Twenty-five years ago research had already established a firm biochemical and physiological understanding of the CO 2 -concentrating mechanism that creates a high CO 2 environment (1,000-3,000 bar) in bundle-sheath cells in leaves of C 4 plants and accounts for most of their distinctive photosynthetic properties (5). It was then clear that the minimum requirements for this CO 2 concentrating mechanism included: (a) cell-specific amplification of enzymes of C 4 photosynthesis (i.e. phosphoenolpyruvate carboxylase [PEPC] in mesophyll, and C 4 acid decarboxylases and Rubisco in bundle-sheath cells), with complementary adjustments of photosystem and electron transport activities; (b) novel cell-specific organelle metabolite translocators; (c) symplastic connections of the spatially separated sources and sinks of 4C-dicarboxylic acid transport metabolites; and (d) barriers to CO 2 diffusion between the site of CO 2 fixation by PEPCase in mesophyll cells and sites of CO 2 release and refixation by Rubisco in bundle-sheath cells.These requirements have been met in a great variety of ways during the evolution of C 4 plants, through diverse cooperative pathways of carbon metabolism and integrated photoreactions in adjacent, differentiated photosynthetic cells. Perhaps the most simple, highly evolved system is that in Sorghum (detailed in the legend of Fig. 1), but it is in the diversity of other systems that we can expect to discover clues as to what it takes to be C 4 .