Construction of a New -Mediated Transformation System based on a Dual Auxotrophic Approach in .

is a significant edible fungus that produces a variety of bioactive compounds. We have previously established a uridine/uracil auxotrophic mutant and a corresponding -mediated transformation (ATMT) system for genetic characterization in using as a screening marker. In this study, we constructed an ATMT system based on a dual and auxotrophic mutant of . Using the uridine/uracil auxotrophic mutant as the background and as a selection marker, the gene encoding imidazole glycerophosphate dehydratase, required for histidine biosynthesis, was knocked out by homologous recombination to construct a histidine auxotrophic mutant. Then, in the histidine auxotrophic mutant was deleted to construct a Δ Δ dual auxotrophic mutant. Further, we established an ATMT transformation system based on the dual auxotrophic by using and as reporter genes. Finally, to demonstrate the application of this dual transformation system for studies of gene function, knock out and complementation of the photoreceptor gene Cm in the dual auxotrophic were performed. The newly constructed ATMT system with histidine and uridine/uracil auxotrophic markers provides a promising tool for genetic modifications in the medicinal fungus .