The embodiment of the invention provides a flexible display substrate and a manufacturing method of the flexible display substrate and relates to the field of display technologies. Film layers on the flexible substrate can be prevented from cracking or falling off, and warping generated when a bearing substrate and a flexible substrate located on the bearing substrate are separated is further relieved; the flexible display substrate comprises a first buffering layer, a second buffering layer, thin film transistors and electrode structures, wherein the first buffering layer and the second buffering layer are arranged on the upper surface and the lower surface of the flexible substrate respectively, and the thin film transistors and the electrode structures are at least arranged on the first buffering layer; display structures are formed at least by the thin film transistors and the electrode structures. The manufacturing method for the flexible display substrate is used for manufacturing the flexible display substrate where the film layers arranged on the flexible substrate are prevented from cracking or falling off, and the warping generated when the bearing substrate and the flexible substrate are separated needs to be relieved.