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Journal of Plant Growth Regulation

, Volume 33, Issue 1, pp 106–118 | Cite as

Senescence Networking: WRKY18 is an Upstream Regulator, a Downstream Target Gene, and a Protein Interaction Partner of WRKY53

  • Maren Potschin
  • Silke Schlienger
  • Stefan Bieker
  • Ulrike Zentgraf
Article

Abstract

Transcriptional reprogramming is a central feature of senescence regulation, implying an essential role for transcription factors. A regulatory function has already been attributed to different members of the plant-specific NAC and WRKY families in Arabidopsis but also in other plant species. WRKY53 is one important senescence regulator of the Arabidopsis WRKY family that is tightly regulated on different levels. In this study we show that WRKY18, which was formerly characterized as a downstream target of WRKY53 in the WRKY network, also regulates the expression of WRKY53. WRKY18 is able to bind directly to different W-boxes in the WRKY53 promoter region and to repress expression of a WRKY53 promoter-driven reporter gene in a transient transformation system using Arabidopsis protoplasts. Consistent with its repressing function on WRKY53 as a positive senescence regulator, WRKY18 overexpression led to delayed senescence, whereas wrky18 mutant plants exhibited a clearly accelerated senescence. In addition, a direct interaction between WRKY53 and WRKY18 proteins could be detected in yeast using the split ubiquitin system and in planta in transiently transformed tobacco epidermal cells via FRET-FLIM. In contrast to WRKY18/18 homodimers, WRKY18/53 heterodimers positively influenced WRKY53 promoter-driven reporter gene expression but appear to act only on a shorter 1.1 kbp promoter fragment but not on a 2.8 kbp longer fragment, indicating a more complex protein-protein-DNA interaction on the longer WRKY53 promoter, most likely also triggered by the accessibility of the promoter on the chromatin level.

Keywords

WRKY transcription factors Senescence DPI-ELISA Feedback regulation Yeast split ubiquitin FRET-FLIM 

Notes

Acknowledgments

We are grateful for the excellent technical assistance of Gabriele Eggers-Schumacher. We thank the NASC for supplying seeds of the WRKY18 T-DNA insertion lines. This work was financially supported by the DFG (ZE 313, 9-1).

Supplementary material

344_2013_9380_MOESM1_ESM.doc (8.8 mb)
Supplementary material 1 (DOC 9057 kb)

References

  1. Ay N, Irmler K, Fischer A, Uhlemann R, Reuter G, Humbeck K (2009) Epigenetic programming via histone methylation at WRKY53 controls leaf senescence in Arabidopsis thaliana. Plant J 58(2):333–346CrossRefPubMedGoogle Scholar
  2. Balazadeh S, Siddiqui H, Allu AD, Matallana-Ramirez LP, Caldana C, Mehrnia M, Zanor MI, Köhler B, Mueller-Roeber B (2010) A gene regulatory network controlled by the NAC transcription factor ANAC092/AtNAC2/ORE1 during salt-promoted senescence. Plant J 62(2):250–264CrossRefPubMedGoogle Scholar
  3. Balazadeh S, Kwasniewski M, Caldana C, Mehrnia M, Zanor MI, Xue GP, Mueller-Roeber B (2011) ORS1, an H2O2-responsive NAC transcription factor, controls senescence in Arabidopsis thaliana. Mol Plant 4(2):346–360PubMedCentralCrossRefPubMedGoogle Scholar
  4. Besseau S, Li J, Palva ET (2012) WRKY54 and WRKY70 co-operate as negative regulators of leaf senescence in Arabidopsis thaliana. J Exp Bot 63:2667–2679PubMedCentralCrossRefPubMedGoogle Scholar
  5. Bieker S, Riester L, Stahl M, Franzaring J, Zentgraf U (2012) Senescence-specific alteration of hydrogen peroxide levels in Arabidopsis thaliana and oilseed rape spring variety Brassica napus L. cv. Mozart. J Integr Plant Biol 54(8):540–554CrossRefPubMedGoogle Scholar
  6. Brand LH, Kirchler T, Hummel S, Chaban C, Wanke D (2010) DPI-ELISA: a fast and versatile method to specify the binding of plant transcription factors to DNA in vitro. Plant Methods 25:6–25Google Scholar
  7. Breeze E, Harrison E, McHattie S, Hughes L, Hickman R, Hill C, Kiddle S, Kim YS, Penfold CA, Jenkins D, Zhang C, Morris K, Jenner C, Jackson S, Thomas B, Tabrett A, Legaie R, Moore JD, Wild DL, Ott S, Rand D, Beynon J, Denby K, Mead A, Buchanan-Wollaston V (2011) High-resolution temporal profiling of transcripts during Arabidopsis leaf senescence reveals a distinct chronology of processes and regulation. Plant Cell 23(3):873–894PubMedCentralCrossRefPubMedGoogle Scholar
  8. Brusslan JA, Rus Alvarez-Canterbury AM, Nair NU, Rice JC, Hitchler MJ, Pellegrini M (2012) Genome-wide evaluation of histone methylation changes associated with leaf senescence in Arabidopsis. PLoS ONE 7(3):e33151PubMedCentralCrossRefPubMedGoogle Scholar
  9. Buchanan-Wollaston V, Page T, Harrison E, Breeze E, Lim PO, Nam HG, Lin JF, Wu SH, Swidzinski J, Ishizaki K, Leaver CJ (2005) Comparative transcriptome analysis reveals significant differences in gene expression and signalling pathways between developmental and dark/starvation-induced senescence in Arabidopsis. Plant J 42(4):567–585CrossRefPubMedGoogle Scholar
  10. Chen C, Chen Z (2002) Potentiation of developmentally regulated plant defense response by AtWRKY18, a pathogen-induced Arabidopsis transcription factor. Plant Physiol 129(2):706–716PubMedCentralCrossRefPubMedGoogle Scholar
  11. Chen H, Lai Z, Shi J, Xiao Y, Chen Z, Xu X (2010) Roles of Arabidopsis WRKY18, WRKY40 and WRKY60 transcription factors in plant responses to abscisic acid and abiotic stress. BMC Plant Biol 10:281PubMedCentralCrossRefPubMedGoogle Scholar
  12. Chi Y, Yang Y, Zhou Y, Zhou J, Fan B, Yu JQ, Chen Z (2013) Protein-protein interactions in the regulation of WRKY transcription factors. Mol Plant 6(2):287–300CrossRefPubMedGoogle Scholar
  13. Eulgem T, Rushton PJ, Robatzek S, Somssich IE (2000) The WRKY superfamily of plant transcription factors. Trends Plant Sci 5:199–206CrossRefPubMedGoogle Scholar
  14. Gregersen PL, Culetic A, Boschian L, Krupinska K (2013) Plant senescence and crop productivity. Plant Mol Biol 82(6):603–622CrossRefPubMedGoogle Scholar
  15. Guo Y, Cai Z, Gan S (2004) Transcriptome of Arabidopsis leaf senescence. Plant Cell Environ 27(5):521–549CrossRefGoogle Scholar
  16. Hinderhofer K, Zentgraf U (2001) Identification of a transcription factor specifically expressed at the onset of leaf senescence. Planta 213(3):469–473CrossRefPubMedGoogle Scholar
  17. Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907PubMedCentralPubMedGoogle Scholar
  18. Miao Y, Zentgraf U (2007) The antagonist function of Arabidopsis WRKY53 and ESR/ESP in leaf senescence is modulated by the jasmonic and salicylic acid equilibrium. Plant Cell 19(3):819–830PubMedCentralCrossRefPubMedGoogle Scholar
  19. Miao Y, Zentgraf U (2010) A HECT E3 ubiquitin ligase negatively regulates Arabidopsis leaf senescence through degradation of the transcription factor WRKY53. Plant J 63(2):179–188CrossRefPubMedGoogle Scholar
  20. Miao Y, Laun T, Zimmermann P, Zentgraf U (2004) Targets of the WRKY53 transcription factor and its role during leaf senescence in Arabidopsis. Plant Mol Biol 55(6):853–867CrossRefPubMedGoogle Scholar
  21. Miao Y, Laun TM, Smykowski A, Zentgraf U (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(1–2):63–76CrossRefPubMedGoogle Scholar
  22. Miao Y, Smykowski A, Zentgraf U (2008) A novel upstream regulator of WRKY53 transcription during leaf senescence in Arabidopsis thaliana. Plant Biol 10(Suppl 1):110–120CrossRefPubMedGoogle Scholar
  23. Negrutiu I, Shillito RD, Potrykus I, Biasini G, Sala F (1987) Hybrid genes in the analysis of transformation conditions I. Setting up a simple method for direct gene transfer in plant protoplasts. Plant Mol Biol 8:363–373CrossRefPubMedGoogle Scholar
  24. Panchuk II, Zentgraf U, Volkov RA (2005) Expression of the APX gene family during leaf senescence of Arabidopsis thaliana. Planta 222:926–932CrossRefPubMedGoogle Scholar
  25. Pandey SP, Roccaro M, Schön M, Logemann E, Somssich IE (2010) Transcriptional reprogramming regulated by WRKY18 and WRKY40 facilitates powdery mildew infection of Arabidopsis. Plant J 64(6):912–923CrossRefPubMedGoogle Scholar
  26. Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29(9):e45PubMedCentralCrossRefPubMedGoogle Scholar
  27. Robatzek S, Somssich IE (2002) Targets of AtWRKY6 regulation during plant senescence and pathogen defense. Genes Dev 16:1139–1149PubMedCentralCrossRefPubMedGoogle Scholar
  28. Rushton PJ, Somssich IE, Ringler P, Shen QJ (2010) WRKY transcription factors. Trends Plant Sci 15:247–258CrossRefPubMedGoogle Scholar
  29. Uauy C, Distelfeld A, Fahima T, Blechl A, Dubcovsky J (2006) A NAC gene regulating senescence improves grain protein, zinc, and iron content in wheat. Science 314(5803):1298–1301CrossRefPubMedGoogle Scholar
  30. Ulker B, Shahid Mukhtar M, Somssich IE (2007) The WRKY70 transcription factor of Arabidopsis influences both the plant senescence and defense signaling pathways. Planta 226:125–137CrossRefPubMedGoogle Scholar
  31. Wang D, Amornsiripanitch N, Dong X (2006) A genomic approach to identify regulatory nodes in the transcriptional network of systemic acquired resistance in plants. PLoS Pathog 2(11):e123PubMedCentralCrossRefPubMedGoogle Scholar
  32. Welner DH, Lindemose S, Grossmann JG, Møllegaard NE, Olsen AN, Helgstrand C, Skriver K, Lo Leggio L (2012) DNA binding by the plant-specific NAC transcription factors in crystal and solution: a firm link to WRKY and GCM transcription factors. Biochem J 444(3):395–404CrossRefPubMedGoogle Scholar
  33. Wenke K, Wanke D, Kilian J, Berendzen K, Harter K, Piechulla B (2012) Volatiles of two growth-inhibiting rhizobacteria commonly engage AtWRKY18 function. Plant J 70(3):445–459CrossRefPubMedGoogle Scholar
  34. Wenkel S, Turck F, Singer K, Gissot L, Le Gourrierec J, Samach A, Coupland G (2006) CONSTANS and the CCAAT box binding complex share a functionally important domain and interact to regulate flowering of Arabidopsis. Plant Cell 18(11):2971–2984PubMedCentralCrossRefPubMedGoogle Scholar
  35. Wu A, Allu AD, Garapati P, Siddiqui H, Dortay H, Zanor MI, Asensi-Fabado MA, Munné-Bosch S, Antonio C, Tohge T, Fernie AR, Kaufmann K, Xue GP, Mueller-Roeber B, Balazadeh S (2012) JUNGBRUNNEN1, a reactive oxygen species-responsive NAC transcription factor, regulates longevity in Arabidopsis. Plant Cell 24(2):482–506PubMedCentralCrossRefPubMedGoogle Scholar
  36. Xu X, Chen C, Fan B, Chen Z (2006) Physical and functional interactions between pathogen-induced Arabidopsis WRKY18, WRKY40, and WRKY60 transcription factors. Plant Cell 18:1310–1326PubMedCentralCrossRefPubMedGoogle Scholar
  37. Zentgraf U, Laun T, Miao Y (2010) The complex regulation of WRKY53 during leaf senescence of Arabidopsis thaliana. Eur J Cell Biol 89(2–3):133–137CrossRefPubMedGoogle Scholar
  38. Zhou X, Jiang Y, Yu D (2011) WRKY22 transcription factor mediates dark-induced leaf senescence in Arabidopsis. Mol Cells 31(4):303–313PubMedCentralCrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Maren Potschin
    • 1
  • Silke Schlienger
    • 1
  • Stefan Bieker
    • 1
  • Ulrike Zentgraf
    • 1
  1. 1.Center for Plant Molecular BiologyUniversity of TuebingenTuebingenGermany

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