Barberry (Berberis spp.) is considered beneficial in the treatment of metabolic abnormalities. Tiny barberries with unique taste and flavor make them ideal for the food industry. This study investigated the morphological, nutritional, and biochemical traits of 12 Berberis genotypes. Non-metric multidimensional scaling (NMDS) and principal component analysis (PCA) were used to classify the genotypes. Based on the results, genotypes #8‑3 and #2‑2 showed the highest length and width of berries, respectively. Pulp percentages ranged from 51.93 to 98.5%. The lowest fresh and dried weights of 100 berries were found in #11‑1 and seedless, respectively. The highest levels of ascorbic acid and protein contents were obtained in #14‑2. The potential genotype for total soluble sugar (TSS) was seedless (59.61%). Genotype #10‑1 showed the fiber content and average nutritive value (ANV) with the highest levels of Fe and Ca. Genotype #5‑1 showed the maximum amounts of total phenolic compound (TPC), while the highest levels of total flavonoid compound (TFC) and total monomeric anthocyanin content (TMAC) were related to #14‑2. Consequently, the IC50 value of #14‑2 was significantly lower than the other genotypes. Genotype #14‑2 with a unique black color and a bluish wax showed 29.44, 2.31, and −3.23 lightness (L*), greenness [−] to redness [+] (a*), and blueness [−] to yellowness [+] (b*), respectively. In terms of TPC, the results of the NMDS ordination showed the separation of #12‑1, while the TFC and TMAC results showed the separation of #14‑2. The stress values were low, which indicated the high goodness of fit of the ordination distances to the observed distances.

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