Cucumber (Cucumis sativus L.) is an economically important vegetable crop with increasing demand in Nigeria due to its nutritional and economic significance. However, challenges such as limited genetic diversity, climate change, and pest pressures hinder its optimal production. Understanding the genetic diversity and relationships of cucumber accessions is critical for effective breeding and crop improvement strategies. This study investigated the molecular relationships among three cucumber accessions using Random Amplified Polymorphic DNA (RAPD) markers to evaluate their genetic diversity and relatedness. Twelve primers were screened, out of which four (OPA04, OPA09, OPAE14, and OPAC11) produced 29 clear, scorable DNA bands ranging from 350 to 1500 base pairs. The percentage polymorphism recorded was 76.5%, with primer OPA04 exhibiting the highest polymorphic information content (PIC). Cluster analysis and genetic distance measurements revealed that the Poinsett accession was genetically distinct from Marketer and Poinsett++. These findings provide foundational genetic data for future cucumber breeding programs and for enhancing germplasm management.