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Conclusion Method Assessment Guadeloupe Martinique

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Conclusion Method Assessment Guadeloupe Martinique Huanglongbing: new epidemic situation in the French West Indies, caused by Candidatus Liberibacter asiaticus G. Cellier, A. Moreau, N. Cassam, B. Hostachy, Anses - Plant Health Laboratory (LSV), Tropical Pests and Diseases Unit, Saint Pierre, Reunion Island; P. Ryckewaert, Cirad, Le Lamentin, Martinique; L. Aurela, FREDON, Les Abymes, Guadeloupe; R. Picard, FREDON, Ducos, Martinique; K. Lombion, B. Marie, Direction of Food, Agriculture and Forestry, Basse-Terre, Guadeloupe; A. L. Rioualec, P. Huguenin, C. Dalibard, J. Iotti, Direction of Food, Agriculture and Forestry, Fort de France, Martinique. Corresponding author: G. Cellier, [email protected] Presentation Huanglongbing is an unculturable vascular citrus bacterium transmitted from infected to healthy plants through grafting or by citrus psyllids Diaphorina citri and Trioza erytreae. Three species are associated based on the 16S rDNA sequence analysis: • Candidatus Liberibacter asiaticus (Las) Lobsided Tahiti lime with curved columella • Ca. L. africanus (Laf) (Photo by Caroline Sylvaniélo) Blotchy mottle pattern symptoms • Ca. L. americanus (Lam) (Photos by Gilles Cellier) Severe leaf and fruit drop (Photos by Gilles Cellier) The Asian citrus psyllid, D. citri, is a sap-sucking, hemipteran bug. Methodology It is one of only two confirmed Since 2012, a total of 450 sites were prospected in Martinique and Guadeloupe, where 20 leaves vectors of Huanglongbing, widely from 10-30 trees were analyzed. DNA extraction was performed (DNeasy Plant Mini Kit, Qiagen) distributed in southern Asia and on fresh or dried leaf midribs, along with negative control midribs (Citrus paradisi “Star Rubis”) has spread to other citrus growing and polymerase chain reaction amplification was done with the species-specific primers, A2/J5 regions: Middle East, America, and (Hocquellet et al. 1999) and GB1/GB3 (Teixeira et al. 2005). the Caribbean. Aduld psyllid Diaphorina citri along with nymph producing waxy exudate Only Las-specific 703 bp amplicons were obtained and 20 (out of 43) were sequenced (Beckman Currently absent in Europe. (Photo by Michael E. Rogers) Coulter Genomics, United Kingdom; Genbank accession : KF699074 to KF699093) and blasted against NCBI-nr. Guadeloupe Results Martinique Based on MOA033 Anse-Bertrand Grand'Rivière Basse-Pointe Detection and Le Prêcheur L'Ajoupa-Bouillon Port Louis caracterisation of Candidatus Le Lorrain Le Marigot Petit Canal Liberibacter asiaticus. Saint-PierreLe Morne-Rouge Le Moule La Trinité Morne-à-l'Eau Désirade Deshaies BLAST analysis revealed Fonds-Saint-Denis Sainte-Rose Gros-Morne Saint-François Le Carbet Abymes 100% identity with the 50S Baie-Mahault Sainte-Anne Saint Joseph Lamentin Pointe-à-Pître Bellefontaine Le Robert Le Gosier Pointe-Noire ribosomal protein subunit Case-Pilote Fort de France Forest Petit-Bourg L1 (rplA) and L10 (rplJ) and Le Lamentin On-site HLB detection 2012 Positives (42) Le François Bouillante Negatives (33) Goyave Observations (75) no significant homology Ducos Type Vieux-Habitants Orchard (76) to other organisms. Saint-Esprit Private garden (59) Le Vauclin Baillif Capesterre-Belle-Eau Isolated tree (3) Growing facilities (12) Les Trois-¤lets Sequence assembly Rivière-Salée 0 4 8 12 Basse-Terre Rivière-Pilote Trois-Rivières Les Anses-d'Arlet Km Sainte-Luce Scale: 1/400 100 on a reference genome Le Diamant Saint-Louis Le Marin Vieux-Fort On-site HLB Detection 2013: (NC_012985) showed Pending analyses (21) Absence (151) Capesterre-de-Marie-Galante Presence (27) 100% homology. Sainte-Anne Terre-de-Haut 0 5 10 15 Terre-de-Bas Km Scale: 1:235 500 Guadeloupe: HLB Situation in 2012 (source: DAAF 971) Martinique: HLB Situation in 2013 (source: DAAF 972) Las was detected in Guadeloupe on March 2012 Las was detected in Martinique on May 2013 on at Le Moule (East) in a Tahiti lime orchard (C. Tahiti lime (C. latifolia) latifolia) and crossed the island in six months. at Bellefontaine (NW / private garden) D. citri was detected in 1998, where the and at Le Lorrain (NE / orchard). control of the the psyllid population has D. citri was detected in 2012 and been effective with Tamarixia radiata. presents a mean parasitism rate of 70% by Tamarixia radiata. HLB Occurence Epidemiologicaly active French West Indies Distribution map of Candidatus Liberibacter asiaticus (source: EPPO) HLB detection scheme from MOA033 Method Assessment Anses-LSV unit for tropical pest is the Conclusion National Reference Laboratory on tropical • First report of Candidatus Liberibacter asiaticus in the French West Indies phytopathogenic bacteria and viruses. It has • Disease management needs to especially target the vector psyllid produced and validated HLB detection methods Midrib excision (Photo by University of Florida) (MOA033), under ISO 17025 accreditation. • Synergy between local NPPO & LSV-RAPT Reference laboratory Based on Li & al. duplex PCR (A2/J5 & This report raises the importance of plant certification, psyllid population control, GB1/GB3) was assessed on several criteria and surveillance of territories close to the French West Indies, with regards to to detect Las, but also Lam & Laf: the risk that HLB presents to citrus production worldwide. • Sensitivity • Detectability • Specificity • Processed speed • Accuracy • Transferability • Repetability • Final score: 6.47 / 7.
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