First Report of Broad bean wilt virus-2 in Pepper in the Czech Republic.

In August 2012, field-grown pepper (Capsicum annuum) plants were inspected for the presence of viral diseases in Moravia, Czech Republic. Leaves taken from plants showing virus-like symptoms were examined for the presence of pepper viruses by double antibody sandwich (DAS)-ELISA. Of the 80 collected samples, 22 were infected with Cucumber mosaic virus, 17 with Potato virus Y, 13 with Broad bean wilt virus-1, and 11 with Broad bean wilt virus-2 (BBWV-2). The BBWV-2 positive samples were collected near the villages of Hrádek and Tasovice in Znojmo District and Loděnice in Brno- Country District. Six samples positive for BBWV-2 were also infected with one or two other viruses. BBWV-2 was mechanically transmitted from the fieldcollected samples where it was present alone onto Chenopodium quinoa plants. After systemic mottling emerged on indicator plants, symptomatic leaves were collected and the presence of BBWV-2 was confirmed by electron microscopy and reverse transcriptase (RT)-PCR. Isometric particles of about 25 nm corresponding in size and shape to the BBWV-2 virions (2) were observed. Total RNA was extracted from infected C. quinoa leaves, and RTPCRs with primer pairs specific to four BBWV-2 genome regions (1) were carried out using oligo-dT and random hexamer mixtures for the cDNA synthesis. Amplicons of expected sizes, ~400 bp for Fab5'R1, ~460 bp for Pol2, ~500 bp for MP4, and ~250 bp for SCP were obtained for all samples. To the best of our knowledge, this is the first report of a natural occurrence of BBWV-2 in the Czech Republic. © The American Phytopathological Society.