Cation amelioration of aluminum toxicity in wheat

Aluminum is a major constituent of most soils and limits crop productivity in many regions. Amelioration is of theoretical as well as practical interest because understanding amelioration may contribute to an understanding of the mechanisms of toxicity. In the experiments reported here 2-day-old wheat (Triticum aestivum L. cv Tyler) seedlings with 15-millimeter roots were transferred to solutions containing 0.4 millimolar CaCl₂ at pH 4.3 variously supplemented with AlCl₃ and additional amounts of a chloride salt. Root lengths, measured after 2 days in the test solutions, were a function of both Al activity and the cation activity of the added salt. Percent inhibition = 100 {Al³⁺}/({Al³⁺} + K_m + α{C}^β) where {Al³⁺} is the activity of Al³⁺ expressed in micromolar, {C} is the activity of the added cation expressed in millimolar, and K_m (= 1.2 micromolar) is the {Al³⁺} required for 50% inhibition in the absence of added salt. For Ca²⁺, Mg²⁺, and Na⁺ the values of α were 2.4, 1.6, and 0.011, respectively, and the values for β were 1.5, 1.5, and 1.8, respectively. With regard to relative ameliorative effectiveness, Ca²⁺ > Mg²⁺ ≈ Sr²⁺ K⁺ ≈ Na⁺. Other cations were tested, but La³⁺, Sc³⁺, Li⁺, Rb⁺, and Cs⁺ were toxic at potentially ameliorative levels. The salt amelioration is not solely attributable to reductions in {Al³⁺} caused by increases in ionic strength. Competition between the cation and Al for external binding sites may account for most of the amelioration.     

Lignin deposition induced by aluminum in wheat (Triticum aestivum) roots

We investigated the relation between the toxic effect of aluminum (Al) on root growth and the lignin deposition in wheat ( Triticum aestivum L. cvs Atlas 66 and Scout 66). In the Al-tolerant cultivar Atlas 66, control treatment without AlCl₃ at pH 4.75, cell length increased dramatically in the portion of the root that was 0.6 to 3.2 mm from the root cap junction (approximately 1.0 to 3.6 mm from the root tip). However, treatment with 20 μ M AlCl₃ for 24 and 48 h completely inhibited root elongation and markedly decreased the length and increased the diameter of the cells in the same portion of the root. Moreover, marked deposition of lignin was observed in the cells that corresponded to the portion 1.5 to 4.5 mm from the root tip in Atlas 66 roots treated with 20 μ M AlCl₃, while no deposition of lignin was detected in control roots. Treatment with 5 μ M AlCl₃ slightly inhibited root growth and there was no deposition of lignin in the root. On the other hand, in roots of the Al-sensitive cultivar Scout 66, treatment with 5 μ M AlCl₃ completely inhibited root growth and markedly induced deposition of lignin. These results suggest that lignification in the elongating region coincided with the extent of inhibition of root growth by Al in two wheat cultivars that differed in their sensitivity to Al.     

Malate-permeable channels and cation channels activated by aluminum in the apical cells of wheat roots

Aluminum (Al(3+))-dependent efflux of malate from root apices is a mechanism for Al(3+) tolerance in wheat (Triticum aestivum). The malate anions protect the sensitive root tips by chelating the toxic Al(3+) cations in the rhizosphere to form non-toxic complexes. Activation of malate-permeable channels in the plasma membrane could be critical in regulating this malate efflux. We examined this by investigating Al(3+)-activated channels in protoplasts from root apices of near-isogenic wheat differing in Al(3+) tolerance at a single locus. Using whole-cell patch clamp we found that Al(3+) stimulated an electrical current carried by anion efflux across the plasma membrane in the Al(3+)-tolerant (ET8) and Al(3+)-sensitive (ES8) genotypes. This current occurred more frequently, had a greater current density, and remained active for longer in ET8 protoplasts than for ES8 protoplasts. The Al(3+)-activated current exhibited higher permeability to malate(2-) than to Cl(-) (P(mal)/P(Cl) > or = 2.6) and was inhibited by anion channel antagonists, niflumate and diphenylamine-2-carboxylic acid. In ET8, but not ES8, protoplasts an outward-rectifying K(+) current was activated in the presence of Al(3+) when cAMP was included in the pipette solution. These findings provide evidence that the difference in Al(3+)-induced malate efflux between Al(3+)-tolerant and Al(3+)-sensitive genotypes lies in the differing capacity for Al(3+) to activate malate permeable channels and cation channels for sustained malate release.     

Protein carbonylation in barley seedling roots caused by aluminum and proton toxicity is suppressed by salicylic acid

Proton (H⁺) and aluminum (Al³⁺) toxicities are major factors limiting crop production on acid soils. To study whether salicylic acid (SA) is functional in alleviating protein damage caused by H⁺ and Al³⁺ toxicities, an investigation of the antioxidant defense response regulated by SA was carried out on barley (Hordeum vulgare L.) seedlings under H⁺, Al³⁺, and combined stresses. It was found that the relative root elongation of seedlings, which grew in the solutions supplemented with SA, was significantly higher than that of seedlings without SA treatment after 24-h treatments with H⁺, Al³⁺, and combined stresses. The lesser amount of carbonylated proteins with molecular weights ranging from 14.4 to 97 kD, was accumulated in seedlings treated with SA than that in the seedlings without SA treatment. The higher activities of antioxidant enzymes and lesser content of MDA were observed in seedlings treated with SA compared with the seedlings without SA treatment. Moreover, the nitroblue tetrazolium staining of roots showed that ROS accumulation was decreased by SA treatments. This study suggested that SA could alleviate cell damage caused by H⁺ and Al³⁺ toxicities on acid soils by both activating antioxidant defense responses and reducing the contents of carbonylated proteins caused by ROS in barley seedlings.     

Early events responsible for aluminum toxicity symptoms in suspension-cultured tobacco cells

We investigated the aluminum (Al)-induced alterations in zeta potential, plasma membrane (PM) potential and intracellular calcium levels to elucidate their interaction with callose production induced by Al toxicity. A noninvasive confocal laser microscopy has been used to analyse the live tobacco (Nicotiana tabacum) cell events by means of fluorescent probes Fluo-3 acetoxymethyl ester (intracellular calcium) and DiBAC4 (PM potential) as well as to monitor callose accumulation. Log-phase cells showed no detectable changes in the PM potential during the first 30 min of Al treatment, but sustained large depolarization from 60 min onwards. Measurement of zeta potential confirmed the depolarization effect of Al, but the kinetics were different. The Al-treated cells showed a moderate increase in intracellular Ca2+ levels and callose production in 1 h, which coincided with the time course of PM depolarization. Compared with the Al treatment, cyclopiazonic acid, an inhibitor of endoplasmic reticulum Ca(2+)-ATPase, facilitated a higher increase in intracellular Ca2+ levels, but resulted in accumulation of only moderate levels of callose. Calcium channel modulators and Al induced similar levels of callose in the initial 1 h of treatment. Callose production induced by Al toxicity is dependent on both depolarization of the PM and an increase in intracellular Ca2+ levels.     

Nitric oxide reduces aluminum toxicity by preventing oxidative stress in the roots of Cassia tora L

Nitric oxide (NO) as a key signaling molecule has been involved in mediation of various biotic and abiotic stress-induced physiological responses in plants. In the present study, we investigated the effect of NO on Cassia tora L. plants exposed to aluminum (Al). Plants pre-treated for 12 h with 0.4 mM sodium nitroprusside (SNP), an NO donor, and subsequently exposed to 10 microM Al treatment for 24 h exhibited significantly greater root elongation as compared with the plants without SNP treatment. The NO-promoted root elongation was correlated with a decrease in Al accumulation in root apexes. Furthermore, oxidative stress associated with Al treatment increased lipid peroxidation and reactive oxygen species, and the activation of lipoxygenase and antioxidant enzymes was reduced by NO. Such effects were confirmed by the histochemical staining for the detection of peroxidation of lipids and loss of membrane integrity in roots. The ameliorating effect of NO was specific, because the NO scavenger cPTIO [2-(4-carboxy-2-phenyl)-4,4,5,5-tetramethylinidazoline-1-oxyl-3-oxide] completely reversed the effect of NO on root growth in the presence of Al. These results indicate that NO plays an important role in protecting the plant against Al-induced oxidative stress.     

Ammonium under solution culture alleviates aluminum toxicity in rice and reduces aluminum accumulation in roots compared with nitrate

Al stress and ammonium–nitrogen nutrition often coexist in acidic soils due to their low pH and weak nitrification ability. Rice is the most Al-resistant species among small grain cereal crops and prefers NH₄ ⁺ as its major inorganic nitrogen source. This study investigates the effects of NH₄ ⁺ and NO₃ ^? on Al toxicity and Al accumulation in rice, and thereby associates rice Al resistance with its NH₄ ⁺ preference. Two rice subspecies, indica cv. Yangdao6 and japonica cv. Wuyunjing7, were used in this study. After treatment with or without Al under conditions of varying NH₄ ⁺ and NO₃ ^? supply, rice seedlings were harvested for the determination of root elongation, callose content, biomass, Al concentration and medium pH. The results indicated that Wuyunjing7 was more Al-resistant and NH₄ ⁺-preferring than Yangdao6. NH₄ ⁺ alleviated Al toxicity in two cultivars compared with NO₃ ^?. Both NH₄ ⁺-Al supply and pretreatment with NH₄ ⁺ reduced Al accumulation in roots and root tips compared with NO₃ ^?. NH₄ ⁺ decreased but NO₃ ^? increased the medium pH, and root tips accumulated more Al with a pH increase from 3.5 to 5.5. Increasing the NO₃ ^? concentration enhanced Al accumulation in root tips but increasing the NH₄ ⁺ concentration had the opposite effect. These results show NH₄ ⁺ alleviates Al toxicity for rice and reduces Al accumulation in roots compared with NO₃ ^?, possibly through medium pH changes and ionic competitive effects. Making use of the protective effect of NH₄ ⁺, in which the Al resistance increases, is advised for acidic soils, and the hypothesis that rice Al resistance is associated with the preferred utilization of NH₄ ⁺ is suggested.     

A protein similar to PR (pathogenesis-related) proteins is elicited by metal toxicity in wheat roots

An acidic, low molecular weight protein called TA1-18 (T for Triticum. Al for aluminium and 18 for its approximate molecular weight) is induced in wheat roots that are exposed to growth-inhibiting concentrations of Al. Enhanced biosynthesis of TA1-18 began during the period 3 to 6 h after exposure to Al, and reached a maximum after 9 to 12 h of treatment. A protein with the same molecular weight and pl was also elicited during toxicity associated with Cu and Cd, with calcium deprivation, and low (3. 5) pH, but not by heat shock. TA1-18 was formed in small amounts in triticale, but was not detected in rye during exposure to growth-inhibiting levels of Al. Amino acid sequencing of trypsin fragments of TA1-18 revealed strong homology to pathogenesis-related protein PR2 from parsley cultures, with which TA1-18 also shares similar molecular weight and pl. Aluminium toxicity appears to have features in common with pathogenesis such that similar proteins are formed in response to both types of stress.     

Oxidative stress triggered by aluminum in plant roots

Aluminum (Al) is a major growth-limiting factor for plants in acid soils. The primary site of Al accumulation and toxicity is the root meristem, and the inhibition of root elongation is the most sensitive response to Al. Al cannot catalyze redox reactions but triggers lipid peroxidation and reactive oxygen species (ROS) production in roots. Furthermore, Al causes respiration inhibition and ATP depletion. Comparative studies of Al toxicity in roots with that in cultured plant cells suggest that Al causes dysfunction and ROS production in mitochondria, and that ROS production, but not lipid peroxidation, seems to be a determining factor of root-elongation inhibition by Al.     

Proton extrusion and attendant transport phenomena in Candida utilis induced by ethanol

Summary Ion fluxes after ethanol addition to Candida utilis depend crucially on aeration (air versus oxygen). In O₂-aerated non-growing cells ethanol causes an H ⁺ / K ⁺ exchange and an extrusion of acetate and lactate accompanied mostly by K ⁺, and their subsequent reimportation together with H ⁺. Cells from continuous culture display generally stronger acidification and more marked K ⁺ movements than non-growing ones. Offprint requests to: A. Prell     

Nitric oxide (as sodium nitroprusside) supplementation ameliorates Cd toxicity in hydroponically grown wheat roots

Cadmium (Cd) is a non-redox toxic heavy metal present in the environment and induces oxidative stress in plants. We investigated whether exogenous nitric oxide (NO) supplementation as sodium nitroprusside (SNP) has any ameliorating action against Cd-induced oxidative damage in plant roots and thus protective role against Cd toxicity. Cd treatment (50 or 250 μM) alone or in combination with 200 μM SNP was given to hydroponically grown wheat roots for a short time period of 24 h and then these were shifted to distilled water to observe changes in levels of oxidative markers (lipid peroxidation, H₂O₂ content and electrolyte leakage). Supplementation of Cd with SNP significantly reduced the Cd-induced lipid peroxidation, H₂O₂ content and electrolyte leakage in wheat roots. It indicated a reactive oxygen species (ROS) scavenging activity of NO. However, even upon removal of Cd-treatment solution, the levels of oxidative markers increased during 24 h recovery stage and later at 48 h these decreased. Cd treatment resulted in an upregulation of activities of antioxidant enzymes—superoxide dismutase (SOD, 1.15.1.1), guaiacol peroxidase (GPX, 1.11.1.7), catalase (CAT, 1.11.1.6), and glutathione reductase (GR, 1.6.4.2). SNP supply resulted in a reduction in Cd-induced increased activities of scavenging enzymes. The protective role of exogenous NO in decreasing Cd-induced oxidative damage was also evident from the histochemical localization of lipid peroxidation, plasma membrane integrity and superoxides. The study concludes that an exogenous supply of NO protects wheat roots from Cd-induced toxicity.     

Organic acids enhance the uptake of lead by wheat roots

The uptake and bioavailability of lead (Pb) in soil–plant systems remain poorly understood. This study indicates that acetic and malic acids enhance the uptake of Pb by wheat (Triticum aestivum L.) roots under hydroponic conditions. The net concentration-dependent uptake influx of Pb in the presence and absence of organic acids was characterized by Michaelis–Menten type nonsaturating kinetic curves that could be resolved into linear and saturable components. Fitted maximum uptake rates (V _max) of the Michaelis–Menton saturable component in the presence of acetic and malic acids were, respectively, 2.45 and 1.63 times those of the control, while the Michaelis–Menten K _m values of 5.5, 3.7 and 2.2 μM, respectively, remained unchanged. Enhanced Pb uptake by organic acids was partially mediated by Ca²⁺ and K⁺ channels, and also depended upon the physiological function of the plasma membrane P-type ATPase. Uptake may have been further enhanced by an effectively thinner unstirred layer of Pb adjacent to the roots, leading to more rapid diffusion towards roots. X-ray absorption spectroscopic studies provided evidence that the coordination environment of Pb in wheat roots was similar to that of Pb(CH₃COO)₂·3H₂O in that one Pb atom was coordinated to four oxygen atoms via the carboxylate group.