The outside of the Arabidopsis thaliana fruit consists of three principal tissues: the valves or seedpod walls, the replum or central ridge between the valves, and the valve margins where the valves separate from the replum to disperse the seeds. Previous studies have shown that valve margin formation is specified by the SHATTERPROOF MADS-box transcription factors and that valve development is controlled by the FRUITFULL MADS-box transcription factor. FRUITFULL negatively regulates SHATTERPROOF to prevent the valves from adopting a valve margin cell fate. Here we identify a gene called REPLUMLESS that is required for replum development. REPLUMLESS encodes a homeodomain protein that prevents replum cells from adopting a valve margin cell fate by negatively regulating expression of the SHATTERPROOF genes. Both REPLUMLESS and FRUITFULL are required to limit SHATTERPROOF expression to a narrow stripe of cells so that the valve margin differentiates precisely at the valve/replum boundary.