A display substrate and a manufacturing method thereof, a display panel, a display motherboard and a testing method thereof, and a display device are provided in the present disclosure. The display substrate is provided with a display region and at least a wiring region outside the display region. The display substrate includes a base substrate and a plurality of connection structures on the base substrate. The plurality of connection structures are connected to a plurality of edge signal terminals through a control module. All of the plurality of connection structures, the plurality of edge signal terminals and the control module are located in the wiring region. The plurality of edge signal terminals are located on an edge of the wiring region. The control circuit is configured to control the connection or disconnection between the plurality of signal terminals and the plurality of connection structures.