Detection of ‘Candidatus Phytoplasma pini’ in Pinus sylvestris trees in Poland

Abnormal shoot branching was observed in Pinus sylvestris trees in Poland. These abnormalities resulted in the formation of dense, ball-like structures with dwarfed needles. The presence of phytoplasma in the needles of branched and surrounding symptomless shoots was demonstrated using nested-polymerase chain reaction (PCR) with universal primer pairs that amplified phytoplasma 16S rDNA, as well as using restriction fragment length polymorphic analysis of PCR products. Comparison of nucleotide sequences of DNA samples from three P. sylvestris trees with balllike structures revealed that their fragments of 16S rDNA were identical. The nucleotide sequence showed more than 99% similarity with the corresponding fragments of sequence of Candidatus phytoplasma pini