Nitrogen and phosphorus economy of a legume tree-cereal intercropping system under controlled conditions.

Considerable amounts of nitrogen (N) and phosphorus (P) fertilizers have been mis-used in agroecosystems, with profound alteration to the biogeochemical cycles of these two major nutrients. To reduce excess fertilizer use, plant-mediated nutrient supply through N(2)-fixation, transfer of fixed N and mobilization of soil P may be important processes for the nutrient economy of low-input tree-based intercropping systems. In this study, we quantified plant performance, P acquisition and belowground N transfer from the N(2)-fixing tree to the cereal crop under varying root contact intensity and P supplies. We cultivated Acacia senegal var senegal in pot-culture containing 90% sand and 10% vermiculite under 3 levels of exponentially supplied P. Acacia plants were then intercropped with durum wheat (Triticum turgidum durum) in the same pots with variable levels of adsorbed P or transplanted and intercropped with durum wheat in rhizoboxes excluding direct root contact on P-poor red Mediterranean soils. In pot-culture, wheat biomass and P content increased in relation to the P gradient. Strong isotopic evidence of belowground N transfer, based on the isotopic signature (δ(15)N) of tree foliage and wheat shoots, was systematically found under high P in pot-culture, with an average N transfer value of 14.0% of wheat total N after 21 days of contact between the two species. In the rhizoboxes, we observed limitations on growth and P uptake of intercropped wheat due to competitive effects on soil resources and minimal evidence of belowground N transfer of N from acacia to wheat. In this intercrop, specifically in pot-culture, facilitation for N transfer from the legume tree to the crop showed to be effective especially when crop N uptake was increased (or stimulated) as occurred under high P conditions and when competition was low. Understanding these processes is important to the nutrient economy and appropriate management of legume-based agroforestry systems.

Stable isotopes of Cu and Zn in higher plants: evidence for Cu reduction at the root surface and two conceptual models for isotopic fractionation processes.

Recent reports suggest that significant fractionation of stable metal isotopes occurs during biogeochemical cycling and that the uptake into higher plants is an important process. To test isotopic fractionation of copper (Cu) and zinc (Zn) during plant uptake and constrain its controls, we grew lettuce, tomato, rice and durum wheat under controlled conditions in nutrient solutions with variable metal speciation and iron (Fe) supply. The results show that the fractionation patterns of these two micronutrients are decoupled during the transport from nutrient solution to root. In roots, we found an enrichment of the heavier isotopes for Zn, in agreement with previous studies, but an enrichment of isotopically light Cu, suggesting a reduction of Cu(II) possibly at the surfaces of the root cell plasma membranes. This observation holds for both graminaceous and nongraminaceaous species and confirms that reduction is a predominant and ubiquitous mechanism for the acquisition of Cu into plants similar to the mechanism for the acquisition of iron (Fe) by the strategy I plant species. We propose two preliminary models of isotope fractionation processes of Cu and Zn in plants with different uptake strategies.

Rhizosphere pH gradient controls copper availability in a strongly acidic soil.

Using a root mat approach, we quantified how root-induced alkalization controlled the establishment of copper (Cu) gradients in the rhizosphere of durum wheat (Triticum turgidum durum L.) cropped in a strongly acidic, Cu-contaminated soil. Rhizosphere pH increased over 6 mm in soil, reaching up to +2.8 units close to root mat surface. Conversely, free Cu2+ activity decreased by 3 orders of magnitude and total Cu concentration by 3-fold in the rhizosphere solution, while labile Cu assessed by DGT (diffusive gradients in thin films) was halved. The DIFS (DGT-induced flux in soils and sediments) model failed to adequately simulate Cu depletion in the rhizosphere solution, showing that root-induced alkalization almost entirely explained Cu depletion while plant uptake had little impact. We modeled the observed pH gradient to recalculate its radial extension around a single root. The gradient of free Cu2+ activity in solution, deduced from pH modeling, extended over 1-4 mm in the rhizosphere depending on root radius and OH- efflux from root. Rhizosphere alkalization dramatically decreased root exposure to Cu, substantiating that root-induced chemical changes in the rhizosphere should be better accounted for to assess metal bioavailability to plants.

A biotest for evaluating copper bioavailability to plants in a contaminated soil.

This work aimed at defining the optimal conditions for a novel ecotoxicological test designed for evaluating the bioavailability and phytotoxicity of metals to plants. This biotest, which provided easy access to roots, shoots, and rhizosphere soil, was applied to a vineyard calcareous soil that had been contaminated by the application of Cu fungicides. A preliminary hydroponic experiment comparing various levels of solution Cu concentration enabled us to determine the no observable adverse effects concentration (NOAEC), which was in the range 5 to 20 microM total Cu (0.01-0.06 microM free Cu ion) for rape (Brassica napus L. cv. Goeland). For the biotest, rape was grown in hydroponic conditions for 21 d in pots designed so that plants developed a planar mat of roots at the surface of a polyamide mesh. By then, the plants were transferred for 4 or 8 d onto a 1- or 3-mm-thick soil layer that was separated from the root mat by the mesh and connected to a reservoir of nutrient solution or deionized water via a filter paper wick. An 8-d period was the best option as it enabled plant growth to be significant. The use of 1-mm soil thickness was recommended if the biotest aimed at investigating root-induced changes in the rhizosphere. Although it may cause some artifacts, compared with deionized water, nutrient solution provided better standardized conditions for comparing widely differing soil samples. The studied soil did not induce any Cu phytotoxicity in spite of its fairly large total Cu content.

Copper bioavailability and extractability as related to chemical properties of contaminated soils from a vine-growing area.

Vineyard soils have been contaminated by Cu as a consequence of the long-term use of Cu salts as fungicides against mildew. This work aimed at identifying which soil parameters were the best related to Cu bioavailability, as assessed by measuring the concentrations of Cu in shoots and roots of tomato cropped (in lab conditions) over a range of 29 (24 calcareous and five acidic) Cu-contaminated topsoils from a vine-growing area (22-398 mg Cu kg(-1)). Copper concentrations in tomato shoots remained in the adequate range and were independent of soil properties and soil Cu content. Conversely, strong, positive correlations were found between root Cu concentration, total soil Cu, EDTA- or K-pyrophosphate-extractable Cu and organic C contents in the 24 calcareous soils, suggesting a prominent role of organic matter in the retention and bioavailability of Cu. Such relations were not observed when including the five acidic soils in the investigated population, suggesting a major pH effect. Root Cu concentration appeared as a much more sensitive indicator of soil Cu bioavailability than shoot Cu concentration. Simple extractions routinely used in soil testing procedures (total and EDTA-extractable Cu) were adequate indicators of Cu bioavailability for the investigated calcareous soils, but not when different soil types were considered (e.g. acidic versus calcareous soils).

Evaluation of copper availability to plants in copper-contaminated vineyard soils.

The repeated use of copper (Cu) fungicides to control vine downy mildew has led to long-term accumulation of Cu in vineyard soils which now raises the issue of the potential bioavailability of Cu for various living organisms including plant species. The bioavailable Cu can be defined as the portion of soil Cu that can be taken up by roots, for a given plant species. In order to evaluate the bioavailability of Cu to plants, a pot experiment was conducted in glasshouse conditions with a crop species (maize) and 12 soils sampled in the upper horizon of 10 vineyard plots (total Cu ranging from 38 to 251 mg kg-1) and two woodland plots (control soils that had not received any Cu application; total Cu amounting to 20-26 mg kg-1). These soils were selected for their diverse physical (large range of particle size distribution) and chemical (from acid to calcareous soils) properties. After 35 days of growth, plant shoots were harvested for analysis. The roots were separated from soil particles for further analysis. The concentrations of Cu in the roots and aerial parts of the maize were then compared with the amounts of Cu extracted from the soil by a range of conventional extractants. Observed Cu concentrations in maize roots which have grown in contaminated vineyard soils were very high (between 90 and 600 mg kg-1), whereas Cu concentrations in the aerial parts varied only slightly and remained low (< 18 mg kg-1). Root Cu concentrations observed for maize increased with increasing total Cu content in the soil and with decreasing soil CEC. Cu accumulation in maize roots may be as high in calcareous soils as in acid soils, suggesting that soil pH had little influence. In the case of the vineyard soils studied, the lack of correlation found for maize between Cu concentrations in roots and in the aerial parts, suggests that an analysis of the aerial parts would not be a good indicator of plant Cu uptake, as it provides no insight into the real amount of Cu transferred from the soil to the plant. For maize, our results show that extraction with organic complexing agents (EDTA, DTPA) and extraction with ammonium acetate seem to provide a reasonably good estimate of root Cu concentration.