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Arabidopsis thaliana WRKY25, WRKY26, and WRKY33 coordinate induction of plant thermotolerance

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Abstract

Limited information is available regarding the exact function of specific WRKY transcription factors in plant responses to heat stress. We analyzed the roles of WRKY25, WRKY26, and WRKY33, three types of group I WRKY proteins, in the regulation of resistance to heat stress. Expression of WRKY25 and WRKY26 was induced upon treatment with high temperature, whereas WRKY33 expression was repressed. Heat-treated WRKY single mutants exhibited small responses, while wrky25wrky26 and wrky25wrky33 double mutants and the wrky25wrky26wrky33 triple mutants showed substantially increased susceptibility to heat stress, showing reduced germination, decreased survival, and elevated electrolyte leakage, compared with wild-type plants. In contrast, constitutive expression of WRKY25, WRKY26, or WRKY33 enhanced resistance to heat stress. Expression studies of selected heat-defense genes in single, double, and triple mutants, as well as in over-expressing lines, were correlated with their thermotolerance phenotypes and demonstrated that the three WRKY transcription factors modulate transcriptional changes of heat-inducible genes in response to heat treatment. In addition, our findings provided evidence that WRKY25, WRKY26, and WRKY33 were involved in regulation of the heat-induced ethylene-dependent response and demonstrated positive cross-regulation within these three genes. Together, these results indicate that WRKY25, WRKY26, and WRKY33 positively regulate the cooperation between the ethylene-activated and heat shock proteins-related signaling pathways that mediate responses to heat stress; and that these three proteins interact functionally and play overlapping and synergetic roles in plant thermotolerance.

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Abbreviations

ABA:

Abscisic acid

ACC:

1-Aminocyclopropane-1-carboxylic acid

APX:

Cytosolic ascorbate peroxidase

EL:

Electrolyte leakage

HS:

Heat shock

Hsf:

Heat shock transcription factor

Hsp:

Heat shock protein

MBF1c:

Coactivator multiprotein bridging factor 1c

SA:

Salicylic acid

References

  • Alonso JM, Stepanova AN, Leisse TJ, Kim CJ, Chen HM, Shinn P, Stevenson DK, Zimmerman J, Barajas P, Cheuk R et al (2003) Genome-wide insertional mutagenesis of Arabidopsis thaliana. Science 301:653–657

    Article  PubMed  Google Scholar 

  • Andreasson E, Jenkins T, Brodersen P, Thorgrimsen S, Petersen NHT, Zhu S, Qiu JL, Micheelsen P, Rocher A, Petersen M (2005) The MAP kinase substrate MKS1 is a regulator of plant defense responses. EMBO J 24:2579–2589

    Article  PubMed  CAS  Google Scholar 

  • Asai T, Tena G, Plotnikova J, Willmann MR, Chiu WL, Gomez-Gomez L, Boller T, Ausubel FM, Sheen J (2002) MAP kinase signalling cascade in Arabidopsis innate immunity. Nature 415:977–983

    Article  PubMed  CAS  Google Scholar 

  • Baniwal SK, Bharti K, Chan KY, Fauth M, Ganguli A, Kotak S, Mishra SK, Nover L, Port M, Scharf KD et al (2004) Heat stress response in plants: a complex game with chaperones and more than twenty heat stress transcription factors. J Biosci 29:471–487

    Article  PubMed  CAS  Google Scholar 

  • Busch W, Wunderlich M, Schöffl F (2005) Identification of novel heat shock factor-dependent genes and biochemical pathways in Arabidopsis thaliana. Plant J 41:1–14

    Article  PubMed  CAS  Google Scholar 

  • Chen CH, Chen ZX (2002) Potentiation of developmentally regulated plant defense response by AtWRKY18, a pathogen-induced Arabidopsis transcription factor. Plant Physiol 129:706–716

    Article  PubMed  CAS  Google Scholar 

  • Chen H, Hwang JE, Lim CJ, Kim DY, Lee SY, Lim CO (2010a) Arabidopsis DREB2C functions as a transcriptional activator of HsfA3 during the heat stress response. Biochem Biophys Res Commun 401:238–244

    Article  PubMed  CAS  Google Scholar 

  • Chen LG, Zhang LP, Yu DQ (2010b) Wounding-induced WRKY8 is involved in basal defense in Arabidopsis. Mol Plant Microbe Interact 23:558–565

    Article  PubMed  CAS  Google Scholar 

  • Clarke SM, Mur LAJ, Wood JE, Scott IM (2004) Salicylic acid dependent signaling promotes basal thermotolerance but is not essential for acquired thermotolerance in Arabidopsis thaliana. Plant J 38:432–447

    Article  PubMed  CAS  Google Scholar 

  • Clarke SM, Cristescu SM, Miersch O, Harren FJM, Wasternack C, Mur LAJ (2009) Jasmonates act with salicylic acid to confer basal thermotolerance in Arabidopsis thaliana. New Phytol 182:175–187

    Article  PubMed  CAS  Google Scholar 

  • Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743

    Article  PubMed  CAS  Google Scholar 

  • Dellagi A, Heilbronn J, Avrova AO, Montesano M, Palva ET, Stewart HE, Toth IK, Cooke DEL, Lyon GD, Birch PRJ (2000) A potato gene encoding a WRKY-like transcription factor is induced in interactions with Erwinia carotovora subsp. atroseptica and Phytophthora infestans and is coregulated with class I endochitinase expression. Mol Plant Microbe Interact 13:1092–1101

    Article  PubMed  CAS  Google Scholar 

  • Dong JX, Chen CH, Chen ZX (2003) Expression profiles of the Arabidopsis WRKY gene superfamily during plant defense response. Plant Mol Biol 51:21–37

    Article  PubMed  CAS  Google Scholar 

  • Eulgem T, Somssich IE (2007) Networks of WRKY transcription factors in defense signaling. Curr Opin Plant Biol 10:366–371

    Article  PubMed  CAS  Google Scholar 

  • Eulgem T, Rushton PJ, Robatzek S, Somssich IE (2000) The WRKY superfamily of plant transcription factors. Trends Plant Sci 5:199–206

    Article  PubMed  CAS  Google Scholar 

  • Gadjev I, Vanderauwera S, Gechev TS, Laloi C, Minkov IN, Shulaev V, Apel K, Inze D, Mittler R, Van Breusegem F (2006) Transcriptomic footprints disclose specificity of reactive oxygen species signaling in Arabidopsis. Plant Physiol 141:436–445

    Article  PubMed  CAS  Google Scholar 

  • Hara K, Yagi M, Kusano T, Sano H (2000) Rapid systemic accumulation of transcripts encoding a tobacco WRKY transcription factor upon wounding. Mol Gen Genet 263:30–37

    Article  PubMed  CAS  Google Scholar 

  • Hong SW, Lee U, Vierling E (2003) Arabidopsis hot mutants define multiple functions required for acclimation to high temperatures. Plant Physiol 132:757–767

    Article  PubMed  CAS  Google Scholar 

  • Howarth CJ, Pollock CJ, Peacock JM (1997) Development of laboratory-based methods for assessing seedling thermotolerance in pearl millet. New Phytol 137:129–139

    Article  Google Scholar 

  • Huang T, Duman JG (2002) Cloning and characterization of a thermal hysteresis (antifreeze) protein with DNA-binding activity from winter bittersweet nightshade, Solanum dulcamara. Plant Mol Biol 48:339–350

    Article  PubMed  CAS  Google Scholar 

  • Jiang YQ, Deyholos MK (2009) Functional characterization of Arabidopsis NaCl-inducible WRKY25 and WRKY33 transcription factors in abiotic stresses. Plant Mol Biol 69:91–105

    Article  PubMed  CAS  Google Scholar 

  • Jiang W, Yu D (2009) Arabidopsis WRKY2 transcription factor mediates seed germination and postgermination arrest of development by abscisic acid. BMC Plant Biol 9:96

    Article  PubMed  Google Scholar 

  • Jing S, Zhou X, Song Y, Yu D (2009) Heterologous expression of OsWRKY23 gene enhances pathogen defense and dark-induced leaf senescence in Arabidopsis. Plant Growth Regul 58:181–190

    Article  CAS  Google Scholar 

  • Johnson CS, Kolevski B, Smyth DR (2002) TRANSPARENT TESTA GLABRA2, a trichome and seed coat development gene of Arabidopsis, encodes a WRKY transcription factor. Plant Cell 14:1359–1375

    Article  PubMed  CAS  Google Scholar 

  • Journot-Catalino N, Somssich IE, Roby D, Kroj T (2006) The transcription factors WRKY11 and WRKY17 act as negative regulators of basal resistance in Arabidopsis thaliana. Plant Cell 18:3289–3302

    Article  PubMed  CAS  Google Scholar 

  • Kalde M, Barth M, Somssich IE, Lippok B (2003) Members of the Arabidopsis WRKY group III transcription factors are part of different plant defense signaling pathways. Mol Plant Microbe Interact 16:295–305

    Article  PubMed  CAS  Google Scholar 

  • Kotak S, Larkindale J, Lee U, von Koskull-Döring P, Vierling E, Scharf KD (2007) Complexity of the heat stress response in plants. Curr Opin Plant Biol 10:310–316

    Article  PubMed  CAS  Google Scholar 

  • Lai Z, Vinod KM, Zheng Z, Fan B, Chen Z (2008) Roles of Arabidopsis WRKY3 and WRKY4 transcription factors in plant responses to pathogens. BMC Plant Biol 8:68

    Article  PubMed  Google Scholar 

  • Larkindale J, Huang BR (2005) Effects of abscisic acid, salicylic acid, ethylene and hydrogen peroxide in thermotolerance and recovery for creeping bentgrass. Plant Growth Regul 47:17–28

    Article  CAS  Google Scholar 

  • Larkindale J, Knight MR (2002) Protection against heat stress-induced oxidative damage in Arabidopsis involves calcium, abscisic acid, ethylene, and salicylic acid. Plant Physiol 128:682–695

    Article  PubMed  CAS  Google Scholar 

  • Larkindale J, Vierling E (2008) Core genome responses involved in acclimation to high temperature. Plant Physiol 146:748–761

    Article  PubMed  CAS  Google Scholar 

  • Larkindale J, Hall JD, Knight MR, Vierling E (2005) Heat stress phenotypes of Arabidopsis mutants implicate multiple signaling pathways in the acquisition of thermotolerance. Plant Physiol 138:882–897

    Article  PubMed  CAS  Google Scholar 

  • Li SJ, Fu QT, Huang WD, Yu DQ (2009) Functional analysis of an Arabidopsis transcription factor WRKY25 in heat stress. Plant Cell Rep 28:683–693

    Article  PubMed  CAS  Google Scholar 

  • Li SJ, Zhou X, Chen LG, Huang WD, Yu DQ (2010) Functional characterization of Arabidopsis thaliana WRKY39 in heat stress. Mol Cells 29:475–483

    Article  PubMed  CAS  Google Scholar 

  • Lichtenthaler HK (1987) Chlorophylls and carotenoids pigments of photosynthetic biomembranes. Methods Enzymol 148:350–382

    Article  CAS  Google Scholar 

  • Lim CJ, Hwang JE, Chen H, Hong JK, Yang K, Choi MS, Lee KO, Chung WS, Lee SY, Lim CO (2007) Over-expression of the Arabidopsis DRE/CRT-binding transcription factor DREB2C enhances thermotolerance. Biochem Biophys Res Commun 362:431–436

    Article  PubMed  CAS  Google Scholar 

  • Lippok B, Birkenbihl RP, Rivory G, Brümmer J, Schmelzer E, Logemann E, Somssich IE (2007) Expression of AtWRKY33 encoding a pathogen- or PAMP-responsive WRKY transcription factor is regulated by a composite DNA motif containing W box elements. Mol Plant Microbe Interact 20:420–429

    Article  PubMed  CAS  Google Scholar 

  • Liu HT, Liu YY, Pan QH, Yang HR, Zhan JC, Huang WD (2006) Novel interrelationship between salicylic acid, abscisic acid, and PIP2-specific phospholipase C in heat acclimation-induced thermotolerance in pea leaves. J Exp Bot 57:3337–3347

    Article  PubMed  CAS  Google Scholar 

  • Liu HT, Gao F, Li GL, Han JL, Liu DL, Sun DY, Zhou RG (2008) The calmodulin-binding protein kinase 3 is part of heat-shock signal transduction in Arabidopsis thaliana. Plant J 55:760–773

    Article  PubMed  CAS  Google Scholar 

  • Miller G, Suzuki N, Rizhsky L, Hegie A, Koussevitzky S, Mittler R (2007) Double mutants deficient in cytosolic and thylakoid ascorbate peroxidase reveal a complex mode of interaction between reactive oxygen species, plant development, and response to abiotic stresses. Plant Physiol 144:1777–1785

    Article  PubMed  CAS  Google Scholar 

  • Miller G, Shulaev V, Mittler R (2008) Reactive oxygen signaling and abiotic stress. Physiol Plant 133:481–489

    Article  PubMed  CAS  Google Scholar 

  • Mishra SK, Tripp J, Winkelhaus S, Tschiersch B, Theres K, Nover L, Scharf KD (2002) In the complex family of heat stress transcription factors, HSfA1 has a unique role as master regulator of thermotolerance in tomato. Genes Dev 16:1555–1567

    Article  PubMed  CAS  Google Scholar 

  • Mittler R (2006) Abiotic stress, the field environment and stress combination. Trends Plant Sci 11:15–19

    Article  PubMed  CAS  Google Scholar 

  • Panchuk II, Volkov RA, Schöffl F (2002) Heat stress- and heat shock transcription factor-dependent expression and activity of ascorbate peroxidase in Arabidopsis. Plant Physiol 129:838–853

    Article  PubMed  CAS  Google Scholar 

  • Pnueli L, Liang H, Rozenberg M, Mittler R (2003) Growth suppression, altered stomatal responses, and augmented induction of heat shock proteins in cytosolic ascorbate peroxidase (Apx1)-deficient Arabidopsis plants. Plant J 34:185–201

    Article  Google Scholar 

  • Qiao H, Chang KN, Yazaki J, Ecker JR (2009) Interplay between ethylene, ETP1/ETP2 F-box proteins, and degradation of EIN2 triggers ethylene responses in Arabidopsis. Genes Dev 23:512–521

    Article  PubMed  CAS  Google Scholar 

  • Qiu Y, Yu D (2009) Over-expression of the stress-induced OsWRKY45 enhances disease resistance and drought tolerance in Arabidopsis. Environ Exp Bot 65:35–47

    Article  CAS  Google Scholar 

  • Rizhsky L, Liang H, Mittler R (2002) The combined effect of drought stress and heat shock on gene expression in tobacco. Plant Physiol 130:1143–1151

    Article  PubMed  CAS  Google Scholar 

  • Rizhsky L, Davletova S, Liang HJ, Mittler R (2004) The zinc finger protein Zat12 is required for cytosolic ascorbate peroxidase 1 expression during oxidative stress in Arabidopsis. J Biol Chem 279:11736–11743

    Article  PubMed  CAS  Google Scholar 

  • Sakuma Y, Maruyama K, Qin F, Osakabe Y, Shinozaki K, Yamaguchi-Shinozaki K (2006) Dual function of an Arabidopsis transcription factor DREB2A in water-stress-responsive and heat-stress-responsive gene expression. Proc Natl Acad Sci USA 103:11827–18822

    Article  Google Scholar 

  • Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual. Cold spring harbor laboratory press, New York

    Google Scholar 

  • Seki M, Narusaka M, Ishida J, Nanjo T, Fujita M, Oono Y, Kamiya A, Nakajima M, Enju A, Sakurai T (2002) Monitoring the expression profiles of 7000 Arabidopsis genes under drought, cold and high-salinity stresses using a full-length cDNA microarray. Plant J 31:279–292

    Article  PubMed  CAS  Google Scholar 

  • Suzuki N, Rizhsky L, Liang HJ, Shuman J, Shulaev V, Mittler R (2005) Enhanced tolerance to environmental stress in transgenic plants expressing the transcriptional coactivator multiprotein bridging factor 1c. Plant Physiol 139:1313–1322

    Article  PubMed  CAS  Google Scholar 

  • Suzuki N, Bajad S, Shuman J, Shulaev V, Mittler R (2008) The transcriptional co-activator MBF1c is a key regulator of thermotolerance in Arabidopsis thaliana. J Biol Chem 283:9269–9275

    Article  PubMed  CAS  Google Scholar 

  • Taki N, Sasaki-Sekimoto Y, Obayashi T, Kikuta A, Kobayashi K, Ainai T, Yagi K, Sakurai N, Suzuki H, Masuda T, Takamiya K, Shibata D, Kobayashi Y, Ohta H (2005) 12-oxo-phytodienoic acid triggers expression of a distinct set of genes and plays a role in wound-induced gene expression in Arabidopsis. Plant Physiol 139:1268–1283

    Article  PubMed  CAS  Google Scholar 

  • Turck F, Zhou A, Somssich IE (2004) Stimulus-dependent, promoter-specific binding of transcription factor WRKY1 to its native promoter and the defense-related gene PcPR1-1 in parsley. Plant Cell 16:2573–2585

    Article  PubMed  CAS  Google Scholar 

  • Ülker B, Somssich IE (2004) WRKY transcription factors: from DNA binding towards biological function. Curr Opin Plant Biol 7:491–498

    Article  PubMed  Google Scholar 

  • von Koskull-Döring P, Scharf KD, Nover L (2007) The diversity of plant heat stress transcription factors. Trends Plant Sci 12:452–457

    Article  Google Scholar 

  • Wu X, Shiroto Y, Kishitani S, Ito Y, Toriyama K (2009) Enhanced heat and drought tolerance in transgenic rice seedlings overexpressing OsWRKY11 under the control of HSP101 promoter. Plant Cell Rep 28:21–30

    Article  PubMed  CAS  Google Scholar 

  • 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 18:1310–1326

    Article  PubMed  CAS  Google Scholar 

  • Yoshida T, Sakuma Y, Todaka D, Maruyama K, Qin F, Mizoi J, Kidokoro S, Fujita Y, Shinozaki K, Yamaguchi-Shinozaki K (2008) Functional analysis of an Arabidopsis heat-shock transcription factor HsfA3 in the transcriptional cascade downstream of the DREB2A stress-regulatory system. Biochem Biophys Res Commun 368:515–521

    Article  PubMed  CAS  Google Scholar 

  • Zheng Z, Qamar SA, Chen Z, Mengiste T (2006) Arabidopsis WRKY33 transcription factor is required for resistance to necrotrophic fungal pathogens. Plant J 48:592–605

    Article  PubMed  CAS  Google Scholar 

  • Zheng ZY, Mosher SL, Fan BF, Klessig DF, Chen ZX (2007) Functional analysis of Arabidopsis WRKY25 transcription factor in plant defense against Pseudomonas syringae. BMC Plant Biol 7:2

    Article  PubMed  Google Scholar 

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Acknowledgments

We thank Dr. Zhixiang Chen (Department of Botany and Plant Physiology, Purdue University, West Lafayette, Indiana, USA) for Arabidopsis wrky33-2, wrky25-1wrky26-2wrky33-1, coi1, ein2, and sid2. This work was supported by the Science Foundation of the Ministry of Agriculture of the People’s Republic of China (2009ZX08009-066B) and the Natural Science Foundation of China (Nos. 90817003 and 30871747).

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Authors and Affiliations

  1. Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, Kunming, 650223, People’s Republic of China

    Shujia Li, Qiantang Fu, Ligang Chen & Diqiu Yu

  2. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083, People’s Republic of China

    Shujia Li & Weidong Huang

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  1. Shujia Li
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  2. Qiantang Fu
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  3. Ligang Chen
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  4. Weidong Huang
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  5. Diqiu Yu
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Correspondence to Weidong Huang or Diqiu Yu.

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Chinese Academy of Sciences, People’s Republic of China and China Agricultural University, People’s Republic of China contributed equally to this work.

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Fig. S1 Comparison of WRKY25, WRKY26, and WRKY33 sequences (TIFF 2,753 kb)

Fig. S2 Identification of homozygous mutant lines (TIFF 2,125 kb)

Fig. S3 Northern blot analysis of WRKY26 and WRKY33 expression in wild-type and transgenic plants (TIFF 979 kb)

Fig. S4 Characterization of wrky25wrky26wrky33 and wrky25wrky26-2wrky33 triple mutants (TIFF 2,973 kb)

425_2011_1375_MOESM5_ESM.tif

Fig. S5 Expression of heat defense-related genes in 21-day-old wrky33, wild-type, and 35S:W33 plants during heat treatment (TIFF 1,004 kb)

425_2011_1375_MOESM6_ESM.tif

Fig. S6 Expression of heat defense-related genes in 21-day-old wrky26, wild-type, and 35S:W26 plants during heat treatment (TIFF 3,239 kb)

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Li, S., Fu, Q., Chen, L. et al. Arabidopsis thaliana WRKY25, WRKY26, and WRKY33 coordinate induction of plant thermotolerance. Planta 233, 1237–1252 (2011). https://doi.org/10.1007/s00425-011-1375-2

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