Abstract
Jasmonates (JAs) are a class of phytohormones that play a crucial role in plant growth, development, and environmental stress responses. Central to JA signaling are the MYC2-type transcription factors, as they activate the expression of JA-responsive genes. We previously used CRISPR-Cas9-based genome editing to engineer rice OsMYC2 and yielded a mutant (myc2-5) with a single amino acid (aa) deletion (75I) outside the known functional domains of the protein. This myc2-5 mutant also showed some JA-deficient phenotypes, promoting us to investigate how 75I deletion affects JA responses. The mutation is found in the α2 helix element at the N-terminal of OsMYC2. The deletion of 75I in OsMYC2 rendered plants deficient in most of the JA responses, including root growth, leaf senescence, spikelet development, and resistance to pathogens and herbivores. Biochemical assays revealed that the 75I deletion markedly reduced OsMYC2 protein accumulation, subsequently diminishing its transcriptional activity. However, the deletion did not influence the protein's subcellular localization, DNA-binding capability, or its interactions with JAZ transcriptional repressors and the Mediator complex subunit MED25. Additionally, the screening of seven other deletions in the α2 helix further reinforces the importance of this protein element. Our results highlight the significance of the α2 helix in the N-terminus for OsMYC2's functionality, primarily through modulating its protein levels. This insight expands our knowledge of JA signaling and opens new avenues for research into the yet-to-be-explored domains of the MYC2 protein, with the potential to tailor JA responses in rice and other plant species.
Key message
MYC2 transcription factor is the master regulator of plant jasmonate signaling. The functional domains or elements of MYC2 protein determine its activity. Here, we identified a novel element in the N-terminus that is essential for the activity of rice MYC2.
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Acknowledgements
We thank Jing Lv, Leilei Li, Xinjue Wang, Jie Xu and Xinhui Gan for technical assistance, and Jane Marczewski, Lucas Cortés Llorca for editorial assistance.
Funding
This work was supported by the State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products (Grant No. 2021DG700024-KF202324, to R.L.), a Grant from Max Planck Partner Group Program (to R.L.), and the Hundred-Talent Program of Zhejiang University (to R.L.).
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RL designed the research. HZ, GJ, YK, ZL and RL performed experiments. HZ, RL, and YL analyzed data. RL and HZ wrote the manuscript.
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Zu, H., Jin, G., Kong, Y. et al. The N-terminal α2 helix element is critical for the activity of the rice transcription factor MYC2. Plant Mol Biol 114, 2 (2024). https://doi.org/10.1007/s11103-023-01411-y
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DOI: https://doi.org/10.1007/s11103-023-01411-y