REAL-TIME RT-qPCR DETECTION OF CHERRY RASP LEAF VIRUS, CHERRY GREEN RING MOTTLE VIRUS, CHERRY NECROTIC RUSTY MOTTLE VIRUS, CHERRY VIRUS A AND APPLE CHLOROTIC LEAF SPOT VIRUS IN STONE FRUITS

F. Osman, M. Al Rwahnih, A. Rowhani

Abstract


Real-time quantitative RT-PCR (RT-qPCR) was developed for the detection of Cherry rasp leaf virus (CRLV), Cherry green ring mottle virus (CGRMV), Cherry necrotic rusty mottle virus (CNRMV), Cherry virus A (CVA) and Apple chlorotic leaf spot virus (ACLSV) in stone fruits, including apricot, cherry, peach, plum and almond. The RT-qPCR assays were designed after aligning nucleotide sequences derived from the most conserved regions of the coat protein gene of geographically diverse isolates and isolates for which sequence information is available in GenBank. RT-qPCR assays were validated and compared to conventional RT-PCR using 221 isolates. Our data showed that more isolates were detected by RT-qPCR than by RT-PCR; 0.9%, 6.3%, 1.29%, 5% and 8.14% more for CRLV, CGRMV, CNRMV, CVA and ACLSV, respectively. 


Keywords


RT-qPCR; sequence alignments; fruit tree virus detection

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DOI: http://dx.doi.org/10.4454/jpp.v99i1.3789

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