An emergent biofilm program from inactivation of Candida albicans master regulators Efg1 and Ndt80

Biofilm formation by the fungus Candida albicans is a central virulence trait that enables colonization of implanted medical devices and mucosal surfaces. Biofilm formation reflects a complex regulatory network, and depends upon multiple master regulators that include transcription factors Efg1 and Ndt80. It is well established that efg1 Δ/Δ and ndt80 Δ/Δ single gene mutants are defective in biofilm formation. We report here that an efg1 Δ/Δ ndt80 Δ/Δ double mutant of reference strain SC5314 is able to form a robust biofilm in vitro and in vivo. We refer to the efg1 Δ/Δ ndt80 Δ/Δ biofilm as an emergent biofilm because this phenotype could not have been predicted from the phenotypes of efg1 Δ/Δ or ndt80 Δ/Δ single gene mutants. In four additional strain backgrounds, efg1 Δ/Δ ndt80 Δ/Δ mutants do not form biofilms, but in all strain backgrounds the efg1 Δ/Δ ndt80 Δ/Δ mutants can form filamentous cells, which are components of biofilms. Emergent biofilm formation is especially pronounced in YPD + FBS medium at 30°C, and RNA-seq under those conditions reveals altered expression in the efg1 Δ/Δ ndt80 Δ/Δ double mutant of biofilm-related genes: upregulation of BCR1, UME6, and HGC1 , and downregulation of ALS3, BRG1, and HWP1. These gene expression changes suggest that the emergent biofilm program is partially distinct from the conventional biofilm program. This inference is supported by functional analysis: emergent biofilm formation is independent of Brg1, Rob1, Tec1, and Wor3, all of which have positive roles in conventional biofilm formation. Emergent biofilm formation depends upon the hyphal cyclin Hgc1, the biofilm transcription factors Bcr1 and Ume6, and the Bcr1/Ume6-activated adhesin gene FLO9 . The seemingly simple emergent biofilm program may represent a primordial surface colonization strategy.