Abstract
The mechanism by which endogenous salicylic acid (SA) regulates leaf senescence remains elusive. Here we provide direct evidence that an enhancement of endogenous SA level, via chemical-induced upregulation of ISOCHORISMATE SYNTHASE 1 (ICS1), could significantly accelerate the senescence process of old leaves through mediation of the key SA signaling component NON EXPRESSOR OF PATHOGENESIS RELATED GENES 1 (NPR1) in Arabidopsis. Importantly, by taking advantage of this chemically induced leaf senescence system, we identified a mitogen-activated protein kinase (MAPK) cascade MKK4/5-MPK1/2 that is required for the SA/NPR1-mediated leaf senescence. Both MKK4/5 and MPK1/2 exhibited SA-induced kinase activities, with MPK1/2 being the immediate targets of MKK4/5. Double mutants of mkk4 mkk5 and mpk1 mpk2 displayed delayed leaf senescence, while constitutive overexpression of the kinase genes led to premature leaf senescence. Such premature leaf senescence was suppressed when they were overexpressed in an SA synthesis defective mutant (sid2) or signaling detective mutant (npr1). We further showed that MPK1, but not MPK2, could directly phosphorylate NPR1. Meanwhile, MPK1 also mediated NPR1 monomerization. Notably, induction of disease resistance was significantly compromised in the single and double mutants of the kinase genes. Taken together, our data demonstrate that the MKK4/5-MPK1/2 cascade plays a critical role in modulating SA signaling through a complex regulatory network in Arabidopsis.
Key message
We reveal a MAPK cascade, MKK4/5-MPK1, which not only phosphorylates the key SA signaling component NPR1 specifically, but also regulates its oligomer-to-monomer conformational change somehow via mediation of TRX-h3/5.
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Change history
13 July 2020
Due to an unfortunate turn of events, the second co-corresponding author, Dr. Benke Kuai, was omitted from the original publication. The corrected authors��� list and author contribution statement are published here and should be treated as definitive.
References
Asai T et al (2002) MAP kinase signalling cascade in Arabidopsis innate immunity. Nature 415:977–983
Bechtold N, Ellis J, Pelletier G (1993) In-planta agrobacterium-mediated gene-transfer by infiltration of adult Arabidopsis-thaliana plants. Cr. Acad. Sci. III-Vie 316:1194–1199
Benedetti CE, Arruda P (2002) Altering the expression of the chlorophyllase gene ATHCOR1 in transgenic Arabidopsis caused changes in the chlorophyll-to-chlorophyllide ratio. Plant Physiol 128:1255–1263
Cao H, Jane G, Joseph DC, Sigrid V, Dong XN (1997) The Arabidopsis NPR1 gene that controls systemic acquired resistance encodes a novel protein containing ankyrin repeats. Cell 88:57–63
Chai J, Liu J, Zhou J, Xing D (2014) Mitogen-activated protein kinase 6 regulates NPR1 gene expression and activation during leaf senescence induced by salicylic acid. J Exp Bot 65:6513–6528
Delaney TP et al (1994) A central role of salicylic acid in plant disease resistance. Science 266:1247–1250
Després C et al (2003) The Arabidopsis NPR1 disease resistance protein is a novel cofactor that confers redox regulation of DNA binding activity to the basic domain/leucine zipper transcription factor TGA1. Plant Cell 15:2181–2191
Eulgem T, Somssich IE (2007) Networks of WRKY transcription factors in defense signaling. Curr Opin Plant Biol 10:366–371
Gaffney T et al (1993) Requirement of salicylic acid for the induction of systemic acquired resistance. Science 261:754–756
Gan SS, Amasino RM (1997) Making sense of senescence (molecular genetic regulation and 584 manipulation of leaf senescence). Plant Physiol 113:313–319
Gao MH et al (2008) MEKK1, MKK1/MKK2 and MPK4 function together in a mitogen-activated protein kinase cascade to regulate innate immunity in plants. Cell Res 18:1190–1198
Hiroaki A et al (2015) WRKY transcription factors phosphorylated by MAPK regulate a plant immune NADPH oxidase in Nicotiana benthamiana. Plant Cell 27:2645–2663
Ishihama N, Yoshioka H (2012) Post-translational regulation of WRKY transcription factors in plant immunity. Curr Opin Plant Biol 15:431–437
Jibran R, Hunter DA, Dijkwel PP (2013) Hormonal regulation of leaf senescence through integration of developmental and stress signals. Plant Mol Biol 82:547–561
Kazuya I et al (2002) Mitogen-activated protein kinase cascades in plants: a new nomenclature. Trends Plant Sci 7:301–308
Kim SH et al (2011) Arabidopsis MKK4 mediates osmotic-stress response via its regulation of MPK3 activity. Biochem Biophys Res Commun 412:150–154
Kusaba M, Tanaka A, Tanaka R (2013) Stay-green plants: what do they tell us about the molecular mechanism of leaf senescence. Photosynth Res 117:221–234
Lee HJ et al (2015) Systemic immunity requires SnRK28-mediated nuclear import of NPR1 in Arabidopsis. Plant Cell 27:3425–3438
Lim PO, Kim HJ, Nam HG (2007) Leaf senescence. Annu Rev Plant Biol 58:115–136
Malamy J, Carr JP, Klessig DF, Raskin I (1990) Salicylic acid: a likely endogenous signal in the resistance response of tobacco to viral infection. Science 250:1002–1004
Mark K, Weihua F, Dong XN (2000) Nuclear localization of NPR1 is required for activation of PR gene expression. Plant Cell 12:2339–2350
Masclaux-Daubresse C, Chardon F (2011) Exploring nitrogen remobilization for seed filling using natural variation in Arabidopsis thaliana. J Exp Bot 62:2131–2142
Metraux JP et al (1990) Increase in salicylic acid at the onset of systemic acquired resistance in cucumber. Science 250:1004–1006
Miao Y, Thomas ML, Anja S, Ulrike Z (2007) Arabidopsis MEKK1 can take a short cut: it can directly interact with senescence-related WRKY53 transcription factor on the protein level and can bind to its promoter. Plant Mol Biol 65:63–76
Morris K et al (2000) Salicylic acid has a role in regulating gene expression during leaf senescence. Plant J 23:677–685
Mou Z, Fan WH, Dong XN (2003) Inducers of plant systemic acquired resistance regulate NPR1 function through redox changes. Cell 113:935–944
Nakagami H, Pitzschke A, Hirt H (2005) Emerging MAP kinase pathways in plant stress signalling. Trends Plant Sci 10:339–346
Ortiz-Masia D, Perez-Amador MA, Carbonell J, Marcote MJ (2007) Diverse stress signals activate the C1 subgroup MAP kinases of Arabidopsis. Febs Lett 581:1834–1840
Pajerowska-Mukhtar KM et al (2012) The HSF-like transcription factor TBF1 is a major molecular switch for plant growth-to-defense transition. Curr Biol 22:103–112
Park JH, Oh SA, Kim YH, Woo HR, Nam HG (1998) Differential expression of senescence-associated mRNAs during leaf senescence induced by different senescence-inducing factors in Arabidopsis. Plant Mol Biol 37:445–454
Peck SC (2006) Analysis of protein phosphorylation: methods and strategies for studying kinases and substrates. Plant J 45:512–522
Popescu SC et al (2009) MAPK target networks in Arabidopsis thaliana revealed using functional protein microarrays. Gene Dev 23:80–92
Raskin I (1992) Role of salicylic acid in plants. Annu Rev Plant Biol 43:439–463
Singh V, Roy S, Singh D, Nandi AK (2014) Arabidopsis FLOWERING LOCUS D influences systemic-acquired-resistance-induced expression and histone modifications of WRKY genes. J Bioscience 39:119–126
Spoel SH et al (2009) Proteasome-mediated turnover of the transcription coactivator NPR1 plays dual roles in regulating plant immunity. Cell 137:860–872
Tada Y et al (2008) Plant immunity requires conformational changes of NPR1 via S-nitrosylation and thioredoxins. Science 321:952–956
van der Graaff E et al (2006) Transcription analysis of Arabidopsis membrane transporters and hormone pathways during developmental and induced leaf senescence. Plant Physiol 141:776–792
Vlot AC, Dempsey DA, Klessig DF (2009) Salicylic acid, a multifaceted hormone to combat disease. Annu Rev Phytopathol 47:177–206
Wang D, Amornsiripanitch N, Dong XN (2006) A genomic approach to identify regulatory nodes in the transcriptional network of systemic acquired resistance in plants. Plos Pathog 2:e123
Wang H, Ngwenyama N, Liu Y, Walker JC, Zhang S (2007) Stomatal development and patterning are regulated by environmentally responsive mitogen-activated protein kinases in Arabidopsis. Plant Cell 19:63–73
Wang XY et al (2015) TCP transcription factors are critical for the coordinated regulation of ISOCHORISMATE SYNTHASE 1 expression in Arabidopsis thaliana. Plant J 82:151–162
Wildermuth MC, Dewdney J, Wu G, Ausubel FM (2001) Isochorismate synthase is required to synthesize salicylic acid for plant defence. Nature 414:562–565
Yoo SD, Cho YH, Sheen J (2007) Arabidopsis mesophyll protoplasts: a versitile cell system for transient gene expression analysis. Nat Protoc 2:1565–1572
Yoshioka K, Nakashita H, Klessig DF, Yamaguchi I (2001) Probenazole induces systemic acquired resistance in Arabidopsis with a novel type of action. Plant J 25:149–157
Yu J et al (2010) The pathway and regulation of salicylic acid biosynthesis in probenazole-treated Arabidopsis. J Plant Biol 53:417–424
Zhang KW, Halitschke R, Yin C, Liu CJ, Gan SS (2013) Salicylic acid 3-hydroxylase regulates Arabidopsis leaf longevity by mediating salicylic acid catabolism. Proc Natl Acad Sci USA 110:14807–14812
Zhao C et al (2014) EDR1 physically interacts with MKK4/5 and negatively regulates a MAP kinase cascade to modulate plant innate immunity. Plos Genet 10:e10043895
Zhou CJ, Cai ZH, Guo YF, Gan SS (2009) An Arabidopsis mitogen-activated protein kinase cascade, MKK9-MPK6, plays a role in leaf senescence. Plant Physiol 150:167–177
Acknowledgements
This work was supported by the National Natural Science Foundation of China (31700246 to J.G.) and Science and Technology Commission of Shanghai Municipality (15JC1400800).
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J.Z. and J.G. conceived and designed the experiments. J.Z., J.G., Z.Z. and Y.S. performed the experiments, X.W., X.W. and X.Z. contributed new materials, J.G. and J.Z. analyzed the data and wrote the paper. All authors read and improved the manuscript.
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Zhang, J., Gao, J., Zhu, Z. et al. MKK4/MKK5-MPK1/MPK2 cascade mediates SA-activated leaf senescence via phosphorylation of NPR1 in Arabidopsis. Plant Mol Biol 102, 463–475 (2020). https://doi.org/10.1007/s11103-019-00958-z
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DOI: https://doi.org/10.1007/s11103-019-00958-z