Discrimination of Three BYDV Species by One-step RT-PCR-RFLP and Sequence Based Methods in Cereal Plants from the Czech Republic

摘要

One-step RT-PCR followed by a RFLP assay was developed for the typing of Barley yellow dwarf virus (BYDV) member of Luteovirus genera. RFLP analysis performed on 23 samples including one lab isolate showed three types of Hpa II restriction profile. A partial coat protein (CP) gene sequencing was carried out and confirmed the RFLP analysis. Both sequencing and RFLP analysis identified the presence of 3 BYDV-PAV (including the lab isolate Blatno85), 10 BYDV-MAV and 10 BYDV-PAS isolates among the field samples. One-step RT-PCR together with RFLP presents an easy and reliable assay for routine BYDV typing. These methods revealed that PAS and MAV are more dominant than PAV in the Czech Republic.