Characterization of A New Almond Witchesʹ Broom Phytoplasma in Iran

Almond witches broom (AlmWB) is a destructive disease in several provinces in Iran. Association of phytoplasma with the disease has been established previously. In the present work two phytoplasmas from Khafr (KAlmWB) and Neyriz (NAlmWB) in the Fars Province were compared by biological and molecular analysis. Both infected bitter almond, wild almond, peach and nectarine but not apple and pear, by grafting. In bitter almond the symptoms induced by KAlmWB consisted of severe proliferation, internode shortening and leaf size reduction. In contrast, NAlmWB caused leaf necrosis, dieback and death. KAlmWB was transmitted to periwinkle and eggplant and from experimentally infected periwinkle to almond by dodder. It was also transmitted from eggplant to eggplant, ornamental eggplant and tomato by grafting. Under similar test conditions, NAlmWB was not transmitted to herbaceous plants by dodder. Phylogenetic analysis of 16S–23S rDNA spacer region (SR) sequences placed both strains in the pigeon pea witches broom (PPWB) group. However, based on phylogenetic and putative restriction site analyses and sequence homology, NAlmWB was identical with the Lebanese AlmWB phytoplasma, while KAlmWB was closer to the Knautia arvensis phyllody (KAP) agent. Clustering of KAlmWB with KAP was confirmed by analysis of full length 16S rDNA sequence. On the basis of host range, dodder transmission, host range, symptomatology and molecular analyses of 16S rDNA and SR, two different phytoplasmas related to PPWB group were associated with AlmWB disease in Iran. KAlmWB phytoplasma is being reported as a new phytoplasma of AlmWB disease