Peach witches’-broom, an emerging disease associated with ‘Candidatus Phytoplasma phoenicium’ and ‘Candidatus Phytoplasma aurantifolia’ in Iran

During field surveys carried out from 2013 to 2017 in the eight main peach producing provinces of Iran, symptoms of a phytoplasma-like peach witches'-broom disease (PWIB), inducing severe yellowing, little leaf, internode shortening, crown and stem witches’-broom, decline, and death, were observed. The aim of this work was to identify and characterize the agent(s) associated with PWIB by biological assays and molecular analyses. PWIB agents were successfully transmitted under controlled conditions from scions of in field-affected peach trees, exhibiting severe or mild symptoms, to peach and bitter almond seedlings inducing phytoplasma-like symptoms. A 16S rDNA fragment of 1250 bp was amplified by nested-PCR from all PWIB-affected trees and grafted seedlings. Nucleotide sequence identity, presence of species-specific signature sequences, in silico RFLP, single nucleotide polymorphisms, and phylogenetic analyses of 16S rDNA allowed the assignation of the phytoplasma strains identified in seven Iranian provinces in peach trees with severe PWIB symptoms to four SNP genetic lineages of ‘Ca. P. phoenicium’ (subgroup 16SrIX-B and its variant). PWIB phytoplasma strains identified in Abarkooh (Yazd province) in peach trees with mild symptoms were attributed to the species ‘Ca. P. aurantifolia’ (subgroup 16SrII-C). This report of a wide spread of ‘Ca. P. phoenicium’ in association with PWIB in Iran supported its capability of adaptation to a broad range of fruit tree species, such as peach, nectarine, and apricot. As ‘Ca. P. phoenicium’ and ‘Ca. P. aurantifolia’ are the etiological agents of other important plant diseases in Iran and neighbouring countries, further investigations are needed to determine the role played by peach in their epidemiological pathways.