Development and Validation of a New TaqMan Real-Time PCR for Detection of ' Candidatus Phytoplasma pruni'.

Phytoplasmas of the 16SrIII group are wide spread, and have a broad plant host range. Among these, 'Candidatus phytoplasma pruni' ('Ca.P. pruni'; phytoplasmas of 16SrIII subgroup A) can cause serious diseases inPrunusspecies and 'Ca.P. pruni'-related strains can infect other plant species, including grapevines. In this study, a new real-time PCR detection system was developed for 'Ca.P. pruni' using TaqMan chemistry. This test was designed to detect 'Ca.P. pruni', by amplifying the species-specificsecYgene. In addition, a test to amplify the group-specific 16S rRNA gene region was also developed. The performances of both tests were evaluated. The test that amplifies thesecYgene provided reliable and quick detection of 'Ca.P. pruni'. Using the newly developed and validated test, 'Ca.P. pruni' was not found in any of the 434 field samples collected from different plants species grown in different regions of Slovenia.