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Molecular identification of diverse ‘Candidatus Phytoplasma’ species associated with grapevine decline in Iran
Authors:Maryam Ghayeb Zamharir  Samanta Paltrinieri  Shokrollah Hajivand  Mahdi Taheri  Assunta Bertaccini
Affiliation:1. Plant Disease Department, Iranian Research Institute of Plant Protection, AREEO, Tehran, Iran;2. Dipartimento di Scienze Agrarie, Patologia vegetale, Alma Mater Studiorum‐University of Bologna, Bologna, Italy;3. Horticulture Research Department, Qazvin Agricultural and Natural Resources Research and Education Centre, AREEO, Qazvin, Iran;4. TakestanJahadehKeshavarzi Center, Qazvin JahadehKeshavarzi Organization, Qazvin, Iran
Abstract:Grapevine (Vitis vinifera) is one of the most important fruits in Iran where the provinces of Qazvin, Lorestan and Markazi are main producers. During 2013–2015, vineyards located in these provinces were surveyed to verify the presence of phytoplasma. The sample collection was based on symptomatology including decline, leaf yellowing and shortening of internodes. Total DNA was extracted from symptomatic and symptomless grapevine samples and used in nested‐polymerase chain reaction (PCR) assays with phytoplasma ribosomal primers (P1/Tint followed by R16F2n/R2, R16mF1/mR1, R16(I)F1/R1 or 6R758f/16R1232r). Nested‐PCR products were obtained only for symptomatic samples while samples from symptomless plants yielded no PCR products. Restriction fragment length polymorphism (RFLP) analyses with Tru1I, TaqI and Tsp509I and direct sequencing of amplicons followed by phylogenetic analyses indicated the presence of ‘Candidatus Phytoplasma fraxini’, ‘Ca. P. aurantifolia’, ‘Ca. P. solani’ and ‘Ca. P. phoenicium’‐related strains. In Marzaki province, there ‘Ca. P. aurantifolia’ strains were mainly detected, while in the other two provinces, all the four ‘Candidatus species’ were identified with the prevalence of ‘Ca. P. solani’‐related strains. In both provinces in one case, mixed phytoplasma infection was also detected by RFLP analyses. The presence of different phytoplasmas in positive samples indicates great phytosanitary significance due to grapevine economic importance for country. Grapevine phytoplasma infection represents a threat for other crops suggesting grapevine as alternative host species for the phytoplasmas already reported in Iran, while the ‘Ca. P. fraxini’ is for the first time identified in Iran.
Keywords:decline diseases  grapevine  mixed infection  PCR/RFLP analyses  phytoplasmas  sequencing
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