Identification of Turnip mosaic virus isolated from canola in northeast area of Iran

Abstract

During March and April of 2011, 436 samples showing viral disease symptoms were collected from canola fields in the Khorasan Razavi province. The samples were tested by double-antibody sandwich (DAS)-enzyme linked immunosorbent assay (ELISA) for the presence of Turnip mosaic virus (TuMV). Among the 436 samples, 117 samples were found to be infected with TuMV. One of the infected samples from Govareshk region (TuMV-IRN GSK) was selected for biological purification. Total RNA of this isolate were extracted and reverse transcriptase (RT)-PCR was performed with specific primers according to the coat protein gene. PCR products (986 bp) was first purified and then directly sequenced. Phylogenetic analyses based on ClustalW multiple alignments with previously reported 33 isolates indicated 88 to 98% similarity in nucleotide and 94 to 99% in amino acid levels among isolates. TuMV-IRN GSK represented the highest identity to another Iranian isolate (IRN TRa6). Phylogenetic tree clustered all sequences into four groups and IRN GSK fell into the basal-B group. Nucleotide and amino acid distances between IRN GSK and other isolates in the basal-B group showed that this isolate was closely related to another Iranian isolate IRN TRa6, and distinct from other isolates in the basal-B group. These results indicate that TuMV is a common pathogen of canola crops in the Khorasan Razavi province.

Key words: Turnip mosaic virus (TuMV), canola, reverse-transcription polymerase chain reaction (RT-PCR), coat protein gene, sequence analysis.