A display substrate and a manufacturing method thereof, and a display device are provided. The display substrate includes a base substrate, a gate driving circuit, power lines, a first signal line group, and a second signal line group. The gate driving circuit includes cascaded shift register units; the power lines are configured to provide power signals to the shift register units; the first signal line group includes at least one clock signal line, and the clock signal line is configured to provide a clock signal to the shift register units; the second signal line group includes a trigger signal line, and the trigger signal line is configured to provide a trigger signal to a first-stage shift register unit; and the gate driving circuit includes at least one transistor, and an extending direction of a channel of the transistor is parallel to an extending direction of the one clock signal line.