The putative Rho guanine nucleotide exchange factor GerA in Aspergillus flavus regulates growth, development, and aflatoxin synthesis

Abstract

Aspergillus flavus is a saprophytic fungus that infects seed crops before and after harvest, producing potent carcinogens known as aflatoxins. Consequently, the urgent need arises to develop control strategies to mitigate the adverse effects associated with A. flavus. The Ras homologue family member GTPases (Rho GTPases) play a crucial role in regulating the developmental and pathogenic processes of filamentous fungi by controlling various signal transduction pathways. Rho guanine nucleotide exchange factors (GEFs) activate Rho GTPases and regulate their conformational conversion from GDP to GTP. In a previous study, a Rho GEF GerA that is closely regulated by the global regulator VeA under different culture conditions was identified by chromatin immunoprecipitation and sequencing (ChIP-seq), which contains a characteristic Dbl homology (DH) domain. GerA exhibits a high degree of conservation within Aspergillus species but lacks homologues in Saccharomyces cerevisiae. The generation of the ΔgerA mutant using CRISPR/Cas9 gene editing tools significantly impacted the synthesis of AFs, reproductive processes, and pathogenicity in A. flavus. Notably, the ΔgerA mutant exhibited polarity loss and impaired septation. Combined with yeast two-hybrid analysis, there is speculation that GerA may function as a Rho4 GEF. The significant involvement of gerA in the morphological development and secondary metabolism of A. flavus highlights its potential as a target for control measures. Moreover, this study establishes a basis for further investigating the roles of other Rho GEFs in filamentous fungi.