Skip to main content
Log in

Functional analysis of a grape WRKY30 gene in drought resistance

  • Original Article
  • Published:
Plant Cell, Tissue and Organ Culture (PCTOC) Aims and scope Submit manuscript

Abstract

WRKY transcription factors constitute a large protein family in plants and take part in a variety of stress response processes, such as drought resistance. In this study, the drought resistant function of a WRKY transcription factor gene WRKY30 from a wine grape cultivar ‘Vidal Blanc’ was investigated. Quantitative real-time PCR (qRT-PCR) analysis indicated that the expression of VvWRKY30 was induced by drought stress and stress response related signal molecules, such as abscisic acid (ABA), nitric oxide and salicylic acid. Phenotypic analysis of transgenic Arabidopsis indicated that over-expression of VvWRKY30 conferred increased drought resistance, demonstrated by enhancements in seed germination rate and root development, as well as a decrease in water loss and stomatal aperture under drought conditions. In further study, we found that VvWRKY30 resistance to drought occurred mainly by promoting the expression of regulators of an ABA-dependent and independent signal pathway. Over-expression of VvWRKY30 also up-regulated the expressions of proline synthetase gene P5CS1, sucrose synthase gene SS4, beta-amylase gene BAM4, glucose-6-phosphate dehydrogenase gene G6PDH, and antioxidant enzyme synthesis genes Cu/ZnSOD, CAT2 and POD2. The content of proline and soluble sugar, as well as the activities of superoxide dismutase, peroxidase, and catalase were increased. Overall, our results demonstrate that VvWRKY30 positively regulates drought response by regulating ABA signal pathway, modulating proline and soluble sugar metabolism, as well as activating the scavenging systems of reactive oxygen species.

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

Access this article

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

Similar content being viewed by others

Abbreviations

ABA:

Abscisic acid

BAM:

Beta-amylase

CAT:

Catalase

DAB:

Diaminobenzidine

G6PDH:

Glucose-6-phosphate dehydrogenase

NBT:

Nitroblue tetrazolium

NO:

Nitric oxide

P5CS:

1-Pyrroline-5-carboxylate synthetase

POD:

Peroxidase

SA:

Salicylic acid

SNP:

Sodium nitroprusside

SOD:

Superoxide dismutase

SS:

Sucrose synthase

References

  • Agarwal P, Jha B (2010) Transcription factors in plants and ABA dependent and independent abiotic stress signaling. Biol Plant 54(2):201–212

    Article  CAS  Google Scholar 

  • Agarwal PK, Agarwal P, Reddy MK, Sopory SK (2006) Roles of DREB transcription factors in abiotic and biotic stress tolerance in plants. Plant Cell Rep 25:1263–1274

    Article  CAS  PubMed  Google Scholar 

  • Bakshi M, Oelmüller R (2014) WRKY transcription factors. Plant Signal Behav 9:e27700

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Che Y, Liu X, Xiao P, Yin P, Liu X (2015) Evaluation of salt tolerance of wine grape. North Hortic 15(23):18–22

    Google Scholar 

  • Chen Y, Yang M, Ding W, Zhao Y, Li X, Xiao K (2017) Wheat ZFP gene TaZFP593;l mediates the N-starvation adaptation of plants through regulating N acquisition and the ROS metabolism. Plant Cell Tissue Organ Cult 129(2):271–288

    Article  CAS  Google Scholar 

  • Chu X, Wang C, Chen X, Lu W, Li H, Wang X, Hao L, Guo X (2015) The cotton WRKY gene GhWRKY41 positively regulates salt and drought stress tolerance in transgenic Nicotiana benthamiana. Plos ONE 10:e0143022

    Article  PubMed  PubMed Central  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 

  • Corso M, Vannozzi A, Maza E, Vitulo N, Meggio F, Pitacco A, Telatin A, D’Angelo M, Feltrin E, Negri AS, Prinsi B, Valle G, Ramina A, Bouzayen M, Bonghi C, Lucchin M (2015) Comprehensive transcript profiling of two grapevine rootstock genotypes contrasting in drought susceptibility links the phenylpropanoid pathway to enhanced tolerance. J Exp Bot 66(19):5739–5752

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  • Fan Q, Song A, Jiang J, Zhang T, Sun H, Wang Y, Chen S, Chen F (2016) CmWRKY1 Enhances the dehydration tolerance of chrysanthemum through the regulation of ABA-associated genes. PloS ONE 11(3):e0150572

    Article  PubMed  PubMed Central  Google Scholar 

  • Gong X, Zhang J, Hu J, Wang W, Wu H, Zhang Q, Liu ZH (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(11):2248–2262

    Article  CAS  PubMed  Google Scholar 

  • Guo X, Zhang L, Zhu J, Wang A, Liu H (2017) Christolea crassifolia HARDY gene enhances drought stress tolerance in transgenic tomato plants. Plant Cell Tiss Organ Cult 129(3):469–481

    Article  CAS  Google Scholar 

  • Hao J, Ma Q, Hou L, Zhao F, Liu X (2017) VvWRKY13 enhances ABA biosynthesis in Vitis vinifera. Acta Soc Bot Pol. https://doi.org/10.5586/asbp.3546

    Google Scholar 

  • Jaradat MR, Feurtado JA, Huang D, Lu Y, Cutler AJ (2013) Multiple roles of the transcription factor AtMYBR1/AtMYB44 in ABA signaling, stress responses, and leaf senescence. BMC Plant Biol 13:192

    Article  PubMed  PubMed Central  Google Scholar 

  • Jiang YJ, Liang G, Yu DQ (2012) Activated expression of WRKY57 confers drought tolerance in Arabidopsis. Mol Plant 5(6):1375–1388

    Article  CAS  PubMed  Google Scholar 

  • Li ZG, Jin JZ (2016) Hydrogen sulfide partly mediates abscisic acid-induced heat tolerance in tobacco (Nicotiana tabacum L.) suspension cultured cells. Plant Cell Tiss Organ Cult 125(2):207–214

    Article  CAS  Google Scholar 

  • Li D, Li Y, Zhang L, Wang X, Zhao Z, Tao Z, Wang J, Wang J, Lin M, Li X, Yang Y (2014) Arabidopsis ABA receptor RCAR1/PYL9 interacts with an R2R3-Type MYB transcription factor, AtMYB44. Int J Mol Sci 15(5):8473–8490

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liu H, Yang W, Liu D, Han Y, Zhang A, Li S (2011) Ectopic expression of a grapevine transcription factor VvWRKY11 contributes to osmotic stress tolerance in Arabidopsis. Mol Biol Rep 38:417–427

    Article  CAS  PubMed  Google Scholar 

  • Liu ZQ, Yan L, Wu Z, Mei C, Lu K, Yu YT, Liang S, Zhang XF, Wang XF, Zhang DP (2012) Cooperation of three WRKY-domain transcription factors WRKY18, WRKY40, and WRKY60 in repressing two ABA-responsive genes ABI4 and ABI5in Arabidopsis. J Exp Bot 63(18):6371–6392

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7:405–410

    Article  CAS  PubMed  Google Scholar 

  • Nanjo T, Kobayashi M, Yoshiba Y, Sanada Y, Wada K, Tsukaya H, Kakubari Y, Yamaguchi-Shinozaki K, Shinozaki K (1999) Biological functions of proline in morphogenesis and osmotolerance revealed in antisense transgenic Arabidopsis thaliana. Plant J 18:185–193

    Article  CAS  PubMed  Google Scholar 

  • Reid KE, Olsson N, Schlosser J, Peng F, Lund ST (2006) An optimized grapevine RNA isolation procedure and statistical determination of reference genes for real-time RT-PCR during berry development. BMC Plant Biol 6:27

    Article  PubMed  PubMed Central  Google Scholar 

  • Ren XZ, Chen ZZ, 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  CAS  PubMed  PubMed Central  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  CAS  PubMed  Google Scholar 

  • Tamirisa S, Reddy VD, Rao KV (2014) Ectopic expression of pigeonpea cold and drought regulatory protein (CcCDR) in yeast and tobacco affords multiple abiotic stress tolerance. Plant Cell Tiss Organ Cult 119(3):489–499

    Article  CAS  Google Scholar 

  • Wang J, Sun PP, Chen CL, Wang Y, Fu XZ, Liu JH (2011) An arginine decarboxylase gene PtADC from Poncirus trifoliata confers abiotic stress tolerance and promotes primary root growth in Arabidopsis. J Exp Bot 62:2899–2914

    Article  CAS  PubMed  Google Scholar 

  • Wang LX, Ma XY, Che YM, Hou LX, Xin L, Zhang W (2015a) Extracellular ATP mediates H2S-regulated stomatal movements and guard cell K+ current in a H2O2-dependent manner in Arabidopsis. Sci Bull 60(4):419–427

    Article  CAS  Google Scholar 

  • Wang Y, Feng L, Zhu Y, Li Y, Yan H, Xiang Y (2015b) Comparative genomic analysis of the WRKY III gene family in populus, grape, Arabidopsis and rice. Biol Direct 10:48

    Article  PubMed  PubMed Central  Google Scholar 

  • Wang K, Zhong M, Wu YH, Bai ZY, Liang QY, Liu QL, Pan YZ, Zhang L, Jiang BB, Jia Y, Liu GL (2017) Overexpression of chrysanthemum transcription factor gene DgNAC1 improves the salinity tolerance in chrysanthemum. Plant Cell Rep 36(4):571–581

    Article  CAS  PubMed  Google Scholar 

  • Willems E, Leyns L, Vandesompele J (2008) Standardization of real-time PCR gene expression data from independent biological replicates. Anal Biochem 379:127–129

    Article  CAS  PubMed  Google Scholar 

  • Xiao W, Hu S et al (2017) A glucuronokinase gene in Arabidopsis, AtGlcAK, is involved in drought tolerance by modulating sugar metabolism. Plant Mol Biol Rep 35(2):298–311

    Article  CAS  Google Scholar 

  • Yamaguchi-Shinozaki K, Shinozaki K (2005) Organization of cis-acting regulatory elements in osmotic- and cold-stress-responsive promoters. Trends Plant Sci 10:88–94

    Article  CAS  PubMed  Google Scholar 

  • Yang Z, Xu LX, Yu JJ, DaCosta M, Huang BR (2013) Changes in carbohydrate metabolism in two kentucky bluegrass cultivars during drought stress and recovery. J Am Soc Hortic Sci 138:24–30

    CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant Nos. 31572107, 31501331and 31401844). High-level Talent Research Fund of Qingdao Agricultural University (Grant No. 6631115032). Shandong “Bohai Granary” Science and Technology Demonstration Project (Grant No. 2017BHLC021).

Author information

Authors and Affiliations

Authors

Contributions

DZ and XL conceived and designed research. YC and DZ conducted experiments. PX and YG performed the experiments. XL contributed new reagents or analytical tools. DZ and LH analyzed data. DZ wrote the manuscript. All authors read and approved the manuscript.

Corresponding author

Correspondence to Xin Liu.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Communicated by Klaus Eimert.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhu, D., Che, Y., Xiao, P. et al. Functional analysis of a grape WRKY30 gene in drought resistance. Plant Cell Tiss Organ Cult 132, 449–459 (2018). https://doi.org/10.1007/s11240-017-1341-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-017-1341-1

Keywords

Navigation