The present invention provides a display substrate, a mother substrate for display substrates and a display device. The display substrate comprises a display area and a non-display area surrounding the display area, wherein an alignment layer is disposed in the display area and the non-display area. In the display substrate according to the present invention, by providing the same pattern of alignment layer in both of the non-display area and the display area of the display substrate, the Mura defect caused by the difference of surface roughness between the display area and the non-display area of the display substrate may be significantly reduced, and a large amount of small foreign matters gathered due to the rubbing between the rubbing cloth and metal residuals may also be significantly reduced. Therefore, the yield of production and the quality of products may be improved.