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

查看全文 作　　者：Chen [1]Zhang;Xue [1,2]Zhong;Shaoya [1,2,3]Li;Lei [1]Yan;Jingying [1,2,3]Li;Yubing [1,2,3]He;Yong [4]Lin;Yangjun [4]Zhang;Lanqin [1,2,3]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》索引2023年第65卷第9期,共10页

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

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

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