Submitted to: American Phytopathology Society
Publication Type: Abstract Only
Publication Acceptance Date: 3/10/2004
Publication Date: 7/31/2004
Citation: Li, R., Mock, R.G. 2004. An improved rt-pcr assay for the detection of two cherry foveaviruses in prunus spp.. American Phytopathology Society. 94(6):S60.
Technical Abstract: A one-step reverse transcription-polymerase chain reaction (RT-PCR) procedure was developed to detect Cherry green ring mottle virus (CGRMV) and Cherry necrotic rusty mottle virus (CNRMV) in indicator plants and infected Prunus spp. Viral RNA suitable for RT-PCR was obtained by a simple method that did not require extraction of dsRNA and total RNA, availability of antibodies to virus or purification of viral particles. Consensus primers and degenerate primers, whose designs were based on alignments of published sequences of foveaviruses infecting cherry, were tested to amplify viral genomic fragments of five CGRMV isolates and one CNRMV isolate. RT-PCR using a pair of consensus primers allowed CGRMV detection in viral RNA and total RNA preparations equivalent to 80 ng and 400 micrograms of infected leaf tissue, respectively. CGRMV was detected in leaves, tender shoots, petioles, and young bark, whereas the strongest bands were obtained using young leaves and tender shoots. Detection was less consistent in summer when temperature was elevated and plant tissues were old. The direct comparison of RT-PCR and bioassay indicates that the RT-PCR assay is sensitive, rapid and reliable. The method developed should allow for the rapid diagnosis of cherry foveaviruses in Prunus spp.