Diversification of plant agronomic traits by genome editing of brassinosteroid signaling family genes in rice
Diversification of plant agronomic traits by genome editing of brassinosteroid signaling family genes in rice
复制标题
通过对水稻油菜素类固醇信号家族基因进行基因组编辑使植物农艺性状多样化
DOI：
10.1093/plphys/kiab394
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发表时间：
2021-08-16
期刊：
影响因子：
7.4
通讯作者：
Tong, Hongning
中科院分区：
文献类型：
--
作者：
Liu, Dapu;Yu, Zhikun;Tong, Hongning
油菜素类固醇 (BR) 调节水稻 (Oryza sativa L.) 的各种农艺性状，如株高、叶角和籽粒大小；因此，BR信号传导成分是分子合理设计的有希望的目标。然而，水稻中 BR 信号基因或家族成员的遗传物质仍然有限。在这里，通过使用成簇规则间隔短回文重复序列 (CRSPR)/Cas9 工具进行基因组编辑，我们在三个 BR 信号基因家族中生成了一组单、双、三或四突变体，包括 GSK3/SHAGGY-LIKE KINASE1 (GSK1)-GSK4、BRASSINAZOLE-RESISTANT1 (OsBZR1)-OsBZR4 和蛋白磷酸酶kelch-like (PPKL)1-PPKL3，在相同背景下（Zhonghua11，粳稻）。高阶突变体是通过同时靶向一个家族（GSK 和 PPKL）的不同基因上的多个位点或靶向家族成员（OsBZR）的重叠序列来产生的。突变体表现出株高、叶角和籽粒形态的多样性。表型的比较分析以及 BR 敏感性测试表明每个家族内的成员之间存在功能冗余、分化或优势。此外，我们还生成了一组分别以野生型或融合不同标签的激活形式过表达 GSK2、OsBZR1/2 和 PPKL2 的转基因植物，并验证了对 BR 应用的蛋白质反应。总的来说，这些植物极大地丰富了水稻重要农艺性状的多样性。我们认为编辑BR相关家族基因可能是筛选所需植物以满足不同要求的可行方法。这些材料以及相关信息的发布也为BR的进一步研究和利用提供了宝贵的资源。
Brassinosteroids (BRs) regulate various agronomic traits such as plant height, leaf angle, and grain size in rice (Oryza sativa L.); thus, BR signaling components are promising targets for molecular rational design. However, genetic materials for BR-signaling genes or family members remain limited in rice. Here, by genome editing using clustered regularly interspaced short palindromic repeats (CRSPR)/Cas9 tools, we generated a panel of single, double, triple, or quadruple mutants within three BR signaling gene families, including GSK3/SHAGGY-LIKE KINASE1 (GSK1)-GSK4, BRASSINAZOLE-RESISTANT1 (OsBZR1)-OsBZR4, and protein phosphatases with kelch-like (PPKL)1-PPKL3, under the same background (Zhonghua11, japonica). The high-order mutants were produced by either simultaneously targeting multiple sites on different genes of one family (GSKs and PPKLs) or targeting the overlapping sequences of family members (OsBZRs). The mutants exhibited a diversity of plant height, leaf angle, and grain morphology. Comparison analysis of the phenotypes together with BR sensitivity tests suggested the existence of functional redundancy, differentiation, or dominancy among the members within each family. In addition, we generated a set of transgenic plants overexpressing GSK2, OsBZR1/2, and PPKL2, respectively, in wild-type or activated forms with fusion of different tags, and also verified the protein response to BR application. Collectively, these plants greatly enriched the diversity of important agronomic traits in rice. We propose that editing of BR-related family genes could be a feasible approach for screening of desired plants to meet different requirements. Release of these materials as well as the related information also provides valuable resources for further BR research and utilization.