THE PROMOTER OF VACUOLAR METAL TRANSPORTER IN ARABIDOPSIS HALLERI: AN EXAMPLE OF EVOLUTION FOR METAL HYPERTOLERANCE/HYPERACCUMULATION

In the metal hyperaccumulator Arabidopsis halleri, the metal vacuolar transporter (VMT) isinvolved in hyperaccumulation and hypertolerance. Three VMT promoter sequences have beenidentified in the genome of A. halleri; expression analysis, conducted by GUS assay, showed thateach promoter sequence of A. halleri induced higher expression than the promoter sequence of theVMT orthologous gene in A. thaliana. Moreover, it was observed that A. halleri VTM members areexpressed in leaf trichomes, whereas VTM of A. thaliana is not present in these organs. In silicoanalysis revealed that the 5’ UTR region of VTM is highly conserved between the two Arabidopsisspecies, although a dimer of MYB-binding motifs harboured by the A. halleri promoters is mutatedin a single nucleotide in the sequence of A. thaliana. Site-specific mutagenesis of these motifs in thepromoter of A. halleri indicates that they are likely involved in trichome-specific expression. Therole of these MYB-binding motifs in the A. halleri VMT promoter was tested in A. thaliana,considering their effect in both metal tolerance and accumulation. Transgenic A. thaliana plantsexpressing VMT under the control of the native A. halleri VMT promoter are more tolerant, in termsof root length, biomass production and chlorophyll content, to high metal concentrations, than theones bearing the same construct with the mutated MYB-binding motifs. Differences in shoots andin roots were also observed regarding metal accumulation. These results underline the divergentevolution of the VMT promoter, conferring higher metal tolerance, accompanied by expression intrichomes, in the hyperaccumulator A. halleri and protection to heavy metal stress in transgenic A.thaliana. Furthermore, a possible biotechnological approach might be applied in phytoremediationor biofortification programs to modulate metal content in plants.