Gα3 regulates growth and development, stress response, patulin synthesis and pathogenicity of Penicillium expansum by mediating cAMP/PKA and PKC signaling

G proteins are signaling proteins in eukaryotes that transduce extracellular signals into the intracellular space. There are three classes of Gα proteins in fungi, among which class III Gα proteins play crucial roles for fungal growth, secondary metabolism, and pathogenicity. In this study, we found that the Gα3 protein from Penicillium expansum belongs to the P-loop NTPase superfamily and contains a G-alpha conserved domain and an adenylate cyclase binding site that can bind to guanine nucleotides. Deletion of Gα3 downregulated the expression of the cAMP/PKA signaling pathway genes AC, PkaC, and PkaR, and the PKC signaling related genes PKC1, PKC2, and PLC of P. expansum. The Gα3-deleted strain showed slow colony growth, irregular depression at the colony edge, and increased branching at the mycelial tip. Gα3 deletion decreased spore production, inhibited spore germination, and reduced the expression levels of genes that regulate conidiospore formation, including abaA, wetA, vosA and brlA. In addition, the Gα3-deleted strain grew faster in carbon and NO3- medium, and was more sensitive to salt and oxidative stress, but less sensitive to cell wall stress. Gα3 deletion significantly downregulated the expression of patulin biosynthesis-related genes except for PatF and PatM, and reduced the patulin levels in vivo and in vitro. In addition, loss of Gα3 reduced the pathogenicity of the strain on fruit, with down-regulated expression of the extracellular enzyme genes PG, PL, and PLA. In conclusion, Gα3 can regulate growth and development, carbon and nitrogen response, stress response, patulin biosynthesis and pathogenicity by mediating cAMP/PKA and PKC signaling pathways of P. expansum.