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In this study, eighteen olive varieties, originating from Spain, France, Italy, Greece and Algeria, and maintained at the olive experimental station of Boughrara (arid region of Tunisia) were evaluated for their oil yield and fatty acid composition. The analysis of variance revealed significant differences among varieties for all traits (p < 0.01). The Hierarchical Cluster Analysis (HCA) classified the varieties into three main groups. The first group included a subgroup which is composed of seven olive varieties (Cornezuelo, Verdial de Velez-Malaga, Leccino, Coratina, Koroneiki, Lechin de Granada and Changlot Real) characterized by high oil yield with high oleic, low palmitic and linoleic acid contents. The fatty acid compositions of the oils from these varieties comply with international standards and show more beneficial characteristics than the oil obtained from Chemlali: the most abundant olive cultivar in Tunisia. Finally, the main fatty acids (palmitic (C16:0), oleic (C18:1) and linoleic (C18:2)) of nine of the studied virgin olive oils were compared to those sampled from their traditional areas. Except for Koroneiki and Oliviere oils which showed an unchanged fatty acid composition and for Cornezuelo oil in which the level of oleic acid raised and the level of linoleic acid decreased, most of oils showed a decrease in oleic acid rates and an increase in palmitic and linoleic acid percentages as compared to those from their original sites.