文献类型：期刊文献

英文题名：Genome-wide hypermutation-engineered Synechocystis sp. PCC 6803 reveals membrane-mediated triclosan resistance

作者：Wu, Ping[1,2];Wei, Kaixin[1,2];Hu, Tianyouzi[1,2];Chen, Jianfeng[2];Luan, Guodong[3];Sun, Liyun[1,2];Fan, Jianhua[1,2,4]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol, Dept Appl Biol, 130 Meilong Rd, Shanghai 200237, Peoples R China;[3]Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, 189 Songling Rd, Qingdao 266101, Peoples R China;[4]Shihezi Univ, Sch Chem & Chem Engn, 221 Beishi Rd, Shihezi 832003, Peoples R China

年份：2026

卷号：200

期号：3

外文期刊名：PLANT PHYSIOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:001716025800001)】；

基金：This work was sponsored by the National Key Research and Development Project of China 2025YFA0921100, Shanghai Science and Technology Innovation Action Plan 24HC2820800, Natural Science Foundation of Shanghai 23ZR1415100 and 24ZR1490800, Tianchi Talent Project BT-2025-TCYC-0060, and Shanghai Collaborative Innovation Center for Biomanufacturing Technology.

语种：英文

摘要：Cyanobacteria represent an ancient group of photosynthetic microorganisms that offer unparalleled insights into evolutionarily conserved stress adaptation mechanisms essential for plant resilience. To investigate how photosynthetic organisms mitigate chemical stressors, we employed Synechocystis sp. PCC 6803-a keystone model for photosynthetic research due to its plant-like electron transport chain and stress-responsive plasticity. By implementing a genomic hypermutation strategy, we synergistically knocked out DNA replication fidelity genes and overexpressed error-prone replication elements, generating hypermutable strains HM24 and HM33 with relative mutation rates of 97 and 116-fold, respectively. Following triclosan (TCS) stress screening, the CRISPR-Cpf1 strategy was used to complement mutations and yielded transformants R-HM24 and R-HM33 that exhibited 96 h EC50 values of 4.963 and 5.238 mg/L representing 322- and 340-fold increases over wild-type levels, respectively. The strains demonstrated enhanced TCS and multidrug antibiotic tolerance. Whole-genome resequencing identified consistent missense mutation in fabI across resistant strains. Mechanistic analyses revealed that the hypermutated Synechocystis strains acquired resistance primarily by mutating the essential fabI protein to decrease its affinity for TCS. This study establishes the application of hypermutation-driven evolution for rapid dissection of pollutant resistance in photosynthetic microbes, thereby advocating for stricter regulation of antimicrobial pollutants in aquatic environments.