Transformation of Fungal Grapevine Trunk Disease Pathogens with the Green Fluorescent Protein Gene

Abstract

Eutypa lata and Phaeomoniella chlamydospora are important fungal grapevine trunk disease pathogens. Studies of their epidemiology and disease progression are hampered by the slow growth rates of the fungi and a lack of molecular tools. A protoplast-based transformation system was developed using hygromycin resistance as a positive selectable marker. The system was used to introduce a green fluorescent protein reporter gene that was expressed in the hyphae of both species. The reporter strains will be useful tools for further epidemiological studies. Furthermore the transformation system will allow for genetic dissection of candidate pathogenicity or virulence genes by targeted replacement or disruption, in order to discern the role of these genes in the disease process.