ABSTRACTThe genus Crataegus, belonging to the Rosaceae family, exhibits widespread distribution across Iran, comprising 17 species. Hawthorn has garnered significant attention in recent years as a prominent herbal remedy in phytotherapy and culinary applications. Various plant parts, including flowers, leaves, and fruits, have been traditionally employed to address cardiovascular conditions such as hypertension, hypotension, palpitations, and cardiac arrhythmias. The presence of bioactive phytochemicals, notably polyphenols and anthocyanins, confers antispasmodic and analgesic properties to hawthorn. This study investigated the bioactive capacity of mature fruits from five Crataegus persica ecotypes indigenous to Iran. High‐performance liquid chromatography (HPLC) was utilized to elucidate the phenolic profile, although morphological characteristics and antimicrobial properties were also assessed. The ecotypes under investigation were sourced from elevations ranging from 1205 to 1681 m above sea level. Morphological analysis revealed that fruit weight varied from 0.84 to 2.53 g, whereas pericarp weight ranged from 0.21 to 1.19 g. The number of seeds per fruit fluctuated between 1 and 3. HPLC analysis identified the primary phenolic compounds in C. persica as catechin, p‐coumaric acid, quercetin, chlorogenic acid, and rutin. Biochemical characterization of the ecotypes yielded the following ranges: total soluble solids (TSS) 10.31°Brix–18.53°Brix, total soluble carbohydrates (TSC) 8.34%–16.65%, vitamin C content 4.24–12.38 mg g−1 DW, total carotenoid content 185.32–294.4 μg g−1 DW, and total anthocyanin content 43.21–85.34 mg C3G g−1 DW. Total phenolic compounds ranged from 170.52 to 254.31 mg GAE g−1 DW, whereas total flavonoid content varied between 59.28 and 84.41 mg RE g−1 DW. Antioxidant activity, as determined by IC50 values, ranged from 14.47 to 37.51 μg mL−1. Antimicrobial assays demonstrated that the C. persica extract exhibited maximum efficacy against Staphylococcus aureus, with the CP3 ecotype extract displaying a minimum inhibitory concentration (MIC) of 0.125 mg mL−1. In conclusion, this comprehensive analysis of five distinct C. persica ecotypes (CP1, CP2, CP3, CP4, and CP5) revealed substantial diversity in terms of morphological traits, functional bioactive compounds, and antimicrobial potential. These findings contribute to the growing body of knowledge regarding the phytochemical composition and potential therapeutic applications of Crataegus persica.