A display substrate includes a substrate; a first pixel defining layer on the substrate, wherein the first pixel defining layer has a first container portion therein; a planarization pattern filled in the first container portion, wherein a surface of the planarization pattern distal to the substrate is flush with a surface of the first pixel defining layer distal to the substrate; a second pixel defining layer on a side of the planarization pattern distal to the substrate, wherein a second container portion is in the second pixel defining layer and penetrates through the second pixel defining layer along a stacking direction of the first pixel defining layer and the substrate, and an orthographic projection of the second container portion on the substrate falls within an orthographic projection of the first container portion on the substrate; and an organic functional layer in the second container portion.