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Inter Simple Sequence Repeat (ISSR) markers were used to study the genetic diversity of 18 spring growth type landraces from Iran and selected 9 cultivars grown in Iran out of 15 ISSR primers .11 primers were found to have enough polymorphism and they were used for assessment. These primers are composed of di, tetra and penta-nucleotide sequences. From a total of 108 DNA fragments produced, 78 (72.22%) fragments were polymorphic. The UPGMA clustering algorithm classified the varieties into three major groups .Majority of landraces located in each group were originated from common locations. The results revealed that ISSR markers could be efficiently used to evaluate genetic variation in the wheat germplasm. Genetic similarity and dissimilarities among genotypes will be useful for genetic differentiation of wheat accessions, selection strategies and genetic development of crop plants.