Abstract: The present disclosure relates to a three-dimensional (3D) printing ink for Pleurotus eryngii and a preparation method for and use of the 3D printing ink and a preparation method for 3D printed food of Pleurotus eryngii. The present disclosure adopts Pleurotus eryngii freeze-dried powder which reduces a moisture content of Pleurotus eryngii. In addition, the problem that the Pleurotus eryngii with high fiber content blocks a nozzle for 3D printing can be solved by making the Pleurotus eryngii into small particle powder, and bioactive substances in the Pleurotus eryngii can be retained as much as possible. The addition of hydrocolloid can improve rheological properties of the Pleurotus eryngii freeze-dried powder, which increases viscosity of the 3D printing ink for Pleurotus eryngii, and enhances mechanical strength of the 3D printing ink, such that the ink can maintain the print shape and is not easy to collapse.