ISHS Acta Horticulturae 234: VII International Symposium on Virus Diseases of Ornamental Plants
SENSITIVE AND SPECIFIC DETECTION OF TWO FILAMENTOUS VIRUSES FROM NERINES USING CLONED CDNA PROBES |
| Authors: | G. Balasingam, N. Ellison, K.S. Milne, R.L.S. Forster |
| Abstract:
Nerine virus X (NeVX), a potexvirus, was purified from Nerine fothergilli 'Major'. Double-stranded cDNA to the 6.3 kb genomic RNA was cloned into the Pst 1 site of the plasmid vector pBR322. 'Nick - translated pBR322 DNA containing a 1.8 kb insert, representing 28.5% of the viral genome, was found to be highly specific and sensitive to nerine isolates NeVX in dot-blot assays. The cDNA probe did not hybridize to seven other potexivruses, including cymbidium mosaic virus, narcissus mosaic virus, tulip virus X and the agapanthus strain of NeVX. An 800 nm potyvirus was purified from Nerine sarniensis hybrids. This hitherto unnamed virus was partially characterized and the 10.0 kb RNA was cDNA cloned into the lambda vector gt10. A 1.5 kb cDNA clone representing 15% of the viral genome was subcloned into the plasmid vector pGEM3. The probe was highly specific and sensitive to the homologous virus in infected nerine tissue. It did not hybridize in dot-blot assays of nerine tissue infected with nerine yellow stripe virus from England, or with three other potyviruses. Nerine virus Y (NeVY) is tentatively proposed as the name for this virus. The cDNA probes provided an effective, sensitive diagnostic technique for the detection of NeVX and NeVY in commercially cultivated nerines. | |
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