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GhWRKY39, a member of the WRKY transcription factor family in cotton, has a positive role in disease resistance and salt stress tolerance

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Abstract

WRKY transcription factors have been suggested to play crucial roles in the response to biotic and abiotic stresses. However, previous studies concerning WRKYs have primarily focused on model plants, and fairly limited research has been performed with cotton. In the present study, we functionally characterized a stress-responsive IId WRKY gene (GhWRKY39) from cotton. GhWRKY39 is present as a single copy gene, and subcellular localization analysis indicated that GhWRKY39 localizes to the nucleus. Additionally, some cis-acting elements associated with the environmental stress response were observed in the promoter region of this gene. Consistently, a β-glucuronidase activity assay and quantitative PCR analysis revealed that GhWRKY39 expression could be induced by bacterial and fungal infection or NaCl treatment. Furthermore, the constitutive overexpression of GhWRKY39 in Nicotiana benthamiana conferred greater resistance to bacterial and fungal pathogen infections, and the expression of several pathogenesis-related (PR) genes was significantly increased. The transgenic plants also exhibited less H2O2 accumulation than wild-type plants following pathogen infection. Moreover, GhWRKY39-overexpressing plants displayed enhanced tolerance to salt and oxidative stress and increased transcription of antioxidant enzyme genes, including ascorbate peroxidase (APX), catalase (CAT), glutathione-S-transferase (GST) and superoxide dismutase (SOD). Importantly, overexpression of GhWRKY39 improved the activities of the antioxidant enzymes SOD, POD and CAT after pathogen infection and salt stress treatment. Overall, our data suggest that the overexpression of GhWRKY39 may positively regulate the plant response against pathogen infection and salt stress, likely through the regulation of the reactive oxygen species system via multiple signaling pathway.

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Abbreviations

APX:

Ascorbate peroxidase

CAT:

Catalase

Ca:

Capsicum annuum

CTAB:

Cetyltrimethyl ammonium bromide

DAB:

3,3′-Diaminobenzidine

GFP:

Green fluorescent protein

Gh:

Gossypium hirsutum

GST:

Glutathione-S-transferase

hiTAIL-PCR:

High-efficiency TAIL-PCR

MeJA:

Methyl jasmonate

MS medium:

Murashige and Skoog medium

MV:

Methyl viologen

Nb:

Nicotiana benthamiana

NPR1:

Non-expression of PR1

OE:

Overexpression

ORF:

Open reading frame

PR:

Pathogenesis-related

RACE:

Rapid amplification of cDNA ends

ROS:

Reactive oxygen species

R. solanacearum :

Ralstonia solanacearum

R. solani :

Rhizoctonia solani

qPCR:

Quantitative PCR

SA:

Salicylic acid

SOD:

Superoxide dismutase

UTR:

Untranslated region

WT:

Wild-type

References

  • Alvarez ME, Pennell RI, Meijer PJ, Ishikawa A, Dixon RA, Lamb C (1998) Reactive oxygen intermediates mediate a systemic signal network in the establishment of plant immunity. Cell 92:773–784

    Article  CAS  PubMed  Google Scholar 

  • Baumann K, De Paolis A, Costantino P, Gualberti G (1999) The DNA binding site of the Dof protein NtBBF1 is essential for tissue-specific and auxin-regulated expression of the rolB oncogene in plants. Plant Cell 11:323–334

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Boland JF, Linthorst HJM, Cornelissen BJC (1990) Plant pathogenesis-related protein induced by virus infection. Annu Rev Phytopathol 28:113–138

    Article  Google Scholar 

  • Borsani O, Zhu J, Verslues PE, Sunkar R, Zhu JK (2005) Endogenous siRNAs derived from a pair of natural cis-antisense transcripts regulate salt tolerance in Arabidopsis. Cell 123:1279–1291

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Bostock RM (2005) Signal crosstalk and induced resistance: straddling the line between cost and benefit. Annu Rev Phytopathol 43:545–580

    Article  CAS  PubMed  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  CAS  PubMed  Google Scholar 

  • Dang FF, Wang YN, Yu L, Eulgem T, Lai Y, Liu ZQ, Wang X, Qiu AL, Zhang TX, Lin J, Chen YS, Guan DY, Cai HY, Mou SL, He SL (2013) CaWRKY40, a WRKY protein of pepper, plays an important role in the regulation of tolerance to heat stress and resistance to Ralstonia solanacearum infection. Plant, Cell Environ 36:757–774

    Article  CAS  Google Scholar 

  • Durrant WE, Dong X (2004) Systemic acquired resistance. Annu Rev Phytopathol 42:185–209

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  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  CAS  PubMed  Google Scholar 

  • Gao L, Xiang CB (2008) The genetic locus At1g73660 encodes a putative MAPKKK and negatively regulates salt tolerance in Arabidopsis. Plant Mol Biol 67:125–134

    Article  CAS  PubMed  Google Scholar 

  • Hammond-Kosack KE, Jones JD (1997) Plant disease resistance genes. Annu Rev Plant Physiol Plant Mol Biol 48:575–607

    Article  CAS  PubMed  Google Scholar 

  • Hasegawa PM, Bressan RA, Zhu JK, Bohnert HJ (2000) Plant cellular and molecular responses to high salinity. Annu Rev Plant Physiol Plant Mol Biol 51:463–499

    Article  CAS  PubMed  Google Scholar 

  • Higashi K, Ishiga Y, Inagaki Y, Toyoda K, Shiraishi T, Ichinose Y (2008) Modulation of defense signal transduction by flagellin-induced WRKY41 transcription factor in Arabidopsis thaliana. Mol Genet Genomics 279:303–312

    Article  CAS  PubMed  Google Scholar 

  • Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1231

    Article  CAS  Google Scholar 

  • Hu J, Barlet X, Deslandes L, Hirsch J, Feng DX, Somssich I, Marco Y (2008) Transcriptional responses of Arabidopsis thaliana during wilt disease caused by the soil-borne phytopathogenic bacterium, Ralstonia solanacearum. PLoS One 3:e2589

    Article  PubMed Central  PubMed  Google Scholar 

  • Hu YR, Dong QY, Yu DQ (2012) Arabidopsis WRKY46 coordinates with WRKY70 and WRKY53 in basal resistance against pathogen Pseudomonas syringae. Plant Sci 185–186:288–297

    Article  PubMed  Google Scholar 

  • Jabs T, Dietrich RA, Dangl JL (1996) Initiation of runaway cell death in an Arabidopsis mutant by extracellular superoxide. Science 273:1853–1856

    Article  CAS  PubMed  Google Scholar 

  • Jiang YQ, Deyholos MK (2006) Comprehensive transcriptional profiling of NaCl-stressed Arabidopsis roots reveals novel classes of responsive genes. BMC Plant Biol 6:25

    Article  PubMed Central  PubMed  Google Scholar 

  • Jones JD, Dangl JL (2006) The plant immune system. Nature 444:323–329

    Article  CAS  PubMed  Google Scholar 

  • Kotchoni SO, Gachomo EW (2006) The reactive oxygen species network pathways: an essential prerequisite for perception of pathogen attack and the acquired disease resistance in plants. J Biosci 31:389–404

    Article  CAS  PubMed  Google Scholar 

  • Lamb C, Dixon RA (1997) The oxidative burst in plant disease resistance. Annu Rev Plant Physiol Plant Mol Biol 48:251–275

    Article  CAS  PubMed  Google Scholar 

  • Lescot M, Dehais P, Thijs G, Marchal K, Moreau Y, Peer VY, Rouze P, Rombauts S (2002) PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences. Nucleic Acids Res 30:325–327

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Levine A, Tenhaken R, Dixon R, Lamb C (1994) H2O2 from the oxidative burst orchestrates the plant hypersensitive disease resistance response. Cell 79:583–593

    Article  CAS  PubMed  Google Scholar 

  • Li SJ, Fu QT, Chen LG, Huang WD, Yu DQ (2011) Arabidopsis thaliana WRKY25, WRKY26, and WRKY33 coordinate induction of plant thermo tolerance. Planta 233:1237–1252

    Article  CAS  PubMed  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

    Google Scholar 

  • Liu YG, Chen YL (2007) High-efficiency thermal asymmetric interlaced PCR for amplification of unknown flanking sequences. Biotechniques 43:649–656

    Article  CAS  PubMed  Google Scholar 

  • Liu QL, Zhong M, Li S, Pan YZ, Jiang BB, Jia Y, Zhong HQ (2013) Overexpression of a chrysanthemum transcription factor gene, DgWRKY3, in tobacco enhances tolerance to salt stress. Plant Physiol Biochem 69:27–33

    Article  CAS  PubMed  Google Scholar 

  • Lu WJ, Chu XQ, Li YZ, Wang C, Guo XQ (2013) Cotton GhMKK1 induces the tolerance of salt and drought stress, and mediates defense responses to pathogen infection in transgenic Nicotiana benthamiana. PLoS One 8:e68503

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Mason G, Provero P, Vaira AM, Accotto GP (2002) Estimating the number of integrations in transformed plants by quantitative real-time PCR. BMC Biotechnol 2:20

    Article  PubMed Central  PubMed  Google Scholar 

  • Menke FL, Kang HG, Chen Z, Park JM, Kumar D, Klessig DF (2005) Tobacco transcription factor WRKY1 is phosphorylated by the MAP kinase SIPK and mediates HR-like cell death in tobacco. Mol Plant Microbe Interact 18:1027–1034

    Article  CAS  PubMed  Google Scholar 

  • Munns R, Tester M (2008) Mechanisms of salinity tolerance. Annu Rev Plant Biol 59:651–681

    Article  CAS  PubMed  Google Scholar 

  • Mur LAJ, Kenton P, Atzorn R, Miersch O, Wasternack C (2006) The outcomes of concentration-specific interactions between salicylate and jasmonate signaling include synergy, antagonism, and oxidative stress leading to cell death. Plant Physiol 140:249–262

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Murray SL, Ingle RA, Petersen LN, Denby KJ (2007) Basal resistance against Pseudomonas syringae in Arabidopsis involves WRKY53 and a protein with homology to a nematode resistance protein. Mol Plant Microbe Interact 20:1431–1438

    Article  CAS  PubMed  Google Scholar 

  • Niu CF, Wei W, Zhou QY, Tian AG, Hao YJ, Zhang WK, Ma B, Lin Q, Zhang ZB, Zhang JS, Chen SY (2012) Wheat WRKY genes TaWRKY2 and TaWRKY19 regulate abiotic stress tolerance in transgenic Arabidopsis plants. Plant, Cell Environ 35:1156–1170

    Article  CAS  Google Scholar 

  • Pandey SP, Somssich IE (2009) The role of WRKY transcription factors in plant immunity. Plant Physiol 150:1648–1655

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Park CY, Lee JH, Yoo JH, Moon BC, Choi MS, Kang YH, Lee SM, Kim HS, Kang KY, Chung WS, Lim CO, Cho MJ (2005) WRKY group IId transcription factors interact with calmodulin. FEBS Lett 579:1545–1550

    Article  CAS  PubMed  Google Scholar 

  • Ross CA, Liu Y, Shen QJ (2007) The WRKY gene family in rice (Oryza sativa). J Integr Plant Biol 49:827–842

    Article  CAS  Google Scholar 

  • Rushton PJ, Somssich IE, Ringler P, Shen QJ (2010) WRKY transcription factors. Trends Plant Sci 15:247–258

    Article  CAS  PubMed  Google Scholar 

  • Rushton DL, Tripathi P, Rabara RC, Lin J, Ringler P, Boken AK, Langum TJ, Smidt L, Boomsma DD, Emme NJ, Chen X, Finer JJ, Shen QJ, Rushton PJ (2012) WRKY transcription factors: key components in abscisic acid signaling. Plant Biotechnol J 10:2–11

    Article  CAS  PubMed  Google Scholar 

  • Shah J, Kachroo P, Nandi A, Klessig DF (2001) A recessive mutation in the Arabidopsis SSI2 gene confers SA- and NPR1-independent expression of PR genes and resistance against bacterial and oomycete pathogens. Plant J 25:563–574

    Article  CAS  PubMed  Google Scholar 

  • Shi J, Zhang L, An HL, Wu CA, Guo XQ (2011) GhMPK16, a novel stress-responsive group D MAPK gene from cotton, is involved in disease resistance and drought sensitivity. BMC Mol Biol 12:22

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Shinozaki K (1999) Plant response to drought and salt stress: overview. Tanpakushitsu Kakusan Koso 44:2186–2187

    CAS  PubMed  Google Scholar 

  • Sun C, Palmqvist S, Olsson H, Boren M, Ahlandsberg S, Jansson C (2003) A novel WRKY transcription factor, SUSIBA2, participates in sugar signaling in barley by binding to the sugar-responsive elements of the iso1 promoter. Plant Cell 15:2076–2092

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Tausz M, Sircelj H, Grill D (2004) The glutathione system as a stress marker in plant ecophysiology: is a stress-response concept valid? J Exp Bot 55:1955–1962

    Article  CAS  PubMed  Google Scholar 

  • Thordal-Christensen H, Zhang ZG, Wei YD, Collinge DB (1997) Subcellular localization of H2O2 in plants. H2O2 accumulation in papillae and hypersensitive response during the barley-powdery mildew interaction. Plant J 11:1187–1194

    Article  CAS  Google Scholar 

  • Tuteja N (2007) Abscisic acid and abiotic stress signaling. Plant Signal Behav 2:135–138

    Article  PubMed Central  PubMed  Google Scholar 

  • Varagona MJ, Schmidt RJ, Raikhel NV (1992) Nuclear localization signal(s) required for nuclear targeting of the maize regulatory protein Opaque-2. Plant Cell 4:1213–1227

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Wang H, Miyazaki S, Kawai K, Deyholos M, Galbraith DW, Bohnert HJ (2003a) Temporal progression of gene expression responses to salt shock in maize roots. Plant Mol Biol 52:873–891

    Article  CAS  PubMed  Google Scholar 

  • Wang W, Vinocur B, Altman A (2003b) Plant responses to drought, salinity and extreme temperatures: towards genetic engineering for stress tolerance. Planta 218:1–14

    Article  CAS  PubMed  Google Scholar 

  • Wang HH, Hao JJ, Chen XJ, Hao ZN, Wang X, Lou Y, Peng Y, Guo Z (2007) Overexpression of rice WRKY89 enhances ultraviolet B tolerance and disease resistance in rice plants. Plant Mol Biol 65:799–815

    Article  CAS  PubMed  Google Scholar 

  • Wang M, Li SW, Yang HF, Gao Z, Wu CA, Guo XQ (2012) Characterization and functional analysis of GhRDR6, a novel RDR6gene from cotton (Gossypium hirsutum L.). Biosci Rep 32:139–151

    Article  CAS  PubMed  Google Scholar 

  • Wang C, Deng PY, Chen LL, Wang XT, Ma H, Hu W, Yao NC, Feng Y, Chai RH, Yang GX, He GY (2013a) A wheat WRKY transcript ion factor TaWRKY10 confers tolerance to multiple abiotic stresses in transgenic tobacco. PLoS One 8:e65120

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Wang Y, Dang FF, Liu ZQ, Wang X, Eulgem T, Lai Y, Yu L, She JJ, Shi YL, Lin JH, Chen CC, Guan DY, Qiu AL, He SL (2013b) CaWRKY58, encoding a group I WRKY transcription factor of Capsicum annuum, negatively regulates resistance to Ralstonia solanacearum infection. Mol Plant Pathol 14:131–144

    Article  PubMed  Google Scholar 

  • Xing DH, Lai ZB, Zheng ZY, Vinod KM, Fan BF, Chen ZX (2008) Stress- and pathogen-induced Arabidopsis WRKY48 is a transcriptional activator that represses plant basal defense. Mol Plant 1:459–470

    Article  CAS  PubMed  Google Scholar 

  • Yang YO, Shah J, Klessig DF (1997) Signal perception and transduction in defense responses. Genes Dev 11:1621–1639

    Article  CAS  PubMed  Google Scholar 

  • Yang PZ, Chen CN, Wang ZP, Fan BF, Chen ZX (1999) A pathogen- and salicylic acid-induced WRKY DNA-binding activity recognizes the elicitor response element of the tobacco class I chitinase gene promoter. Plant J 18:141–149

    Article  CAS  Google Scholar 

  • Yang L, Tang R, Zhu J, Liu H, Mueller-Roeber B, Xia H, Zhang H (2008) Enhancement of stress tolerance in transgenic tobacco plants constitutively expressing AtIpk2β, an inositol polyphosphate 6-/3-kinase from Arabidopsis thaliana. Plant Mol Biol 66:329–343

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Yoshioka H, Numata N, Nakajima K, Katou S, Kawakita K, Rowland O, Jones JD, Doke N (2003) Nicotiana benthamiana gp91phox homologs NbrbohA and NbrbohB participate in H2O2 accumulation and resistance to Phytophthora infestans. Plant Cell 15:706–718

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Yu D, Chen C, Chen Z (2001) Evidence for an important role of WRKY DNA binding proteins in the regulation of NPR1 gene expression. Plant Cell 13:1527–1540

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Yu FF, Huaxia YF, Lu WJ, Wu C, Guo XQ (2012) GhWRKY15, a member of the WRKY transcription factor family identified from cotton (Gossypium hirsutum L.), is involved in disease resistance and plant development. BMC Plant Biol 12:144

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Zheng Z, Mosher SL, Fan B, Klessig DF, Chen Z (2007) Functional analysis of Arabidopsis WRKY25 transcription factor in plant defense against Pseudomonas syringae. BMC Plant Biol 7:2–15

    Article  PubMed Central  PubMed  Google Scholar 

  • Zheng L, Liu GF, Meng XN, Liu YJ, Ji XY, Li YB, Nie XG, Wang YC (2013) A WRKY gene from Tamarix hispida, ThWRKY4, mediates abiotic stress responses by modulating reactive oxygen species and expression of stress-responsive genes. Plant Mol Biol 82:303–320

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This work was financially supported by the Genetically Modified Organisms Breeding Major Projects of China [Grant Numbers 2009ZX08009-092B] and the National Natural Science Foundation of China [Grant Number 31171837].

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Authors and Affiliations

  1. State Key Laboratory of Crop Biology, College of Life Sciences, Shandong Agricultural University, Taian, 271018, Shandong, People’s Republic of China

    Weina Shi, Dongdong Liu, Lili Hao, Chang-ai Wu, Xingqi Guo & Han Li

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  1. Weina Shi
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  2. Dongdong Liu
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  3. Lili Hao
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  4. Chang-ai Wu
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  5. Xingqi Guo
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Correspondence to Xingqi Guo or Han Li.

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Shi, W., Liu, D., Hao, L. et al. GhWRKY39, a member of the WRKY transcription factor family in cotton, has a positive role in disease resistance and salt stress tolerance. Plant Cell Tiss Organ Cult 118, 17–32 (2014). https://doi.org/10.1007/s11240-014-0458-8

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