叶片衰老启动具有高度时序性，受内外环境影响，并整合多基因精细调控。其中，拟南芥转录因子WRKY53发挥重要作用，但WRKY53具有的自身负反馈抑制受何种直接动态调控从而精准协调叶片衰老启动，仍亟待解析。VQ家族是WRKYs潜在调节者，但叶片衰老相关的VQs尚不明确。本研究前期发现，拟南芥VQ25加速叶片衰老发生，且VQ25在未衰老叶片中即可促进WRKY53表达。同时，VQ25可能不依赖其VQ基序与WRKY53发生互作，并影响WRKY53结合自身启动子，且该过程可能涉及MAPKs途径。在此基础上，本研究将明确VQ25与WRKY53关键互作位点；阐明叶片衰老启动中VQ25影响WRKY53负反馈的具体机制；分析VQ25上游衰老相关MAPKs及二者磷酸化关系；揭示磷酸化修饰对于VQ25调控WRKY53的影响。最终解析VQ25-WRKY53精细调控叶片衰老适时发生的分子机制，具有重要意义和创新性。

The precise initiation timing of leaf senescence is a highly ordered and sophisticated process. It integrates many environmental signals into developmental age-dependent senescence, and is finely tuned by multiple gene families under multi-level. Transcriptional factor AtWRKY53 has been regarded as a key node of many leaf senescence inducible signaling pathways. It has been demonstrated that AtWRKY53 possesses a negative feed-back auto-regulaiton. However, the functional role of this auto-regulation in onset of leaf senescence is unclear and needs to be uncover. VQs are potential regulators of WRKYs, but senescence-related VQs are still unknown. Our previous data manifested that a VQ family member，AtVQ25, promotes leaf senescence，and AtVQ25 elevates AtWRKY53 transcription level even at juvenile stage . Moreover, AtVQ25 interacts with AtWRKY53 independent on VQ motif，which is potentially involved in regulation of the negative feed-back module of AtWRKY53 and related with MAPKs. In this study, we will confirm the interaction domains between AtVQ25 and AtWRKY53 during leaf senescence onset，and how AtVQ25 influences the negative feed-back loop of AtWRKY53. Meanwhile，we attempt to screen senescence-associated upper stream MAPKs of AtVQ25 and analyze the phosphorylation between candidate MAPKs and AtVQ25，and investigate the impact of AtVQ25 phosphorylation level on regulation of WRKY53. Ultimately, with significantly importance and innovation, the underlying molecular mechanism of the VQ25-WRKY53 regulatory module in fine-tuning the initiation timing of leaf senescence will be unpuzzled.