Detection and partial molecular characterization of soil borne viruses infecting wheat and barley fields in Yazd province, Iran

Soil-borne viruses, belonging to the genera Furovirus and Bymovirus, are serious pathogens of autumn-sown wheat and barley fields in Iran and transmitted by Polymyxa graminis. These viruses are of great economic importance and causing significant quantitative and qualitative losses in yield. There are a few studies about soil borne cereal viruses and in conclusion there is not sufficient information about their distribution and diversity in cereal fields in Iran. During 2014-15, sampling was carried out on wheat and barley fields in Taft, Yazd, Meybod and Dehno (Yazd province, Iran). Disease symptoms were yellowing, mosaic and stunting and total 164 symptomatic and 14 asymptomatic samples were collected. Using double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) with Soil borne wheat mosaic virus (SBWMV) and Barley mild mosaic virus (BaMMV) polyclonal antibodies, SBWMV and BaMMV were detected in 34.3 and 17.4 % of collected samples respectively. Direct and nested PCR was carried out on positive samples in DAS-ELISA using the external (Wb1/Wb2, Bm1/Bm2) and internal (Wns1/Wns2, Bns1/Bns2) designed primer pairs for SBWMV and BaMMV respectively. The direct PCR amplified two fragments size of 1150 and 960 bp and nested PCR successfully amplified two products with the predicted size of 410 and 430 bp in SBWMV and BaMMV diseased plants respectively. No fragments were amplified with asymptomatic wheat and barley plants. Based on the results of serological and molecular analysis in this study, SBWMV and BaMMV are distributed in wheat and barley fields of Yazd province.

Keywords:

ELISA , Nested PCR , Polyclonal antibody , Primer