Maintaining genetic diversity for crop improvement and improving the quality of genetic resource management is both an inevitable part of nowadays maize breeding. ISSR markers brings the potential of finding a marker system to discriminate Iowa Stiff Stalk Synthetic and Iodent Reid populations of Zea mays, L. and on the base on coefficients of genetic distance and UPGMA grouping appropriate maize genotypes for the best potential for hybridization can be selected. The work presents the potential of ISSR markers in analysis of length polymorphism when a specific ISSR pattern can be used for fast screening of differences among maize genotypes and based of these differences lines can be used in breeding programms in a very specific mode.