A display substrate includes a substrate, first electrodes on the substrate, a first pixel defining layer on the substrate and having first openings, a second pixel defining layer on the first pixel defining layer and having second openings, and light-emitting functional layers on the first electrodes. Each first opening exposes at least a portion of a corresponding first electrode. A sidewall of the first opening is recessed in a direction away from a center of the first opening. On the substrate, an orthographic projection of each second opening at least partially overlaps with an orthographic projection of a corresponding first opening. On the substrate, a border of an orthographic projection of an end of the second opening proximate to the substrate is within a border of an orthographic projection of an end of the second opening away from the substrate. A light-emitting functional layer is in a first opening.