Russian Journal of Plant Physiology

, Volume 64, Issue 6, pp 899–905 | Cite as

Antioxidative enzymes, calcium, and ABA signaling pathway are required for the stress tolerance of transgenic wheat plant by the ectopic expression of harpin protein fragment Hpa110–42 under heat stress

  • D. F. Wang
  • X. J. Pang
  • F. Yang
  • L. S. Kou
  • X. Zhang
  • P. X. Yu
  • Y. B. Niu
Research Papers


Genetic engineering for heat stress tolerance can promote crop growth and improve yield. One wheat (Triticum aestivum L.) line Y16 (wild type) and two transgenic plants (Y16-3 and Y16-46) that express Hpa110-42, a functional fragment of harpin protein, were used in this study to investigate their possible abiotic stress tolerance under heat stress. Results showed that enhanced thermotolerance was observed in the Y16-3 and Y16-46 lines over the control wheat under stress conditions. However, this increased stress tolerance was significantly abolished by specific inhibitors such as fluridone or sodium tungstate (i.e., arrests abscisic acid (ABA) biosynthesis) and EGTA or La3+ (i.e., arrests Ca2+ signaling pathway) under heat exposure. By contrast, high activities of antioxidant enzymes such as superoxide dismutase, catalase, and ascorbate peroxidase (but not peroxidase) and low levels of oxidative damage (thiobarbituric acid reactive substance (TBARS) and chlorophyll) were detected in transgenic wheat lines compared with the control plant under stress exposure. However, this significant difference diminished after the addition of these specific inhibitors. Furthermore, a slight increase of H2O2 was observed in the transgenic plant, instead of the control, without the addition of chemicals under heat stress. These results suggested that antioxidant enzymes, calcium, and ABA signaling pathways were involved in this Hpa110–42-mediated thermotolerance of transgenic wheat plants under stress exposure. Finally, a hypothetical model based on H2O2 signaling was proposed to illustrate the possible mechanism of this enhanced heat stress tolerance.


Triticum aestivum abscisic acid antioxidative enzyme harpin protein high temperature hydrogen peroxide 



ascorbate peroxidase




hydrogen peroxide




superoxide dismutase


thiobarbituric acid reactive substance


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Porter, J., Rising temperatures are likely to reduce crop yields, Nature, 2005, vol. 436, p.174.CrossRefPubMedGoogle Scholar
  2. 2.
    Gill, S. and Tuteja, N., Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants, Plant Physiol. Biochem., 2010, vol. 48, pp. 909–930.CrossRefPubMedGoogle Scholar
  3. 3.
    Mittler, R., Oxidative stress, antioxidants and stress tolerance, Trends Plant Sci., 2002, vol. 7, pp. 405–410.CrossRefPubMedGoogle Scholar
  4. 4.
    Sairam, R., Srivastava, G., and Saxena, D., Increased antioxidant activity under elevated temperatures: a mechanism of heat stress tolerance in wheat genotypes, Biol. Plant., 2000, vol. 43, pp. 245–251.CrossRefGoogle Scholar
  5. 5.
    Gong, M., Chen, S., Song, Y., and Li, Z., Effect of calcium and calmodulin on intrinsic heat tolerance in relation to antioxidant systems in maize seedlings, Aust. J. Plant Physiol., 1997, vol. 24, pp. 371–379.CrossRefGoogle Scholar
  6. 6.
    Larkindale, J. and Knight, M., Protection against heat stress-induced oxidative damage in Arabidopsis involves calcium, abscisic acid, ethylene, and salicylic acid, Plant Physiol., 2002, vol. 128, pp. 682–695.PubMedGoogle Scholar
  7. 7.
    Larkindale, J. and Huang, B., Thermotolerance and antioxidant systems in Agrostis stolonifera: involvement of salicylic acid, abscisic acid, calcium, hydrogen peroxide, and ethylene, J. Plant Physiol., 2004, vol. 161, pp. 405–413.PubMedGoogle Scholar
  8. 8.
    Ton, J., Jakab, G., Toquin, V., Flors, V., Lavicoli, A., Maeder, M., Métraux, J., and Mauch-Mani, B., Dißsecting the β-aminobutyric acid-induced priming phenomenon in Arabidopsis, Plant Cell, 2005, vol. 17, pp. 987–999.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Bruce, T., Matthes, M., Napier, J., and Pickett, J., Stressful “memories” of plants: evidence and possible mechanisms, Plant Sci., 2007, vol. 173, pp. 603–608.CrossRefGoogle Scholar
  10. 10.
    Iba, K., Acclimative response to temperature stress in higher plants: approaches of gene engineering for temperature tolerance, Annu. Rev. Plant Biol., 2002, vol. 53, pp. 225–245.CrossRefPubMedGoogle Scholar
  11. 11.
    Wang, W., Vinocur, B., and Altman, A., Plant responses to drought, salinity and extreme temperatures: towards genetic engineering for stress tolerance, Planta, 2003, vol. 218, pp. 1–14.PubMedGoogle Scholar
  12. 12.
    Wei, Z., Lacy, R., Zumoff, C., Bauer, D., He, S., Collmer, A., and Beer, S., Harpin, elicitor of the hypersensitive response produced by the plant pathogen Erwinia amylovora, Science, 1992, vol. 257, pp. 85–88.PubMedGoogle Scholar
  13. 13.
    Dong, H., Delaney, T., Bauer, D., and Beer, S., Harpin induces disease resistance in Arabidopsis through the systemic acquired resistance pathway mediated by salicylic acid and the NIM1 gene, Plant J., 1999, vol. 20, pp. 207–215.CrossRefPubMedGoogle Scholar
  14. 14.
    Wang, D., Wang, Y., Fu, M., Mu, S., Han, B., Ji, H., Cai, H., Dong, H., and Zhang, C., Transgenic expression of the functional fragment Hpa110–42 of the harpin protein Hpa1 imparts enhanced resistance to powdery mildew in wheat, Plant Dis., 2014, vol. 98, pp. 448–455.CrossRefGoogle Scholar
  15. 15.
    Dong, H., Yu, H., Bao, Z., Guo, X., Peng, J., Yao, Z., Chen, G., Qu, S., and Dong, H., The ABI2-dependent abscisic acid signalling controls HrpN-induced drought tolerance in Arabidopsis, Planta, 2005, vol. 221, pp. 313–327.CrossRefPubMedGoogle Scholar
  16. 16.
    Zhang, L., Xiao, S., Li, W., Feng, W., Li, J., Wu, Z., Gao, X., Liu, F., and Shao, M., Overexpression of a harpin-encoding gene hrf1 in rice enhances drought tolerance, J. Exp. Bot., 2011, vol. 62, pp. 4229–4238.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Kunkel, B. and Brooks, D., Cross talk between signaling pathways in pathogen defense, Curr. Opin. Plant Biol., 2002, vol. 5, pp. 325–331.CrossRefPubMedGoogle Scholar
  18. 18.
    Yoshioka, T., Endo, T., and Satoh, S., Restoration of seed germination at supraoptimal temperatures by fluridone, an inhibitor of abscisic acid biosynthesis, Plant Cell Physiol., 1998, vol. 39, pp. 307–312.Google Scholar
  19. 19.
    Jiang, M. and Zhang, J., Cross-talk between calcium and reactive oxygen species originated from NADPH oxidase in abscisic acid-induced antioxidant defence in leaves of maize seedlings, Plant Cell Environ., 2003, vol. 26, pp. 929–939.CrossRefPubMedGoogle Scholar
  20. 20.
    Deng, B., Jin, X., Yang, Y., Lin, Z., and Zhang, Y., The regulatory role of riboflavin in the drought tolerance of tobacco plants depends on ROS production, Plant Growth Regul., 2013, vol. 72, pp. 269–277.CrossRefGoogle Scholar
  21. 21.
    Heath, R. and Packer, L., Photoperoxidation in isolated chloroplast. I. Kinetics and stoichiometry of fatty acid peroxidation, Arch. Biochem. Biophys., 1968, vol. 125, pp. 180–198.CrossRefGoogle Scholar
  22. 22.
    Gay, C., Collins, J., and Gebicki, J., Hydroperoxide assay with the ferric-xylenol orange complex, Anal. Biochem., 1999, vol. 273, pp. 149–155.CrossRefPubMedGoogle Scholar
  23. 23.
    Lichtenthaler, H., Chlorophyll and carotenoids: pigments of photosynthetic biomembranes, Meth. Enzymol., 1987, vol. 148, pp. 349–382.Google Scholar
  24. 24.
    Jiang, M. and Zhang, J., Water stress-induced abscisic acid accumulation triggers the increased generation of reactive oxygen species and up-regulates the activities of antioxidant enzymes in maize leaves, J. Exp. Bot., 2002, vol. 53, pp. 2401–2410.CrossRefPubMedGoogle Scholar
  25. 25.
    Maehly, P. and Chance, M., The assay of catalase and peroxidases, in Methods of Biochemical Analysis, Gluck, D, Ed., New York: Wiley, 1954, pp. 357–424.CrossRefGoogle Scholar
  26. 26.
    Bradford, M., A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding, Anal. Biochem., 1976, vol. 72, pp. 248–254.CrossRefPubMedGoogle Scholar
  27. 27.
    Dat, J., Vandenabeele, S., Vranová, E., Van, M.M., Inzé, D., and Van, B.F., Dual action of the active oxygen species during plant stress responses, Cell Mol. Life Sci., 2000, vol. 57, pp. 779–795.CrossRefPubMedGoogle Scholar
  28. 28.
    Suzuki, N. and Mittler, R., Reactive oxygen species and temperature stresses: a delicate balance between signaling and destruction, Physiol. Plant., 2006, vol. 126, pp. 45–51.CrossRefGoogle Scholar
  29. 29.
    Hu, X., Zhang, A., Zhang, J., and Jiang, M., Abscisic acid is a key inducer of hydrogen peroxide production in leaves of maize plants exposed to water stress, Plant Cell Physiol., 2006, vol. 47, pp. 1484–1495.CrossRefPubMedGoogle Scholar
  30. 30.
    Desikan, R., Burnett, E., Hancock, J., and Neill, S., Harpin and hydrogen peroxide induce the expression of a homologue of gp91-phox in Arabidopsis thaliana suspension cultures, J. Exp. Bot., 1998, vol. 49, no. 327, pp. 1767–1771.CrossRefGoogle Scholar

Copyright information

© Pleiades Publishing, Ltd. 2017

Authors and Affiliations

  • D. F. Wang
    • 1
  • X. J. Pang
    • 1
  • F. Yang
    • 1
  • L. S. Kou
    • 1
  • X. Zhang
    • 1
  • P. X. Yu
    • 1
  • Y. B. Niu
    • 1
  1. 1.College of Life ScienceShanxi Agricultural UniversityTaigu, ShanXi ProvinceP.R. China

Personalised recommendations