Identification of herbicide resistance OsACC1 mutations via in planta prime-editing-library screening in rice
Identification of herbicide resistance OsACC1 mutations via in planta prime-editing-library screening in rice
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DOI：
10.1038/s41477-021-00942-w
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发表时间：
2021-06
期刊：
影响因子：
18
通讯作者：
R. Xu;Xiaoshuang Liu;Juan Li;R. Qin;P. Wei
中科院分区：
文献类型：
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作者：
R. Xu;Xiaoshuang Liu;Juan Li;R. Qin;P. Wei
碱基编辑文库诱导的高密度核苷酸替换已被应用于植物功能突变的筛选。然而，由于碱基编辑的范围和转换特异性的限制，关键蛋白位点上的许多所需突变可能会被忽略。在这里，我们开发了一种编辑文库介导的饱和突变(PLSM)方法，以显著增加目标位点氨基酸替换的多样性，用于植物内筛选。在OsACC1的6个保守残基上，鉴定出16种类型的除草剂抗性突变。这些突变大多对芳氧苯氧丙酸类除草剂表现出可靠的耐受性，在水稻育种中尚未见报道或应用。此外，通过比较碱基编辑介导的突变在所选靶点的作用，进一步显示了PLSM的优势。本研究建立的PLSM方法对于直接进化作物重要农艺性状相关基因具有很大的潜力。
Base-editing-library-induced high density nucleotide substitutions have been applied to screen functional mutations in plants. However, due to limitations in the scope and conversion specificity of base editors, many desired mutations at pivotal protein sites may be overlooked. Here, we developed a prime-editing-library-mediated saturation mutagenesis (PLSM) method to substantially increase the diversity of amino acid substitutions at target sites for in planta screening. At six conserved residues of OsACC1, 16 types of herbicide-resistance-endowing mutations were identified. Most of these mutations exhibit reliable tolerance to aryloxyphenoxypropionate herbicides and have not been reported or applied in rice breeding. In addition, the advantage of PLSM was further shown by comparing the base-editing-mediated mutagenesis at the selected targets. The PLSM method established in this study has great potential for the direct evolution of genes related to agronomically important traits for crop improvement.