Zinc distribution and speciation in roots of various genotypes of tobacco exposed to Zn

Cell walls of roots have a great reactivity towards metals, and may act as a barrier limiting the entry of metals, especially in non-hyperaccumulating species. The aim of this study was to determine the localization and speciation of Zn in roots of tobacco (Nicotiana tabacum) grown in Zn-contaminated substrates. Chemical extractions and EXAFS spectroscopy were applied on whole roots and on isolated cell walls of roots. Our results show that cell walls of roots exhibited a distribution of Zn affinity sites, from water-soluble to non-exchangeable Zn. In whole roots, Zn was bound with oxalate and other COOH/OH groups: the first species was probably intracellular while the second was attributed to Zn bound to the cell walls and, to a lesser extent, to intracellular organic acids. Moreover, Zn-phosphate was also identified, and this species was CuSO4-extractable. It probably resulted from chemical precipitation in the apoplasm, and explained the steady increase in exchangeable root Zn observed in root of tobacco during the culture. This study shows the strength of combining EXAFS and chemical extractions for studying localization and speciation of metals in plants.