A 16SrII-D Phytoplasma strain associated with Tomato Witchesʹʹ- Broom in Bushehr province, Iran

In 2010- 2012 surveys, witchesʹ- boom disease of tomato was observed in Borazjan area (Bushehr province, Iran). Agent of the disease was transmitted from tomato to tomato and eggplant by grafting and to Madagascar periwinkle via dodder inoculation, inducing phytoplasma-type symptoms in inoculated plants. Presence of phytoplasma in naturally affected tomatoes and all symptomatic graft and dodder inoculated plants was confirmed by direct and nested polymerase chain reactions (PCR) using primer pairs P1/P7 and R16F2n/R16R2. BLAST search and phylogenetic analysis of 16SrDNA showed that detected phytoplasma belonged to peanut witchesʹ- broom (16SrII) group. Phylogenetic analysis, percent homology and virtual RFLP indicated that, as a member of 16SrII group, Borazjan tomato witches’- broom (BTWB) phytoplasma together with Bushehr eggplant and alfalfa witches’- broom (BEWB and BAWB, respectively) phytoplasmas were classified with Candidatus Phytoplasma australasia, a phytoplasma related to 16SrII-D subgroup. Based on the same analysis, BTWB, BEWB and BAWB phytoplasmas were differentiable from three other Iranian 16SrII related phytoplasmas associated with alfalfa witchesʹ- broom diseases in Yazd and Fars provinces and lime witchesʹ- broom disease in southern Iran.This is the first report of tomato witchesʹ- broom disease and characterization of its associated phytoplasma in Iran.