J Phytopathol ORIGINAL ARTICLE Occurrence and Characterization of a 16SrII-D Subgroup Phytoplasma Associated with Parsley Witches’ Broom Disease in Iran Mohammad Salehi1, Seyyed Alireza Esmailzadeh Hosseini2, Elham Salehi1 and Assunta Bertaccini3 1 Plant Protection Research Department, Fars Agricultural and Natural Resources Research and Education Center, AREEO, Shiraz, Iran 2 Plant Protection Research Department, Yazd Agricultural and Natural Resources Research and Education Center, AREEO, Yazd, Iran 3 Department of Agricultural Sciences, Plant Pathology, Alma Mater Studiorum - University of Bologna, Bologna, Italy Keywords Abstract 16S rRNA, ‘Candidatus Phytoplasma australasia’, dodder and graft transmission, During 2010–2013 surveys for the presence of phytoplasma diseases in molecular analysis, parsley diseases Yazd province (Iran), a parsley witches’ broom (PrWB) disease was observed. Characteristic symptoms were excessive development of short Correspondence spindly shoots from crown buds, little leaf, yellowing, witches’ broom, M. Salehi, Plant Protection Research stunting, flower virescence and phyllody. The disease causative agent was Department, Fars Agricultural and Natural dodder transmitted from symptomatic parsley to periwinkle and from Resources Research and Education Center, AREEO, Shiraz, Iran. periwinkle to periwinkle by grafting inducing phytoplasma-type symp- E-mail:
[email protected] toms. Expected length DNA fragments of nearly 1800 and 1250 bp were, respectively, amplified from naturally infected parsley and experimentally Received: June 12, 2016; accepted: August 9, inoculated periwinkle plants in direct polymerase chain reaction (PCR) 2016. using phytoplasma primer pair P1/P7 or nested PCR using the same pri- mer pair followed by R16F2n/R16R2 primers. Restriction fragment length doi: 10.1111/jph.12520 polymorphism and phylogenetic analyses of 16S rRNA gene sequences showed that the phytoplasma associated with PrWB disease in Yazd pro- vince belong to 16SrII-D phytoplasma subgroup.