Noccaea caerulescens (Nc) exhibits a very high constitutive expression of the heavy metal transporting ATPase, HMA4, as compared to the non-hyperaccumulator Arabidopsis thaliana (At), due to copy number expansion and altered cis-regulation. We screened a BAC library for HMA4 and found that HMA4 is triplicated in the genome of a N. caerulescens accession from a former Zn mine near La Calamine (LC), Belgium. We amplified multiple HMA4 promoter sequences from three calamine N. caerulescens accessions, and expressed AtHMA4 and different NcHMA4 cDNAs under At and Nc HMA4 promoters in the A. thaliana (Col) hma2hma4 double mutant. Transgenic lines expressing HMA4 under the At promoter were always fully complemented for root-to-shoot Zn translocation and developed normally at a 2-μM Zn supply, whereas the lines expressing HMA4 under Nc promoters usually showed only slightly enhanced root to shoot Zn translocation rates in comparison with the double mutant, probably owing to ectopic expression in the roots, respectively. When expression of the Zn deficiency responsive marker gene ZIP4 was tested, the transgenic lines expressing AtHMA4 under an NcHMA4-1-LC promoter showed on average a 7-fold higher expression in the leaves, in comparison with the double hma2hma4 mutant, showing that this construct aggravated, rather than alleviated the severity of foliar Zn deficiency in the mutant, possible owing to expression in the leaf mesophyll.

The problem of metal-induced toxicity is proliferating with an increase in industrialization and urbanization. The buildup of metals results in severe environmental deterioration and harmful impacts on plant growth. In this study, we investigated the potential of two ornamental plants, Catharanthus roseus (L.) G.Don and Celosia argentea L., to tolerate and accumulate Ni, Cr, Cd, Pb, and Cu. These ornamental plants were grown in Hoagland’s nutrient solution containing metal loads (50 µM and 100 µM) alone and in combination with a synthetic chelator, ethylenediaminetetraacetic acid (EDTA) (2.5 mM). Plant growth and metal tolerance varied in both plant species for Ni, Cr, Cd, Pb, and Cu. C. roseus growth was better in treatments without EDTA, particularly in Ni, Cr, and Pb treatments, and Pb content increased in all parts of the plant. In contrast, Cd content decreased with EDTA addition. In C. argentea, the addition of EDTA resulted in improved plant biomass at both doses of Cu. In contrast, plant biomass reduced significantly in the case of Ni. In C. argentea, without EDTA, root length in Cd and Cu treatments was significantly lower than the control and other treatments. However, the addition of EDTA resulted in improved growth at both doses for Pb and Cu. Metal accumulation in C. argentea enhanced significantly with EDTA addition at both doses of Cu and Cd. Hence, it can be concluded that EDTA addition resulted in improved growth and better metal uptake than treatments without EDTA. Metal accumulation increased with EDTA addition compared to treatments without EDTA, particularly for Pb in C. roseus and Cu and Cd in C. argentea. Based on the present results, C. roseus showed a better ability to phytostabilize Cu, Cd, and Ni, while C. argentea worked better for Ni, Cd, Cu, and Pb.