The present disclosure relates to a technical field of OLED display and discloses an OLED backboard, a method of manufacturing the same, an alignment system and an alignment method thereof, which aims to solve a problem of low efficiency in forming an organic light-emitting material layer on the OLED backboard. The OLED backboard comprises: a transparent substrate; at least two functional layers disposed on the upper surface of the transparent substrate and stacked on each other; at least two via holes formed in predefined areas of the at least two functional layers and extending through at least the upper one of the at least two functional layers; and at least two alignment modules located in the at least two via holes respectively, wherein each of the at least two alignment modules comprises at least three first alignment structures arranged in a predefined trace, and a virtual alignment point is allowed to be determined based on the relative position among all the first alignment structures in each of the alignment modules. The efficiency of forming the organic light-emitting material layer on the OLED backboard is thus increased by using the above OLED backboard.