pH adjustment and the use of ion exchange resins as pH buffers were unsuitable methods of pH control. However, pH control by automated titration had little effect on solution composition while maintaining constant pH. The system described is suitable for studies in which the pH of the bulk nutrient solution must be maintained. The system was used to examine OH⁻ toxicity in mungbeans (Vigna radiata (L.) Wilczek cv. Emerald), with root length reduced at a bulk solution pH of 8.5 and greater.     

Effects of aluminium on growth and cation uptake in seedlings of Eucalyptus mannifera and Pinus radiata

This experiment was designed to examine the effects of aluminium (Al) on the growth of Pinus radiata (D. Don) and Eucalyptus mannifera subsp. mannifera (Mudie) seedlings in culture solutions in a glasshouse to help explain the failure of radiata pine trees on some acid, low fertility soils in Australia on which the native eucalypts flourish. Aluminium (Al) in culture solution increased the growth of roots and shoots of seedlings of both species but while growth of the eucalypt continued to increase with increases in Al to 2.222 μM, growth of the pine was largest at 370 μM Al. In addition to total root length, specific root length (length per unit dry weight), a measure of fineness of the root, increased in the eucalypt seedlings as the substrate Al increased. Growth of the shoots and roots of the pine in the absence of any added Al was extremely poor suggesting that Al, in low concentrations, may be an essential element or ameliorate some other factors in solution culture at low pH. Root and shoot concentrations of K increased with increasing Al, whilst Ca and Mg Concentrations decreased and Mn concentrations were unaffected in both species. Tissue Ca and Mg concentrations were 2 to 3 times higher in the eucalypt seedlings than the pine at all levels of added Al due to greater uptake of these elements by the eucalypt. In contrast, at the highest concentration of Al in the medium, shoot Al concentrations were lower in the cucalypt than in the pine due to a greater proportion of Al being retained in the eucalypt roots. These differences between the seedlings in terms of root growth and tissue cation concentrations may help explain the ability of eucalypt species to maintain vigorous growth on acid soils high in Al and low in Ca and P, where growth of the pines failed.     

Solution pH modifies the response of Norway spruce seedlings to aluminium

Norway spruce (Picea abies) was exposed to nutrient solutions containing a range of aluminium (Al) concentrations at several pH levels (3.2, 4 and 5). Root growth was reduced by 100 µM and 400 µM Al at pH 4 and 5, but at pH 3.2 only by 400 µM Al. The Al content of the roots increased with increasing pH. The Al content of the roots was higher at the root tips than at the older root parts at all pH values. Using X-ray microanalysis it could be shown that higher levels of Al at increased pH were mainly due to increased Al contents in root cortex cell walls. In seedlings, mycorrhizal with Pisolithus tinctorius or Lactarius rufus, the Al concentration of cortex cell walls was higher when nitrate (NO₃) rather than ammonium (NH₄) was the nitrogen (N) source.     

Identity of the rhizotoxic aluminium species

The aluminium (III) released from soil minerals to the soil solution under acid conditions may appear as hexaaquaaluminium (Al(H₂O)₆ ³⁺, or Al³⁺ for convenience) or may react with available ligands to form additional chemical species. That one or more of these species is rhizotoxic (inhibitory to root elongation) has been known for many decades, but the identity of the toxic species remains problematical for the following reasons. 1. Several Al species coexist in solution so individual species cannot be investigated in isolation, even in artificial culture media. 2. The activities of individual species must be calculated from equilibrium data that may be uncertain. 3. The unexpected or undetected appearance of the extremely toxic triskaidekaaluminium (AlO₄Al₁₂(OH)₂₄(H₂O)₁₂ ⁷⁺ or Al₁₃) may cause misatribution of toxicity to other species, especially to mononuclear hydroxy-Al. 4. If H⁺ ameliorates Al³⁺ toxicity, or vice versa, then mononuclear hydroxy-Al may appear to be toxic when it is not. 5. The identity and activities of the Al species contacting the cell surfaces are uncertain because of the H⁺ currents through the root surface and because of surface charges. This article considers the implications of these problems for good experimental designs and critically evaluates current information regarding the relative toxicities of selected Al species. It is concluded that polycationic Al (charge >2) is rhizotoxic as are other polyvalent cations.     

Inhibition of Al-induced root elongation and enhancement of Al-induced peroxidase activity in Al-sensitive and Al-resistant barley cultivars are positively correlated

The quantitative changes in peroxidase activity and composition of anionic and cationic isoperoxidases were investigated in roots of two barley cultivars differing in Al resistance. Root growth of Al-resistant cv. Bavaria was in lesser extent reduced by Al treatment (23% after 24 h Al-treatment), whereas 40% reduction of the root growth was observed in Al-sensitive cv. Alfor. The strong root growth inhibition in Al-sensitive cv. Alfor correlated with a 6-fold enhancement of peroxidase activity by Al treatment. Al-induced enhancement of peroxidase activity was found also in roots of Al-resistant cv. Bavaria, but this increase was only half of the Al-sensitive cv. Alfor. Comparison of peroxidase isoenzyme composition of Al-treated and non-treated roots revealed that activity of at least five anionic and four cationic isoperoxidases was stimulated by Al treatment. Three of anionic isoperoxidases (aPOD2-4) were selectively induced only in the Al-sensitive cv. Alfor. A possible involvement of peroxidases in root-growth inhibition is discussed.     

The trans labilization of cis-[PtCl2(13CH3NH2)2] by glutathione can be monitored at physiological pH by [1H,13C] HSQC NMR

In order to monitor the trans labilization of cisplatin at physiological pH we have prepared the complex cis-[PtCl₂(¹³CH₃NH₂)₂] and studied its interactions with excess glutathione in aqueous solution at neutral pH by two-dimensional [1H,13C] heteronuclear single-quantum correlation (HSQC) NMR spectroscopy. [1H,13C] HSQC spectroscopy is a good method for following the release of ¹³CH₃NH₂ but is not so good for characterizing the Pt species in solution. In the reaction of cisplatin with glutathione, Pt–S bonds are formed and Pt–NH₃ bonds are broken. The best technique for following the formation of Pt–S bonds of cisplatin is by UV spectroscopy. [1H,13C] HSQC spectroscopy is the best method for following the breaking of the Pt–N bonds. [1H,15N] HSQC spectroscopy is the best method for characterizing the different species in solution. However, the intensity of the peaks in the ¹⁵NH₃–Pt–S region, in [1H,15N] HSQC, reflects a balance between the formation of Pt–S bonds, which increases the signal intensity, and the trans labilization, which decreases the signal intensity. [1H,15N] HSQC spectroscopy and [1H,13C] HSQC spectroscopy are complementary techniques that should be used in conjunction in order to obtain the most accurate information on the interaction of platinum complexes with sulfur-containing ligands.     

Gradients in maize roots: local elongation and pH

The elongation rate, the gradient of the local elongation rate and the surface pH of maize roots were measured over 12 h. A data bank was constituted by storing these values. By sorting these results on the basis of different elongation rates, different classes of root were obtained. Two classes were chosen: the low-growth roots and the high-growth roots. The mean growth of these two root classes was stable with time and differed significantly from one another. The surface pH of the elongation zone was the same for the roots of these two classes, but the roots selected for their higher growth rate had a larger acid efflux in this zone.     

Interaction between aluminium and phosphorus in sorghum plants

A solution culture experiment was carried out to study the effects of interactions between aluminium (Al) and phosphorus (P) on Al-toxicity under conditions of suboptimal P supply. The experiment was conducted in a growth chamber with seedlings of the Al-sensitive sorghum genotype TAM428 (Sorghum bicolor (L.) Moench). Phosphorus deficiency differed from Al toxicity in its effect on shoot/root ratio and root morphological charateristics. Results indicated that there were positive effects of Al on the uptake and assimilation of P. Therefore, it was unlikely that an Al-induced P deficiency could account for the observed reduction in plant biomass. Plants suffered more from Al toxicity at very low P supply. Moreover, decreasing P supply resulted in increased root H-ion efflux density. In the soil, where a rhizosphere can be formed, this would make the plant even more susceptible to Al. Dry matter yield of the plants was affected more severely by Al at the first harvest (14 days) than at the second (35 days), but the opposite was true for P. Aluminium-inhibited root development and reduced uptake of N, K and Mg (but not Ca) may be partly responsible for the growth depression. Increasing the P supply exerted certain roles in eliminating Al phytotoxicity, possibly through improved root development and nutrient uptake. The detrimental influence of Al on biomass could be overcome by doubling the P supply.     

Acute toxicity of Cd in stream invertebrates in relation to pH and test design

The acute toxicity of Cd to three stream invertebrates (Baetis rhodani Pict., Leptophlebia marginata (L.) and Pisidium sp.) was tested at pH 5 and 7 simultaneously in static (ST) and flow through (FT) systems. In the static design, the animals were kept individually in small boxes containing aerated stream water. In the flow through system, the three species were kept together in circular aquaria simulating stream ecosystems with patches of gravel and recirculating aerated stream water. The nominal Cd concentrations ranged from 0 to 5 mg 1^–1 during an exposure period of 120 h.The graphically obtained LC₅₀ (120 h) values for B. rhodani were: pH 7: 2.3 mg 1^–1 (ST), 2.5 mg 1^–1 (FT) and pH 5: 3 mg 1^–1 (ST), 1 mg 1^–1 (FT). For L. marginata the following values were found: pH 7: > 5 mg 1^–1 (ST), 4.4 mg 1^–1 (FT) and pH 5: > 5 mg 1^–1 (ST), 3.6 mg 1^–1 (FT). Pisidium sp. showed a 100% survival in all Cd- and pH-treatments. From thee values it can be concluded that L. marginata is more tolerant than B. rhodani and that especially both species tolerated Cd better in the ST system than in the FT system, especially at pH 5.At neutral pH the Cd-concentrations in the animals were higher than at low pH, probably due to surface adsorption of Cd onto the body. B. rhodani tended to molt more at low than at neutral pH.     

Peroxidase catalysed formation of cytotoxic prooxidant phenothiazine free radicals at physiological pH

The antipsychotic phenothiazines may have other therapeutic applications because of their ability to kill bacteria, plasmids and tumor cells. They are also known to undergo a peroxidase-catalysed oxidation to form cation radicals that are stable at acid pH, but are not detected at a neutral pH. The objective of this project was to determine whether phenothiazine cation radical metabolites could cause oxidative stress at a neutral pH resulting in cytotoxicity. At a neutral pH, catalytic amounts of phenothiazines were found to be oxidised by a peroxidase/H2O2 system and also caused ascorbate, GSH and NADH cooxidation. NADH and GSH co-oxidation was accompanied by oxygen uptake and was increased by the addition of catalytic amounts of superoxide dismutase, indicating that the superoxide radical was formed. The phenothazines were different from other peroxidase substrates in that the NADH, ascorbate or GSH cooxidation was faster at pH 6.0 than pH 7.4, thereby partly reflecting the cation radical stability. The order of catalytic effectiveness found was promazine > chlorpromazine > trifluoperazine. Peroxidase/H2O2 also markedly increased phenothiazine cytotoxicity towards isolated rat hepatocytes at nontoxic phenothiazine concentrations. At both pH 6.0 and 7.4, the same order of phenothiazine catalytic effectiveness was observed as seen in the co-oxidation experiments. Cytotoxicity to hepatocytes could be attributed to oxidative stress as most hepatocyte glutathione oxidation and lipid peroxidation preceded phenothiazine induced cytotoxicity and that cytotoxicity was prevented by the antioxidant butylated hydroxyanisole. This hepatocyte/peroxidase/H2O2 system could be a useful model for studying drug induced idiosyncratic hepatic injury enhanced by inflammation.     

Immobilization of Zinc and Cadmium in Polluted Soils by Polynuclear Al13 and Al-Montmorillonite

We investigated the suitability of two aluminum-based binding agents, polynuclear Al₁₃ and Al-coated montmorillonite (Al-mont-morillonite), for the immobilization of heavy metals in two contaminated agricultural soils: a loamy luvisol from an arable site in Rafz, Canton Zürich, Switzerland, and a sandy podsol from Szopienice, Upper Silesia, Poland. Both soils were polluted by lead, zinc, and cadmium: the soil from Szopienice by the emissions of a nearby zinc-lead smelter, and the soil from Rafz by sewage sludge applications. While the samples from Szopienice exhibited extremely high loads of these metals, the samples from Rafz were only moderately contaminated. The samples from both soils were slightly acidic. The Rafz soil contained 2.5% organic matter, that from Szopienice only 1.5%. Destruction of the organic matter in the Szopienice samples by H₂O₂ led to a significant release of Zn and Cd into solution. This indicated that organic matter is an important factor for the immobilization of heavy metals in this soil. The treatment of the Szopienice samples with 8?mmol Al₁₃ per kg dry soil resulted in a considerable mobilization of the two metals. As the pH of the samples did not decrease, this effect was presumably due to direct interactions between the applied aluminium and organic matter. After destruction of soil organic matter, the two binding agents exhibited an immobilizing effect on Zn, which, however, was weak compared with the binding of the metal by the organic matter prior to its destruction. In the case of the Rafz samples, metal mobilization was observed only for Al₁₃ if applied in high doses (4 and 8?mmol per kg soil), but not for Al-montmorillonite. In this soil, Al-montmorillonite as well as Al₁₃ at low doses (1.2?mmol per kg soil and less) decreased soluble zinc concentrations significantly. The mobilization of metals at high doses of the applied binding agents and the dependence of this effect on the type of soil show that care has to be taken with this remediation method and that the proper doses of applied binding agents can be crucial for the success of metal immobilization in polluted soils.     

Growth response of coffee tree shoots and roots to subsurface liming

In the greenhouse growth of two coffee-tree varieties, Catuaí (sensitive) and Icatu (tolerant) to aluminum, was evaluated in surface-fertilized and limed soil following subsurface treatment with seven lime levels (0.0; 0.49; 1.7; 2.9; 4.1; 6.6 and 9.3 t/ha). Plants were grown for 6.5 months in soils in PVC columns, subdivided into two horizons. In the lower 12 – 34 cm depth horizon, soil Al saturation varied between 93 and 0%. For both varieties evaluated, shoot dry weight and leaf area remained unchanged following limestone application. This fact shows that surface layer correction permitted normal shoot growth. High Al saturation resulted in decrease of root dry weight percent, root length percent and root surface percent in the 12–34 cm horizon, which were compensated by higher percentages of these properties in the upper 0–12 cm horizon. The ratio between root surface – root dry matter (cm²/g) of Catuaí variety was increased by limestone application to the lower soil horizons, indicating that roots turn longer and thinner, when Al soil saturation decreased. This also shows a great sensitivity to Al of the Catuaí variety. In contrast, in the Icatu variety, all root characteristics remained stable at all levels of Al tested.     

The effect of aluminium on phosphate uptake by three isolated ectomycorrhizal fungi

Three wheat cultivars with different tolerances against free aluminium were grown monoxenically in association with Azospirillum brasilense. In situ nitrogen fixation, measured with the acetylene reduction assay, was higher by the aluminium-tolerant cultivars than by the sensitive cultivar. The transfer of fixed nitrogen to the host plant, determined by the ¹⁵N dilution technique, was also significantly higher in the aluminium-resistant wheat plants. The total accumulation of fixed nitrogen in the host plants due to an A. brasilense inoculation varied from approximately 13% to 17% of the total nitrogen in the root and 2.9% to 3.9% of the nitrogen in the shoot.The quantity and quality of exudates released in liquid nutrient solution were analysed separately for two of the wheat cultivars, one aluminium-tolerant and one aluminium-sensitive. After 29 days of growth the aluminium-tolerant plants exudated significantly higher total amounts of carbon than aluminium-sensitive plants. No differences between the two cultivars existed in the carbon exudation rate per gram dry root.Much higher concentrations of low molecular dicarboxylic acids i.e. succinic, malic and oxalic acid, were found in the exudates of aluminium-tolerant plants. Dicarboxylic acids are potential chelating compounds for positively charged metals such as aluminium and they may play an important role in protecting the plant against aluminium incorporation. They are also very suitable substrates for Azospirillum spp. It is therefore suggested that these factors may be causing the higher associative nitrogen fixation rates which was found in the aluminium-tolerant wheat cultivars.     

Photosynthesis and apparent affinity for dissolved inorganic carbon by cells and chloroplasts of Chlamydomonas reinhardtii grown at high and low CO2 concentrations

Chloroplasts with high rates of photosynthetic O₂ evolution (up to 120 mol O₂· (mg Chl)^-1·h^-1 compared with 130 mol O₂· (mg Chl)^-1·h^-1 of whole cells) were isolated from Chlamydomonas reinhardtii cells grown in high and low CO₂ concentrations using autolysine-digitonin treatment. At 25° C and pH=7.8, no O₂ uptake could be observed in the dark by high- and low-CO₂ adapted chloroplasts. Light saturation of photosynthetic net oxygen evolution was reached at 800 mol photons·m^-2·s^-1 for high- and low-CO₂ adapted chloroplasts, a value which was almost identical to that observed for whole cells. Dissolved inorganic carbon (DIC) saturation of photosynthesis was reached between 200–300 M for low-CO₂ adapted chloroplasts, whereas high-CO₂ adapted chloroplasts were not saturated even at 700 M DIC. The concentrations of DIC required to reach half-saturated rates of net O₂ evolution (K_m(DIC)) was 31.1 and 156 M DIC for low- and high-CO₂ adapted chloroplasts, respectively. These results demonstrate that the CO₂ concentration provided during growth influenced the photosynthetic characteristics at the whole cell as well as at the chloroplast level.Abbreviations Chl chlorophyll - DIC dissolved inorganic carbon - K_m(DIC) coneentration of dissolved inorganic carbon required for the rate of half maximal net O₂ evolution - PFR photon fluence rate - SPGM silicasol-PVP-gradient medium     

Prevention of aluminium toxicity with supplemental boron. I. Maintenance of root elongation and cellular structure

Aluminium toxicity is an important factor limiting plant growth mi acid soils. Symptoms of B deficiency and Al toxicity are very similar and generally associated with impaired membrane Function and root growth. Thus the objective of this study was to determine whether supplemental B prevents Al inhibition of root growth and development. Squash (Cucurbita pepo L. cv. Sunbar) was grown in hydroponic nutrient media with 44 mmol m^?3 free Al and B concentrations extending from 5 to 100 mmol m^?3. Our results establish that B protects against Al inhibition of root growth. Protection was apparent at all levels of organization examined: primary root and lateral root lengths; primary root cell elongation, cell production rate, tissue organization and cell structure; primary root morphology and maturation. Protection against Al inhibition was also apparent for shoot growth. These studies were undertaken in solution culture to limit the variables examined; however, the underlying motivation for this study is the problem of worldwide Al toxicity in soils. Therefore, the effect of adding additional B to a high-Al soil was also investigated and is the subject of the companion paper (Le Noble. Blevins & Miles 1996, Plant, Cell and Environment 19, 1143–1148).