Plants of daisy (Bellis perennis L.) exhibiting disease symptoms characteristic of phytoplasma infection were found in the Botanical Garden of Vilnius University, Lithuania. These symptoms included general yellowing and stunting of plants, proliferation of shoots, phyllody and virescence of flowers. Amplification of a phytoplasmal 16S rRNA gene sequence, in polymerase chain reactions (PCRs) containing phytoplasma universal primer pair R16F2n/R2 and template DNA extracted from diseased plants, confirmed that the plants were infected by phytoplasma. The 1.2 kbp 16S rDNA product was subjected to single enzyme digestions with 10 different restriction endonucleases: AluI, HaeIII, HpaII, HhaI, HinfI, KpnI, MseI, RsaI, Sau3AI, and TaqI. RFLP analysis revealed that the plants were infected by a phytoplasma belonging to group 16SrI (aster yellows phytoplasma group), subgroup A (I-A, tomato big bud phytoplasma subgroup).
Keywords: phytoplasma, 16S rRNA gene, PCR, RFLP.