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<i>Cercospora seminalis</i> is an imperfect fungus causing disease in buffalograss caryopses. The disease is called false smut since the diseased caryopses resemble smut. The disease is spread by asexual spores and contaminated plant debris causing considerable loss of yield and reduced germination of buffalograss seed. No genetic diversity studies have been reported on this pathogen. The objective of this study was to obtain information on genetic diversity of fungal isolates collected from diseased buffalograss burs. A collection of 84 <i>C. seminalis</i> isolates were made from six sites in Nebraska. DNA fingerprinting analysis was performed by amplified fragment length polymorphism (AFLP) using four selective primer combinations. In total, 482 DNA markers were detected across all isolates. Out of that 478 markers were polymorphic while 4 were monomorphic. The unweighted pair-grouped method with arithmetic average (UPGMA) clustering resulted in six lineages (I to VI). AMOVA test supported the significance of these lineages at <i>P</i> = 0.05. However 14 isolates did not fall into any cluster indicating high heterogeneity of the fungus. Most of the isolates (62%) belonged to lineage I (36 isolates) and II (16 isolates). The geographic origin of the isolates and resulted clusters had no relationship. These results show high genetic diversity of <i>C. seminalis</i> isolates that are not site-specific. Analysis of more isolates from different geographic areas is needed to establish the population structure of the pathogen. <p><p>Keywords: Fungus, Turfgrass, Warm-season