Russian Journal of Genetics

, Volume 55, Issue 3, pp 330–336 | Cite as

RT-qPCR Analysis of Host Defense-Related Genes in Nonhost Resistance: Wheat-Bgh Interaction

  • A. RezaeiEmail author
  • S. Mahdian
  • V. Babaeizad
  • S. H. Hashemi-PetroudiEmail author
  • S. M. Alavi


Non-host resistance (NHR) is a plant defense system against the majority of microbial pathogens. Blumeria graminis f. sp. hordei (Bgh), is an obligate biotrophic ascomycete fungal pathogen that can grow and reproduce only on living cells of wild or domesticated barley (Hordeum sp.).‎ In this study, the expression pattern of eight defense-related genes including NPR1, PR2, PR5, PR1-2, Lipase, LTP, PAL and POX was assessed in bread wheat (Triticum aestivum cv. Darya) against the powdery mildew fungus Bgh. Hydrogen peroxide (H2O2) activity in leaf tissues was also detected histochemically with 3,3-diaminobenzidine (DAB) staining. Result showed that in response to virulence agent, Bgh spores, papillae and hypersensitive response (HR) were formed in the 53.9 and 40.1% of infected area, respectively while in the rest of the infected area (6%) spores were not germinated. According to the histochemical analysis, the wheat NHR against Bgh belongs to type IІ NHR and display a multi-layered feature including wax composition, pre-formed barriers, papilla formation, hypersensitive response. According to results of expression analysis, the Pox, PR5 and PR1-2 expression level were clearly upregulated 24 hours post inoculation (hpi) and LTP was identified as an early signaling response to Bgh. In this study NPR1 was down regulated during all given time points, thus we suggested that salicylic acid (SA) signaling pathway may be mediated by a NPR1-independent mechanism(s) in wheat-Bgh interaction. The coordinated induction of a set of so-called systemic acquired resistance (SAR) genes including POX, PR1-2 and PR5 was observed in wheat-Bgh interaction. Expression pattern of defense-related genes at the wheat-Bgh interaction can be taken as an indication of their functional relevance at different time points of tissue infection.


wheat-Bgh interaction nonhost resistance barley powdery mildew qPCR papilla formation NPR1 



This research was supported by the Genetic and Agricultural Biotechnology Institute of Tabarestan (GABIT), Sari Agricultural Sciences and Natural Resources University (SANRU).


The authors declare that they have no conflict of interest. This article does not contain any studies involving animals or human participants performed by any of the authors.


  1. 1.
    Egusa, M., Miwa, T., Kaminaka, H., Takano, Y., and Kodama, M., Nonhost resistance of Arabidopsis thaliana against Alternaria alternata involves both pre- and postinvasive defenses but is collapsed by AAL-toxin in the absence of LOH2, Phytopathology, 2013, vol. 103, pp. 733—740.CrossRefGoogle Scholar
  2. 2.
    Adlung, N., Prochaska, H., Thieme, S., Banik, A., Blüher, D., John, P., Nagel, O., Schulze, S., Gantner, J., and Delker, C., Non-host resistance induced by the Xanthomonas effector XopQ is widespread within the genus Nicotiana and functionally depends on EDS1, Front. Plant Sci., 2016, vol. 7.Google Scholar
  3. 3.
    Cheng, Y., Zhang, H., Yao, J., Wang, X., Xu, J., Han, Q., Wei, G., Huang, L., and Kang, Z., Characterization of non-host resistance in broad bean to the wheat stripe rust pathogen, BMC Plant Biol., 2012, vol. 12, p. 96.Google Scholar
  4. 4.
    Ayliffe, M., Jin, Y., Kang, Z., Persson, M., Steffenson, B., Wang, S., and Leung, H., Determining the basis of nonhost resistance in rice to cereal rusts, Euphytica, 2011, vol. 179, pp. 33—40.CrossRefGoogle Scholar
  5. 5.
    Gill, U.S., Uppalapati, S.R., Nakashima, J., and Mysore, K.S., Characterization of Brachypodium distachyon as a nonhost model against switchgrass rust pathogen Puccinia emaculata, BMC Plant Biol., 2015, vol. 15, p. 113.Google Scholar
  6. 6.
    Lee, H.-A., Lee, H.-Y., Seo, E., Lee, J., Kim, S.-B., Oh, S., Choi, E., Choi, E., Lee, S.E., and Choi, D., Current understandings of plant nonhost resistance, Mol. Plant—Microbe Interact., 2017, vol. 30, pp. 5–15.CrossRefGoogle Scholar
  7. 7.
    Thordal-Christensen, H., Fresh insights into processes of nonhost resistance, Curr. Opin. Plant Biol., 2003, vol. 6, pp. 351—357.CrossRefGoogle Scholar
  8. 8.
    Collinge, D.B., Cell wall appositions: the first line of defence, J. Exp. Bot., 2009, vol. 60, pp. 351—352.CrossRefGoogle Scholar
  9. 9.
    Mysore, K.S. and Ryu, C.-M., Nonhost resistance: how much do we know?, Trends Plant Sci., 2004, vol. 9, pp. 97—104.CrossRefGoogle Scholar
  10. 10.
    Ellis, J., Insights into nonhost disease resistance: can they assist disease control in agriculture?, Plant Cell, 2006, vol. 18, pp. 523—528.CrossRefGoogle Scholar
  11. 11.
    Cohn, J., Sessa, G., and Martin, G.B., Innate immunity in plants, Curr. Opin. Immunol., 2001, vol. 13, pp. 55—62.CrossRefGoogle Scholar
  12. 12.
    Gaudet, D. A., Wang, Y., Penniket, C., Lu, Z., Bakkeren, G., and Laroche, A., Morphological and molecular analyses of host and nonhost interactions involving barley and wheat and the covered smut pathogen Ustilago hordei, Mol. Plant—Microbe Interact., 2010, vol. 23, pp. 1619—1634.CrossRefGoogle Scholar
  13. 13.
    Loehrer, M., Langenbach, C., Goellner, K., Conrath, U., and Schaffrath, U., Characterization of nonhost resistance of Arabidopsis to the Asian soybean rust, Mol. Plant—Microbe Interact., 2008, vol. 21, pp. 1421—1430.CrossRefGoogle Scholar
  14. 14.
    Zhou, F., Kurth, J., Wei, F., Elliott, C., Valè, G., Yahiaoui, N., Keller, B., Somerville, S., Wise, R., and Schulze-Lefert, P., Cell-autonomous expression of barley Mla1 confers race-specific resistance to the powdery mildew fungus via a Rar1-independent signaling pathway, Plant Cell, 2001, vol. 13, pp. 337—350.CrossRefGoogle Scholar
  15. 15.
    Livak, K.J. and Schmittgen, T.D., Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method, Methods, 2001, vol. 25, pp. 402—408.CrossRefGoogle Scholar
  16. 16.
    Leon, J., Lawton, M.A. and Raskin, I., Hydrogen peroxide stimulates salicylic acid biosynthesis in tobacco, Plant Physiol., 1995, vol. 108, pp. 1673—1678.CrossRefGoogle Scholar
  17. 17.
    Saleh, A., Withers, J., Mohan, R., Marqués, J., Gu, Y., Yan, S., Zavaliev, R., Nomoto, M., Tada, Y., and Dong, X., Posttranslational modifications of the master transcriptional regulator NPR1 enable dynamic but tight control of plant immune responses, Cell Host Microbe, 2015, vol. 18, pp. 169—182.CrossRefGoogle Scholar
  18. 18.
    Shah, J., The salicylic acid loop in plant defense, Curr. Opin. Plant Biol., 2003, vol. 6, pp. 365—371.CrossRefGoogle Scholar
  19. 19.
    Kim, D.S. and Hwang, B.K., An important role of the pepper phenylalanine ammonia-lyase gene (PAL1) in salicylic acid-dependent signalling of the defence response to microbial pathogens, J. Exp. Bot., 2014, eru109.Google Scholar
  20. 20.
    Dong, X., NPR1, all things considered, Curr. Opin. Plant Biol., 2004, vol. 7, pp. 547—552.CrossRefGoogle Scholar
  21. 21.
    Wildermuth, M.C., Dewdney, J., Wu, G., and Ausubel, F.M., Isochorismate synthase is required to synthesize salicylic acid for plant defence, Nature, 2001, vol. 414, pp. 562—565.CrossRefGoogle Scholar
  22. 22.
    van Loon, L.C., Rep, M., and Pieterse, C.M., Significance of inducible defense-related proteins in infected plants, Annu. Rev. Phytopathol., 2006, vol. 44, pp. 135—162.CrossRefGoogle Scholar
  23. 23.
    Almagro, L., Ros, L.G., Belchi-Navarro, S., Bru, R., Barceló, A.R., and Pedreño, M., Class III peroxidases in plant defence reactions, J. Exp. Bot., 2009, vol. 60, pp. 377—390.CrossRefGoogle Scholar
  24. 24.
    Vance, C., Kirk, T., and Sherwood, R., Lignification as a mechanism of disease resistance, Annu. Rev. Phytopathol., 1980, vol. 18, pp. 259—288.CrossRefGoogle Scholar
  25. 25.
    Bowles, D.J., Defense-related proteins in higher plants, Annu. Rev. Biochem., 1990, vol. 59, pp. 873—907.CrossRefGoogle Scholar
  26. 26.
    Gonzalez, A. M., Marcel, T.C., Kohutova, Z., Stam, P., van der Linden, C.G., and Niks, R.E., Peroxidase profiling reveals genetic linkage between peroxidase gene clusters and basal host and non-host resistance to rusts and mildew in barley, PLoS One, 2010, vol. 5. e10495Google Scholar
  27. 27.
    Rivière, M.-P., Marais, A., Ponchet, M., Willats, W., and Galiana, E., Silencing of acidic pathogenesis-related PR-1 genes increases extracellular β-(1→ 3)-glucanase activity at the onset of tobacco defence reactions, J. Exp. Bot., 2008, vol. 59, pp. 1225–1239.Google Scholar
  28. 28.
    Van Loon, L. and Van Strien, E., The families of pathogenesis-related proteins, their activities, and comparative analysis of PR-1 type proteins, Physiol. Mol. Plant Pathol., 1999, vol. 55, pp. 85–97.Google Scholar
  29. 29.
    Wu, S.-W., Wang, H.-W., Yang, Z.-D., and Kong, L.-R., Expression comparisons of pathogenesis-related (PR) genes in wheat in response to infection/infestation by Fusarium, Yellow dwarf virus (YDV) aphid-transmitted and hessian fly, J. Integr. Agric., 2014, vol. 13, pp. 926–936.Google Scholar
  30. 30.
    Oide, S., Bejai, S., Staal, J., Guan, N., Kaliff, M., and Dixelius, C., A novel role of PR2 in abscisic acid (ABA) mediated, pathogen-induced callose deposition in Arabidopsis thaliana, New Phytol., 2013, vol. 200, pp. 1187–1199.Google Scholar
  31. 31.
    Lu, Z.-X., Gaudet, D. A., Frick, M., Puchalski, B., Genswein, B., and Laroche, A., Identification and characterization of genes differentially expressed in the resistance reaction in wheat infected with Tilletia tritici, the common bunt pathogen, BMB Rep., 2005, vol. 38, pp. 420–431.Google Scholar
  32. 32.
    Ahangar, L., Ranjbar, G. A., Babaeizad, V., Najafi Zarrini, H., and Biabani, A., Assay of NPR1 gene expression in wheat under powdery mildew stress, J. Crop Protect., 2017, vol. 6, pp. 157–166.Google Scholar
  33. 33.
    Liu, B., Xue, X., Cui, S., Zhang, X., Han, Q., Zhu, L., Liang, X., Wang, X., Huang, L., and Chen, X., Cloning and characterization of a wheat β-1,3-glucanase gene induced by the stripe rust pathogen Puccinia striiformis f. sp. tritici, Mol. Biol. Rep., 2010, vol. 37, p. 1045.Google Scholar
  34. 34.
    Freitas, L.B. d., Koehler-Santos, P., and Salzano, F.M., Pathogenesis-related proteins in Brazilian wheat genotypes: protein induction and partial gene sequencing, Ciênc. Rural, 2003, vol. 33, pp. 497–500.Google Scholar
  35. 35.
    Purwar, S., Sundaram, S., Sinha, S., Gupta, A., Dobriyall, N., and Kumar, A., Expression and in silico characterization of phenylalanine ammonium lyase against karnal bunt (Tilletia indica) in wheat (Triticum aestivum), Bioinformation, 2013, vol. 9, p. 1013.Google Scholar
  36. 36.
    Xue, G.-P., Rae, A.L., White, R.G., Drenth, J., Richardson, T., and McIntyre, C.L., A strong root-specific expression system for stable transgene expression in bread wheat, Plant Cell Rep., 2016, vol. 35, pp. 469–481.Google Scholar

Copyright information

© Pleiades Publishing, Inc. 2019

Authors and Affiliations

  1. 1.Department of Plant Protection, Sari Agricultural Sciences and Natural Resources UniversitySariIran
  2. 2.Genetics and Agricultural Biotechnology Institute of Tabarestan, Sari Agricultural Sciences and Natural Resources UniversitySariIran

Personalised recommendations