Creation of herbicide‐resistance in allotetraploid peanut using CRISPR /Cas9‐meditated cytosine base‐editing

作者：Lei Shi, Xiaona Li, Lu‐Lu Xue, Jin Zhang, Bingyan Huang, Ziqi Sun, Zhongxin Zhang, Xiaodong Dai, Suoyi Han, Wenzhao Dong, Xinyou Zhang · 发表于：Plant Biotechnology Journal · 年份：2023 · DOI：10.1111/pbi.14114 · 被引用次数：20 · 研究领域：CRISPR and Genetic Engineering、Plant Virus Research Studies、Insect Resistance and Genetics

Genome editing has been employed to introduce accurate and predictable genetic mutations in plants, gaining wide acceptance due to its high efficiency, precision, usability and significant potential for crop improvement (Manghwar et al., 2019). The recent discovery of CRISPR-associated base editors, such as the cytidine base editor (CBE) that converts C∙G to T∙A (Komor et al., 2016) and the adenine base editor (ABE) that converts A∙T to G∙C (Gaudelli et al., 2017), has considerably broadened the scope of genome editing by enabling the creation of irreversible point mutations. This base editor approach has facilitated the genetic enhancement of numerous plant species. However, it remains unclear whether the base editing system would function seamlessly in the allotetraploid peanut (Arachis hypogaea), one of the most vital annual oilseed crops. Acetolactate synthase (ALS), which catalyses the initial step in the biosynthetic pathway to valine, isoleucine and leucine, is targeted by over 50 commercial herbicides that are extensively utilized for weed control. A single base mutation at a specific target site in the ALS nucleotide sequence, such as a natural point mutation converting Pro197 to Ser197 or Leu197 in ALS (numbered based on the corresponding Arabidopsis thaliana ALS sequence), leads to resistance to ALS-inhibiting herbicides (Jin et al., 2022). In this study, we generated herbicide-resistant peanut plants by introducing a targeted single-base substitution into the A. h...