Effect of 1-naphthaleneacetic acid on organic acid exudation by the roots of white lupin plants grown under phosphorus-deficient conditions.

The effect of NAA (1-naphthaleneacetic acid) on organic acid exudation in white lupin plants grown under phosphorus deficiency was investigated. Plants were sampled periodically for collecting of organic acids (citrate, malate, succinate), and also were used to study the effect on proton extrusion and release of Na(+), K(+), Ca(2+) and Mg(2+). The tissues were later processed to quantify the organic acids in tissues, the phosphorus content and the effects on plant biomass. The exogenous addition of NAA led to an increase in organic acid exudation, but this response was not proportional to the concentration of the dose applied, noticing the largest increments with NAA 10(-8)M. In contrast the increase in root weight was proportional to the dose applied, which shows that with higher doses the roots produced are not of proteoid type. Proton extrusion and the release of cations were related to the NAA dose, the first was proportional to the dose applied and the second inversely proportional. Regarding the analysis of tissues, the results of citrate and phosphorus content in shoots show that the overall status of these parts are the main responsible of the organic acids exuded. NAA served as an enhancer of the organic acid exudation that occurs under phosphorus deficient conditions, with a response that depends on the dose applied, not only in its magnitude, but also in the mechanism of action of the plant hormone.

Cadmium in white lupin nodules: impact on nitrogen and carbon metabolism.

The aims of this work were to investigate the microlocalisation of cadmium (Cd) in Lupinus albus L. cv. Multolupa nodules, and to determine its effects on carbon and nitrogen metabolism. Nodulated white lupin plants were grown in a growth chamber with or without Cd (150 µM). Energy-dispersive X-ray microanalysis showed the walls of the outer nodule cortex cells to be the main area of Cd retention, helping to reduce the harmful effect Cd might have on the amount of N(2) fixed by the bacteroids. Sucrose synthase activity declined by 33% in the nodules of the Cd-treated plants, and smaller reductions were recorded in glutamine synthetase, aspartate aminotransferase, alkaline invertase and NADP-dependent isocitrate dehydrogenase activities. The Cd treatment also sharply reduced nodule concentrations of malate, succinate and citrate, while that of starch doubled, but that of sucrose experienced no significant change. In summary, the present results show that white lupins accumulate significant amounts of Cd in their root nodules. However, the activity of some enzymes involved in ammonium assimilation did decline, promoting a reduction in the plant N content. The downregulation of sucrose synthase limits the availability of carbon to the bacteroids, which might interfere with their respiration. Carbon metabolism therefore plays a primary role in the impaired function of the white lupin root nodule caused by Cd, while N metabolism appears to have a more secondary involvement.

Effects of cadmium on cork oak (Quercus suber L.) plants grown in hydroponics.

Cork oak (Quercus suber L.) is an autochthonous tree species that is being used for reforestation in heavy-metal-contaminated areas in Spain. A hydroponics experiment was carried out to characterize the effects of Cd on several morphological and physiological parameters in this species, including shoot length, nutrient concentrations and allocation in different organs, leaf pigment concentrations, photosynthetic efficiency, root ferric chelate reductase (FCR) activity and organic acid concentrations in xylem sap. Four different Cd treatments were applied, adding Cd chelated with EDTA or as chloride salt at two different concentrations (10 and 50 µM Cd). After 1 month of Cd treatment, plant growth was significantly inhibited in all treatments. Results indicate that Cd accumulates in all organs 7- to 500-fold when compared with control plants. The highest Cd concentration was found in the 50 µM CdCl(2) treatment, which led to concentrations of ~30, 123 and 1153 µg Cd g(-1) dry weight in leaves, stems and roots, respectively. In the strongest Cd treatments the concentrations of P and Ca decreased in some plant parts, whereas the Mn leaf concentrations decreased with three of the four Cd treatments applied. The concentrations of chlorophyll and carotenoids on an area basis decreased, whereas the (zeaxanthin plus antheraxanthin)/(total violaxanthin cycle carotenoids) ratio and the non-photochemical quenching increased significantly in all Cd treatments. Cadmium treatments caused significant increases in the activity of the enzyme FCR in roots and in the concentrations of organic acids in xylem sap. Some of the physiological changes found support the fact that Cd induces a deficiency of Fe in cork oak, although the plant Fe concentrations were not reduced significantly. At higher concentrations the effects of Cd were more pronounced, and were more marked when Cd was in the free ion form than when present in the form of Cd-EDTA.

Natural attenuation of residual heavy metal contamination in soils affected by the Aznalcóllar mine spill, SW Spain.

Non-amended soils affected by pyritic sludge residues were monitored for 7 years to assess the long-term natural attenuation ability of these soils. The decrease in both the total concentration of elements (particularly As) and (NH(4))(2)SO(4)-extractable fractions of Mn, and Zn, below the maximum permissible levels indicate a successful natural ability to attenuate soil pollution. Soil acidification by pyrite oxidation and rainfall-enhanced leaching were the largest contributors to the reduction of metals of high (Mn, Cu, Zn and Cd) and low (Fe, Al, and As) availability. Periodic use of correlation and spatial distribution analysis was useful in monitoring elemental dispersion and soil property/element relationships.

Interaction and accumulation of manganese and cadmium in the manganese accumulator Lupinus albus.

The effects of the interaction between Mn and Cd on the growth of the white lupin (Lupinus albus), uptake of these metals, their accumulation, and effects on heavy metal stress indicators were studied under glasshouse conditions. Plants were grown with and without Mn and/or Cd for 4 weeks. The absence of Mn and Cd led to lipid peroxidation-induced loss of flavonoids and anthocyanins in the roots, reduced the size of the plant canopy, and led to the appearance of proteoid roots. Sensitivity to Cd in white lupin was enhanced by a low Mn supply, despite lower Cd uptake and accumulation (leaf Mn:Cd concentration ratio <3), as evidenced by increased lipid peroxidation in the leaves and strong inhibition of growth. However, when the Mn supply was adequate, the plants showed few symptoms of Cd toxicity, even though Cd uptake and accumulation increased. A Mn:Cd ratio of up to 20 was enough to minimize Cd stress in the leaf, reflecting the plants' relative tolerance to Cd under such conditions. Irrespective of the Mn supply, the increase in antioxidant compounds observed in the roots of Cd-treated plants might act as a protective mechanism by minimizing the oxidative stress caused by Cd exposure. In summary, high leaf Mn concentrations seem to render white lupins more tolerant to Cd stress.

Comparative analysis of the contribution of phytochelatins to cadmium and arsenic tolerance in soybean and white lupin.

The biosynthesis of phytochelatins (PCs) plays a crucial role in the detoxification and homeostasis of heavy metals and metalloids in plants. However, in an increasing number of plant species metal(loid) tolerance is not well correlated with the accumulation of PCs: tolerant ecotypes frequently contain lower levels of PCs than non-tolerant ecotypes. In this study we have compared the responses of soybean (Glycine max L. cv. Resnik) and white lupin (Lupinus albus L. cv. Marta) to cadmium and arsenate in order to assess the role of homophytochelatins (hPCs) in the tolerance of soybean to these toxic elements. Soybean plants treated with Cd and As showed a high contribution of homo-glutathione (hGSH) to the pool of thiols in shoots in comparison to white lupin. Higher levels of hPCs in Cd-treated soybeans compared to PCs in lupins did not prevent growth inhibition. In contrast, the role of hPCs in the detoxification mechanism to arsenate in soybean seems to be clearer, showing higher thiol concentrations and lower growth reductions than those present in lupin plants.

Evolution of arsenate toxicity in nodulated white lupine in a long-term culture.

White lupine is an As-resistant legume that is of interest for phytoremediation of As-contaminated soils. To achieve successful phytoremediation, monitoring of the nutritional status of the selected plant species during the entire culture cycle is required to maintain a plant cover with high biomass production. A long-term pot experiment was carried out with nodulated lupine grown on perlite with 10 and 100 microM As concentrations. The reproductive period (from 10 weeks) was the most sensitive phenologic stage of white lupine to long-term As exposure. The 10 microM As treatment increased the uptake and translocation of micronutrients, except for Cu, mainly at flowering with As levels in pods below the statutory limit (1 mg kg (-1) fresh weight). However, the 100 microM As treatment induced significant differences compared to the control. These findings confirm the relatively high resistance of white lupine to arsenate and support the use of this species in phytoremediation and/or revegetation of As-contaminated sites, with special attention on P and Cu nutrition at flowering.

Bioavailability of metals and As from acidified multicontaminated soils: use of white lupin to validate several extraction methods.

White lupin is an annual crop that has been used for phytostabilization of acidified multicontaminated (heavy metals and As) soils from the Aznalcóllar spill-affected area, Southern Spain. One of the most important factors for successful phytostabilization is monitoring the pollutant bioavailability in the soil. The aim of this work was to determine the best-suited method for assessing the bioavailability of heavy metals together with As in the Aznalcóllar spill-affected area, by means of a systematic comparison between different extraction methods (Ammonium bicarbonate-diethylenetriamine pentaacetic acid (AB-DTPA), CaCl2, NaNO3, BCR, (NH4)2SO4 and rhizo). Both AB-DTPA and the first step of the BCR method were found to be unsuitable for assessing the bioavailability of heavy metals and As to plants growing in acidic soils. However, CaCl2-extractable As, Cu, and Zn and NaNO3-extractable As and Zn were well correlated with their concentrations in plant organs. Rhizo and (NH4)2SO4, with the highest determination coefficients, were the most recommended simple extraction methods to assess the bioavailability of As, Cu, Fe, Mn, and Zn in acidified multicontaminated soils using white lupin as an excluder model plant.

Subcellular compartmentalisation of cadmium in white lupins determined by energy-dispersive X-ray microanalysis.

The microlocalisation of cadmium (Cd) at the tissue-cellular level in Lupinus albus L. cv. Multolupa was determined by energy-dispersive X-ray microanalysis (EDXMA). Experimental plants were grown on Cd-treated (0 and 150 microM) perlite for 35 days. In leaves, Cd was found inside cells (cytoplasm or vacuoles), especially in the vascular bundle cells. Cd-induced damage of the chloroplast structure was also detected. EDXMA of the roots showed the cell wall to be the main area of Cd binding at the cellular level; only a small amount of Cd was found in the vacuoles. At the tissue level, a decreasing Cd gradient was seen from the outer to the inner root cortical parenchyma. Cd and S were found co-localised in the vascular cylinder.

Chelate-assisted phytoextraction of heavy metals in a soil contaminated with a pyritic sludge.

The occurrence of many polluted areas as that affected by the accident of the Aznalcóllar pyrite mine has promoted phytoremediation as a technology able to reduce the risk of heavy metal contamination at low cost. White lupin plant has been considered a good candidate for phytoremediation. We studied the capacity of several complexing agents to improve the ability of white lupin for heavy metal phytoremediation in soils with multi-elemental pollution from acid pyritic sludge. Solution-soil interaction was studied and pot experiments with sludge-affected soil were carried out to this end. The interaction experiments indicated that EDTA and NTA were more efficient than malate and citrate in solubilizing metals (Fe, Mn, Cu, Zn, Cd), with minimum differences between EDTA and NTA. The pot trial showed that NTA was able to mobilize toxic elements from sludge-polluted soil and hence increasing their concentrations in plant (Mn, Cu, Zn, As, Cd). However, the NTA treatment promoted an increase of toxic elements concentrations, especially for As, Cd, Pb, in the lixiviates exceeding the maximum permissible levels, so a careful management of chelate is necessary.