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Editing miR482b and miR482c Simultaneously by CRISPR/Cas9 Enhanced Tomato Resistance to Phytophthora infestans

文献类型： 外文期刊

作者： Hong, Yuhui 1 ; Meng, Jun 2 ; He, Xiaoli 1 ; Zhang, Yuanyuan 1 ; Liu, Yarong 1 ; Zhang, Chengwei 3 ; Qi, Hongyan 4 ; Luan 1 ;

作者机构： 1.Dalian Univ Technol, Sch Bioengn, Dalian 116024, Peoples R China

2.Dalian Univ Technol, Sch Comp Sci & Technol, Dalian 116024, Peoples R China

3.Beijing Acad Agr & Forestry Sci, Beijing Key Lab Maize DNA Fingerprinting & Mol Br, Beijing 100000, Peoples R China

4.Shenyang Agr Univ, Coll Hort, Key Lab Protected Hort, Minist Educ,Northern Natl & Local Joint Engn Res, Shenyang 110866, Peoples R China

关键词： CRISPR/Cas9; disease resistance; late blight; miRNA; plant immune responses; tomato

期刊名称：PHYTOPATHOLOGY （ 影响因子：4.025； 五年影响因子：4.394 ）

ISSN： 0031-949X

年卷期： 2021 年 111 卷 6 期

页码：

收录情况： SCI

摘要： Late blight, caused by Phytophthora infestans, is severely damaging to the global tomato industry. Micro-RNAs (miRNAs) have been widely demonstrated to play vital roles in plant resistance by repressing their target genes. Recently, the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) method has been continuously improved and extensively applied to edit plant genomes. However, editing multiplex miRNAs by CRISPR/Cas9 in tomato has not been studied yet. We knocked out miR482b and miR482c simultaneously in tomato through the multiplex CRISPR/Cas9 system. Two transgenic plants with silenced miR482b and miR482c simultaneously and one transgenic line with silenced miR482b alone were obtained. Compared with wild-type plants, the disease symptoms of three transgenic plants upon infection were reduced, accompanied by increased expression of their common target nucleotide binding site-leucine-rich repeat genes and decreased levels of reactive oxygen species. Furthermore, silencing miR482b and miR482c simultaneously was more resistant than silencing miR482b alone in tomato. More importantly, we found that knocking out miR482b and miR482c can elicit expression perturbation of other miRNAs, suggesting cross-regulation between miRNAs. Our study demonstrated that editing miR482b and miR482c simultaneously with CRISPR/Cas9 is an efficient strategy for generating pathogen-resistant tomatoes, and cross-regulation between miRNAs may reveal the novel mechanism in tomato-P. infestans interactions.

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