Abstract: Orthotropic membranes are more and more widely used in various inflatable structures. The membrane is easy to wrinkle under the action of non-uniform deformation, so it is necessary to study the wrinkle deformation of orthotropic membranes. For the planar orthotropic membranes, the wrinkling criterion of orthotropic materials and the stress extremum method were combined, the stress extremum condition was used to determine the half wavelength of wrinkles, and the amplitude of the wrinkles was determined using strain conditions. The directions of the maximum principal stress and the maximum principal strain were both used to determine the range of the wrinkle direction angle of the orthotropic membrane, and the variation law was studied. The comparison between theoretical analysis and numerical simulation results verifies the reliability of the proposed research method. For the spatial orthotropic membrane, the finite element model was used to analyze the evolution process of the wrinkles, and the difference between the membrane model and the shell model was compared. On the basis of the above research, the air pressure of a waterproof air cushion made by the spatial orthotropic membrane under the coupling of pressure air and water was analyzed and verified by experimental results. The research results provide a reference for the engineering application of orthotropic materials.