Morphological and Molecular Genetic Analysis of Epigenetic Switching of the Human Fungal Pathogen Candida albicans

Candida albicans is a pleiomorphic fungal pathogen whose morphogenetic plasticity has long been considered as a major virulence factor. In addition to the yeast-filament transition, C. albicans cells also have the unique ability to switch between two epigenetic phases referred to as white and opaque. White and opaque cells harbor identical genomes yet they differ in cellular morphologies, gene expression profiles, mating abilities, and virulence properties. The switching process is regulated by a small network of transcription factors and is suggested to be driven by stochastic fluctuations of the regulatory components, which correlates with altered switching frequencies. Traditionally, phase variants have been identified based on cellular morphologies and expression levels of a few marker transcripts, yet it has recently become clear that several other criteria are also essential and relevant, because phase markers are regulated at multiple branching sites of transcriptional circuitry regulating switching. Here, we describe basic methods to discriminate between white and opaque switching variants, based on cellular and macroscopic morphologies, expression levels of phase-specific transcripts, Wor1 protein levels, as well as quantitative mating assays.