Zhang Jia, Hu Wei, Wen Qingli, Fan Xiaowei, Hu Yong, He Qin, Lu Li, Li Jinfeng, Xing Yongzhong
National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Hongshan Laboratory, Wuhan, 430070, China.
College of Life Science, Jiangxi Normal University, Nanchang, 330022, China.
Rice (N Y). 2024 Nov 15;17(1):70. doi: 10.1186/s12284-024-00750-8.
Ghd7 is a central regulator to multiple growth and development processes in rice. While it is not clear how Ghd7 is regulated by upstream factors. To identify its upstream regulator, the truncated Ghd7 promoter fragments were used to screen cis elements binding to rice total nuclear proteins. Electrophoretic mobility shift assays screened one truncated fragment f3 binding to the proteins. Subsequently, the fragment f3 was employed to screen a yeast one-hybrid library, and a transcription factor OsIAA23 was screened as a direct upstream regulator of Ghd7. Dual-luciferase transient assay demonstrated the transcriptional repression effect of OsIAA23 on the activity of Ghd7, and the location of the cis elements binding to OsIAA23 in the region 1264 to 1255 bp upstream of ATG. Genetic analysis between the wild type Ghd7-OsIAA23 and single/double mutants further verified that OsIAA23 downregulated Ghd7 expression and led to a delayed heading under long day conditions. Moreover, natural variations in fragment f3 were associated with heading and geographic distribution in rice. This study sheds light on the direct regulatory mechanism of OsIAA23 on Ghd7, which enriches the understanding of the Ghd7 involved flowering regulatory network in rice.
Ghd7是水稻多种生长发育过程的核心调控因子。然而,目前尚不清楚Ghd7是如何被上游因子调控的。为了鉴定其上游调控因子,利用截短的Ghd7启动子片段筛选与水稻总核蛋白结合的顺式元件。电泳迁移率变动分析筛选出一个与蛋白质结合的截短片段f3。随后,利用片段f3筛选酵母单杂交文库,并筛选出转录因子OsIAA23作为Ghd7的直接上游调控因子。双荧光素酶瞬时分析证明了OsIAA23对Ghd7活性的转录抑制作用,以及与OsIAA23结合的顺式元件在ATG上游1264至1255 bp区域的位置。野生型Ghd7 - OsIAA23与单/双突变体之间的遗传分析进一步证实,OsIAA23下调Ghd7表达并导致长日照条件下抽穗延迟。此外,片段f3的自然变异与水稻的抽穗和地理分布有关。本研究揭示了OsIAA23对Ghd7的直接调控机制,丰富了对水稻中参与开花调控网络的Ghd7的理解。