The invention provides a preparation method of multilayer anti-tensile non-woven fabric. The preparation method includes the steps of preparation of a film; preparation of fiber layers, wherein long fibers are prepared into the net-shaped long fiber layer, and short fibers are prepared into the short fiber layer; fiber hardening; preparation of glue; synthesis of the non-woven fabric, wherein the long fiber layer is attached to the upper portion of the film, the short fiber layer is attached to the lower portion of the film, the joints are coated with the glue, and through hot pressing, synthesis is completed. The preparation method has the advantages that an adopted starch source is rich, price is low, and environment friendliness is achieved; the prepared non-woven fabric is high in strength, low in production cost, biodegradable, long in storage time and resistant to high temperature and water, the white pollution problem can be effectively relieved, the service life is prolonged, white pollution is reduced, and the environment is protected.