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bioRxiv · 10.1101/2025.07.27.667079

Engineering light robustness: Adaptive evolution uncovers new genetic determinants of HL tolerance in Synechocystis

Sun, T.·Pan, K.·Xie, Y.·Li, S.·Li, C.·Liu, D.·Zhu, X.·Zhang, W.·Chen, L.
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Abstract

Excess light absorption causes severe photo-oxidative damage and limits photosynthetic efficiency. Enhancing high-light (HL) tolerance and energy utilization in photosynthetic cyanobacteria holds great potential for sustainable biomanufacturing. Here, using Synechocystis sp. PCC 6803 (Syn6803) as a model, we generated eight independently evolved strains tolerant to hyper-HL (2000 mol photons/m2/s) through approximately two years of natural adaptive laboratory evolution (ALE). Remarkably, these strains circumvented common evolutionary trade-offs; all demonstrated markedly enhanced quantum yields and photosynthetic performance, with four strains exhibiting a 121.71%-168.36% increase in biomass accumulation under HL. When equipped with the heterologous sucrose transporter CscB, sucrose productivity under HL increased by up to 208.33%. Whole-genome resequencing unveiled a total of 77 mutations across the eight lineages, uncovering a diverse polygenic landscape for HL adaptation. Reverse genetics combined with structural modeling highlighted pivotal roles for regulatory and structural loci, including slr0758 (KaiC1) and slr1513 (SbtB), in modulating circadian rhythmicity and inorganic carbon uptake to prevent photo-oxidative overload. Overall, our findings uncover diverse evolutionary routes toward high-light acclimation and provide superior cyanobacterial chassis for photosynthetic biotechnology.

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Sun, T., Pan, K., Xie, Y., Li, S., Li, C., Liu, D., Zhu, X., Zhang, W., Chen, L.. 2025-07-28. Engineering light robustness: Adaptive evolution uncovers new genetic determinants of HL tolerance in Synechocystis. https://doi.org/10.1101/2025.07.27.667079

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