Typing and distribution of Plum pox virus isolates in Romania

Plum pox or Sharka, caused by Plum pox virus (PPV) is considered the most destructive disease of plum. Although PPV is widespread in all plum growing areas of Romania and causes serious yield losses, little is known about the variability of its isolates at a country level. For this reason, a large-scale study was performed with the aim of obtaining a picture of the prevalence and distribution of PPV strains in plum. During a three year survey, 200 PPV isolates collected from 23 different plum orchards from Transylvania, Moldavia and Muntenia areas were investigated. DASELISA 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/RT-PCR 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 the RsaI polymorphism located in (Cter)CP. To confirm the presence of PPV-Rec strain, 13 PCR products spanning the (Cter)NIb/(Nter)CP were sequenced. Overall results showed that in Romania the predominant strain is PPV-D (73%), followed, with a much lower frequency, by PPV-Rec (14%). Mixed infections (PPV-D+PPVRec), which might generate additional variation by recombination, are also frequent (13%).Keywords: Romania, PPV strains, DAS/TAS-ELISA, IC/RT-PCR, RFLP, sequencin