P-type Ca2+-ATPase (Pmr1) is a core element in calcium-calcineurin pathway and evidence for its cross-talk with other signalling pathways in filamentous fungi is of scarcity. Here, we characterized the striking functions of a Golgi Ca2+-ATPase (Bbpmr1) in Beauveria bassiana (fungal entomopathogen) by multi-phenotypic and transcriptional analyses under normal and stressful conditions. Bbpmr1 inactivation caused severe defects in nutritional uptake, growth, conidiation and germination under normal conditions, drastic reductions in cell tolerances to oxidative, hyperosmotic, cell wall disturbing and fungicidal stresses and toxic metal ions during colony growth and/or conidial germination, and half loss of the fungal biocontrol potential represented by conidial virulence, thermotolerance and UV-B resistance. Accompanied with the multi-phenotypic defects, four important genes associated with asexual development were repressed by ≥ 75% in ΔBbpmr1 versus wild type, and all or most of stress-responsive genes encoding 14 cascaded proteins in MAPK pathways, two Ras GTPases, two protein kinases, Ssk1-type response regulator, TOR signalling protein, and many downstream enzymes and proteins were greatly downregulated in ΔBbpmr1 under the chemical stresses. Conclusively, Bbpmr1 regulates positively fundamental aspects on B. bassiana biology and environmental adaptation through wide cross-talk with cellular signalling networks including MAPK cascades and those upstream or independent of the cascades.