This study focuses on characterizing the macro-biochemical composition of leaves of Pistacia lentiscus L from 11 natural populations in Morocco, to allow genetic differentiation of this species. This study population characterization allows research and identification of the most informative markers and analysis of relatedness between populations and their origin sites. The results of ANOVA showed that there were significant differences (p=0.05) in total sugar content, dry matter content, and fiber content between the studied populations. Organic matter, dry matter, and total nitrogen contents were positively and significantly correlated with energy values (r = 0.92 and 0.962). In contrast, negative correlations were found between dry matter, minerals, fat content, and carbohydrates (r = -0.217, -0.379). The results of the principal component analysis showed that the 11 studied populations were dispersed among four groups, regardless of their geographical proximity. This grouping is confirmed by hierarchical classification.