Red maple (Acer rubrum) species is one of the most widespread deciduous (hardwood) trees of eastern North America. It is among the dominant tree species in the Northern Ontario after land reclamation. To date, the effects of heavy metal contamination from the mining activities on terrestrial ecosystems are not well understood. The main objectives of the present study are (1) to determine the level of phytoavailable metal in soil and accumulation in A. rubrum, and (2) to compare the levels of genetic variation among and within A. rubrum populations from areas with different metal contents in a Northern Ontario region. The total heavy metal levels were found to be high but the availability of these metals were much lower. We found that red maple does not accumulate heavy metals in their leaves as other hardwood species. The translocation factors were 0.05, 0.21, 0.38, 0.90, and 2.8 for Cu, Ni, Fe, Zn, and Mg, respectively. The levels of genetic variation in red maple populations from reclaimed lands in Northern Ontario were moderate to high since the percentage of polymorphic loci varied between 51 and 67%. The mean values for observed number of alleles (Na), effective number of alleles (Ne), Nei's gene diversity (h), and Shannon's information index (I) were 1.60, 1.24, 0.15 and 0.24, respectively. The population differentiation (GST) among the fragmented populations was high (0.28) despite a high level of gene flow (Nm = 1.28). Nevertheless, all the populations within the targeted region were genetically closely related. A specific ISSR marker that was identified in all the samples from the reference sites was absent in most samples from metal contaminated. This specific band was cloned and sequenced. Overall, the present study confirms that red maple populations in Northern Ontario are genetically sustainable despite the high level of total metal content in soil.