| ELISA systems were developed for the detection of Hydrangea ringspot virus (HdRSV) and Nandina mosaic virus (NaMV) in hydrangea and nandina respectively, using established antisera. The two viruses were isolated, purified, and cloned. Sequence data from clones proximal to the 3' termini of the viruses were used to establish reverse transcriptase polymerase chain reaction (RT-PCR) systems for the detection of the two viruses. The initial two-step RT-PCR was modified to a one-step format which was used successfully for the sensitive detection of the viruses. Sequence data for NaMV indicated that it is an isolate of the previously sequenced Plantago asiatica mosaic virus (PlAMV). As sequence data was generated first for Plantago asiatica mosaic virus its name takes precedence over the name Nandina mosaic virus which must be considered to be a synonym of Plantago asiatica mosaic virus. This information promotes NaMV from the status of a tentative species within the genus Potexvirus to a full member of this genus.;Other one-step PCR tests were developed for the detection of Alfalfa mosaic virus (AMV), Cucumber mosaic virus (CMV), and Tomato ringspot virus (ToRSV) in buddleia. Using cloned PCR fragments of the respective viruses, cRNA probes were synthesized and detected using non-radioactive chemiluminescent substrates.;All detection systems were developed as part of a larger program to identify virus-free individuals of buddleia, hydrangea, and nandina and use these as sources of material for experiments to demonstrate the effects of viruses on the asexual propagation of these three widely grown species. |
