title: Generic detection of poleroviruses using an RT-PCR assay targeting the RdRp coding sequence.
abstract: In this study a two-step RT-PCR assay was developed for the generic detection of poleroviruses. The RdRp coding region was selected as the primers' target, since it differs significantly from that of other members in the family Luteoviridae and its sequence can be more informative than other regions in the viral genome. Species specific RT-PCR assays targeting the same region were also developed for the detection of the six most widespread poleroviral species (Beet mild yellowing virus, Beet western yellows virus, Cucurbit aphid-borne virus, Carrot red leaf virus, Potato leafroll virus and Turnip yellows virus) in Greece and the collection of isolates. These isolates along with other characterized ones were used for the evaluation of the generic PCR's detection range. The developed assay efficiently amplified a 593bp RdRp fragment from 46 isolates of 10 different Polerovirus species. Phylogenetic analysis using the generic PCR's amplicon sequence showed that although it cannot accurately infer evolutionary relationships within the genus it can differentiate poleroviruses at the species level. Overall, the described generic assay could be applied for the reliable detection of Polerovirus infections and, in combination with the specific PCRs, for the identification of new and uncharacterized species in the genus.
authors.affiliation: Aristotle University of Thessaloniki, School of Agriculture, Laboratory of Plant Pathology, 54124 Thessaloniki, Greece.
authors.forename: Leonidas
authors.initials: L
authors.lastname: Lotos
authors.affiliation: Aristotle University of Thessaloniki, School of Agriculture, Laboratory of Plant Pathology, 54124 Thessaloniki, Greece.
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Efthimiou
authors.affiliation: Aristotle University of Thessaloniki, School of Agriculture, Laboratory of Plant Pathology, 54124 Thessaloniki, Greece. Electronic address: vmaliogk@agro.auth.gr.
authors.forename: Varvara I
authors.initials: VI
authors.lastname: Maliogka
authors.affiliation: Aristotle University of Thessaloniki, School of Agriculture, Laboratory of Plant Pathology, 54124 Thessaloniki, Greece.
authors.forename: Nikolaos I
authors.initials: NI
authors.lastname: Katis
mesh_terms: D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D016679:Genome, Viral / Q000235:genetics; D006115:Greece; D052679:Luteoviridae / Q000235:genetics*; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics*; D010802:Phylogeny; D012367:RNA, Viral / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D016415:Sequence Alignment
chemical_list: D017931:DNA Primers; D012367:RNA, Viral
keywords: Generic RT-PCR; Luteoviridae; Phylogeny; Polerovirus; RdRp; Species-specific RT-PCR