Development of a simplified NASBA protocol for detecting viable cells of the citrus pathogen Xanthomonas citri subsp. citri under different treatments |
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Authors: | G Scuderi M Golmohammadi J Cubero M M López G Cirvilleri P Llop |
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Institution: | 1. Department of Phytosanitary Sciences and Technologies (DISTEF) – Section Plant Pathology, University of Catania, 95123 Catania, Italy;2. Instituto Valenciano de Investigaciones Agrarias (IVIA), 46113 Moncada, Valencia;3. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Ctra de La Coru?a Km 7,5, Madrid, Spain |
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Abstract: | Nucleic acid sequence based amplification (NASBA) is a method of amplifying RNA, for the detection of RNA viruses and human pathogenic bacteria. Recently, NASBA has also been employed for the detection of plant diseases caused by viruses and quarantine bacteria. A major citrus pathogen, Xanthomonas citri subsp. citri (Xcc), causal agent of citrus bacterial canker, is being studied in depth due to its economic importance, with recent focus concentrating on its viability and survival under different stress conditions and control treatments. In this work, a NASBA protocol using primers for gumD mRNA has been developed to assess the viability of this pathogen under different bacteriocidal treatments. This method is rapid, specific and sensitive, and is able to detect viable bacterial cells, using a hybridization device which allows the visualization of the results in only 30 min. The usefulness of the method has been confirmed with bacterial suspensions subjected to different heat treatments and to sodium orthophenylphenate. |
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Keywords: | citrus bacterial canker Citrus spp nucleic acid sequence based amplification RNA hybridization Xanthomonas citri subsp citri |
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