Guo Siyi, Dai Shaojun, Singh Prashant K, Wang Hongyan, Wang Yanan, Tan Jeanie L H, Wee Wanyi, Ito Toshiro
State Key Laboratory of Cotton Biology, Department of Biology, Institute of Plant Stress Biology, Henan University, Kaifeng, China.
Temasek Life Sciences Laboratory, National University of Singapore, Singapore, Singapore.
Front Plant Sci. 2018 May 3;9:555. doi: 10.3389/fpls.2018.00555. eCollection 2018.
In spite of short-day (SD) nature, rice () shares a conserved photoperiodic network for flowering control with long-day plants like Flowering or heading is an important agronomic trait in rice. NAC transcription factors (TFs) are well-conserved and one of the largest families of plant TFs. However, their function in flowering or heading time is not well-known yet. A preferential expression of a membrane-bound NAC-like TF OsNTL5 in developing leaves and panicles of rice indicated to us its putative role in flowering. To examine its function, three independent constructs was generated, one with a deletion in the C terminus membrane-spanning domain (OsNTL5∆C), OsNTL5∆C fused with the SRDX transcriptional repressor motif and OsNTL5∆C used with the VP16 activation domain under the promoter to produce the overexpressing lines ∆C-, and , respectively in rice. The ∆ line showed an early-flowering phenotype. In contrast to this, the plants with ∆C and ∆C- showed a very strong late-flowering phenotype, suggesting that OsNTL5 suppresses flowering as a transcriptional repressor. The protein subcellular localization assay suggested that N-terminal part of the OsNTL5 is localized to the nucleus after the protein is cleaved from its membrane-spanning domain at the C-terminal end and functions as a TF. Expression of flowering genes responsible for day length signals such as (), (), and () was significantly changed in the overexpression lines of ∆∆, and ∆ as analyzed by Quantitative Real-time PCR. ChIP-qPCR and rice protoplasts assays indicate that OsNTL5 directly binds to the promoter of and negatively regulates the expression of , which shows antagonistic photoperiodic expression patterns of in a 24-h SD cycle. Hence in conclusion, the NAC-like TF OsNTL5 functions as a transcriptional repressor to suppress flowering in rice as an upstream factor of .
尽管水稻具有短日照(SD)特性,但它与拟南芥等长日照植物在开花控制方面共享一个保守的光周期网络。开花或抽穗是水稻的一个重要农艺性状。NAC转录因子(TFs)高度保守,是植物TFs中最大的家族之一。然而,它们在开花或抽穗时间方面的功能尚不清楚。水稻中一种膜结合NAC样TF OsNTL5在发育中的叶片和穗中优先表达,这向我们表明了它在开花中的假定作用。为了研究其功能,构建了三个独立的构建体,一个在C末端跨膜结构域有缺失(OsNTL5∆C),OsNTL5∆C与SRDX转录抑制基序融合,以及在启动子下与VP16激活结构域一起使用的OsNTL5∆C,以分别在水稻中产生过表达系∆C-、和。∆系表现出早花表型。与此相反,带有∆C和∆C-的植株表现出非常强烈的晚花表型,这表明OsNTL5作为转录抑制因子抑制开花。蛋白质亚细胞定位分析表明,OsNTL5的N末端部分在蛋白质从其C末端的跨膜结构域切割后定位于细胞核,并作为TF发挥作用。通过定量实时PCR分析,在∆∆、和∆的过表达系中,负责日长信号的开花基因如()、()和()的表达发生了显著变化。ChIP-qPCR和水稻原生质体分析表明,OsNTL5直接结合到的启动子上,并负调控的表达,在24小时SD周期中显示出拮抗的光周期表达模式。因此,总之,NAC样TF OsNTL5作为转录抑制因子,作为的上游因子抑制水稻开花。