In 2018, a total of 58 mustard samples from two different localities in Serbia (Rimski Šančevi and Senta) were collected and tested for the presence of Turnip yellow virus (TuYV), Cauliflower mosaic virus (CaMV) and Turnip mosaic virus (TuMV) by DAS-ELTSA. TuYV was serologically detected in 42 tested samples while all collected samples were negative for CaMV and TuMV. By using aphid transmission tests, five test plants were inoculated with two ELTSApositive TuYV samples of naturally infected mustard plants. Virus species identification was performed by amplification of a 780 bp fragment in all tested samples using the specific primers TuYVorf0F/TuYVorf0R. The RTPCR products from two isolates 88Sal (MK144816) and 98Bni (MK144817) were sequenced and compared with the GenBank sequences of TuYV. Serbian isolates showed the highest identity with Poland TuYV isolate (EU022489). Phylogenetic analysis showed that TuYV isolates from Serbia were clustered with other TuYV sequences retrieved from the GenBank.