N-acetylglucosamine　(GlcNAc),　a　key　component　of　fungal　cell　walls　and　insect　cuticles,　is　an　important　signal　to　activate　fungal　response　during　entomopathogen-insect　interactions.　Research　on　Ngs1,　the　only　identified　GlcNAc　sensor　and　transducer,　has　been　primarily　restricted　to　Candida　species.　Although　our　previous　work　identified　an　Ngs1　homology　in　Beauveria　bassiana,　its　physiological　functions　in　entomopathogenic　fungi　remain　largely　unexplored.　In　this　study,　we　unveiled　the　sub-localization　of　Ngs1　in　the　nucleolus.　Further　transcriptomic　analysis　revealed　that　Ngs1　plays　a　crucial　role　in　vegetative　growth,　fungal　development,　and　cell-wall　construction　by　acting　as　a　transcriptional　mediator,　particularly　influencing　carbon　metabolism　in　response　to　insect　cuticle　stimulation.　The　absence　of　Ngs1　compromised　vegetative　growth　across　various　carbon　sources　by　downregulating　expressions　of　key　catalytic　enzymes.　Conversely,　Ngs1　deficiency　enhanced　transcription　levels　of　oxidative　phosphorylation,　leading　to　increased　ATP　and　reactive　oxygen　species　(ROS)　production.　Despite　higher　ATP　levels,　Ngs1-deletion　mutants　exhibited　reduced　asexual　development　and　hyphal　germination,　primarily　due　to　the　function　of　Ngs1　in　the　central　developmental　pathway　and　Brg1/Nrg1-dependent　pathway.　Additionally,　the　downregulation　of　N-glycan　biosynthesis　in　ΔNgs1　diminished　cell　wall　components,　resulting　in　decreased　cell　wall　resistance　to　lysis　and　impaired　fungal　development.　These　findings　advance　our　understanding　of　the　regulatory　role　of　Ngs1　in　B.　bassiana　during　host　interactions　and　provide　a　theoretical　foundation　for　engineering　fungi　to　maintain　or　even　enhance　pesticidal　activity.　©　2024