Aims Recent advances in mass spectrometry have demonstrated that higher plants discriminate stable Zn isotopes during uptake and translocation depending on environmental conditions and physiological status of the plant. Stable Zn isotopes have emerged as a promising tool to characterize the plants response to inadequate Zn supply. The aim of this review is to build a comprehensive model linking Zn homeostasis and Zn isotopic fractionation in plants and advance our current view of Zn homeostasis and interaction with other micronutrients. Methods The distribution of stable Zn isotopes in plants and the most likely causes of fractionation are reviewed, and the interactions with micronutrients Fe, Cu, and Ni are discussed. Results The main sources of Zn fractionation in plants are i) adsorption, ii) low- and high-affinity transport phenomena, iii) speciation, iv) compartmentalization, and v) diffusion. We propose a model for Zn fractionation during uptake and radial transport in the roots, root-to-shoot transport, and remobilization. Conclusions Future work should concentrate on better understanding the molecular mechanisms underlying the fractionations as this will be the key to future development of this novel isotope system. A combination of stable isotopes and speciation analyses might prove a powerful tool for plant nutrition and homeostasis studies

Stable isotope signatures of Zn have shown great promise in elucidating changes in uptake and translocation mechanisms of this metal in plants during environmental changes. Here this potential was tested by investigating the effect of high Zn concentrations on the isotopic fractionation patterns of Phragmites australis (Cav.) Trin. ex Steud. Plants were grown for 40 d in a nutritive solution containing 3.2 μM (sufficient) or 2 mM (toxic) Zn. The Zn isotopic composition of roots, rhizomes, shoots, and leaves was analysed. Stems and leaves were sampled at different heights to evaluate the effect of long-distance transport on Zn fractionation. During Zn sufficiency, roots, rhizomes, and shoots were isotopically heavy (δ66ZnJMC Lyon=0.2‰) while the youngest leaves were isotopically light (–0.5‰). During Zn excess, roots were still isotopically heavier (δ66Zn=0.5‰) and the rest of the plant was isotopically light (up to –0.5‰). The enrichment of heavy isotopes at the roots was attributed to Zn uptake mediated by transporter proteins under Zn-sufficient conditions and to chelation and compartmentation in Zn excess. The isotopically lighter Zn in shoots and leaves is consistent with long-distance root to shoot transport. The tolerance response of P. australis increased the range of Zn fractionation within the plant and with respect to the environment.

The mercury (Hg) accumulation mechanism was studied in rape (Brassica napus) plants grown under a Hg concentration gradient (0 mM-1,000 mM). Hg mainly accumulated in roots. Therefore, the presence of phytochelatins (PCs) was studied in the roots of the plants. The high stability of the PC-Hg multicomplexes (mPC-nHg) seems to be the main reason for the lack of previous Hg-PC characterization studies. We propose a modification of the method to detect and quantify unbound PC of Hg in plant extracts via high-performance liquid chromatography coupled to electrospray tandem mass spectrometry and inductively coupled plasma mass spectrometry in parallel. We separated the PC from the Hg by adding the chelating agent sodium 2,3-dimercaptopropanesulfonate monohydrate. We only detected the presence of PC after the addition of the chelating agent. Some multicomplexes mPC-nHg could be formed but, due to their large sizes, could not be detected. In this study, only PC 2 was observed in plant samples. Hg accumulation was correlated with PC 2 concentration (r 2 5 0.98).

The aim of the present study was to examine the ability of I. pseudacorus L., an ornamental macrophyte of great potential for phytoremediation, to tolerate and accumulate Cr and Zn. Plants were grown in nutritive solution with ZnCl 2 or CrCl 3 Á6H 2 O at 0, 10, 50, 100, and 200 lg ml -1 for 5 weeks; all survived and continued growing. The accumulation of Cr and Zn increased with increasing supply in all plant tissues, to reach 59.97 mg Cr and 25.64 mg Zn in roots. Leaves retained a remarkable amount of Zn (14.2 mg). Growth inhibition reached 65% and 31% (dry weight) in response to Cr and Zn, respectively. The root:shoot dry matter partitioning (R/S) increased 80% at 100 lg ml -1 CrCl 3 . The most marked alterations in mineral content were in roots, where both metals decreased Al, Ca, Mg, Mn and S, and increased P concentration. No effect was noted on either leaf chlorophyll fluorescence kinetics (F v /F m and U PSII ), or photosynthetic pigment content, signifying that the light phase of photosynthesis was not impaired. Carbon isotope composition (d 13 C) was only slightly heavier, indicating that the reduction of carbon fixation was not the main cause for growth decrease. This was attributed to the restricted mineral uptake and to the increased demand of carbohydrates of damaged roots. Biomass allocation to rhizomes (Cr) or roots (Zn) contributes to heavy metal tolerance by limiting transpiration and increasing metal-storing tissues and the surface for water and cation uptake. This species is a good candidate for Cr rhizofiltration and Zn phytoextraction.