The selection of drought-tolerant sour cherry genotypes is essential for developing sustainable fruit production in today’s climate-change conditions. The phenotypic heterogenic population of sour cherry Oblačinska, with high and regular yield suitable for mechanical harvesting and industrial processing, is a traditional and predominant cultivar in northern Croatia (Pannonian region) and Serbia commercial orchards. In this context, 2-year old virus-free sour cherry plants of 4 isolated Oblačinska sour cherry ecotypes (OS, 18, D6, and BOR) produced by micropropagation were exposed to severe drought in a greenhouse under semi-controlled conditions to evaluate its photosynthetic intra-varietal variability. Relative water content (RWC), chlorophyll fluorescence (ChlF), and photosynthetic pigments were evaluated during the ten days of the experiment. As a visible symptom of stress, the withering of plants was followed by a diminution of RWC and photosynthetic pigments in the drought exposed leaves of sour cherry ecotypes compared to the control treatment. ChlF elucidated variability in the photosynthetic efficiency within studied sour cherry ecotypes, highlighting PIABS, PItotal, and ψE0 as the most sensitive and thus the most informative JIP parameters for drought screening. Among the investigated ecotypes, BOR proved to be the most sensitive. The Oblačinska sour cherry ecotype OS showed the highest tolerance to drought conditions and, therefore, can be used as a source of tolerance in sour cherry breeding programs.