Acidovorax citrulli is a seed-borne pathogen and the causal agent of bacterial fruit blotch of cucurbits. It is listed as an A1 quarantine pathogen by EPPO. Seed certification is based on the availability of a sensitive and specific pathogen detection in seed lots: this is a must for an effective disease management strategy. Therefore, an effective DNA extraction and purification procedure is a critical issue to ensure a robust PCR analysis. Pathogen detection in seed lots has been implemented by testing different known contamination levels by Acidovorax citrulli. Initially, two different sample preparation methods have been tested: a) Overnight soaking; b) Hammering of dry seeds, followed by three different manual DNA extraction. Each DNA sub-sample obtained has been analysed with two different primers sets, SEQID3/SEQID4 and WFB1/WFB2, to evaluate the capability to detect the pathogen. Results showed that a DNA extraction and purification procedure, based on soaking the seeds, followed by the use of the DNeasy Plant Mini kit (Qiagen) on the washing fluids gave the highest amount of DNA, sufficient to increase the detection threshold of the pathogen. This will allow the improvement of current detection procedures. Furthermore, naturally contaminated watermelon seeds were treated through different methods, in order to achieve a possible sanitation or eradication of Acidovorax citrulli: a bacterial antagonist, a microbial consortium, a plant polyphenol. Our results showed that treated seeds were only partially disinfected, and the pathogen was not eradicated after any of the methods used.
|Data di pubblicazione:||2015|
|Titolo:||Seed transmission of Acidovorax citrulli: implementation of detection in watermelon seeds and development of disinfection methods|
|Autori:||Giovanardi, Davide; Ferrari, Michele; Stefani, Emilio|
|Data del convegno:||24-28 November 2014|
|Nome del convegno:||VII Congress on Plant Protection|
|Luogo del convegno:||Zlatibor, Serbia|
|Titolo del libro:||Integrated Plant Protection: Knowledge-based Steps towards Sustainable Agriculture, Forestry and Landscape Architecture. Proceedings of the 7th Congress on Plant Protection|
|Appare nelle tipologie:||Relazione in Atti di Convegno|
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