Plant Molecular Biology Reporter

, Volume 34, Issue 4, pp 845–853 | Cite as

Expression of ZmHDZ4, a Maize Homeodomain-Leucine Zipper I Gene, Confers Tolerance to Drought Stress in Transgenic Rice

Original Paper


Climate change is predicted to be a major threat to crop yield due to increasing global temperatures and periods of unpredictable rainfall. Therefore, it is important to identify some new genes that can help plants cope with drought stress. In this study, a drought-induced homeodomain-leucine zipper (HD-Zip) I gene, ZmHDZ4, was isolated from maize and characterized for its role in drought stress. Transient expression experiments showed that ZmHDZ4 is localized to the nucleus. Yeast one-hybrid assays demonstrated that ZmHDZ4 has trans-activation activity, and that the minimal activation domain was ZmHDZ4-1. We found that overexpression of ZmHDZ4 in rice can enhance tolerance to drought and increase sensitivity to abscisic acid (ABA). Compared to wild-type plants, ZmHDZ4-expressing transgenic plants had lower relative electrolyte leakage (REL), lower malondialdehyde (MDA) levels, and increased proline contents under drought stress conditions, all of which may contribute to enhanced drought tolerance. Taken together, these results suggest that ZmHDZ4 functions as a transcriptional regulator that can positively affect plant drought tolerance. Thus, ZmHDZ4 is an excellent candidate gene with potential applications in molecular breeding to improve crop drought tolerance.


Drought Stress tolerance ZmHDZ4 Transcription factor ABA 



This work was supported by the National Natural Science Foundation of China (31501321), China Postdoctoral Science Foundation (2014M561811), the Anhui University Natural Science Research Projects (KJ2015A100), and the Anhui Postdoctoral Science Foundation.


  1. Agalou A, Purwantomo S, Overna¨s E, Johannesson H, Zhu X, Estiati A, de Kam RJ, Engstro¨m P, Slamet-Loedin IH, Zhu Z, Wang M, Xiong L, Meijer AH, Ouwerkerk PBF (2008) A genome-wide survey of HD-Zip genes in rice and analysis of drought-responsive family members. Plant Mol Biol 66:87–103CrossRefPubMedGoogle Scholar
  2. Ariel FD, Manavella PA, Dezar CA, Chan RL (2007) The true story of the HD-Zip family. Trends Plant Sci 12:419–426CrossRefPubMedGoogle Scholar
  3. Ariel F, Diet A, Verdenaud M, Gruber V, Frugier F, Chan R (2010) Environmental regulation of lateral root emergence in Medicago truncatula requires the HD-Zip I transcription factor HB1. Plant Cell 22:2171–2183CrossRefPubMedPubMedCentralGoogle Scholar
  4. Bates L, Waldren R, Teare I (1973) Rapid determination of free proline for water-stress studies. Plant Soil 39:205–207CrossRefGoogle Scholar
  5. Dai X, Xu Y, Ma Q, Xu W, Wang T, Xue Y (2007) Overexpression of an R1R2R3 MYB gene, OsMYB3R-2, increases tolerance to freezing, drought, and salt stress in transgenic Arabidopsis. Plant Physiol 143:1739–1751CrossRefPubMedPubMedCentralGoogle Scholar
  6. Deng X, Phillips J, Bräutigam A, Engström P, Johannesson H, Ouwerkerk PBF, Ruberti I, Salinas J, Vera P, Iannacone R, Meijer AH, Bartels D (2006) A homeodomain leucine zipper gene from Craterostigma plantagineum regulates abscisic acid responsive gene expression and physiological responses. Plant Mol Biol 61:469–489CrossRefPubMedGoogle Scholar
  7. Dezar CA, Gago GM, Gonzalez DH, Chan RL (2005) HAHB-4, a sunflower homeobox-leucine zipper gene, confers drought tolerance to Arabidopsis thaliana plants. Transgenic Res 14:429–440CrossRefPubMedGoogle Scholar
  8. Gago GM, Almoguera C, Jordano J, Gonzales DH, Chan RL (2002) Hahb-4, a homeobox-leucine zipper gene potentially involved in abscisic acid-dependent responses to water stress in sunflower. Plant Cell Environ 25:633–640CrossRefGoogle Scholar
  9. Gao T, Wu Y, Zhang Y, Liu L, Ning Y, Wang D (2011) OsSDIR1 overexpression greatly improves drought tolerance in transgenic rice. Plant Mol Biol 76:145–156CrossRefPubMedGoogle Scholar
  10. Harris JC, Hrmova M, Lopato S, Langridge P (2011) Modulation of plant growth by HD-Zip class I and II transcription factors in response to environmental stimuli. New Phytol 190:823–837CrossRefPubMedGoogle Scholar
  11. Henriksson E, Olsson AS, Johannesson H, Johansson H, Hanson J, Engstrom P (2005) Homeodomain leucine zipper class I genes in Arabidopsis. Expression patterns and phylogenetic relationships. Plant Physiol 139:509–518CrossRefPubMedPubMedCentralGoogle Scholar
  12. Kim S, Kang JY, Cho DI, Park JH, Kim SY (2004) ABF2, an ABRE-binding bZIP factor, is an essential component of glucose signaling and its overexpression affects multiple stress tolerance. Plant J 40:75–87CrossRefPubMedGoogle Scholar
  13. Kong XP, Pan JW, Zhang MY, Xing X, Zhou Y, Liu Y, Li DP, Li DQ (2011) ZmMKK4, a novel group C mitogen-activated protein kinase kinase in maize (Zea mays), confers salt and cold tolerance in transgenic Arabidopsis. Plant Cell Environ 34(8):1291–1303CrossRefPubMedGoogle Scholar
  14. Lee YH, Oh HS, Cheon CI, Hwang IT, Kim YJ, Chun JY (2001) Structure and expression of the Arabidopsis thaliana homeobox gene Athb-12. Biochem Biophys Res Commun 284:133–141CrossRefPubMedGoogle Scholar
  15. Li MR, Lin XJ, Li HQ, Pan XP, Wu GJ (2011) Overexpression of AtNHX5 improves tolerance to both salt and water stress in rice (Oryza sativa L.). Plant Cell Tiss Org 107(2):283–293CrossRefGoogle Scholar
  16. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(T)(−Delta Delta C) method. Methods 25:402–408CrossRefPubMedGoogle Scholar
  17. Manavella PA, Arce AL, Dezar CA, Bitton F, Renou FP, Crespi M, Chan RL (2006) Cross-talk between ethylene and drought signaling pathways is mediated by the sunflower Hahb-4 transcription factor. Plant J 48:125–137CrossRefPubMedGoogle Scholar
  18. Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7:405–410CrossRefPubMedGoogle Scholar
  19. Shan H, Chen S, Jiang J, Chen F, Chen Y, Gu C, Li P, Song A, Zhu X, Gao H, Zhou G, Li T, Yang X (2011) Heterologous expression of the Chrysanthemum R2R3-MYB transcription factor CmMYB2 enhances drought and salinity tolerance, increases hypersensitivity to ABA and delays flowering in Arabidopsis thaliana. Mol Biotech 51:160–173CrossRefGoogle Scholar
  20. Shinozaki K, Yamaguchi-Shinozaki K (2000) Molecular responses to dehydration and low temperature: differences and cross-talk between two stress signaling pathways. Curr Opin Plant Biol 3:217–223CrossRefPubMedGoogle Scholar
  21. Shinozaki K, Yamaguchi-Shinozaki K, Seki M (2003) Regulatory network of gene expression in the drought and cold stress responses. Curr Opin Plant Biol 6:410–417CrossRefPubMedGoogle Scholar
  22. Soderman E, Hjellstrom M, Fahleson J, Engstrom P (1999) The HD-Zip gene ATHB6 in Arabidopsis is expressed in developing leaves, roots and carpels and up-regulated by water deficit conditions. Plant Mol Biol 40:1073–1083CrossRefPubMedGoogle Scholar
  23. Song SY, Chen Y, Zhao MG, Zhang WH (2012) A novel Medicago truncatula HD-Zip gene, MtHB2, is involved in abiotic stress responses. Environ Exp Bot 80:1–9CrossRefGoogle Scholar
  24. Tran LSP, Nakashima K, Sakuma Y, Simpson SD, Fujita Y, Maruyama K, Fujita M, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2004) Isolation and functional analysis of Arabidopsis stress-inducible NAC transcription factors that bind to a drought-responsive cis-element in the early responsive to dehydration stress 1 promoter. Plant Cell 16:2481–2498CrossRefPubMedPubMedCentralGoogle Scholar
  25. Xiong L, Schumaker KS, Zhu JK (2002) Cell signaling during cold, drought, and salt stress. Plant Cell 14:165–183CrossRefGoogle Scholar
  26. Xiong L, Wang RG, Mao G, Koczan JM (2006) Identification of drought tolerance determinants by genetic analysis of root response to drought stress and abscisic acid. Plant Physiol 142:1065–1074CrossRefPubMedPubMedCentralGoogle Scholar
  27. Yamaguchi-Shinozaki K, Shinozaki K (2005) Organization of cis-acting regulatory elements in osmotic- and cold-stress-responsive promoters. Trends Plant Sci 10:88–94CrossRefPubMedGoogle Scholar
  28. Zhang SJ, Li N, Gao F, Yang AF, Zhang JR (2010) Over-expression of TsCBF1 gene confers improved drought tolerance in transgenic maize. Mol Breed 26(3):455–465CrossRefGoogle Scholar
  29. Zhang S, Haider I, Kohlen W, Jiang L, Bouwmeester H, Meijer AH (2012) Function of the HD-Zip I gene Oshox22 in ABA-mediated drought and salt tolerances in rice. Plant Mol Biol 80:571–585CrossRefPubMedGoogle Scholar
  30. Zhao Y, Zhou Y, Jiang H, Li X, Gan D, Peng X (2011) Systematic analysis of sequences and expression patterns of drought-responsive members of the HD-Zip gene family in maize. PLoS One 6:e28488CrossRefPubMedPubMedCentralGoogle Scholar
  31. Zhao Y, Ma Q, Jin X, Peng X, Liu J, Deng L, Yan H, Sheng L, Jiang H, Cheng B (2014) A novel maize homeodomain-leucine zipper (HD-Zip) I gene, Zmhdz10, positively regulates drought and salt tolerance in both rice and Arabidopsis. Plant Cell Physiol 55:1142–1156CrossRefPubMedGoogle Scholar
  32. Zhu JK (2002) Salt and drought stress signal transduction in plants. Ann Rev Plant Biol 53:247–273CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Jiandong Wu
    • 1
  • Wei Zhou
    • 1
  • Xuefeng Gong
    • 1
  • Beijiu Cheng
    • 1
  1. 1.Key Laboratory of Crop Biology of Anhui Province, School of Life SciencesAnhui Agricultural UniversityHefeiChina

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