Cassava (Manihot esculenta) is a traditional crop in tropical and subtropical regions that is used for human consumption and in animal feed. This studied cassava landraces cultivated in northern Mato Grosso State, Brazil, to detect the variability in family farms, aiming at the preservation and use potential of these genetic resources. A total of 120 M. esculenta individuals were evaluated at the rate of 40 per location (population), where each landrace was represented by 10 plants. The 14 microsatellite markers examined showed genetic diversity. The average polymorphic information content (PIC) was 0.621 and the average number of alleles per locus was seven. Genetic diversity parameters indicated that H o was higher than H e for the three populations, showing negative fixation indices and a lack of inbreeding. AMOVA revealed greater molecular variation within the populations (92%). Bayesian analysis and the ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 18 (3): gmr18315 E.C.M. Pedri et al. 2UPGMA clustering method resulted in two main groups formed with the individuals distributed randomly; i.e., regardless of collection site (location). The gene flow found in this study is a consequence of the introduction and exchange of genetic material (landraces) performed by the farmers, who act on their farms as maintainers of local diversity. Manihot esculenta cultivation in northern Mato Grosso State ensure the on-farm conservation of genetic variability of the species and constitutes a source of genetic resources such as genes of resistance and adaptation, which can be exploited and utilized in breeding programs.