Apple rubbery wood is an important disease occurring worldwide. Typical symptoms have been observed since 80' decade in orchards located in the South part of Brazil. In previous studies, grafting has evidenciated that the disease had infeccious etiology and visualization of filamentous bodies inside phloem had indicated that a phytoplasma could be associated with the disease. In the present study, plants with symptoms of rubbery wood were sampled in a commercial orchard in order to demonstrate the presence of phytoplasma in infected tissue and to identify molecularly that the organism. Using nested PCR with universal primers pairs R16mF2/R1 and R16F2n/R2, phytoplasma was consistently detected in symptomatic plants. Identification conducted with specific primer pair R16(III)F2/R1 in nested PCR demonstrated that the phytoplasma was a member of group 16SrIII. RFLP analyses with endonucleases AluI, KpnI, HinfI, HpaII, MseI, RsaI e SauIIIA confirmed that the phytoplasma was a representative of the group 16SrIII. The detection and molecular identification are strong evidences that a phytoplasma is associated with apple rubbery wood in Brazil and agrees with previous evidences using grafting and electron microscopy.
Mollicutes; phytophathogenic prokaryontes; phytoplasmal diseases