The European corn borer, Ostrinia nubilalis Hübner, is one of the most important economic pests that could be found in all parts of Iran. In order to study the geographical diversity of this pest, larvae were collected from Ardabil, Golestan, Kordestan, East-Azarbayjan and Sistan-Baluchestan provinces during 2012-2013. Subject to polymerase chain reaction (PCR) using 10 different SSR primers that produced 10 polymorphic bands with 100 percent polymorphism were determined. Genetic diversity analysis of the European corn borer showed significant differences within and among geographical populations and their genetic diversity based on Nei’s gene index accounted 19.2 and 80.8 percent, respectively. Maximum and minimum genetic diversity and number of polymorphic loci were in Gorgan population 0.277, 9 and Iranshahr 0.104, 6, respectively. Maximum and minimum heterozygosity were in Os-N3 and Os-N10 markers by 0.748 and 0.203, respectively. Genetic similarity of studied geographical populations observed from 38 to 82 percent. Maximum and minimum genetic distances were observed between Sanandaj - Gorgan and Mogan - Shabstar populations 0.917, 0.198, respectively. Based on UPGMA method Mogan and Shabstar populations settled as unique in one cluster and Gorgan, Sanandaj, Iranshahr populations in another cluster. To test the correlation genetic and geographical populations distances was used Mantel test that indicated low or negative correlation (0.243 P