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Role of an Arabidopsis mitogen-activated protein kinase kinase kinase in ROS-mediated leaf senescence

Fengbo Yang1†, Kun Li1,2†, José Ramón Botella3, Xiaohong Zhu1, Yuchen Miao1,2, Yun Zhou1, Yuan Li4, Dongtao Ren4 and Chun‐Peng Song1,2*   

  1. 1. State Key Laboratory of Crop Stress Adaptation and Improvement, National Key Laboratory of Cotton Bio‐breeding and Integrated Utilization, Henan Joint International Laboratory for Crop Multi‐Omics Research, School of Life Sciences, Henan University, Kaifeng 475004, China

    2. Sanya Institute of Henan University, Sanya 572025, China

    3. Plant Genetic Engineering Laboratory, School of Agriculture and Food Sciences, The University of Queensland, Brisbane 4072, Australia

    4. State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing 100193, China

    †These authors contributed equally to this work.

    *Correspondence: Chun‐Peng Song (songcp@henu.edu.cn)

  • Received:2025-06-24 Accepted:2026-01-22 Online:2026-02-24
  • Supported by:
    This work was supported by the National Natural Science Foundation of China (Projects 32300308, U21A20206, and 32230079), Postdoctoral start‐up Funding of Henan University (CJ3050A0670659), and the Key Scienti?c Research Projects of the Higher Education Institutions of Henan Province (24A180007 and 24A180004).

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Abstract: Mitogen-activated protein kinase (MAPK) cascades regulate growth, development, stress responses, and immunity in plants by transmitting signals from upstream regulators to downstream components. In this study, we identify a MAPK cascade composed of MAPK kinase kinase 19 (MKKK19), three MAPK kinases (MKK3/MKK5/MKK9), and MAPK 6 (or MPK6) that is involved in Arabidopsis leaf senescence. The kinase oxidative signal-inducible 1 (OXI1) functions upstream of the MKKK19–MKK3/MKK5/MKK9–MPK6 cascade to promote leaf senescence, whereas two of RESPIRATORY BURST OXIDASE HOMOLOGS (RBOHs), RBOHD and RBOHF, act downstream to mediate the accumulation of reactive oxygen species (ROS). Loss-of-function mutation of OXI1, MKKK19, MKK3/MKK5/MKK9, or MPK6 resulted in delayed leaf senescence associated with reduced ROS levels, whereas transgenic lines overexpressing OXI1, MKKK19, MKK3/MKK5/MKK9, or MPK6 displayed the opposite phenotypes. Epistatic analyses supported the involvement of OXI1, MKKK19, MKK3/MKK5/MKK9, MPK6, and RBOHD/RBOHF in the same signaling pathway for leaf senescence. In conclusion, genetic and biochemical analysis of the Arabidopsis MKKK19–MKK3/MKK5/MKK9–MPK6 cascade through OXI1 and RBOHD/RBOHF revealed a vital role for the MAPK cascade and ROS in natural leaf senescence.

Key words: Arabidopsis thaliana, leaf senescence, MAPK cascade signal, OXI1, phosphorylation, ROS

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