Sylgard-184 silicone elastomer negative replica and resorcinol-formaldehyde (RF) positive replica were made by biomimicking the patterns of natural Trifolium and three other kinds of leaves using the micromolding lithography. An effective antifouling (AF) polymer, poly(3-sulfopropyl methacrylate) (PSPMA), was then grafted on these replica surfaces via the surface-initiated atom transfer radical polymerization (SI-ATRP). The AF property of the modified biomimetic surfaces was tested via the settlement assay with two microalgae in different sizes, and their fouling-release (FR) property was evaluated by the removal assay. The results indicate that the structure of microspines on Trifolium leaf can inhibit settlement of microalgae and facilitate the cell release. The AF property was improved by modification with PSPMA brushes.