Les émergences de maladies des

Transcription

1 Les émergences de maladies des plantes : Le cas de Xylella fastidiosa et de candidatus Liberobacter solanacearum 1

2 Détecter et anticiper les émergences Veille phytosanitaire OEPP Promed mail list Fiche d alerte LSV 2

3 Gestion des alertes phytosanitaires nématodes BVO, DMA, ERB Angers Rennes Champignons Quarantaine Clermont Ferrand Nancy Plantes tropicales Saint Pierre de la Réunion Montpellier Arthropodes, plantes invasives 3

4 Gestion des alertes phytosanitaires 4

5 Alert in France: 2012 Positive ELISA tests on coffee plants Chlorotic leaves Marginal scorches on Coffea spp. Source : Anses LSV Source : Anses LSV Positive results confirmed for 4 plants Coffea arabica from Equator Coffea canephora from Mexico 5 5

6 Processing symptomatic samples in PBS Isolation IF (antiserum INRA/LSV) Typical colonies Typical cells PCR (Minsavage et al / Pooler & Hartung 1995 / Firrao & Bazzi 1994 ) Real Time PCR and LAMP (Harper et al. 2010) PCR multiplex (Hernandez-Martinez et al., 2007) RST31/RST int/272-2-int XF1-F/XF6-R X.fastidiosa detected and identified Diagnosis scheme 6 6

7 - Associated data - Biodiversity Plant-associated bacteria CFBP - Plant associated bacteria To valorize resources Enhance data / Resource quality /Access / Expertise ( ) To permit access to ressources For international scientific community Research / Development / Teaching To preserve 7 12/2013 Molecular typing for all accessions (in progress) 6700 accessions 60 genera (all plant-pathogens) Strategic resources for plant protection CIRM-CFBP French Collection for Plant-associated Bacteria

8 Partial sequences of 7 housekeeping genes: cysg, gltt, holc, leua, malf, nuol, petc Phylogenetic concataned tree X. f. subsp. fastidiosa CFBP 8083 ATCC ATCC Ƭ 95-2 Strain PD5-2 Taïwan CFBP 8069 ATCC Temecula CIT 0221 M 23 COF0400 COF0223 COF0227 CFBP 8073 ATCC Strains isolated from coffee plants 2012 CVC 145 9a5c COF COF 0239 X. f. subsp. pauca ATCC ATCC CFBP 8072 CFBP 8074 ATCC CFBP 8070 ATCC Ƭ ATCC Dixon M 12 ATCC Ann 1 OLS 9 OLS 20 OLS 19 X. f. subsp. sandyi X. f. subsp. multiplex 8 8

9 New emergence of Xylella fastidiosa reported in 2013 Taïwan : 1 st description of X. f. subsp. fastidiosa on grapevine intaïwan (Chiou Chu Su et al., 2013) Europe : 1 st report of X. fastidiosa on olive in Italy (Apulia county) Geographical distribution Source : EPPO PQR 9 9

10 History 10 10

11 Photo : F. VALENTINI Photo : F. VALENTINI

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14 Biology-History Diagnostic Identification Detection Conclusion Selected methods for evaluation on plant samples: DNA extraction with DNeasy Plant Mini Kit (Qiagen) End point PCR (Minsavage et al., 1994) Real-time PCR (Harper et al., 2010) 19th Australasian Plant Pathology Conference Auckland NZ november

15 Biology-History Diagnostic Identification Detection Conclusion Evaluation of methods on plant sample Test plan : spiked macerate preparation Photo : Kamonrat Meunklad Photo : Nito500 Bacterial suspension 10 X dilutions Photo : Leekris Petioles and midribs of 4 matrix Crushing with AP1 lysis buffer (Qiagen) 1g/5mL 395 µl /tube Photo : Teri Virbickis + SM D1 D2 D3 D4 D5 D6 D (bact/ml) Spiking with 5 µl / tube for each dilution Enumeration by microspcopy IF X 3 assay for each pair plant/bacterium X 3 dilution series per assay DNA Extraction with DNA Plant Mini Kit (Qiagen) 1, , , , (bact/ml) 1, , NEG 19th Australasian Plant Pathology Conference Auckland NZ november

16 Biology-History Diagnostic Identification Detection Conclusion Molecular methods intralaboratory validated for early detection coffee tree (Coffea arabica and C. canephora) peach tree (Prunus persica) grapevine (Vitis vinifera) sweet orange (Citrus sinensis) (ongoing) Call for candidate laboratories to participate in a ring test for interlaboratory validation (2014) Photo : Fernando Rebelo - Wikipédia Photo : Schäbin Wikimedia Photo : Ministère en charge de l agriculture Photo : Ellen Levy Finch Wikipédia 19th Australasian Plant Pathology Conference Auckland NZ november

17 The European Commission has adopted the following decisions The movement, out of the province of Lecce, region of Apulia, Italy, of plants for planting, other than seeds, shall be prohibited. Member States shall conduct official annual surveys for the presence of the specified organism on plants and plant products in their territory. Member States shall ensure that where anyone becomes aware of the presence of the specified organism or has reason to suspect such a presence, that person shall notify, in writing, the competent authority within ten calendar days. Member States shall immediately inform the Commission of the measures they have taken to comply with this Decision