Engineering and implementation of synthetic molecular tools in the basidiomycete fungus Ustilago maydis
Heucken, N.; Tang, K.; Huesemann, L.; Hessler, N.; Müntjes, K.; Feldbrügge, M.; Göhre, V.; Zurbriggen, M. D.
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The basidiomycete Ustilago maydis is a well-characterized model organism for studying pathogen-host interactions and of great interest for a broad spectrum of biotechnological applications. To facilitate research and enable applications, in this study, three luminescence-based and one enzymatic quantitative reporter were implemented and characterized. We generated dual-reporter constructs for ratiometric normalization that can be used as a fast-screening platform for reporter gene expression, applicable to in vitro and in vivo detection. Furthermore, we constructed and implemented synthetic bi-directional promoters, that enable bicisitronic expression for gene expression studies and engineering strategies. These non-invasive, quantitative reporters and expression tools will widen significantly the application range of biotechnology in U. maydis and enable in planta detection of fungal infection.
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