Phytoremediation offers owners and managers of metal-contaminated sites an innovative and cost-effective option to address recalcitrant environmental contamination. The use of plants to restore or stabilize contaminated sites, known as phytoremediation, takes advantage of the natural abilities of plants to take up, accumulate or store metals. This includes the use of plants that tolerate and accumulate metals at high levels for phytoextraction and the use of plants growing under conditions that are toxic to other plants, for preventing, for example, soil erosion (phytostabilisation). Rapeseed (Brassica napus L.) was shown to be able to accumulate substantial amounts of metals combined with high biomass. Brassica napus was therefore selected for heavy metal (HM) tolerance and accumulation through in vitro selection. A selective pressure applied during the neoformation process from transversal thin cell layers (tTCLs) allowed us to select tolerant cells and tissues. Toxic metals (such as Zn) were added to the culture media in order to select zinc-tolerant plants. Exerting a selective pressure during tTCLs regeneration aimed at selecting plants with exceptional zinc tolerance and/or accumulating capacity. The morphological and physicochemical characteristics of regenerated plants cultivated in greenhouse appeared to depend very significantly on the concentration of ZnSO(4) applied during the neoformation process. Plants regenerated in the presence of ZnSO(4) at 100 microM exhibited a greater size and a higher biomass together with flowering precocity. The contents of zinc, chlorophyll, and proline were modified in the regenerated plants. Pre-treatment with an excess of ZnSO(4) (>500 microM) was responsible for a percentage of tTCLs intolerance above 96%. With lower Zn concentrations (100-250 microM), the survival rates (33-15%) were higher.