Edward Berkelaar

Coupling the use of computer chemical speciation models and culture techniques in laboratory investigations of trace metal toxicity

Abstract The bioavailability and toxicity of a dissolved metal are closely linked to the metal’s chemical speciation in solution. A variety of inorganic and organic ligands are often used in laboratory toxicity tests to control the concentration of labile trace metal in solution. Computerised chemical speciation models based on thermodynamic principles can be used to estimate metal speciation under such experimental conditions. However, these models are sensitive to the quality of their thermodynamic databases. Detailed protocols for the incorporation of reliable equilibrium formation constants into widely available computer chemical speciation programs (e.g., MINEQL+ and MINTEQ) are provided. The examples demonstrate both the benefits and the potential pitfalls involved in the use of chemical speciation models. The application of chemical speciation modelling to metal toxicity studies is discussed and guidelines are proposed for its proper use. Both defined media and chemical speciation programs have co-existed for two decades but the combined use of these techniques has been reserved for those possessing in-depth knowledge of both chemistry and biology. The techniques presented should enable an investigator with basic biological, chemical and computing skills to design an aqueous medium and incorporate correct thermodynamic constants into a computer chemical speciation program, starting from a standardised database, thereby providing a sound framework for critically assessing the biological response of a particular test organism to a given metal.

The biotic ligand model for plants and metals: Technical challenges for field application†

To improve predictions of phytoavailable metal, the mechanistic bases of bioaccumulation and toxicity of metals to plants can be integrated into a biotic ligand model (BLM). There are a number of significant challenges to the application of the BLM to plants in soils, including reliable measurements of free ion concentrations for the metals of interest in rhizospheric soil solution, as well as other free ions, and concentrations of ligands to which the ions could bind; identification of the simplest model that can adequately predict root accumulation, and the potential for more complex models to add accuracy to the predictions; incorporating the dissociation of labile metal complexes (i.e., nonequilibrium processes) into a BLM, which is an equilibrium model; application of factors in a BLM that adequately describe translocation, in order to estimate metal concentration and speciation in plant shoots. The review concluded that the ability to estimate trace metal speciation in samples of soil solution are not likely to be better than within one order of magnitude of actual, thus this would be an additional source of uncertainty to the predictions of toxicity. Further, regulatory use of the BLM would require mechanistic bases; and, until root ligands associated with toxicity are well characterized, incorporating the ameliorative effects of competitive cations cannot be mechanistically based. As well, a functional BLM for soils with lower metal free ion activities will have to include kinetic data for metal-ligand complexes, as their association/disassociation may constitute a greater metal supply to roots than what would be predicted by the free ion concentration in soil solution. To apply the BLM to trophic transfer where metal concentration in plant shoots is the main focus, a probabilistic approach using experimentally determined root-shoot partitioning of metals might permit estimates of shoot accumulation from root data, to within one or two orders of magnitude.

The influence of photosynthetically active radiation on the effects of ultraviolet-B radiation on Arabidopsis thaliana

Ultraviolet‐B (UVB;280–320 nm) radiation is a small but biologically significant portion of the solar spectrum reaching the earths surface. Research interests have been fostered because UVB has been increasing in recent years due to depletion of stratospheric ozone. Ultraviolet‐B that penetrates into plant tissue may damage important cellular macromolecules. Although there has been considerable research on the effects of UVB on plants, the influence of the level of photosynthetically active radiation (PAR;400–700 nm) on effects of UVB requires further definition as a prelude to studies of UVB sensitivity and defense mechanisms. Arabidopsis thaliana wildtype ecotype Landsberg erecta (LER), which is relatively insensitive to UVB, and the relatively sensitive LER‐based mutant transparent testa‐5 (tt5), were grown under 100 or 250 μmol m−2 s−1 PAR and then exposed to O or 7 kJ m−2 day −1 UVBBE under these PAR levels. Plants exposed to UVB had reduced dry weight and leaf area and higher levels of UV‐absorbing compounds in leaf tissue. The level of PAR did influence the effects of UVB, with the higher level of PAR prior to UVB exposure reducing sensitivity of LER to UVB. In contrast to other studies, higher PAR supplied simultaneously with UVB increased rather than decreased sensitivity of both genotypes to UVB. These results demonstrate the importance of controlling and comparing PAR levels when undertaking studies of UVB sensitivity, as effects of UVB on plants are influenced by the PAR levels plants are growing under prior to and during exposure to UVB.