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上游开放阅读框-HsfA1a-WOX11模块调控水稻冠根发育。

The uORF-HsfA1a-WOX11 module controls crown root development in rice.

作者信息

Zhang Ting, Xiang Yimeng, Ye Miaomiao, Yuan Meng, Xu Guoyong, Zhou Dao-Xiu, Zhao Yu

机构信息

National Key Laboratory of Crop Genetic Improvement, Hubei Hongshan Laboratory, Huazhong Agricultural University, Wuhan, 430070, China.

College of Food and Biology, Jingchu University of Technology, Jingmen, Hubei, 448000, China.

出版信息

New Phytol. 2025 Jul;247(2):760-773. doi: 10.1111/nph.70214. Epub 2025 May 21.

Abstract

OsWOX11 is a key essential determinant of crown root development in rice. However, either overexpression or downregulation of OsWOX11 results in pleiotropic developmental defects, including dwarfism and reduced yield. Therefore, it is necessary to ensure an optimal level of OsWOX11 expression for balancing the subterranean root system and aerial organ development. OsHsfA1a activates OsWOX11 expression by directly binding to heat stress element-like elements within its promoter. Genetic evidence demonstrated that OsHsfA1a overexpressing or knockout transgenic plants phenocopied the crown root growth in OsWOX11 transgenic plants. Additionally, increased expression of OsWOX11 in OsHsfA1a RNAi background could partially complement the defective crown root phenotypes. A uORF (uORF) was identified within the 5'-untranslated region of OsHsfA1a. Transient expression assays coupled with ribosome profiling revealed that uORF attenuated the translation efficiency of OsHsfA1a mRNA. Furthermore, HsfA1a:uORF-HsfA1a-GFP plants exhibited wild-type crown root phenotypes, whereas uORF knockout transgenic plants displayed similar crown root phenotypes to OsWOX11 overexpressing plants. These findings suggest that uORF fine-tunes the crown root development by repressing OsHsfA1a translation, thereby indirectly modulating OsWOX11 transcript levels. Our study demonstrated a novel uORF-HsfA1a-WOX11 regulatory module that coordinated transcriptional and translational control to maintain optimal OsWOX11 expression. This mechanism ensures the trade-off between root and shoot development. Importantly, targeting uORF regulatory elements provided a new strategy for engineering robust root system architecture without compromising agronomic traits, thereby addressing a critical challenge in cereal crop improvement.

摘要

OsWOX11是水稻冠状根发育的关键必需决定因素。然而,OsWOX11的过表达或下调都会导致多效性发育缺陷,包括矮化和产量降低。因此,有必要确保OsWOX11表达处于最佳水平,以平衡地下根系和地上器官的发育。OsHsfA1a通过直接结合其启动子内的热应激元件样元件来激活OsWOX11的表达。遗传证据表明,过表达或敲除OsHsfA1a的转基因植物表现出与OsWOX11转基因植物冠状根生长相似的表型。此外,在OsHsfA1a RNA干扰背景下增加OsWOX11的表达可以部分弥补有缺陷的冠状根表型。在OsHsfA1a的5'非翻译区内鉴定出一个上游开放阅读框(uORF)。结合核糖体分析的瞬时表达试验表明,uORF减弱了OsHsfA1a mRNA的翻译效率。此外,HsfA1a:uORF-HsfA1a-GFP植物表现出野生型冠状根表型,而uORF敲除转基因植物表现出与OsWOX11过表达植物相似的冠状根表型。这些发现表明,uORF通过抑制OsHsfA1a的翻译来微调冠状根的发育,从而间接调节OsWOX11的转录水平。我们的研究证明了一种新的uORF-HsfA1a-WOX11调控模块,该模块协调转录和翻译控制以维持最佳的OsWOX11表达。这种机制确保了根和地上部发育之间的权衡。重要的是,靶向uORF调控元件为构建强大的根系结构提供了一种新策略,同时不影响农艺性状,从而解决了谷类作物改良中的一个关键挑战。

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