FIRST DETECTION AND MOLECULAR CARACTERIZATION OF PLUM POX VIRUS RECOMBINANT STRAIN IN ROMANIA
Abstract
Plum pox virus (PPV) is the most dangerous viral pathogen of stone fruits causing serious yield losses. Although PPV is widespread in Romanian plum orchards, very limited information about the variability of isolates is known. In this study we collected and investigated twenty PPV isolates from two experimental plots from Fruit Research and Development Station Bistrita. PPV strains were serologically determined by TAS-ELISA using PPV-D and PPV-M specific monoclonal antibodies. Molecular strain typing was performed by IC-RT-PCR targeting three genomic regions (Cter)CP, (Cter)NIb - (Nter)CP and CI. RFLP analysis was used to distinguish the two major strains, D and M based on a Rsa I polymorphism located in (Cter) CP. PCR products spanning (Cter)CP and (Cter)NIb - (Nter)CP regions were sequenced. All PPV isolates typed as PPV-M by serological analysis and also by molecular differentiation in (Cter)CP region proved to be recombinants between PPV-D and PPV-M when the molecular analysis were performed in (Cter) NIb – (Nter)CP region. The sequencing results revealed a high homology with different sequences of PPV recombinant (PPV-Rec) previously reported. All these recombinant isolates share the same recombination breakpoint and have conserved the DAG motif, which is considered essential for potyvirus aphid transmission. This genetic similarity confirms that PPV-Rec may represent an ancestral group with a common evolutionary origin. This is the first report of naturally infected plum trees with recombinant PPV strain (PPV-Rec) in Romania and predicted a widespread in the other areas where plum is growing.
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