First Report of Plum Pox Potyvirus in Ontario, Canada.

Abstract

Plum pox potyvirus (PPV) causes plum pox (sharka) disease, which is considered the most serious disease of stone fruits including peach, plum, nectarine, and apricot (2). The disease may cause losses as high as 80 to 100% of some crops (2). A survey was initiated in the Niagara region of Ontario, Canada, after it was reported that PPV was detected in Pennsylvania (1). The initial survey focused on Prunus material imported into Canada from the Pennsylvania region. Where imported trees could be identified, every tree was sampled. In cases where the imported trees were growing in mixed blocks with plants from other sources, 25% of the trees were sampled and tested as composites of four trees. PPV was detected in three symptomless Fantasia nectarine (Prunus persica var. nectarina) trees by triple-antibody sandwich (TAS) ELISA using the REAL Durviz kit (Valencia, Spain), which contains the universal PPV monoclonal 5B. PPV infection was confirmed by western blot analyses (a PPV polyclonal antibody and PPV 5B monoclonal were used as primary antibodies), reverse transcription polymerase chain reaction (RT-PCR), and TC/RT-PCR. In western blot analyses, the coat protein subunit sizes of the Canadian PPV isolates were estimated at 32 kDa based on electrophoretic mobility in 12% SDS-PAGE. RFLP analysis of the 243-bp fragment amplified using PPV specific primers P1 and P2 (4) indicated the presence of RsaI and AluI enzyme restriction sites, which is characteristic of PPV D strains. In RT-PCR analysis using D and M specific primers (3), only the D specific primers amplified a fragment 198 bp in size. This data provided conclusive evidence that the PPV isolates detected in Canada were PPV D, similar to the strain detected in Pennsylvania. The survey is continuing and is being expanded to determine the extent of spread and the exact distribution of the virus.