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Surveillance and monitoring of the invasive forest pathogen Heterobasidion irregulare in Europe through an optimized LAMP assay

Paolo Gonthier: University of Torino / DISAFA

<div>The North American pine pathogen<em> Heterobasidion irregulare </em>was introduced and became invasive in Italy. The pathogen is recommended for regulation in Europe by the European and Mediterranean Plant Protection Organisation (EPPO). Here, we report on the development and optimization of an efficient molecular diagnostic tool based on Loop-mediated isothermal AMPlification (LAMP) coupled with two different DNA extraction methods for the detection of <em>H. irregulare</em>. The LAMP assay showed a limit of detection of approximately 20 picograms of target DNA and detection time of about 40 minutes. The assay was successfully validated both in controlled conditions and in the field on a plethora of different samples, including fungal fruiting bodies, infected plants, wood colonized by the pathogen, and woody spore traps. The process of pathogen detection through LAMP on woody spore traps was optimized both in terms of methods and in terms of length of incubation period of traps. Practical applications of this molecular diagnostic tool developed and optimized in the frame of the EU Project EMPHASIS (grant agreement 634179) include the surveillance in the ports of entry of wood imported from North America, the laboratory or in field analyses of samples from symptomatic pines, and the routinely monitoring of pine forests by the aid of woody spore traps.</div>