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Consensus Detection and Identification Protocol for Acidovorax citrulli on cucurbit seeds (DIP-ACIT)

Stefani, Emilio; Giovanardi, Davide; Cunty, Amandine; Olivier, Valérie; Grimault, Valérie; Pucci, Nicoletta; Bergsma-Vlami, Maria; Kornev, Konstantin; Holeva, Maria; Yildiz, Raziye Cetinkaya; Popovic, Tatjana

Acidovorax citrulli (A. citrulli) is the causal agent of bacterial fruit blotch (BFB) of cucurbits. Since there are no resistant commercial cultivars, successful management of BFB depends on exclusion of primary inoculum by using pathogen-free seeds and seedlings. Seed health testing reduces the risk of outbreaks. For this purpose, the Euphresco project had the goal to implement media and methods, validate them and, finally, develop a consensus protocol for the detection of A. citrulli on cucurbit seeds. Based on project’s results, two validated agar media for direct isolation of the pathogen are recommended, together with an implemented protocol for the pathogenicity test. Additionally, MALDI-TOF mass spectrometry was proven to be a robust method for pathogen identification. A TaqMan real-time PCR protocol was also selected, validated and ring-tested. All together, the results of this project will allow the update of the EPPO Diagnostic Protocol PM7/127(1) with robust detection and identification procedures and validated data.

Report of the Euphresco project 2015-D-142 'Consensus detection and Identification protocol for Acidovorax citrulli on cucurbit seeds (DIP-ACIT)'
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