The obligate symbiotic nature of arbuscular mycorrhizal (AM) fungi makes extremely difficult their genetic manipulation or transformation. For this reason, a heterologous system has been traditionally used for functional analysis of AM fungal genes, being the budding yeast Saccharomyces cerevisiae an organism suitable for this purpose. Here we present the yeast methods required for the functional analysis of AM fungal genes, including protocols for yeast transformation, heterologous gene expression, functional complementation assays, preparation of yeast extracts, and subcellular localization of the encoded protein.
Keywords: Arbuscular mycorrhizal fungus; Functional characterization; Heterologous gene expression; Iron transport; Saccharomyces cerevisiae; Subcellular localization; Yeast transformation.