The use of healthy planting materials is of paramount importance for an effective management of Verticillium wilt, a vascular disease caused by the soil-borne pathogen Verticillium dahliae. Moreover, the use of pathogen-free propagative materials is crucial to prevent the introduction of the fungus into disease-free areas. The certification of healthy plant propagative materials depends on the availability of sensitive and specific diagnostic tools. Fluorescence of Loop Primer Upon Self Dequenching loop-mediated isothermal amplification (FLOS LAMP), is an emerging diagnostic real time LAMP technique initially developed for the diagnosis of human viruses. Recently, it has been successfully applied for the detection of a plant pathogenic bacterium in its insect vector. The main objective of this work was therefore to develop a FLOS LAMP essay to detect V. dahliae in infected olive plants.
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Two previously described LAMP primer sets were evaluated in order to select the most efficient ones for use in routine testing. Genomic DNA extracted from 41 isolates of V. dahliae was used during the trial. Thirty-six isolates were specifically detected but a non-specific signal was also recognised in the negative control. Preliminary results of this study highlighted the need to design a new set of primers, targeting the conserved IGS genomic sequence in order to develop an effective real time LAMP essay for V. dahliae detection.
