COST-Action 864: PomeFruitHealth. Combining traditional and advanced strategies for plant protection in pome fruit growin

Cambra Alvarez, Miguel Angel

Gorris, M.T.

López, M.M.

Montesinos, E.

Palacio Bielsa, Ana

Peñalver, J.

Roselló Pérez, Montserrat

citaREA Repositorio Electrónico Agroalimentario

Efficiency of the EPPO protocolfor preventing the introduction anddissemination of fire blight in two Spanish areasPalacio-Bielsa A., Cambra M.A., Roselló M., Gorris M.T.,Peñalver J., Montesinos E., López M.M. FRANCEBasqueCountry(1995)Surveys, analyses of samples and eradication measures have been taken in Spanish regions in order to prevent introduction and/or spread of fire blightAragonValencianCommunity   Quick Search:   You are here: Science / Areas of Study / Plant Health / DIAGPRO (diagnostic protocols)   DIAGPRO (DIAGNOSTIC PROTOCOLS)  http://www.csl.gov.uk/science/organ/ph/diagpro/Diagnosis of E. amylovoraSCHEME FOR SYMPTOMATIC FIRE BLIGHT SAMPLESISOLATION, ENRICHMENT- ISOLATIONPlants with symptomsRAPID SCREENING TESTSIF, Enrichment DASI-ELISA,nested PCRE. amylovoranot detectedE. amylovoradetectedTest allnegative PositiveConfirm pathogenicity E. amylovora confirmedIDENTIFICATION TESTSColonies with typical morphologyYesYes NoNoAsymptomatic sampleSCREENING TESTSIF, enrichment-isolation, enrichment DASI-ELISA,enrichment-PCRPathogen extraction, optative concentration and enrichmentSCHEME FOR ASYMPTOMATIC FIRE BLIGHT SAMPLESAt least onetest positiveIsolationand/or enrichment isolationColonies with typical morphologyYesNoIDENTIFICATION TESTSYesNoE. amylovora detected Confirm pathogenicityE. amylovora confirmedTests all negativeE. amylovora not detectedVALENCIAN COMMUNITY:PREVENTIVE MEASURESSurveys and analyses of asymptomatic and suspicious samples in nurseries, orchards and forests every yearFRANCEValencianCommunityANALYZED SAMPLES (1996-2008)HOST Nurseries Orchards/Forests TOTALPear 1.213 364 1.577Apple 1.116 101 1.217Loquat 695 158 853Medlar 15 - 15Quince 290 7 297Crataegus spp. 193 63 256Pyracantha spp. 2.298 1 2.299Cotoneaster spp.              2.090 - 2.090Sorbus spp. 116 - 116Photinia spp. 25 - 25Wild pear - 1 1TOTAL 8.051 695 8.746E. amylovora HAS NOT BEEN DETECTED Bacterial isolates with colony morphology resembling E. amylovorawere obtained from necrotic pear blossomsISOLATES CHARACTERIZATION AND IDENTIFICATIONPseudomonas sp.2007ApplePseudomonas syringae pv. syringae2006ApplePseudomonas syringae pv. syringae2001, 2003, 2005, 2006PearIDENTIFICATIONYEARCROP Bacterial isolates with colony morphology resembling Pseudomonaswere obtained from suspicious samples:NECROTIC SYMPTOMS SIMILAR TO E. amylovora ONLYPEAR BLOSSOMS  ARE AFFECTED           (No progress to other parts of the tree)Colony morphology (48h after 25 ºC incubation)E. amylovoraNecrotic pear blossoms isolatesKing´s B Levan CCTPolyphasic approach characterization:• Phenotypic (biochemical and physiological, fatty acid profiles, serological tests)• Genetic (PCR, rep-PCR, plasmids profile,  16S rRNA, G+C content and DNA-DNA hybridization)Identification of an Erwinia sp., different from E. amylovora, and responsible for necrosis on pear blossoms (proposed as Erwinia piriflorinigrans)(Roselló et al., 2008)Interest of the use of an integrated approach, with specific tools for accurate detection of E. amylovora, to avoid false positive detectionsARAGON:ERADICATION MEASURES OF FIRE BLIGHT AND PREVENTION OF THE SPREADARAGONFRANCE**********... 2001 *2000 2002 **#### 2003 .Jalon river fertile plainAll detections inside a 5 km radius areaYearNo. affected orchards(measures adopted)Area (ha)* No. infected trees2000 12 ( E ) 19.6  ( E ) 2492001 2  ( E )2  ( SE )7  ( E )12.4  ( SE )227392002 1  ( E )2  ( SE )3.2  ( E )7.9  ( SE )15252003 3 ( E ) 9.2  ( SE ) 4TOTAL15 ( E )7 ( SE )( 22 )29.829.5( 59.3 )49168( 559 )FIRE BLIGHT ON FRUIT TREES IN ARAGON(E): total eradication; (SE):  selective eradication; *Aprox. 1.000 trees/haNO NEW DETECTIONS SINCE 2004 UP TO DATE30/12.921AbateFételRed CrimsonWilliams BlanquillaConference74/3.130153/4.171Weekly inspections2002 - 2008No new detectionsYear 2001(25 ha)143298765111013121514161718192021222324Analyses performed in years 2002 and 2003ANALYSES PERFORMED IN A PEAR ORCHARD AFTER SELECTIVE ERADICATION (3 laboratories)Date Materialb No. samples No. positive samplesE-ELISA    Isolation PCR24 plotsaMarch 2002June 2002June 2002BlossomsShootsAirSoil720 (240 x 3)720 (240 x 3)24241 0 02 0            00               0            00               0            013 plotsaMay 2003June 2003ShootsShoots390 (130 x 3)390 (130 x 3)0               0            00               0            0TOTAL                                                           2.268a Ten trees per plotb Three samples per tree(4 corymbs, shoots / sample)VERY LOW INOCULUM AVAILABLE(Maryblyt predicted high risk)OTHER ANALYSES PERFORMED IN THE ORCHARDMaterial Date No. samples No. positive samples*E-ELISA   Isolation PCRDiverse plant material(Shoots, blossoms)200220031.1641.6509 1 10Honeybees(Full bloom)200220032003600                0              00                0Beehive pollen 2002200330400                0              00                0              0Flies 2003 90 5 0              0TOTAL 3.606VERY, VERY INOCULUM AVAILABLE IN THE PLOT, NO SYMPTOMS*One positive tree (only in 2002,  no detection in further analyses)CONCLUSIONSThe EPPO protocol for diagnosis of E. amylovora has proved to give accurate data for fireblight prevention and eradication programsThe efficient detection of E. amylovora in asymptomatic plant material requires an integrated approachThe combination of intensive surveys and fast elimination of infected plant material has been successful in long- term eradication of fire blight in Aragon region (Spain)María M. LópezMª Teresa GorrisJavier PeñalverPablo LlopEster Marco-NoalesMónica OrdaxIVIA, Valencia, SpainMiguel A. CambraCPV, Zaragoza, SpainEmilio MontesinosIsidre LlorenteJordi CabrefigaJesús FrancésAnna BonaterraEsther BadosaIsidre LlorenteJordi CabrefigaLidia RuzNuria GajuConcepció MoragregaCIDSAV-INTEA, Universidad Gerona Gerona, SpainMontserrat RosellóConselleria Agricultura, Valencia, SpainElena G. BioscaUniversidad de Valencia, Valencia, SpainAna Palacio-BielsaCITA, Zaragoza, SpainThanks for your attention

Efficiency of the EPPO protocol for preventing the introduction and dissemination of fire blight in two Spanish areas

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Efficiency of the EPPO protocol for preventing the introduction and dissemination of fire blight in two Spanish areas

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