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Characteristics of the C4 bundle sheath emerge in C3 rice after editing a plasma membrane proton ATPase

Preprint Created on 23 May 2026 bioRxiv

C4photosynthesis improves light, water and nitrogen-use efficiencies and can raise yieldby50% compared with the ancestral C3pathway.Engineering C4traits into C3crops could substantially boost food production but requires coordinated modifications to leaf anatomy and cell-specific photosynthetic function. For example, C4leaves contain morenumerous, shorter bundle sheath cells that are photosynthetically active. In searching for transcriptional regulators of bundle sheath development in C3 rice, we unexpectedly found OSA3, a plasma membrane H-ATPase that is expressed in bundle sheath cells as they elongate, and when knocked out reduces their length due to reduced apoplastic acidification. Bundle sheath cell number and chloroplast occupancy are increased. Thus, switching between C3 and C4 bundle sheath identity is controlled by acid growth, and OSA3 represents a simple tool for C4 engineering.

Hua, L., Plackett, A. R. G., Wang, N., Hibberd, J. M.

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