Molecular characterisation of phytoplasmas infecting roses in Poland

Abstract

Symptoms of shoot dieback and leaf yellowing followed by leaf chlorosis were observed in naturally infected roses 'Frisco' and 'Suela', cultivated in a commercial greenhouse in Poland. The presence of phytoplasma was demonstrated in affected plants by nested polymerase chain reaction (PCR) with R16Fl/RO and Pl/P7 primer pairs in the first round followed by a second one with R16F2n/R2, fA/rA, Pc399/P1694, R16(I)Fl/Rl and Pl/fArev primer pairs. Restriction fragment length polymorphism (RFLP) analysis of PCR products (primed with primers R16F2n/R2) was done using enzymes AluI, MseI, RsaI and HpaII. Restriction profiles obtained with these enzymes were identical to those of reference strain AY1 belonging to aster yellows phytoplasma group, subgroup I-B (16SrI-B). Nested PCR products from roses 'Frisco' and 'Suela' were sequenced. Analysis of sequences confirmed that the phytoplasma infecting those roses could be classified to aster yellows phytoplasma group, subgroup B.