Low-quality water and soil salinization are increasingly becoming limiting factors for food production, including olive – a major fruit crop in several parts of the world. Identifying putative salinity-stress tolerance in olive would be helpful in the future development of new tolerant varieties. In this study, novel salinity-responsive biomarkers (SRBs) were characterized in the species, namely, monooxygenase 1 (OeMO1), cation calcium exchanger 1 (OeCCX1), salt tolerance protein (OeSTO), proteolipid membrane potential modulator (OePMP3), universal stress protein (OeUSP2), adaptor protein complex 4 medium mu4 subunit (OeAP-4), WRKY1 transcription factor (OeWRKY1) and potassium transporter 2 (OeKT2). Unique structural features were highlighted for encoded proteins as compared with other plant homologues. The expression of olive SRBs was investigated in leaves of young plantlets of two cultivars, ‘Nabali’ (moderately tolerant) and ‘Picual’ (tolerant). At 60 mM NaCl stress level, OeMO1, OeSTO, OePMP3, OeUSP2, OeAP-4 and OeWRKY1 were up-regulated in ‘Nabali’ as compared with ‘Picual’. On the other hand, OeCCX1 and OeKT2 were up-regulated at three stress levels (30, 60 and 90 mM NaCl) in ‘Picual’ as compared to ‘Nabali’. Salinity tolerance in olive presumably engages multiple sets of responsive genes triggered by different stress levels.