Enhancing Photosynthesis under Salt Stress via Directed Evolution in Cyanobacteria

Abstract

A key aspect of enhancing photosynthesis is improving its recycling kinetics, enabling swift resumption of photochemical quenching following environmental disruptions or stress. Salt stress exacerbates high light stress in cyanobacteria and leads to severe yield losses in crop plants. Genetic traits that confer salt tolerance without compromising photosynthetic performance are essential for improving photosynthesis under these conditions. Here we applied accelerated evolution in Synechococcus elongatus PCC 7942 by conditionally suppressing its methyl-directed mismatch repair system to obtain beneficial genetic traits for enhanced photosynthesis under salt stress. Screening over 10,000 mutants, we isolated eight strains with increased biomass or sucrose productivity under salt stress. Genome sequencing revealed an average of 8-20 single nucleotide polymorphisms (SNPs) or indels per genome. Notably, mutations in the photosystem II (PSII) reaction center D1 gene, resulting in the amino acid changes L353F, I358N, and H359N at the carboxyl terminus of the pre-D1 (pD1) protein, improve photosynthesis under salt and combined salt and light stress by potentially accelerating D1 maturation during PSII repair. Phylogenetic analysis of pD1 across cyanobacteria and red algae highlights the broad significance of these adaptive genetic traits, underscoring the importance of leveraging evolutionary insights to improve photosynthesis under stress or fluctuating environments.