The present disclosure relates to the field of manufacturing displays, and provides a method for manufacturing a display substrate, a method for manufacturing a mask plate, and a display device. The method for manufacturing a display substrate comprises: providing a first substrate; providing a mask plate opposite to the first substrate, the mask plate comprising one or more light-transmissive regions, and an electrically conductive material is provided on a surface of the mask plate facing the first substrate; and irradiating a surface of the mask plate facing away from the first substrate with light rays, such that the electrically conductive material is transferred to a surface of the first substrate facing the mask plate, thereby forming an electrically conductive layer having one or more electrically conductive portions, wherein a projection of each of the one or more electrically conductive portions on the mask plate coincides with a respective light-transmissive region.