Background Ficus johannis subsp. afghanistanica (Warb.) Browicz is an important plant species belonging to the Moraceae family and is part of the Ficus genus. In this study, the phenotypic diversity of 47 accessions of F. johannis subsp. afghanistanica was investigated using 61 morphological traits, evaluated from the Rachedr area in Sarbaz county, Sistan-va-Baluchestan province, Iran. Results One-way ANOVA (p < 0.05) revealed significant differences among the accessions. The coefficient of variation (CV) ranged from 4.54% (seed thickness) to 75.60% (trunk color), with 34 of the 50 traits (68%) having CVs greater than 20.00%, indicating a high degree of variation among the accessions. Leaf blade lengths ranged from 55.17 to 91.50 mm and leaf areas from 3125.93 to 7434.38 mm². Additionally, fruit length varied from 15.61 to 27.50 mm and fruit width from 13.65 to 25.16 mm. The study highlights significant morphological diversity of F. johannis subsp. afghanistanica, with key traits, including erect growth habits (25 accessions), intermediate and high vigor (21 accessions), and varied fruit ripening times (35 early, 12 late). Correlation analysis revealed a high correlation between leaf blade length and width (r = 0.835**) and fruit length and width (r = 0.987**). The results of principal component analysis showed that PC1 (6.63%), PC2 (5.06%), and PC3 (3.64%) highlighted leaf sizes, fruit sizes, and branch characteristics, respectively. The scatter plot showed that a total of 46 accessions fell within the 95% confidence ellipse, while the ‘Rachedr-39’ accession was located outside of it. Hierarchical cluster analysis revealed 4 accessions in subgroup A1, 9 in A2, 21 in B1, and 13 in B2. Conclusions As a result, the first 10 accessions of F. johannis subsp. afghanistanica were detected (‘Rachedr-1’, ‘Rachedr-39’, ‘Rachedr-16’, ‘Rachedr-31’, ‘Rachedr-37’, ‘Rachedr-12’, ‘Rachedr-15’, ‘Rachedr-3’, ‘Rachedr-27’, and ‘Rachedr-2’), with ‘Rachedr-39’ standing out due to its significant morphological differences. This study highlights the importance of morphological variation in understanding the biological diversity of the species, contributing valuable insights for conservation strategies aimed at ensuring sustainability and adaptability. These findings provide a crucial reference point for future research in plant biology and conservation.
