Co-operational PCR coupled with dot blot hybridization for detection and 16SrX grouping of phytoplasmas |
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| Authors: | E Bertolini E Torres A Olmos M P Martín A Bertaccini M Cambra |
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| Institution: | Departamento de Protección Vegetal y Biotecnología, Instituto Valenciano de Investigaciones Agrarias (IVIA), Carretera Moncada-Náquera km 4,5. 46113 Moncada, Valencia;;Departament d'Agricultura, Alimentaciói AccióRural, Generalitat de Catalunya, Via CirculacióNord, 08004 Barcelona;;Departamento de Micologia, Real Jardín Botánico, Consejo Superior de Investigaciones Científicas (CSIC), Plaza de Murillo 2, 28014 Madrid, Spain;and;Dipartimento di Scienze e tecnologie Agroambientali (DiSTA), Patología Vegetale, Alma Mater Studiorum, Universitàdi Bologna, Bologna, Italy |
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| Abstract: | A protocol based on Co-operational PCR has been successfully applied to the detection of phytoplasmas. A triprimer reaction coupled with hybridization using general and specific probes permitted detection of ' Candidatus Phytoplasma mali', ' Ca . Phytoplasma prunorum' and ' Ca . Phytoplasma pyri', and their identification as members of 16S ribosomal quarantine group X. The sensitivity of this method was at least one hundred times greater than conventional PCR and similar to that achieved by nested PCR and real-time PCR. The method was validated by testing field samples collected from Malus , Prunus and Pyrus spp. and Olea europaea and compared with seven phytoplasmas maintained in Catharanthus roseus . |
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| Keywords: | 'Ca Phytoplasma mali' 'Ca Phytoplasma prunorum' 'Ca Phytoplasma pyri' DNA targets molecular detection and diagnostics Nested PCR |
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