Abstract：
Knockout mutants provide definitive information about the functions of genes related to agronomic traits, including seed dormancy. However, it takes many years to produce knockout mutants using conventional techniques in polyploid plants such as hexaploid wheat. Genome editing with sequence-specific nucleases is a promising approach for obtaining knockout mutations in all targeted homoeologs of wheat simultaneously. Here, we describe a procedure to produce a triple recessive mutant in wheat via genome editing. This protocol covers the evaluation of gRNA and Agrobacterium-mediated transformation to obtain edited wheat seedlings.
摘要：
敲除突变体提供了有关农艺性状相关基因的功能的明确信息，包括种子休眠。然而,使用常规技术在多倍体植物如六倍体小麦中产生敲除突变体需要很多年。使用序列特异性核酸酶进行基因组编辑是同时在小麦的所有靶向同源物中获得敲除突变的有前途的方法。这里,我们描述了通过基因组编辑在小麦中产生三隐性突变体的方法。该方案涵盖了gRNA和农杆菌介导的转化的评估，以获得编辑的小麦幼苗。
