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Genetic diversity of Plum pox virus Isolates in Muntenia, Romania

Romanian Biotechnological Letters

Authors:

Ion Zagrai, Luminita Zagrai
Fruit Research and Development Station Bistrita, Romania

Silvia Preda, Maria Isac
Fruit Research and Development Station Valcea, Romania

Breatrice Kelemen, Octavian Popescu
Babes Bolyai University, Faculty of Biology and Geology, Cluj-Napoca, Romania

Ioana Petricele, Doru Pamfil
University of Agricultural Science and Veterinary Medicine, Cluj-Napoca, Romania

Abstract:
Plum pox virus (PPV) is considered the most devastating viral pathogen of stone fruits causing serious yield losses. Although PPV is endemic in Romania, limited data about the occurrence of its strains in some plum regions like Muntenia are available. Considering this fact, fifty PPV isolates collected from five different commercial plum orchards from Muntenia were investigated. PPV detection was made by DAS-ELISA and by IC-RT-PCR. PPV strains were serologically determined by TAS-ELISA using PPV-D and PPV-M specific monoclonal antibodies. Molecular strain typing was done by IC/RTPCR targeting three genomic regions corresponding to (Cter)CP, (Cter)NIb/(Nter)CP and CI. RFLP analysis at the C-ter of CP cistron was used to distinguish the two major strains, D and M based on a RsaI polymorphism located in this genomic section. All PCR products targeting (Cter)CP and two PCR product spanning the (Cter)NIb/(Nter)CP were sequenced. All PPV isolates typed as PPV-M by serological analysis and by molecular differentiation in the genomic region corresponding to (C-ter)CP proved to be PPV recombinant (PPV-Rec) when the molecular analysis were performed in the region corresponding to NIb/CP. Sequencing results confirmed a high similarity with different sequences of PPV-Rec previously reported. Overall results provided that in Muntenia the prevalent strain is PPV-D. PPV-Rec is also present with a much lower frequency both in singular and mixed infections (PPV-D+PPV-Rec).