Plant identification and genotyping within a germplasm collection are important elements in planning a breeding program that can improve the development of plants with desirable market characteristics. In the current study, 20 squash (Cucurbita pepo L.) landraces were collected and identified from different areas in Egypt to investigate the plant diversity and genetic variations based on differences in plant morphology and the International Union for Protection of New Varieties of Plants (UPOV) descriptors. A total of 24 quantitative and qualitative morphological differences characterized the genetic variability among genotypes. The demonstrated variability among all the tested landraces suggested a large number of genotypes within the squash lines. The results showed the potential of morphological markers for the identification and assessment of genetic variations among squash landraces and provided a number of choices for planning a successful breeding program to improve squash landraces. The obtained results on the 20 studied genotypes established that the agronomic traits based on morphological traits and UPOV descriptor data provide cost-effective preliminary tools for genotypic varietal characterization and evaluation for breeding purposes. Moreover, the utilization of these neglected local squash landraces and the subsequent improvement can increase the diversity in the food marketplace at the local level and enhance agricultural biodiversity.