NDH-mediated cyclic electron flow around photosystem I is crucial for C₄ photosynthesis

C₄ photosynthesis exhibits efficient CO₂ assimilation in ambient air by concentrating CO₂ around ribulose 1,5- bisphosphate carboxylase/oxygenase (Rubisco) through a metabolic pathway called the C₄ cycle. It has been suggested that cyclic electron flow (CEF) around PSI mediated by chloroplast NADH dehydrogenase-like complex (NDH), an alternative pathway of photosynthetic electron transport (PET), plays a crucial role in C₄ photosynthesis, although the contribution of NDH-mediated CEF is small in C3 photosynthesis. Here, we generated NDH-suppressed transformants of a C₄ plant, Flaveria bidentis, and showed that the NDH-suppressed plants grow poorly, especially under lowlight conditions. CO₂ assimilation rates were consistently decreased in the NDH-suppressed plants under low and medium light intensities. Measurements of non-photochemical quenching (NPQ) of Chl fluorescence, the oxidation state of the reaction center of PSI (P700) and the electrochromic shift (ECS) of pigment absorbance indicated that proton translocation across the thylakoid membrane is impaired in the NDH-suppressed plants. Since proton translocation across the thylakoid membrane induces ATP production, these results suggest that NDH-mediated CEF plays a role in the supply of ATP which is required for C₄ photosynthesis. Such a role is more crucial when the light that is available for photosynthesis is limited and the energy production by PET becomes rate-determining for C₄ photosynthesis. Our results demonstrate that the physiological contribution of NDH-mediated CEF is greater in C₄ photosynthesis than in C3 photosynthesis, suggesting that the mechanism of PET in C₄ photosynthesis has changed from that in C3 photosynthesis accompanying the changes in the mechanism of CO₂ assimilation.