AbstractBACKGROUNDThe Lim‐domain binding protein PtaB, a homolog of Mfg1, governs conidiation and biofilm formation in several fungi. PtaB includes a conserved Lim‐binding domain and two predicted nuclear localization sequences at its C terminus, and is co‐regulated with the transcription factor Som1 downstream of the cyclic AMP‐dependent protein kinase A (cAMP/PKA) pathway. However, the function of PtaB in entomopathogenic fungi remain poorly understood.RESULTSInactivation of PtaB in Metarhizium acridum resulted in delayed conidial germination, reduced conidial yield and increased sensitivities to cell wall disruptors, ultraviolet B irradiation and heat shock. In addition, the fungal virulence was significantly decreased after deletion of MaPtaB because of impairments in appressorium formation, cuticle penetration and evasion of insect immune responses in M. acridum. The MaPtaB‐deletion and MaSom1‐deletion strains showed similar phenotypes supporting that MaSom1/MaPtaB complex controls M. acridum normal conidiation and pathogenic progress. Upon loss of MaPtaB or MaSom1, the fungal sporulation mode in M. acridium shifted from microcycle conidiation to normal conidiation on SYA, a microcycle conidiation medium. Transcriptional analysis showed that more differentially expression genes were identified in MaSom1 RNA sequencing, and MaSom1 and MaPtaB may regulate the expression of genes for conidiation, nutrient metabolism and the cell cycle to control conidiation pattern shift.CONCLUSIONThese data corroborate a complex control function for MaPtaB as an important central factor interacting with MaSom1 in the cAMP/PKA pathway, which links stress tolerance, conidiation and virulence in the entomopathogenic fungus M. acridum. © 2024 Society of Chemical Industry.