Plants have been proved as a novel production platform for a wide range of biologically important compounds such as enzymes, therapeutic proteins, antibiotics, and proteins with immunological properties. In this context, plastid genetic engineering can be potentially used to produce recombinant proteins. However, several challenges still remain to be overcome if the full potential of plastid transformation technology is to be realized. They include the development of plastid transformation systems for species other than tobacco, the expression of transgenes in non-green plastids, the increase of protein accumulation and the appearance of pleiotropic effects. In this paper, we discuss the novel tools recently developed to overcome some limitations of chloroplast transformation.