Key Laboratory of Tropical Plant Resources and Sustainable Use, Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, Kunming, Yunnan 650223, China.
Key Laboratory of Tropical Plant Resources and Sustainable Use, Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, Kunming, Yunnan 650223, China; University of Chinese Academy of Sciences, Beijing 100049, China.
Mol Plant. 2017 Sep 12;10(9):1174-1189. doi: 10.1016/j.molp.2017.07.008. Epub 2017 Jul 19.
Leaf senescence can be triggered and promoted by various environmental stressors, developmental cues, and endogenous hormone signals. Several lines of evidence have suggested the involvement of WRKY transcription factors in regulating leaf senescence, but the underlying mechanisms and signaling pathways involved remain elusive. In this study, we identified Arabidopsis thaliana WRKY DNA-binding protein 45 (WRKY45) as a positive regulator of age-triggered leaf senescence. Loss of WRKY45 function resulted in increased leaf longevity in age-triggered senescence, whereas overexpression of WRKY45 significantly accelerated age-triggered leaf senescence. Consistently, expression of SENESCENCE-ASSOCIATED GENEs (SAGs) was significantly reduced in wrky45 mutants but markedly enhanced in transgenic plants overexpressing WRKY45. Chromatin immunoprecipitation assays revealed that WRKY45 directly binds the promoters of several SAGs such as SAG12, SAG13, SAG113, and SEN4. Both in vivo and in vitro biochemical analyses demonstrated that WRKY45 interacts with the DELLA protein RGA-LIKE1 (RGL1), a repressor of the gibberellin (GA) signaling pathway. We found that RGL1 repressed the transcription activation function of WRKY45, thereby attenuating the expression of its regulon. Consistent with this finding, overexpression of RGL1 resulted in significantly increased leaf longevity in age-triggered senescence. Taken together, our results provide compelling evidence that WRKY45 functions as a critical component of the GA-mediated signaling pathway to positively regulate age-triggered leaf senescence.
叶片衰老可以被各种环境胁迫、发育信号和内源激素信号所触发和促进。有几条证据表明,WRKY 转录因子参与调节叶片衰老,但涉及的潜在机制和信号通路仍不清楚。在这项研究中,我们鉴定出拟南芥 WRKY DNA 结合蛋白 45(WRKY45)是一个正调控年龄触发叶片衰老的因子。WRKY45 功能缺失导致年龄触发的衰老中叶片寿命延长,而 WRKY45 的过表达则显著加速年龄触发的叶片衰老。一致地,wrky45 突变体中 SENESCENCE-ASSOCIATED GENES(SAGs)的表达显著降低,但在过表达 WRKY45 的转基因植物中显著增强。染色质免疫沉淀分析显示,WRKY45 直接结合几个 SAGs 的启动子,如 SAG12、SAG13、SAG113 和 SEN4。体内和体外生化分析表明,WRKY45 与 DELLA 蛋白 RGA-LIKE1(RGL1)相互作用,RGL1 是赤霉素(GA)信号通路的抑制剂。我们发现 RGL1 抑制 WRKY45 的转录激活功能,从而减弱其调控基因的表达。与这一发现一致的是,RGL1 的过表达导致年龄触发的衰老中叶片寿命显著延长。总之,我们的结果提供了令人信服的证据,表明 WRKY45 作为 GA 介导的信号通路的关键组成部分,正向调节年龄触发的叶片衰老。