The sweet potato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a major agricultural pest that causes economic damages worldwide. In particular, B. tabaci MED (Mediterranean) has resulted in serious economic losses in tomato production of Korea. In this study, 1,145 B. tabaci MED females from 35 tomato greenhouses in different geographic regions were collected from 2016 to 2018 (17 populations in 2016, 13 in 2017, and five in 2018) and analyzed to investigate their population genetic structures using eight microsatellite markers. The average number of alleles per population (NA) ranged from 2.000 to 5.875, the expected heterozygosity (HE) ranged from 0.218 to 0.600, the observed heterozygosity (HO) ranged from 0.061 to 0.580, and the fixation index inbreeding coefficient (FIS) ranged from -0.391 to 0.872 over the three years of the study. Some significant correlation (p < 0.05) was present between genetic differentiations (FST) and geographical distance, and a comparatively high proportion of variation was found among the B. tabaci MED populations. The B. tabaci MED populations were divided into two well-differentiated genetic clusters within different geographic regions. Interestingly, its genetic structures converged into one genetic cluster during just one year. The reasons for this genetic change were speculated to arise from different fitness, insecticide resistance, and insect movement by human activities.