ISHS Acta Horticulturae 657: XIX International Symposium on Virus and Virus-like Diseases of Temperate Fruit Crops - Fruit Tree Diseases
PRELIMINARY MOLECULAR CHARACTERIZATION OF PLUM POX POTYVIRUS ISOLATE W3174: EVIDENCE OF A NEW STRAIN |
| Authors: | D. James, A. Varga |
| Keywords: | Plum pox virus, isolate W3174, strain typing, multiplex RT-PCR, host range |
| Abstract:
Plum pox potyvirus (PPV) isolate W3174 was detected in plum (Prunus domestica) growing on a homeowner property in Ontario, Canada. The virus was detected using the universal PPV monoclonal antibody 5B in TAS-ELISA, and by RT-PCR using primers that amplify a 243 bp fragment in the C-terminus of the coat protein coding region. The restriction sites RsaI and AluI were absent, a feature unique to PPV-C isolates. Attempts to detect the virus by RT-PCR using strain C specific primers were unsuccessful. All attempts at strain typing using published techniques, nucleic acid-based and serological, were unsuccessful. No reaction was observed when D, M, C, or EA specific monoclonal antibodies were used for ELISA or western blot detection of W3174. The level of identity of the 3' non-coding region and the coat protein amino acid (deduced) sequence of W3174 was estimated at 90-94%, and 80-82%, respectively, when compared to selected members of the 4 known strains of PPV. The DAG motif was detected in the amino acid sequence of the coat protein at position 12-14. W3174 grouped distinctly in phylogenetic analysis based on the replicase nucleotide sequences. Nucleic acid analysis and serological assay seem to indicate that PPV W3174 does not belong to any of the currently recognized strains of PPV. A multiplex RT-PCR assay was developed that allows simultaneous detection and typing of W3174. | |
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