Identification of phytoplasmas associated with cultivated and ornamental plants in Kerman Province, Iran

Hossain Kerman Province is a major agricultural centre in south-eastern Iran and an increase in agricultural activities results in an increase in disease. We report phytoplasmal infections in Iran of five plant species; spinach (Spinacia oleracea L.), sunflower (Helianthus annuus L.), canola (Brassica napus L.), cucumber (Cucumis sativus L.) and Aegean wallflower (Erysimum cheiri (L.) Crantz; Family Brassicaceae) using in silico restriction fragment length polymorphism and sequence analysis. Amplicons of approximately 300 bp were amplified using polymerase chain reaction ampli- fication with universal P3 ⁄ P7 primer pair. The ampli- fied products were cloned and sequenced. On the basis of in silico restriction analysis of the amplicon digested with 17 distinct restriction enzymes and 16 ⁄ 23S spacer region sequence, Erysimum and cucumber phyllodies (EPh and CuPh2, respectively) were 100% identical and showed closest similarity with members of the peanut witches-broom group (16SrII). Whereas spinach yellows (SpY) and canola phyllody (CaPh) revealed closest homology with phytoplasmas of the aster yellows group (AY) 16SrI. Mixed infections of the SuWB sample were confirmed in which two different phytoplasmas belonging to 16SrII and 16SrVI groups were found. This is the first report of phytoplasmal infection of Aegean wallflower (EPh) caused by a phytoplasma belonging to the 16SrII group. While spinach phytoplasmas have been isolated in the past; however, our isolate from spinach belonging to the 16SrI group is the first spinach isolate from Iran