Genome‐Scale Model of Rhizopus microsporus: Metabolic integration of a fungal holobiont with its bacterial and viral endosymbionts

Rhizopus microsporus often lives in association with bacterial and viral symbionts that alter its biology. This fungal model represents an example of the complex interactions established among diverse organisms in functional holobionts. We constructed a Genome‐Scale Model (GSM) of the fungal‐bacterial‐viral holobiont (iHol). We employed a constraint‐based method to calculate the metabolic fluxes to decipher the metabolic interactions of the symbionts with their host. Our computational analyses of iHol simulate the holobiont's growth and the production of the toxin rhizoxin. Analyses of the calculated fluxes between R. microsporus in symbiotic (iHol) versus asymbiotic conditions suggest that changes in the lipid and nucleotide metabolism of the host are necessary for the functionality of the holobiont. Glycerol plays a pivotal role in the fungal‐bacterial metabolic interaction, as its production does not compromise fungal growth, and Mycetohabitans bacteria can efficiently consume it. Narnavirus RmNV‐20S and RmNV‐23S affected the nucleotide metabolism without impacting the fungal‐bacterial symbiosis. Our analyses highlighted the metabolic stability of Mycetohabitans throughout its co‐evolution with the fungal host. We also predicted changes in reactions of the bacterial metabolism required for the active production of rhizoxin. This iHol is the first GSM of a fungal holobiont. We constructed the first Genome‐Scale Model (GSM) of a fungal holobiont to decipher the metabolic interactions between Rhizopus microsporus and its bacterial and viral endosymbionts. We discovered that glycerol plays a pivotal role in the fungal–bacterial interaction and identified the metabolic reactions that support this tri‐partite microbial symbiosis.