The invention discloses a lubricating grease filling porous structure anti-fouling surface, a preparation method and application thereof. The preparation method comprises the steps of providing a titanium substrate; adopting sand blasting and/or chemical etching for performing etching treatment on the titanium substrate, and forming a rough porous micro-nanometer structural layer on the titanium substrate; adopting a modifying solution for performing chemical modification on the titanium substrate with the porous micro-nanometer structural layer, and obtaining the lubricating grease filling porous structure anti-fouling surface; filling the porous micro-nanometer structural layer on the surface of the hydrophobic and oleophylic titanium substrate surface with lubricating grease, and obtaining the lubricating grease filling porous structure anti-fouling surface. According to the method for filling the titanium substrate surface porous structure with lubricating grease, the technology iseasy and convenient to implement, operability is high, the cost is low, the practicability is high, the method is the efficient anti-fouling means of new thoughts and new concepts, the technical bottleneck that lubricating oil existing on a lubricating oil filling micro-hole structure surface is likely to be lost, and the anti-fouling effect is short is solved, and the good prospect and application value in the ocean anti-fouling field are achieved.