Acanthodactylus is a widespread lacertid genus including about thirthy species whose systematics remains much debated.The present study is aimed at improving the taxonomic status of the Tunisian species whose geographical distribution and ecological requirements have been studied elsewhere. Allozymic variation at 22 loci was scored in 14 populations for a total of 249 individuals. High genetic variability is confirmed in all samples; it is higher in species displaying a wide bioclimatic habitat range. No reduction of genetic variability was observed in two insular populations.Any interspecific hybrid could be detected from our genetic data, even in sympatric areas arguing in favor of specific status of four taxons in Northern, Central and Southern Tunisia.Phylogenetic relationships are discussed on the basis of UPGMA and Fitch-Margoliash procedures applied to the genetic distances matrix and centered data analysis of allozyme frequencies.