Molecular characterization of ‘Candidatus Phytoplasma mali’ strains in outbreaks of apple proliferation in north eastern Italy, Hungary, and Serbia

During 2005-2008 apple plants of different varieties showing proliferation symptoms were observed in diverse areas of north eastern Italy, Hungary and Serbia. PCR/RFLP analyses showed that all the samples were infected with ‘Candidatus Phytoplasma mali’. In the 16S plus spacer region two phytoplasma profiles (P-I and P-II) were distinguished. P-I profile was detected in reference strains AP, AT1, AT2, in samples from Serbia, and in the majority of samples from Trentino; the P-II profile was prevalent in samples from Veneto; both profiles were identified in samples from Hungary, in some cases together in single samples. The analyses of rpl22-s3 genes allow the identification, in all the samples showing a P-I profile, the presence of phytoplasmas belonging to rpX-A subgroup, while in the samples showing a P-II profile it was possible to distinguish the other three reported rpX subgroups. In the majority of samples from the Veneto region phytoplasmas belonging to rpX-D subgroup were identified, while rpX-B and rpX-C subgroups were identified only in a few samples from Trentino and Veneto regions, respectively. Further RFLP analyses on AP13/AP10 amplicons differentiate among strains belonging to the rpX-A subgroup: the samples from Serbia show AP profiles, while those from Trentino show AT-2 profiles. In the samples from Hungary the presence of AT1, AT2, and AP profiles was identified.