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Highly Efficient and Heritable Targeted Mutagenesis in Wheat via the Agrobacterium tumefaciens-Mediated CRISPR/Cas9 System

文献类型： 外文期刊

作者： Zhang, Shujuan 1 ; Zhang, Rongzhi 1 ; Gao, Jie 1 ; Gu, Tiantian 1 ; Song, Guoqi 1 ; Li, Wei 1 ; Li, Dandan 1 ; Li, Yulian 1 ;

作者机构： 1.Shandong Acad Agr Sci, Crop Res Inst, Jinan 250100, Shandong, Peoples R China

2.Minist Agr, Key Lab Wheat Biol & Genet Improvement North Yell, Jinan 250100, Shandong, Peoples R China

3.Natl Engn Lab Wheat & Maize, Jinan 250100, Shandong, Peoples R China

关键词： Agrobacterium tumefaciens; CRISPR; Cas9; heritable; later generations; mutation; wheat

期刊名称：INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES （ 影响因子：5.923； 五年影响因子：6.132 ）

ISSN：

年卷期： 2019 年 20 卷 17 期

页码：

收录情况： SCI

摘要： The CRISPR/Cas9 system has been successfully used in hexaploid wheat. Although it has been reported that the induced mutations can be passed to the next generation, gene editing and transmission patterns in later generations still need to be studied. In this study, we demonstrated that the CRISPR/Cas9 system could achieve efficient mutagenesis in five wheat genes via Agrobacterium-mediated transformation of an sgRNA targeting the D genome, an sgRNA targeting both the A and B homologues and three tri-genome guides targeting the editing of all three homologues. High mutation rates and putative homozygous or biallelic mutations were observed in the T0 plants. The targeted mutations could be stably inherited by the next generation, and the editing efficiency of each mutant line increased significantly across generations. The editing types and inheritance of targeted mutagenesis were similar, which were not related to the targeted subgenome number. The presence of Cas9/sgRNA could cause new mutations in subsequent generations, while mutated lines without Cas9/sgRNA could retain the mutation type. Additionally, off-target mutations were not found in sequences that were highly homologous to the selected sgRNA sequences. Overall, the results suggested that CRISPR/Cas9-induced gene editing via Agrobacterium-mediated transformation plays important roles in wheat genome engineering.

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