Poster: Diseases of Plants: Disease Detection & Diagnosis
Toward broad detection of emaraviruses: Endpoint RT-PCR
A. OLMEDO-VELARDE (1), F. Ochoa-Corona (2), T. Elbeaino (3) (1) Universidad de las Fuerzas Armadas ESPE, Ecuador; (2) Oklahoma State University, U.S.A.; (3) Istituto Agronomico Mediterraneo di Bari, Italy
Emaravirus has six confirmed species and two unclassified species. Rapid detection and an effective screening of existing and new emaraviruses is needed for detection, identification, plant virus discovery and biosecurity applications. Available sequences of RNA dependent RNA polymerase (RNA 1) for European mountain ash ringspot-associated virus (EMARaV), Fig mosaic virus (FMV), Pigeonpea sterility mosaic virus (PPSMV), Pigeonpea sterility mosaic virus 2 (PPSMV2), Rose rosette virus (RRV), High Plains wheat mosaic virus (HPWMoV) formerly High plains virus (HPV), Raspberry leaf blotch virus (RLBV) and Redbud yellow ringspot-associated virus (RYRSaV), were aligned using Mega 6. A primer set (EMARA F&R7/8) was designed from conserved domains after strand sense normalization. The sensitivity of EMARA F&R7/8 was found to be 100 fg/reaction assessed using a reference positive control of HPWMoV (Agdia). The multiple Emaravirus detection was confirmed with three rose samples showing symptoms caused by RRV from three locations in Stillwater, OK, a HPWMoV reference positive control, and cDNA of EMARaV and FMV. All samples were screened by endpoint RT-PCR and tested positive for HPWMoV, RRV, EMARaV and FMV. Other emaraviruses (RLBV, RYRSaV, PPSMV and PPSMV2) were detected in-silico using Primer-Blast. This method addresses the need for sensitive molecular detection methods and discovery of novel emaraviruses.