Cultivated rocket (Eruca vesicaria) is a leafy vegetable highly appreciated for its health-promoting virtues and consumed both raw and cooked as ready-to-use vegetable. Despite Eruca being cultivated worldwide, only a few cultivars are available and limited breeding activities have been carried out so far. Therefore, the genetic resources available represent an unexploited potential source of variation for breeding. In the present study, 155 E. vesicaria accessions from 30 countries across Europe, Asia, Africa, and America have been characterized for 54 qualitative and quantitative morphological and quality traits. Conventional descriptors and automated tools for the determination of the quality, morphology, and colour of leaves have been used. Genetic diversity was assessed using 15 inter simple sequence repeat and simple sequence repeat markers. A high level of diversity was evidenced in the collection. Significant differences were found in most of the traits with the exception of five pseudo-qualitative descriptors. The first and second dimensions of the principal components analysis with phenotypic traits accounted for 25.69% of total variation showing a stratification of the genotypes according to the European and Asian origins. In total, 75% of the variation was contained in the first 15 components having eigenvalues higher than 1.0. Also, the population structure divided the collection into two main clusters separating European genotypes from the rest. Furthermore, hierarchical cluster analysis confirmed a geographical separation, grouping the accessions into three major clusters, which were differentiated by plant architecture, leaf and flower colour, leaf water status, leaf blade shape and hairiness of the leaves and stem. Our approach has broadened the knowledge of the diversity within the Eruca gene pool, thus contributing to identify sources of variation and to select the best candidates for cultivated rocket breeding programs, as well as to determine the genetic basis of plant and leaf traits in future genome-wide association studies.