Tunisia is known to be a country with poor water resources, and water scarcity is evident in certain regions. In the long term, this situation could worsen, given the increased risk of drought. The olive tree (Olea europaea L.) is one of the plants best adapted to this climate, and numerous studies have been carried out to assess the effects of water stress. The aim of this work was to study and compare the ecophysiological behavior of a main Tunisian olive cultivar (Chetoui) and four rare Tunisian olive cultivars (Chemchali, Besbessi, Sayali and Jarboui) under drought stress and to identify the main parameters while comparing the tolerance of the cultivars studied to this abiotic stress. One-year-old olive trees grown in pots in a greenhouse were subjected to four drought treatments (i.e., 15, 30, 45 and 60 days of drought stress) in comparison with control trees. The evaluation of the response of the olives to this induced stress was based on five parameters: relative water content (RWC), stomatal resistance (SR), photosystem PSII, maximal photochemical efficiency (F_V/F_M), performance index on absorption basis (PI), measured by the handy PEA, and chlorophyll index, measured by SPAD. The relative water content (RWC) of the five cultivars decreased with increasing drought stress. Jarboui showed lower RWC values than Chemchali, especially under severe drought stress. This result was confirmed by changes in fluorescence characteristics. F_V/F_M, PI and SPAD index decreased during the development of drought stress. These observations are discussed in relation to the strategies developed by the cultivars to grow under drought stress. The Principal Component Analysis allowed the parameter with the strongest loading factor, which is F_V/F_M, to be highlighted and the cultivars most tolerant to drought stress to be distinguished. These cultivars, Besbessi and Sayali in the north of Tunisia and Chemchali in the south, can present a possible alternative to replace the local or foreign cultivars most cultivated in the country, which are characterized by high water needs.