Beet necrotic yellow vein virus (BNYVV) is the casual agent of sugar beet rhizomania. BNYVV has a multipartite genome, a limited host range, a single vector species (Polymyxa betae Keskin) and a worldwide geographical distribution. Earlier obtained results show existence of three major subgroups, the A, B and P types. The A type is presented in most European countries, Iran, North America, China and Japan, whereas the B type is detected frequently in France and Germany, and in some cases in Sweden, Poland, China and Japan.

Total RNA was extracted from roots of the symptomatic plants and used in a multiplex reverse transcription-polymerase chain reaction (mRT-PCR) assay. Specific primers were designed by Meunier to amplify a fragment of the RNA2 for BNYVV.

The amplified RT-PCR products were purified using the Qiaex II gel extraction kit (Qiagen). Obtained fragments were ligated into the pGEM-T easy vector (Promega) and transformed E. coli Dh5α (Invitrogen). Three independent purified recombinant plasmids DNA were sequenced on an ABI automatic sequencer. Nucleotide and amino acid sequences were compared with the sequences available in the GenBank database. Phylogenetic trees were constructed to establish relationships between Polish BNYVV isolates and others reported to data. Multiple sequence alignments were performed using ClustalW. The alignments were used to calculate pairwise genetic distances by Kimura’s two-parameter method. Phylogenetic analysis was carried out by the neighbor-joining method implemented with Mega version 3.1. Bootstrap values for phylogenetic comparisons are based on 1000 pseudoreplicates. On the basis of phylogenetic analyses we showed that Polish isolates of BNYVV are very similar to others deposited in the GenBank database. In case of both types of BNYVV we obtained 100% homology with other isolates from the GenBank database.

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