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XVIII Convegno Nazionale Società Italiana di Patologia Vegetale

XVIII Convegno Nazionale Società Italiana di Patologia Vegetale Sassari 24 - 26 Settembre 2012. T he D etection O f Pseudomonas Syringae Patovar Phaseolicola On F rench Bean Seeds Using Official Methods. INRAN Laboratory Exsperience. IstitutoNazionale

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XVIII Convegno Nazionale Società Italiana di Patologia Vegetale

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  1. XVIII Convegno Nazionale Società Italiana di Patologia Vegetale Sassari 24 - 26 Settembre 2012 The Detection Of Pseudomonas SyringaePatovarPhaseolicolaOn French Bean Seeds Using Official Methods. INRAN Laboratory Exsperience IstitutoNazionale di Ricerca per gli Alimenti e la Nutrizione • Veronica Senape1, Loredana Sigillo1, Giovanna Serratore1, Romana Bravi1 1INRAN (ex ENSE) - Sezione di Battipaglia SS 18 – km 77.700 , 84091 Battipaglia (SA) Italia - E-mail: l.sigillo@ense.it Pseudomoassyringaepvphaseolicola (Psp), causing the halo blight disease, is a worldwide spread severe seedborne bacterial pathogen of French bean. In the last years, seed companies used to check the French bean seed lot health to assure the phythosanytary quality of their production. The Laboratory of Phytopathological Analyses of INRAN (ex ENSE) (Battipaglia, Italy) offers to seed companies and growers a service of diagnostic analyses on seeds and growing materials. From 2010 to 2012, the laboratory analysed122 French bean seed samples for Psp detection. The detection were carried out using an internal procedure developed according the validate ISTA protocol no. 7-023 (2008). Symptomsof haloblight • ISTA. International SeedTestingAssociation – www.seedtest.org • Protocol • The samples were separated into 1000 seed sub-samples, soaked overnight in a buffer(photo 1) • The extractswereplated on semi-selective media (MSP, MT)(photo 2) • Typicalcolonieswerepurified on King B medium and the suspectisolatesweretested for tobaccohypersensitivityreaction(photo 3) • Pathogenicitywasconfirmed with in vivo test (photo 4) MSP photo 1. sub-sample MSP negative reaction MT photo 3. HR on tobacco photo 2. isolation on MSP and MT weakreaction of pathogenicstrain. 100 bp • The isolatedcolonieswereidentifyed by PCRusing the primers PS5.1 e PS 3.1 (Schaad et al., 1995) (photo 5) positive reaction photo 4. Pathogenicity test. For the resultinterpretation, the techicianexperienceis a critical control point photo 5. PCRamplification The 30% of the seedsamplesresultedinfected by Psp. Only the isolate from threesampleswere negative to PCRamplificationthoughtheywerepathogenic on frechbean and gave a positive HR reaction on tobacco, they are probablyclassifiableasPseudomonassyringaepvsyringae. On theseisolatesfurtheranalysis are in progress. According to Italian and Europeanlegislations the phytosanitarycertification of vegetableseedsisnotprovided. Moreoveritshould be consideredthatmanydangerouspathogen are trasmitted by seeds and the seedhealthis an importantcondition for a high quality production.

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