?Duplex real-time PCR assay for the simultaneous detection of Ophiostoma novo-ulmi and Geosmithia spp. in elm wood and insect vectors

Dutch elm disease (DED) is a destructive tracheomycosis caused by Ophiostoma novo-ulmi , an ascomycete probably originating in East-Asia that devastating natural populations throughout Europe, North America and Asia. The fungus mainly spread bark beetles complete their life cycle between healthy diseased elms. Recently, it has been highlighted some fungi of the genus Geosmithia which are similarly well associated with beetles, seem to also play role DED pathosystem acting as mycoparasites O. . Although relationship clear, biological spp. within still partly unclear, association beetles. In this work, we tried clarify these aspects developing qPCR duplex TaqMan assay detect quantify DNA both fungi. extremely sensitive showing limit detection low 2 fg ?l –1 for We collected woody samples from infected trees beetle cycle. All were negative DNA. never present infected, but living trees, while they frass (EBB – Scolytus spp.) at each stage EBB much higher quantities than This work provides better understanding interactions occurring amongst main players pathosystem.