Overexpression of the Rieske FeS protein of the Cytochrome b ₆ f complex increases C₄ photosynthesis in Setaria viridis

C₄ photosynthesis is characterised by a CO₂ concentrating mechanism that operates between mesophyll and bundle sheath cells increasing CO₂ partial pressure at the site of Rubisco and photosynthetic efficiency. Electron transport chains in both cell types supply ATP and NADPH for C₄ photosynthesis. Cytochrome b₆f is a key control point of electron transport in C₃ plants. To study whether C₄ photosynthesis is limited by electron transport we constitutively overexpressed the Rieske FeS subunit in Setaria viridis. This resulted in a higher Cytochrome b₆f content in mesophyll and bundle sheath cells without marked changes in the abundances of other photosynthetic proteins. Rieske overexpression plants showed better light conversion efficiency in both Photosystems and could generate higher proton-motive force across the thylakoid membrane underpinning an increase in CO₂ assimilation rate at ambient and saturating CO₂ and high light. Our results demonstrate that removing electron transport limitations can increase C₄ photosynthesis.