Spinach (Spinacia oleracea L.) is an economically important leafy vegetable crop in many countries. This is the first case study of using SRAP markers to analyze genetic diversity of Iranian spinach accessions. Eight SRAP primer combinations generated 88 scorable bands ranging from 50 to 1000 bp, among which 73 were polymorphic, with an average of 82.9 polymorphic bands per primer combination and average polymorphism information content (PIC) of 0.35. Based on the analysis of SRAP data, a similarity matrix was calculated according to the Dice coefficient. Similarity dendrogram was constructed using UPGMA algoritm. Cophenetic correlation between ultrametric similarities of tree and similarity matrix was found to be high (r=0.85, P < 0.0001), suggesting that the cluster analysis strongly represents the dissimilarity matrix. According to the UPGMA dendrogram, the spinach accessions clustered into two groups. This preliminary study demonstrated that SRAP markers are effective for evaluating genetic diversity between and within spinach accessions.