Artificial evolution of OsEPSPS through an improved dual cytosine and adenine base editor generated a novel allele conferring rice glyphosate tolerance  

作　　者：Chen Zhang Xue Zhong Shaoya Li Lei Yan Jingying Li Yubing He Yong Lin Yangjun Zhang Lanqin Xia 

机构地区：[1]Institute of Crop Sciences(ICS),Chinese Academy of Agricultural Sciences(CAAS),Beijing 100081,China [2]CAAS/Hainan Yazhou Bay Seed Laboratory,National Nanfan Research Institute(Sanya),Sanya 572024,China [3]Key Laboratory of Gene Editing Technologies(Hainan),Ministry of Agricultural and Rural Affairs,Sanya 572024,China [4]Beijing Dabeinong Technology Group Co.,Ltd,Beijing 10080,China

出　　处：《Journal of Integrative Plant Biology》2023年第9期2194-2203,共10页植物学报（英文版）

基　　金：partly funded by the National Natural Science Foundation of China (32188102 to LX);Hainan Yazhou Bay Seed Lab (B21HJ0215 to LX);the Central Public-Interest Scientific Institution-Based Research Fund (S2023ZD03 to LX);Key Laboratory of Gene Editing Technologies (Hainan), China;National Engineering Research Center of Crop Molecular Breeding。

摘　　要：Exploiting novel endogenous glyphosate-tolerant alleles is highly desirable and has promising potential for weed control in rice breeding. Here,through fusions of different effective cytosine and adenine deaminases with nCas9-NG, we engineered an effective surrogate two-component composite base editing system, STCBE-2, with improved C-to-T and A-to-G base editing efficiency and expanded the editing window. Furthermore,we targeted a rice endogenous OsEPSPS gene for artificial evolution through STCBE-2-mediated near-saturated mutagenesis. After hygromycin and glyphosate selection, we identified a novel OsEPSPS allele with an Asp-213-Asn(D213N)mutation(OsEPSPS-D213N) in the predicted glyphosate-binding domain, which conferred rice plants reliable glyphosate tolerance and had not been reported or applied in rice breeding. Collectively, we developed a novel dual base editor which will be valuable for artificial evolution of important genes in crops. And the novel glyphosate-tolerant rice germplasm generated in this study will benefit weeds management in rice paddy fields.

关 键 词：artificial evolution dual cytosine and adenine base editor nCas9‐NG rice(Oryza sativa L) near‐saturated mutagenesis 

分 类 号：S511[农业科学—作物学]