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Tef is the most important and well adapted cereal crop in Ethiopia and grows better than other cereals both in biotic and abiotic stress conditions. In this study 93 tef accessions representative of various agro-ecologies of Ethiopia along with four improved varieties, one local check and six wild relatives were studied. The materials were genotyped with 12 simple sequence repeat markers (SSRs) to investigate the extent and structure of the genetic diversity of tef. A total of 152 alleles were detected indicating high polymorphism in the studied accessions. The number of alleles per polymorphic loci ranged from seven for CNLT 538 to 29 for CNLT150 with an average of 12.67. High average polymorphic information content (0.82) and average expected heterozygosity (0.86) also signifies the suitability and efficiency of SSR markers to discriminate among tef genotypes. Cluster, principal coordinate and structure analyses grouped the 98 tef genotypes and six wild relatives into four distinct classes. The groupings showed no special pattern despite the different regions of origins for the accessions; however, the five wild relatives were grouped together. The analysis of molecular variance and fixation index showed low genetic differentiation and high gene flow between regions revealing that much of the variability was attributed to within populations variation. Generally, the present study showed that there is high genetic variability with unstructured genetic pattern among the studied tef genotypes which implies the vitality of conserved tef germplasms at Ethiopian Biodiversity Institute and the impact of the seed exchange system on tef breeding activities in Ethiopia.