Surveys were carried out throughout Turkey and a total of 437 symptomatic samples were collected from winter vegetables and weeds, most of which belong to the Brassicaceae family. Testing the collected samples for the presence of turnip mosaic virus (TuMV) by double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA) and reverse transcription-polymerase chain reaction (RT-PCR) confirmed that 117 samples were infected with TuMV. In total, 20 out of 117 TuMV-infected samples representing different regions and host plants were then selected and further analyzed for biological and molecular characteristics. As a result of biological indexing studies, two host infecting types of TuMV, i.e., B and BR were identified in Turkish TuMV isolates. The partial genomic sequences of the nuclear inclusion b protein and coat protein genes (NIb+CP region) of these isolates were determined. The corresponding sequence of the remaining 61 Turkish TuMV isolates were retrieved from GenBank for further analysis. The phylogenetic analysis showed that Turkish isolates fell into the basal-B, Asian-BR, and world-B phylogenetic groups. The evolutionary divergence value of Turkish isolates in the basal-B, Asian-BR, and world-B phylogroups were determined as 0.0026, 0.0134, and 0.0065 at nucleotide level; 0.0243, 0.0102, and 0.0129 at the amino acid level, respectively. Genetic structure analysis of Turkish TuMV isolates revealed that some isolates were admixed or had migrated among different subgroups. Our analysis provides the first depth study for the biological characteristics and genetic structures of Turkish TuMV populations.