Ancient olives are considered a major resource of gene pool, adapted across ages to ever changing environments. The ancient ‘Mehras’ olive is the only cultivar inscribed on the UNESCO Representative List of the Intangible Cultural Heritage Of Humanity as recognized for its longevity and historical significance. However, detailed genetic and phenotypic analyses are still missing. Nineteen ‘Mehras’ accessions were collected from northern Jordan and subjected to inter-simple sequence repeat (ISSR) marker analysis and to a set of phenotypic parameters (leaf, fruit, and stone). ISSR analysis revealed similarity values ranging between 0.53 and 1.00, indicating moderate to high genetic diversity. Bayesian clustering and dendrogram analyses identified two major genetic clusters with limited admixture, indicating extended clonal propagation in addition to the transport of planting material. Phenotypic parameters revealed tangible variation among accessions, with major influence of fruit and stone traits followed by leaf traits. Strong correlations were observed between key traits, including fruit width and flesh thickness, while an inverse relationship was observed between flesh and stone percentages. Multivariate analysis further revealed clear separation among investigated accessions. The Mantel test showed a moderate correlation between genetic and phenotypic distances. Spatial analysis suggested weak geographic structuring of diversity, indicating exchange of plant material. ‘Mehras’ olive harbors structured genetic and phenotypic diversity influenced mainly by adaptation and traditional cultivation practices. These findings provide a foundation for conservation, breeding, and sustainable utilization of this ancient and culturally significant cultivar.

Ascorbate peroxidase (APX) is a heme-containing enzyme involved in hydrogen peroxide (H₂O₂) detoxification within the ascorbate–glutathione (AsA–GSH) cycle. In this study, the full-length genomic DNA and cDNA of an APX1 gene (VsAPX1) were cloned and characterized from Vicia sativa. The genomic sequence of VsAPX1 is 2425 bp in length and comprises 10 exons separated by nine introns, with the first intron located within the 5′ untranslated region (5′UTR). The corresponding cDNA is 1010 bp long and includes a 61 bp 5′UTR, a 753 bp open reading frame, and a 196 bp 3′UTR. VsAPX1 encodes a predicted cytosolic APX protein of 250 amino acids, with a molecular weight of 27.1 kDa and a theoretical isoelectric point (pI) of 5.60. Bioinformatics analysis revealed that the deduced VsAPX1 protein shares high sequence similarity with cytosolic APX1 proteins from other plant species, contains conserved APX domains, and clusters within the cytosolic APX clade in phylogenetic analysis. Quantitative real-time PCR analysis showed that VsAPX1 expression exhibits transient and moderate changes in response to abiotic stress and phytohormone treatments. Transcript levels increased at early time points following heat stress (42 °C), abscisic acid, and salicylic acid treatments, and after 4 h of jasmonic acid exposure, whereas hydrogen peroxide treatment resulted in a gradual down-regulation of expression. Overall, this study provides the first molecular and expression characterization of a cytosolic APX1 gene from Vicia sativa and establishes a foundation for future functional analyses of antioxidant genes in this species.