National Institute of Plant Genome Research (NIPGR), Aruna Asaf Ali Marg, New Delhi, 110067, India.
Department of Plant Molecular Biology, University of Delhi South Campus, Benito Juarez Marg, New Delhi, 110021, India.
Plant J. 2020 Mar;101(6):1411-1429. doi: 10.1111/tpj.14605. Epub 2019 Dec 12.
Mediator, a multisubunit co-activator complex, regulates transcription in eukaryotes and is involved in diverse processes in Arabidopsis through its different subunits. Here, we have explored developmental aspects of one of the rice Mediator subunit gene OsMED14_1. We analyzed its expression pattern through RNA in situ hybridization and pOsMED14_1:GUS transgenics that showed its expression in roots, leaves, anthers and seeds prominently at younger stages, indicating possible involvement of this subunit in multiple aspects of rice development. To understand the developmental roles of OsMED14_1 in rice, we generated and studied RNAi-based knockdown rice plants that showed multiple effects including less height, narrower leaves and culms with reduced vasculature, lesser lateral root branching, defective microspore development, reduced panicle branching and seed set, and smaller seeds. Histological analyses showed that slender organs were caused by reduction in both cell number and cell size in OsMED14_1 knockdown plants. Flow cytometric analyses and expression analyses of cell cycle-related genes revealed that defective cell-cycle progression led to these defects. Expression analyses of auxin-related genes and indole-3-acetic acid (IAA) immunolocalization study indicated altered auxin level in these knockdown plants. Reduction of lateral root branching in knockdown plants was corrected by exogenous IAA supplement. OsMED14_1 physically interacts with transcription factors YABBY5, TAPETUM DEGENERATION RETARDATION (TDR) and MADS29, possibly regulating auxin homeostasis and ultimately leading to lateral organ/leaf, microspore and seed development.
中介体是一个多亚基共激活复合物,在真核生物中调节转录,通过其不同亚基参与拟南芥的多种过程。在这里,我们探索了水稻中介体亚基基因 OsMED14_1 的一个发育方面。我们通过 RNA 原位杂交和 pOsMED14_1:GUS 转基因分析其表达模式,结果表明该基因在根、叶、花药和种子中在早期阶段表达明显,表明该亚基可能参与水稻发育的多个方面。为了了解 OsMED14_1 在水稻中的发育作用,我们生成并研究了基于 RNAi 的敲低水稻植株,这些植株表现出多种效应,包括株高降低、叶片变窄、维管束减少、侧根分支减少、花粉发育缺陷、穗分支和结实减少、种子变小。组织学分析表明,OsMED14_1 敲低植株中细胞数量和细胞大小的减少导致了细长器官的形成。流式细胞术分析和细胞周期相关基因的表达分析表明,细胞周期进程的缺陷导致了这些缺陷。生长素相关基因的表达分析和吲哚-3-乙酸 (IAA) 免疫定位研究表明,这些敲低植株中的生长素水平发生了改变。在敲低植株中,侧根分支的减少通过外源 IAA 补充得到了纠正。OsMED14_1 与转录因子 YABBY5、TAPETUM DEGENERATION RETARDATION (TDR) 和 MADS29 物理相互作用,可能调节生长素稳态,并最终导致侧生器官/叶片、花粉和种子发育。