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 μM total Cu (0.01–0.06 μM 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.