Embodiments of the present disclosure provide a display substrate motherboard and a manufacturing method and a cutting method thereof, a display substrate and a display device. The display substrate motherboard includes a preset cutting position, a back film and an adhesive layer disposed on the back film, the adhesive layer includes: a first adhesive layer corresponding to the preset cutting position; a second adhesive layer disposed on two sides of the first adhesive layer in a direction parallel to the back film; and a first light blocking layer disposed between the first adhesive layer and the second adhesive layer, wherein the first light blocking layer is configured to reduce light entering the second adhesive layer through the first light blocking layer after being incident from the first adhesive layer.