Skip to main content
SpringerLink
Log in

Overexpression of VaWRKY14 increases drought tolerance in Arabidopsis by modulating the expression of stress-related genes

  • Original Article
  • Published:
Plant Cell Reports Aims and scope Submit manuscript

Abstract

Key message

Overexpression of VaWRKY14 increases drought tolerance in Arabidopsis by modulating the expression of stress-related genes, including COR15A, COR15B, COR413, KIN2, and RD29A.

Abstract

The WRKY family is one of a largest transcription factors in plants, and it is a key component of multiple stress responses. In this study, the drought- and cold-induced WRKY family gene VaWRKY14 was isolated and characterized. Phylogenetic analysis indicated that VaWRKY14 belongs to the WRKY IIa subfamily, of which several members participate in biotic and abiotic stress responses in plants. Fluorescence observation from Arabidopsis mesophyll protoplasts transformed with the VaWRKY14::eGFP fusion vector suggested that VaWRKY14 was localized in the nucleus. The VaWRKY14 in yeast cells did not display any transcriptional activity. The expression of VaWRKY14 could be induced by exogenous phytohormones, including salicylic acid (SA) and abscisic acid (ABA). Overexpression of VaWRKY14 enhanced the drought tolerance of transgenic Arabidopsis. Compared with wild-type Arabidopsis, the VaWRKY14-OE lines exhibited higher water content and antioxidant enzyme activities in leaves after drought treatment. RNA sequencing analysis revealed that several stress-related genes, including COR15A, COR15B, COR413, KIN2, and RD29A, were upregulated in transgenic plants relative to their expression in wild-type Arabidopsis under normal conditions. Several genes (3 upregulated and 49 down-regulated) modulated by VaWRKY14 were also affected by drought stress in wild-type plants. These data suggest that VaWRKY14 responds to drought and cold stresses and that drought tolerance may be enhanced by regulating the expression of stress-related genes in Arabidopsis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  • Besseau S, Li J, Palva ET (2012) WRKY54 and WRKY70 co-operate as negative regulators of leaf senescence in Arabidopsis thaliana. J Exp Bot 63:2667–2679

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • 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 10:443–462

    Google Scholar 

  • Chen L, Song Y, Li S, Zhang L, Zou C, Yu D (2012) The role of WRKY transcription factors in plant abiotic stresses. Biochim Biophys Acta 1819:120–128

    Article  PubMed  CAS  Google Scholar 

  • 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 6:287–300

    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 

  • Cominelli E, Galbiati M, Vavasseur A, Conti L, Sala T, Vuylsteke M, Leonhardt N, Dellaporta SL, Tonelli C (2005) A guard-cell-specific MYB transcription factor regulates stomatal movements and plant drought tolerance. Curr Biol 15:1196–1200

    Article  PubMed  CAS  Google Scholar 

  • Dang FF, Wang YN, Lu YU, Eulgem T, Lai Y, Liu ZQ, Wang XU, Qiu AL, Zhang TX, Lin J (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  PubMed  CAS  Google Scholar 

  • Dempsey DA, Shah J, Klessig DF (1997) Salicylic acid and disease resistance in plants. Crit Rev Plant Sci 18:547–575

    Article  Google Scholar 

  • Ding ZJ, Yan JY, Xu XY, Yu DQ, Li GX, Zhang SQ, Zheng SJ (2014) Transcription factor WRKY46 regulates osmotic stress responses and stomatal movement independently in Arabidopsis. Plant J 79:13–27

    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 

  • Dubouzet JG, Sakuma Y, Ito Y, Kasuga M, Dubouzet EG, Miura S, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2003) OsDREB genes in rice, Oryza sativa L., encode transcription activators that function in drought-, high-salt- and cold-responsive gene expression. Plant J 33:751–763

    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 

  • Fang L, Su L, Sun X, Li X, Sun M, Karungo SK, Fang S, Chu J, Li S, Xin H (2016) Expression of Vitis amurensis NAC26 in Arabidopsis enhances drought tolerance by modulating jasmonic acid synthesis. J Exp Bot 67:2829–2845

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Gong XQ, Hu JB, Liu JH (2014) Cloning and characterization of FcWRKY40, a WRKY transcription factor from Fortunella crassifolia linked to oxidative stress tolerance. Plant Cell Tiss Org 119:197–210

    Article  CAS  Google Scholar 

  • Gong X, Zhang J, Hu J, Wang W, Wu H, Zhang Q, Liu JH (2015) FcWRKY70, a WRKY protein of Fortunella crassifolia, functions in drought tolerance and modulates putrescine synthesis by regulating arginine decarboxylase gene. Plant Cell Environ 38:2248–2262

    Article  PubMed  CAS  Google Scholar 

  • Guillaumie S, Mzid R, Méchin V, Léon C, Hichri I, Destrac-Irvine A, Lauvergeat V (2010) The grapevine transcription factor WRKY2 influences the lignin pathway and xylem development in tobacco. Plant Mol Biol 72:215–234

    Article  PubMed  CAS  Google Scholar 

  • Hall BG (2013) Building phylogenetic trees from molecular data with MEGA. Mol Biol Evol 30:1229–1235

    Article  PubMed  CAS  Google Scholar 

  • Hamada AM (2001) Salicylic acid versus salinity-drought-induced stress on wheat seedlings. Rostl Vyr 47:444–450

    CAS  Google Scholar 

  • He GH, Xu JY, Wang YX, Liu JM, Li PS, Chen M, Ma YZ, Xu ZS (2016) Drought-responsive WRKY transcription factor genes TaWRKY1 and TaWRKY33 from wheat confer drought and/or heat resistance in Arabidopsis. BMC Plant Biol 16:116

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Hubbard KE, Nishimura N, Hitomi K, Getzoff ED, Schroeder JI (2010) Early abscisic acid signal transduction mechanisms: newly discovered components and newly emerging questions. Gene Dev 24:1695–1708

    Article  PubMed  CAS  Google Scholar 

  • Ishiguro S, Nakamura K (1994) Characterization of a cDNA encoding a novel DNA-binding protein, SPF1, that recognizes SP8 sequences in the 5′ upstream regions of genes coding for sporamin and β-amylase from sweet potato. Mol Gen Genet 244:563–571

    Article  PubMed  CAS  Google Scholar 

  • Kang GZ, Zhu ZH, Guo TC, Ren JG (2009) Isolation and expression pattern of COR15b and KIN1 genes in watermelon and pumpkin. Afr J Biotechnol 8:5666–5672

    Article  CAS  Google Scholar 

  • Kim CY, Vo KTX, Cong DN, Jeong DH, Lee SK, Kumar M, Kim SR, Park SH, Kim JK, Jeon JS (2016) Functional analysis of a cold-responsive rice WRKY gene, OsWRKY71. Plant Biotechnol Rep 10:13–23

    Article  CAS  Google Scholar 

  • Kitajima S, Sato F (1999) Plant Pathogenesis-Related Proteins: Molecular Mechanisms of Gene Expression and Protein Function. J Biochem 125:1–8

    Article  PubMed  CAS  Google Scholar 

  • Korkmaz A, Uzunlu M, Demirkiran A (2007) Treatment with acetyl salicylic acid protects muskmelon seedlings against drought stress. Acta Physiol Plant 29:503–508

    Article  CAS  Google Scholar 

  • Lcvan L, Eavan S (2002) The families of pathogenesis-related proteins, their activities, and comparative analysis of PR-1 type proteins. Physiol Mol Plant P 55:85–97

    Google Scholar 

  • Lindemose S, O’Shea C, Jensen MK, Skriver K (2013) Structure, function and networks of transcription factors involved in abiotic stress responses. Int J Mol Sci 14:5842–5878

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Mare C, Mazzucotelli E, Crosatti C, Francia E, Cattivelli L (2004) Hv-WRKY38: a new transcription factor involved in cold-and drought-response in barley. Plant Mol Biol 55:399–416

    Article  PubMed  CAS  Google Scholar 

  • Mittler R, Vanderauwera S, Gollery M, Van Breusegem F (2004) Reactive oxygen gene network of plants. Trends Plant Sci 9:490–498

    Article  PubMed  CAS  Google Scholar 

  • Miura K, Tada Y (2014) Regulation of water, salinity, and cold stress responses by salicylic acid. Front Plant Sci 5:4

    Article  PubMed  PubMed Central  Google Scholar 

  • Mortazavi A, Williams BA, McCue K, Schaeffer L, Wold B (2008) Mapping and quantifying mammalian transcriptomes by RNA-SEq. Nat Methods 5:621–628

    Article  PubMed  CAS  Google Scholar 

  • Msanne J, Lin J, Stone JM, Awada T (2011) Characterization of abiotic stress-responsive Arabidopsis thaliana RD29A and RD29B genes and evaluation of transgenes. Planta 234:97–107

    Article  PubMed  CAS  Google Scholar 

  • Pan Y, Seymour GB, Lu C, Hu Z, Chen X, Chen G (2012) An ethylene response factor (ERF5) promoting adaptation to drought and salt tolerance in tomato. Plant Cell Rep 31:349–360

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Pandey SP, Roccaro M, Schon M, Logemann E, Somssich IE (2010) Transcriptional reprogramming regulated by WRKY18 and WRKY40 facilitates powdery mildew infection of Arabidopsis. Plant J 64:912–923

    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 

  • Ren X, Chen Z, Liu Y, Zhang H, Zhang M, Liu Q, Hong X, Zhu JK, Gong Z (2010) ABO3, a WRKY transcription factor, mediates plant responses to abscisic acid and drought tolerance in Arabidopsis. Plant J 63:417–429

    Article  PubMed  PubMed Central  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Santner A, Estelle M (2009) Recent advances and emerging trends in plant hormone signalling. Nat 459:1071–1078

    Article  CAS  Google Scholar 

  • Schön M, Töller A, Diezel C, Roth C, Westphal L, Wiermer M, Somssich IE (2013) Analyses of wrky18 wrky40 plants reveal critical roles of SA/EDS1 signaling and indole-glucosinolate biosynthesis for Golovinomyces orontii resistance and a loss-of resistance towards Pseudomonas syringae pv. tomato AvrRPS4. Mol Plant Microbe Interact 26:758–767

    Article  PubMed  CAS  Google Scholar 

  • Shen H, Liu C, Zhang Y, Meng X, Zhou X, Chu C, Wang X (2012) OsWRKY30 is activated by MAP kinases to confer drought tolerance in rice. Plant Mol Biol 80:241–253

    Article  PubMed  CAS  Google Scholar 

  • Shinozaki K, Yamaguchi-Shinozaki K, Seki M (2003) Regulatory network of gene expression in the drought and cold stress responses. Curr Opin Plant Biol 6:410–417

    Article  PubMed  CAS  Google Scholar 

  • Singh NK, Bracker CA, Hasegawa PM, Handa AK, Buckel S, Hermodson MA, Pfankoch E, Regnier FE, Bressan RA (1987) Characterization of Osmotin. Plant Physiol 85:529–536

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Thalhammer A, Hundertmark M, Popova AV (2010) Interaction of two intrinsically disordered plant stress proteins (COR15A and COR15B) with lipid membranes in the dry state. BBA-Biomembranes 1798:1812–1820

    Article  PubMed  CAS  Google Scholar 

  • Thalhammer A, Bryant G, Sulpice R, Hincha DK (2014) Disordered cold regulated 15 proteins protect chloroplast membranes during freezing through binding and folding, but do not stabilize chloroplast enzymes in Vivo. Plant Phyol 166:190–194

    Article  CAS  Google Scholar 

  • Thimm O, Bläsing O, Gibon Y, Nagel A, Meyer S, Krüger P, Selbig J, Müller LA, Rhee SY, Stitt M (2004) MAPMAN: a user-driven tool to display genomics data sets onto diagrams of metabolic pathways and other biological processes. Plant J 37:914–939

    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  CAS  Google Scholar 

  • Wang Z, Gerstein M, Snyder M (2009) RNA-Seq: a revolutionary tool for Transcriptomics. Nat Rev Genet 10:57–63

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Wang L, Zhu W, Fang L, Sun X, Su L, Liang Z, Wang N, Londo JP, Li S, Xin H (2014) Genome-wide identification of WRKY family genes and their response to cold stress in Vitis vinifera. BMC Plant Biol 14:103

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Wang F, Chen HW, Li QT, Wei W, Li W, Zhang WK, Ma B, Bi YD, Lai YC, Liu XL, Man WQ, Zhang JS, Chen SY (2015) GmWRKY27 interacts with GmMYB174 to reduce expression of GmNAC29 for stress tolerance in soybean plants. Plant J 83:224–236

    Article  PubMed  CAS  Google Scholar 

  • Wu KL, Guo ZJ, Wang HH, Li J (2005) The WRKY family of transcription factors in rice and Arabidopsis and their origins. DNA Res 12:9–26

    Article  PubMed  CAS  Google Scholar 

  • Wu FH, Shen SC, Lee LY, Lee SH, Chan MT, Lin CS (2009) Tape-Arabidopsis sandwich-a simpler Arabidopsis protoplast isolation method. Plant Meth 5(1):16

    Article  CAS  Google Scholar 

  • Xie Z, Zhang ZL, Zou X, Huang J, Ruas P, Thompson D, Shen QJ (2005) Annotations and functional analyses of the rice WRKY gene superfamily reveal positive and negative regulators of abscisic acid signaling in aleurone cells. Plant Physiol 137:176–189

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Xiong X, James VA, Zhang H, Altpeter F (2010) Constitutive expression of the barley HvWRKY38 transcription factor enhances drought tolerance in turf and forage grass (Paspalumnotatum Flugge). Mol Breeding 25:419–432

    Article  CAS  Google Scholar 

  • Xu YH, Wang JW, Wang S, Wang JY, Chen XY (2004) Characterization of GaWRKY1, a cotton transcription factor that regulates the sesquiterpene synthase gene (+)-delta-cadinene synthase-A. Plant Physiol 135:507–515

    Article  PubMed  PubMed Central  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  PubMed Central  CAS  Google Scholar 

  • Yamaguchi-Shinozaki K, Shinozaki K (2006) Transcriptional regulatory networks in cellular responses and tolerance to dehydration and cold stresses. Annu Rev Plant Biol 57:781–803

    Article  PubMed  CAS  Google Scholar 

  • Yin MZ, Wang YP, Zhang LH, Li JZ, Quan WL, Yang L, Wang QF, Chan ZL (2017) The Arabidopsis Cys2/His2 zinc finger transcription factor ZAT18 is a positive regulator of plant tolerance to drought stress. J Exp Bot 68:2991–3005

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Yokotani N, Sato Y, Tanabe S, Chujo T, Shimizu T, Okada K, Yamane H, Shimono M, Sugano S, Takatsuji H, Kaku H, Minami E, Nishizawa Y (2013) WRKY76 is a rice transcriptional repressor playing opposite roles in blast disease resistance and cold stress tolerance. J Exp Bot 64:5085–5097

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Yoo SD, Cho YH, Sheen J (2007) Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis. Nat Protoc 2:1565–1572

    Article  PubMed  CAS  Google Scholar 

  • Yu Y, Liu Z, Wang L, Kim SG, Seo PJ, Qiao M, Wang N, Li S, Cao X, Park CM, Xiang F (2016) WRKY71 accelerates flowering via the direct activation of FLOWERING LOCUS T and LEAFY in Arabidopsis thaliana. Plant J 85:96–106

    Article  PubMed  CAS  Google Scholar 

  • Zhang S, Klessig DF (2001) MAPK cascades in plant defense signaling. Trends Plant Sci 6:520–527

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the National Science Foundation of China (NSFC Accession No.: 31471857), Youth Innovation Promotion Association of CAS (2015281), Science and Technology Service Network Initiative of CAS (KFJ-STS-ZDTP-025), Grape Breeding Project of Ningxia (NXNYYZ201502) and Open Project Program of State Key Laboratory of Crop Stress Biology for Arid Areas (NWAFU, CSBAA2016009).

Author information

Authors and Affiliations

  1. Key Laboratory of Plant Germplasm Enhancement and Specialty Agriculture, Wuhan Botanical Garden, Chinese Academy of Sciences, Wuhan, 430074, People’s Republic of China

    Langlang Zhang, Jun Cheng, Xiaoming Sun, Tingting Zhao, Qingfeng Wang & Haiping Xin

  2. Beijing Key Laboratory of Grape Sciences and Enology, CAS Key Laboratory of Plant Resources, Institute of Botany, Chinese Academy of Sciences, Beijing, 100093, People’s Republic of China

    Jun Cheng, Xiaoming Sun & Shaohua Li

  3. University of Chinese Academy of Sciences, Beijing, 100049, People’s Republic of China

    Langlang Zhang

  4. State Key Laboratory of Crop Stress Biology for Arid Areas, College of Horticulture, Northwest A&F University, Yangling, 712100, People’s Republic of China

    Mingjun Li

Authors
  1. Langlang Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jun Cheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xiaoming Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Tingting Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mingjun Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Qingfeng Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Shaohua Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Haiping Xin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Haiping Xin.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Data availability statements

The resulting RNA-Seq data were deposited in The National Center for Biotechnology Information (NCBI) GEO database repository (https://www.ncbi.nlm.nih.gov/geo/).

Additional information

Communicated by Ying-Tang Lu.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhang, L., Cheng, J., Sun, X. et al. Overexpression of VaWRKY14 increases drought tolerance in Arabidopsis by modulating the expression of stress-related genes. Plant Cell Rep 37, 1159–1172 (2018). https://doi.org/10.1007/s00299-018-2302-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00299-018-2302-9

Keywords

Search

Navigation