Abstract
The 14-3-3 proteins, a highly conserved class in all eukaryotes, are widely associated with plant growth and stress responses. However, their role in plant immunity and its regulatory mechanisms remains elusive. Here, we show that two homologous rice 14-3-3 proteins, OsGF14f and OsGF14c, function redundantly to enhance rice resistance against Magnaporthe oryzae. The E3 ligase OsPUB20 targets OsGF14f and OsGF14c for ubiquitination and 26S proteasome-mediated degradation, thereby negatively regulating rice immunity. Remarkably, chitin perception activates the receptor-like cytoplasmic kinase OsRLCK185 that phosphorylates OsPUB20 at Thr153, which stabilizes OsGF14f and enhances rice blast resistance. Furthermore, during M. oryzae infection, OsGF14f translocates into the nucleus, where it facilitates the degradation of OsWRKY42, a transcription factor that negatively regulates defense responses. Collectively, our findings reveal a phosphorylation-dependent ubiquitination switch that links cell surface chitin perception to nuclear immune reprogramming during M. oryzae invasion.
Data availability
The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium (https://proteomecentral.proteomexchange.org) via the iProX partner repository with the dataset identifier PXD073757 and PXD073756. All data supporting the findings of this work are available in the paper, Supplementary Information files, and repository platform. Source data are provided with this paper.
References
Couto, D. & Zipfel, C. Regulation of pattern recognition receptor signalling in plants. Nat. Rev. Immunol. 16, 537–552 (2016).
Wang, R. et al. Occurrence and integrated control of major rice diseases in China. N. Plant Prot. 2, e70004 (2025).
Ngou, B. P. M., Ahn, H.-K., Ding, P. & Jones, J. D. G. Mutual potentiation of plant immunity by cell-surface and intracellular receptors. Nature 592, 110–115 (2021).
Liang, X. & Zhou, J. M. Receptor-like cytoplasmic kinases: central players in plant receptor kinase-mediated signaling. Annu. Rev. Plant Biol. 69, 267–299 (2018).
Vij, S., Giri, J., Dansana, P. K., Kapoor, S. & Tyagi, A. K. The Receptor-like cytoplasmic kinase (OsRLCK) gene family in rice-organization, phylogenetic relationship, and expression during development and stress. Mol. Plant 1, 732–750 (2008).
Bai, J. et al. BIK1 protein homeostasis is maintained by the interplay of different ubiquitin ligases in immune signaling. Nat. Commun. 14, 4624 (2023).
Li, L. et al. The FLS2-associated kinase BIK1 directly phosphorylates the NADPH oxidase RbohD to control plant immunity. Cell Host Microbe 15, 329–338 (2014).
Wang, J. et al. A cyclic nucleotide-gated channel mediates cytoplasmic calcium elevation and disease resistance in rice. Cell Res. 29, 820–831 (2019).
Yamaguchi, K. et al. A receptor-like cytoplasmic kinase targeted by a plant pathogen effector is directly phosphorylated by the chitin receptor and mediates rice immunity. Cell Host Microbe 13, 347–357 (2013).
Shinya, T. et al. Selective regulation of the chitin-induced defense response by the Arabidopsis receptor-like cytoplasmic kinase PBL27. Plant J. 79, 56–66 (2014).
Wang, C. et al. OsCERK1-mediated chitin perception and immune signaling requires receptor-like cytoplasmic kinase 185 to activate an MAPK cascade in rice. Mol. Plant 10, 619–633 (2017).
Yamada, K. et al. The Arabidopsis CERK1-associated kinase PBL27 connects chitin perception to MAPK activation. EMBO J. 35, 2468–2483 (2016).
Bigeard, J., Colcombet, J. & Hirt, H. Signaling mechanisms in pattern-triggered immunity (PTI). Mol. Plant 8, 521–539 (2015).
Oh, C.-S. Characteristics of 14-3-3 proteins and their role in plant immunity. Plant Pathol. J. 26, 1–7 (2010).
Ormancey, M., Thuleau, P., Mazars, C. & Cotelle, V. CDPKs and 14-3-3 proteins: emerging duo in signaling. Trends Plant Sci. 22, 263–272 (2017).
Qi, H. et al. 14-3-3 proteins contribute to autophagy by modulating SINAT-mediated degradation of ATG13. Plant Cell 34, 4857–4876 (2022).
Fan, B. et al. Calcium-dependent activation of CPK12 facilitates its cytoplasm-to-nucleus translocation to potentiate plant hypoxia sensing by phosphorylating ERF-VII transcription factors. Mol. Plant 16, 979–998 (2023).
Guo, H. et al. Plastid-nucleus communication involves calcium-modulated MAPK signalling. Nat. Commun. 7, 12173 (2016).
Dong, X. et al. 14-3-3 proteins facilitate the activation of MAP kinase cascades by upstream immunity-related kinases. Plant Cell 35, 2413–2428 (2023).
Yang, X. et al. Arabidopsis 14-3-3 lambda is a positive regulator of RPW8-mediated disease resistance. Plant J. 60, 539–550 (2009).
Elmayan, T. et al. Regulation of reactive oxygen species production by a 14-3-3 protein in elicited tobacco cells. Plant Cell Environ. 30, 722–732 (2007).
Konagaya, K. -i, Matsushita, Y., Kasahara, M. & Nyunoya, H. Members of 14-3-3 protein isoforms interacting with the resistance gene product N and the elicitor of Tobacco mosaic virus. J. Gen. Plant Pathol. 70, 221–231 (2004).
Yan, S. et al. OsGF14b modulates defense signaling pathways in rice panicle blast response. Crop J. 9, 725–738 (2021).
Manosalva, P. M., Bruce, M. & Leach, J. E. Rice 14-3-3 protein (GF14e) negatively affects cell death and disease resistance. Plant J. 68, 777–787 (2011).
Ma, Y. et al. Overexpression of OsGF14f enhances quantitative leaf blast and bacterial blight resistance in rice. Int J. Mol. Sci. 23, 7440 (2022).
Lu, L. et al. The 14-3-3 protein GF14c positively regulates immunity by modulating the protein homoeostasis of the GRAS protein OsSCL7 in rice. Plant Cell Environ. 45, 1065–1081 (2022).
Duplan, V. & Rivas, S. E3 ubiquitin-ligases and their target proteins during the regulation of plant innate immunity. Front Plant Sci. 5, 42 (2014).
Ma, X. et al. Ubiquitylome analysis reveals a central role for the ubiquitin-proteasome system in plant innate immunity. Plant Physiol. 185, 1943–1965 (2021).
Wang, Y. et al. Ubiquitination in the rice blast fungus Magnaporthe oryzae: from development and pathogenicity to stress responses. Phytopathol. Res. 4, 1–10 (2022).
Vierstra, R. D. The ubiquitin-26S proteasome system at the nexus of plant biology. Nat. Rev. Mol. Cell Biol. 10, 385–397 (2009).
Zheng, N. & Shabek, N. Ubiquitin ligases: structure, function, and regulation. Annu. Rev. Biochem. 86, 129–157 (2017).
Yan, Y., Wang, H., Bi, Y. & Song, F. Rice E3 ubiquitin ligases: from key modulators of host immunity to potential breeding applications. Plant Commun. 5, 101128 (2024).
Wang, R. et al. An ORFeome of rice E3 ubiquitin ligases for global analysis of the ubiquitination interactome. Genome Biol. 23, 154 (2022).
Xu, X. et al. A pair of E3 ubiquitin ligases control immunity and flowering by targeting different ELF3 proteins in rice. Dev. Cell 59, 2731–2744.e4 (2024).
Sato, T. et al. Identification of 14-3-3 proteins as a target of ATL31 ubiquitin ligase, a regulator of the C/N response in Arabidopsis. Plant J. 68, 137–146 (2011).
Yasuda, S. et al. Phosphorylation of Arabidopsis ubiquitin ligase ATL31 is critical for plant carbon/nitrogen nutrient balance response and controls the stability of 14-3-3 proteins. J. Biol. Chem. 289, 15179–15193 (2014).
Xu, G. et al. FERONIA phosphorylates E3 ubiquitin ligase ATL6 to modulate the stability of 14-3-3 proteins in response to the carbon/nitrogen ratio. J. Exp. Bot. 70, 6375–6388 (2019).
Cui, L. H. et al. OsATL38 mediates mono-ubiquitination of the 14-3-3 protein OsGF14d and negatively regulates the cold stress response in rice. J. Exp. Bot. 73, 307–323 (2022).
Chen, F., Li, Q., Sun, L. & He, Z. The rice 14-3-3 gene family and its involvement in responses to biotic and abiotic stress. DNA Res. 13, 53–63 (2006).
Qin, M. et al. Transcriptome analysis of oserf922 mutants reveals new insights into rice blast resistance. Rice Sci. 30, 374–378 (2023).
Chen, X.-L. et al. Proteomic analysis of ubiquitinated proteins in rice (Oryza sativa) after treatment with pathogen-associated molecular pattern (PAMP) elicitors. Front Plant Sci. 9, 1064 (2018).
Zeng, L. R., Park, C. H., Venu, R. C., Gough, J. & Wang, G. L. Classification, expression pattern, and E3 ligase activity assay of rice U-box-containing proteins. Mol. Plant 1, 800–815 (2008).
Cheng, H. et al. The WRKY45-2 WRKY13 WRKY42 transcriptional regulatory cascade is required for rice resistance to fungal pathogen. Plant Physiol. 167, 1087–1099 (2015).
Lozano-Durán, R. & Robatzek, S. 14-3-3 Proteins in plant-pathogen interactions. Mol. Plant Microbe Interact. 28, 511–518 (2015).
Fuglsang, A. T. et al. Arabidopsis protein kinase PKS5 inhibits the plasma membrane H+-ATPase by preventing interaction with 14-3-3 protein. Plant Cell 19, 1617–1634 (2007).
Yang, Z. et al. Calcium-activated 14-3-3 proteins as a molecular switch in salt stress tolerance. Nat. Commun. 10, 1199 (2019).
Shiu, S.-H. et al. Comparative analysis of the receptor-like kinase family in arabidopsis and rice. Plant Cell 16, 1220–1234 (2004).
Fan, J. et al. The monocot-specific receptor-like kinase SDS2 controls cell death and immunity in rice. Cell Host Microbe 23, 498–510 (2018).
Wang, J. et al. The kinase OsCPK4 regulates a buffering mechanism that fine-tunes innate immunity. Plant Physiol. 176, 1835–1849 (2018).
Chen, L. et al. Direct attenuation of Arabidopsis ERECTA signalling by a pair of U-box E3 ligases. Nat. Plants 9, 112–127 (2022).
Wang, X. et al. PUB25 and PUB26 promote plant freezing tolerance by degrading the cold signaling negative regulator MYB15. Dev. Cell 51, 222–235.e5 (2019).
Lu, D. et al. Direct ubiquitination of pattern recognition receptor FLS2 attenuates plant innate immunity. Science 332, 1439–1442 (2011).
Wang, J. et al. A regulatory module controlling homeostasis of a plant immune kinase. Mol. Cell 69, 493–504 (2018).
Taoka, K. -i et al. 14-3-3 proteins act as intracellular receptors for rice Hd3a florigen. Nature 476, 332–335 (2011).
Liu, Z. et al. Plasma membrane CRPK1-mediated phosphorylation of 14-3-3 proteins induces their nuclear import to fine-tune cbf signaling during cold response. Mol. Cell 66, 117–128.e5 (2017).
Huang, X. et al. Shade-induced nuclear localization of PIF7 is regulated by phosphorylation and 14-3-3 proteins in Arabidopsis. eLife 7, e31636 (2018).
Wang, H. et al. Dual role of BKI1 and 14-3-3 s in brassinosteroid signaling to link receptor with transcription factors. Dev. Cell 21, 825–834 (2011).
Bai, M.-Y. et al. Functions of OsBZR1 and 14-3-3 proteins in brassinosteroid signaling in rice. Proc. Natl. Acad. Sci. 104, 13839–13844 (2007).
Ma, Y. et al. The 14-3-3 protein OsGF14f interacts with OsbZIP23 and enhances its activity to confer osmotic stress tolerance in rice. Plant Cell 35, 4173–4189 (2023).
He, F. et al. OsATG1 and OsATG8 exhibit autophagy-independent functions to oppositely regulate ROP GTPase-mediated plant immunity in rice. Mol. Plant 18, 1472–1489 (2025).
Zhao, Y. et al. A viral protein orchestrates rice ethylene signaling to coordinate viral infection and insect vector-mediated transmission. Mol. Plant 15, 689–705 (2022).
Tao, H. et al. Phosphorylation and ubiquitination synergistically promote the degradation of OsRbohB to modulate rice immunity. Plant Cell 37, koaf276 (2025).
Acknowledgements
The authors thank Dr. Dongping Lv from Shanghai Jiao Tong University and Dr. Xiangxiu Liang from South China Agricultural University for their insightful comments on the manuscript. This project was supported by grants from the National Natural Science Foundation of China (32161143009 and 32272505), the National Key Research and Development Program of China (2022YFD1401400), and the Innovation Program of Chinese Academy of Agricultural Sciences (CAAS-CSCB-202301) to Y.N., the National Natural Science Foundation of China (U24A20388) to R.W., the National Natural Science Foundation of China (32402396), and the Basic Scientific Research Operating Expenses of Central Universities (FRF-TP-25-087) to C.Z., the National Natural Science Foundation of China (32402476) to Z.H.
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C.Z. and Y.N. designed the experiments. C.Z., P.S., R.W., F.H., H.T., D.W., J.W., L.F., Z.H., X.Y., and W. L. conducted the experiments. C.Z., R.W., G.-L.W., and Y.N. wrote the manuscript.
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Zhang, C., Suttiviriya, P., Wang, R. et al. A phosphorylation-dependent ubiquitination switch orchestrates nuclear immune reprogramming upon chitin perception. Nat Commun (2026). https://doi.org/10.1038/s41467-026-69627-2
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DOI: https://doi.org/10.1038/s41467-026-69627-2
