Chloroplasts evolved from a cyanobacterial endosymbiont, and chloroplast division requires the formation of an FtsZ division ring, which is descended from the cytokinetic machinery of cyanobacteria. As in bacteria, the positioning of the chloroplast FtsZ ring is regulated by the proteins MinD and MinE. However, chloroplast division also involves mechanisms invented by the eukaryotic host cell. Here we show that a plant-specific protein MULTIPLE CHLOROPLAST DIVISION SITE 1 (MCD1) regulates FtsZ ring positioning in Arabidopsis thaliana chloroplasts. Our analyses show that both MCD1 and MinD are required for chloroplast division, localizing at the division sites and punctate structures dispersed on the inner envelope. MinD overexpression inhibited FtsZ ring formation whereas MCD1 overexpression did not. Localization studies suggest that MCD1 is required for MinD localization to regulate FtsZ ring formation. Furthermore, the interaction between MCD1 and MinD in yeast two-hybrid assays suggests that MCD1 recruits MinD by direct interaction. These results point out differences in the MinD localization mechanism between chloroplasts and bacterial model systems and suggest that the plant cell evolved a component to modulate the cyanobacteria-derived Min system so as to regulate chloroplast FtsZ ring positioning.