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油菜籽中的膜结合转录激活因子NTL1通过靶向参与活性氧产生和程序性细胞死亡的基因正向调控叶片衰老。

Membrane-Bound Transcriptional Activator NTL1 from Rapeseed Positively Modulates Leaf Senescence through Targeting Genes Involved in Reactive Oxygen Species Production and Programmed Cell Death.

作者信息

Yan Jingli, Li Yanfei, Zhao Peiyu, Mu Bangbang, Chen Qinqin, Li Xin, Cui Xing, Wang Zhaoqiang, Li Jing, Li Shaojun, Yang Bo, Jiang Yuan-Qing

机构信息

State Key Laboratory of Crop Stress Biology for Arid Areas and, College of Life Sciences, Northwest A & F University, Yangling, Shaanxi 712100, China.

College of Plant Protection, Henan Agricultural University, Zhengzhou, Henan 450002, China.

出版信息

J Agric Food Chem. 2021 May 5;69(17):4968-4980. doi: 10.1021/acs.jafc.1c00182. Epub 2021 Apr 20.

Abstract

Leaf senescence is the last stage of leaf development and is determined by various environmental and endogenous signals. Leaf senescence can determine plant productivity and fitness. Transcription factors (TFs) with the transmembrane domain constitute a special group of regulatory proteins that can translocate from the membrane system into nuclei to exert the transcriptional function upon endogenous or exogenous stimuli. Reactive oxygen species (ROSs) play an important role in numerous processes throughout the life cycle of plants including leaf senescence. Leaf senescence is characterized by massive programmed cell death (PCD) and is a type of developmental PCD. The transcriptional regulatory relationships between membrane-bound TFs and leaf senescence remain largely uncharacterized, especially in rapeseed ( L.), an important oil crop. Here, we show that BnaNTL1 is a membrane-bound NAC (NAM, ATAF, and CUC) TF, which is predominantly expressed in senescent leaves. Expression of , a form of BnaNTL1 devoid of the transmembrane domain, can induce serious HR-like cell death symptoms and ROS accumulation in cells. Plants overexpressing show earlier leaf senescence compared with the control, accompanied by chlorophyll degradation and electrolyte leakage. Genes involved in ROS production (), PCD ( and ), and leaf senescence () are significantly induced and activated by BnaNTL1ΔTM according to the quantitative reverse transcription PCR (qRT-PCR) analysis and dual luciferase reporter (Dual-LUC) assay. Moreover, electrophoretic mobility shift assay revealed that BnaNTL1 directly bound to the NTLBS elements in promoters of , , and . In conclusion, these results demonstrate that BnaNTL1 positively modulates ROS production and HR-like cell death to induce leaf senescence.

摘要

叶片衰老为叶片发育的最后阶段,由多种环境和内源信号决定。叶片衰老能够决定植物的生产力和适应性。具有跨膜结构域的转录因子(TFs)构成了一类特殊的调节蛋白,其可在内源或外源刺激下从膜系统转运至细胞核发挥转录功能。活性氧(ROSs)在植物包括叶片衰老在内的整个生命周期的众多过程中发挥重要作用。叶片衰老的特征是大量程序性细胞死亡(PCD),属于发育性PCD的一种。膜结合转录因子与叶片衰老之间的转录调控关系在很大程度上仍未明确,尤其是在重要油料作物油菜(L.)中。在此,我们表明BnaNTL1是一种膜结合NAC(NAM、ATAF和CUC)转录因子,主要在衰老叶片中表达。缺失跨膜结构域的BnaNTL1形式BnaNTL1ΔTM的表达可诱导细胞出现严重的类过敏反应(HR)样细胞死亡症状和ROS积累。与对照相比,过表达BnaNTL1ΔTM的植株叶片衰老提前,伴有叶绿素降解和电解质渗漏。根据定量逆转录PCR(qRT-PCR)分析和双荧光素酶报告基因(Dual-LUC)检测,参与ROS产生()、PCD(和)以及叶片衰老()的基因受到BnaNTL1ΔTM的显著诱导和激活。此外,电泳迁移率变动分析表明BnaNTL1直接结合至、和启动子中的NTLBS元件。总之,这些结果表明BnaNTL1正向调控ROS产生和类HR细胞死亡以诱导叶片衰老。

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