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Diversity of plum pox virus isolates in Romania

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Author(s): Luminita Zagrai | Ioan. Zagrai | Beatrice Kelemen | Ioana Petricele3 | Doru Pamfil | Beatrice Ferencz | Silvia Preda | O. Popescu | Maria Isac | Alexandru Briciu

Journal: Scientific Papers of the Research Institute for Fruit Growing Pitesti, Romania
ISSN 1584-2231

Volume: XXV;
Start page: 158;
Date: 2009;
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Keywords: PPV strains | DAS/TAS-ELISA | IC/RT-PCR | RFLP | sequencing

ABSTRACT
Plum pox or Sharka, caused by Plum pox virus (PPV) is considered as the most destructive disease of plum. Although PPV is widespread in all plum growing areas from Romania and causes serious yield losses, little is known about the variability of its isolates at country level. For this reason, a large-scale study was performed with the aim to get a picture of the diversity of PPV strains in plum. During three years surveys, 200 PPV isolates collected from 23 different orchards covering the main plum growing areas of Romania were investigated. DAS-ELISA and IC-RT-PCR were used for PPV detection. 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 was used to distinguish D and M strains, based on RsaI polymorphism located in (Cter)CP. Thirteen PCR products spanning the (Cter)NIb/(Nter)CP region were sequenced. Overall results provide that in Romania the predominant strain is PPV-D (73%), followed with a much lower frequency by PPV-Rec (14%). Mixed infections (PPV-D+PPV-Rec), which might generate additional variation by recombination, are also frequent (13%).
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