Identification of a phytoplasma associated with pomegranate little leaf disease in Iran

• Detection of a phytoplasma associated with pomegranate little leaf disease in Iran.
• Molecular characterization of a phytoplasma associated with pomegranate little leaf disease.
• Identification of a 16SrII-D phytoplasma associated with pomegranate little leaf disease.
• Identification of a sesame phyllody phytoplasma as the probable source of pomegranate little leaf disease.

During 2012–2014 surveys for the presence of phytoplasma diseases in Fars province (Iran), pomegranate little leaf symptoms were observed in several orchards in Khafr and Neyriz areas. Samples collected from symptomatic plants positively reacted in nested PCR assays using P1/P7 followed by R16F2n/R16R2 primer pairs producing the expected 1,250 bp DNA fragments. Real and virtual RFLP analysis showed that the sequences of phytoplasma strains from Khafr and Neyriz (KPLL and NPLL strains, respectively) were identical to each other and belong to 16SrII phytoplasma group, subgroup D. Phylogenetic analysis of the R16F2n⁄R16R2 DNA region confirmed that KPLL and NPLL phytoplasmas were enclosed in the same clade as other 16SrII-D subgroup phytoplasmas. This is the first reported occurrence of a 16SrII phytoplasma infecting pomegranate trees.