Simultaneous gene editing of three homoeoalleles in self-incompatible allohexaploid grasses

• 作者: Liang Zhang Tao Wang Guangyang Wang Aoyue Bi Misganaw Wassie Yan Xie Liwen Cao Huawei Xu Jinmin Fu Liang Chen Yang Zhao Tao Hu
• 来源期刊: 植物学报（英文版）
• 年，卷（期）: 2021,（8）
• 所属期刊栏目: Breakthrough Reports
• 页码范围: 1410-1415
• 页数: 6页
• 语种: 英文

摘要

Clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system has been widely used for precise gene editing in plants. However, simultaneous gene editing of multiple homoeoalleles remains challenging, especially in self-incompatible polyploid plants. Here, we simultaneously in-troduced targeted mutations in all three homoeo-alleles of two genes in the self-incompatible allo-hexaploid tall fescue, using both CRISPR/Cas9 and LbCas12a (LbCpf1) systems. Loss-of-function mu-tants of FaPDS exhibited albino leaves, while knockout of FaHSP17.9 resulted in impaired heat resistance in T0 generation of tall fescue. More-over, these mutations were inheritable. Our findings demonstrate the feasibility of generating loss-of-function mutants in T0 generation polyploid per-ennial grasses using CRISPR/Cas systems.