Plant Molecular Biology Reporter

, Volume 31, Issue 2, pp 314–322 | Cite as

Characterization of a Hypersensitive Response-Induced Gene TaHIR3 from Wheat Leaves Infected with Leaf Rust

  • Xiu-Mei Yu
  • Wei-Quan Zhao
  • Wen-Xiang Yang
  • Fen Liu
  • Jia-Ping Chen
  • Claudia Goyer
  • Da-Qun Liu
Original Paper


Hypersensitive-induced reaction (HIR) proteins and more specific members of the proliferation, ion and death superfamily participate in response to pathogen attacks and development of spontaneous hypersensitive lesions in maize and barley leaves. In the present study, a full-length TaHIR3 (1,246-bp) was cloned and characterized from wheat near-isogenic line Thatcher-Lr15 infected by Puccinia triticina isolate 05-19-43② through a homology cloning strategy. A 4,203-bp sequence of TaHIR3 including five exons was also obtained from wheat Thatcher-Lr15 genomic DNA. TaHIR3 shared higher similarity with HIR3 isolated from wheat, barley, and maize. Gene expression of TaHIR3 was spatially measured in young wheat leaves, young roots, young stems, mature seeds, and temporally in wheat leaves inoculated with virulent and avirulent P. triticina. TaHIR3 was expressed in all samples except mature seeds and was upregulated in both combinations. More transcripts accumulated in the incompatible than compatible combination, implying a role in wheat growth and resistance to pathogens attack. A polyclonal antibody was prepared with recombinant protein purified from a prokaryotic expression system with the open reading frame of TaHIR3, and it detected the target protein in wheat leaves by Western blotting. Detection results at the protein level also showed that TaHIR3 was upregulated expression in wheat leaves infected with the leaf rust pathogen. Characterization of TaHIR3 and its expression profiles at the DNA and protein levels suggest that TaHIR3 and its deduced protein play a role in wheat HR causing by the leaf rust pathogen.


Triticum aestivum Hypersensitive-induced reaction protein Transcripts expression profiles Western blotting 



Hypersensitive response


Hypersensitive-induced reaction


Stomatins, prohibitins, flotillins, and HflK/C


Leucine-rich repeat protein


Hours postinoculation


Rapid amplification of cDNA ends


Untranslated regions


Reverse transcription-polymerase chain reaction


Quantitative reverse transcription-polymerase chain reaction


Base pair


Basic local alignment search tool


Open reading frame


Isoelectric point





This study was supported financially by the Natural Science Foundation of Hebei Province (No. C2010000762) and the National Natural Science Foundation of China (No. 30771391).


  1. Alvarez ME (2000) Salicylic acid in the machinery of hypersensitive cell death and disease resistance. Plant Mol Biol 44:429–442PubMedCrossRefGoogle Scholar
  2. Browman DT, Resek ME, Zajchowski LD, Robbins SM (2006) Erlin-1 and erlin-2 are novel members of the prohibitin family of proteins that define lipid-raft-like domain s of the ER. J Cell Sci 119:3149–3160PubMedCrossRefGoogle Scholar
  3. Browman DT, Hoegg MB, Robbins SM (2007) The SPFH domain-containing proteins: more than lipid raft markers. Trends Cell Biol 17:394–402PubMedCrossRefGoogle Scholar
  4. Chen JP, Yu XM, Zhao WQ, Li X, Meng T, Liu F, Yang WX, Zhang T, Liu DQ (2012) Temporal and tissue-specific expression of wheat TaHIR2 gene and resistant role of recombinant protein during interactions between wheat and leaf rust pathogen. Physiol Mol Plant Pathol 79:64–70CrossRefGoogle Scholar
  5. Choi HW, Kim YJ, Hwang BK (2011) The hypersensitive induced reaction and leucine-rich repeat proteins regulate plant cell death associated with disease and plant immunity. Mol Plant Microbe Interact 24:68–78PubMedCrossRefGoogle Scholar
  6. Dangl JL, Jones JD (2001) Plant pathogens and integrated defense responses to infection. Nature 411:826–833PubMedCrossRefGoogle Scholar
  7. Delledonne M, Zeier J, Marocco A, Lamb C (2001) Signal interactions between nitric oxide and reactive oxygen intermediates in the plant hypersensitive disease resistance response. Proc Natl Acad Sci USA 98:13454–13459PubMedCrossRefGoogle Scholar
  8. Heath MC (2000) Hypersensitive response-related death. Plant Mol Biol 44:321–334PubMedCrossRefGoogle Scholar
  9. Jung HW, Hwang BK (2007) The leucine-rich repeat (LRR) protein, CaLRR1, interacts with the hypersensitive induced reaction (HIR) protein, CaHIR1, and suppresses cell death induced by the CaHIR1 protein. Mol Plant Pathol 8:503–514PubMedCrossRefGoogle Scholar
  10. Jung HW, Lim CW, Lee SC, Choi HW, Hwang CH, Hwang BK (2008) Distinct roles of the pepper hypersensitive induced reaction protein gene CaHIR1 in disease and osmotic stress, as determined by comparative transcriptome and proteome analyses. Planta 227:409–425PubMedCrossRefGoogle Scholar
  11. Karrer EE, Beachy RN, Holt CA (1998) Cloning of tobacco genes that elicit the hypersensitive response. Plant Mol Biol 36:681–690PubMedCrossRefGoogle Scholar
  12. Kobayashi Y, Kobayashi I (2007) Depolymerization of the actin cytoskeleton induces defense responses in tobacco plants. J Gen Plant Pathol 73:360–364CrossRefGoogle Scholar
  13. Kolmer JA (2005) Tracking wheat rust on a continental scale. Curr Opin Plant Biol 8:441–449PubMedCrossRefGoogle Scholar
  14. Kozak M (1987) An analysis of 5′-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Res 15:8125–8148PubMedCrossRefGoogle Scholar
  15. Langhorst MF, Reuter A, Stuermer CA (2005) Scaffolding microdomains and beyond: the function of reggie/flotillin proteins. Cell Mol Life Sci 62:2228–2240PubMedCrossRefGoogle Scholar
  16. Liu J, DeYoung SM, Zhang M, Dold LH, Saltiel AR (2005) The stomatin/prohibitin/flotillin/HflK/C domain of flotillin-1 contains distinct sequences that direct plasma membrane localization and protein interactions in 3T3-L1 adipocytes. J Biol Chem 280:16125–16134PubMedCrossRefGoogle Scholar
  17. Marasas CN, Smale M, Singh RP (2004) The economic impact in developing countries of leaf rust resistance breeding in CIMMYT-related spring bread wheat. International Maize and Wheat Improvement Center, Mexico, DFGoogle Scholar
  18. Morrow IC, Parton RG (2005) Flotillins and the PHB domain protein family: rafts, worms and anaesthetics. Traffic 6:725–740PubMedCrossRefGoogle Scholar
  19. Mur LAJ, Kenton P, Lloyd AJ, Ougham H, Prats E (2008) The hypersensitive response; the centenary is upon us but how much do we know. J Exp Bot 59:501–520PubMedCrossRefGoogle Scholar
  20. Nadimpalli R, Yalpani N, Johal GS, Simmons CR (2000) Prohibitins, stomatins, and plant-disease response genes compose a protein superfamily that controls cell proliferation, ion channel regulation, and death. J Biol Chem 275:29579–29586PubMedCrossRefGoogle Scholar
  21. Park JM (2005) The hypersensitive response. A cell death during disease resistance. Plant Pathol J 21:99–101CrossRefGoogle Scholar
  22. Qi YP, Katagiri F (2009) Purification of low-abundance Arabidopsis plasma-membrane protein complexes and identification of candidate components. Plant J 57:932–944PubMedCrossRefGoogle Scholar
  23. Qi YP, Tsuda K, Nguyen LV, Wang X, Lin JS, Murphy AS, Glazebrook J, Thordal-Christensen H, Katagiri F (2011) Physical association of Arabidopsis hypersensitive induced reaction proteins (HIRs) with the immune receptor RPS2. J Biol Chem 286:31297–31307PubMedCrossRefGoogle Scholar
  24. Roelfs AP (1985) Specificity and methods of study. In: Bushnell WR, Roelfs AP (eds) The cereal rusts: origins, specificity, structure and physiology. Academic, Orlando, pp 131–164Google Scholar
  25. Rostoks N, Schmierer D, Kudrna D, Kleinhofs A (2003) Barley putative hypersensitive induced reaction genes: genetic mapping, sequence analyses and differential expression in disease lesion mimic mutants. Theor Appl Genet 107:1094–1101PubMedCrossRefGoogle Scholar
  26. Saghai-Maroof MA, Soliman KM, Jorgesen RA, Allard RW (1984) Ribosomal DNA spacer-length polymorphisms in barley Mendelian inheritance, chromosomal location and population dynamics. Proc Natl Acad Sci USA 81:8014–8018PubMedCrossRefGoogle Scholar
  27. Sambrook J, Fritsch EF, Maniatis T (1998) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring HarborGoogle Scholar
  28. Tavernarakis N, Driscoll M, Kyrpides NC (1999) The SPFH domain: implicated in regulating targeted protein turnover in stomatins and other membrane-associated proteins. Trends Biochem Sci 24:425–427PubMedCrossRefGoogle Scholar
  29. Thordal-Christensen H, Zhang ZG, Wei YD, Collinge DB (2002) Subcellular localization of H2O2 in plants. H2O2 accumulation in papillae and hypersensitive response during the barley-powdery mildew interaction. Plant J 11:1187–1194CrossRefGoogle Scholar
  30. Wang XX, Chen J, Wang B, Liu LJ, Jiang H, Tang DL, Peng DX (2012) Characterization by suppression subtractive hybridization of transcripts that are differentially expressed in leaves of anthracnose-resistant ramie cultivar. Plant Mol Biol Rep 30:547–555CrossRefGoogle Scholar
  31. Xu H, Heath MC (1998) Role of calcium in signal transduction during the hypersensitive response caused by basidiospore derived infection of the cowpea rust fungus. Plant Cell 10:85–97Google Scholar
  32. Yang C, Li AL, Zhao YL, Zhang ZL, Zhu YF, Tan XM, Geng SF, Guo HZ, Zhang XY, Kang ZS (2011) Overexpression of a wheat CCaMK gene reduces ABA sensitivity of Arabidopsis thaliana during seed germination and seedling growth. Plant Mol Biol Rep 29:681–692CrossRefGoogle Scholar
  33. Yu XM, Yu XD, Qu ZP, Huang XJ, Guo J, Han QM, Zhao J, Huang LL, Kang ZS (2008) Cloning of a putative hypersensitive induced reaction gene from wheat infected by stripe rust fungus. Gene 407:193–198PubMedCrossRefGoogle Scholar
  34. Zambounis AG, Kalamaki MS, Tani EE, Paplomatas EJ, Tsaftaris AS (2012) Expression analysis of defense-related genes in cotton (Gossypium hirsutum) after Fusarium oxysporum f. sp. vasinfectum infection and following chemical elicitation using a salicylic acid analog and methyl jasmonate. Plant Mol Biol Rep 30:225–234CrossRefGoogle Scholar
  35. Zhang G, Dong YL, Zhang Y, Li YM, Wang XJ, Han QM, Guo J, Huang LL, Kang ZS (2009) Cloning and characterization of a novel hypersensitive-induced reaction gene from wheat infected by stripe rust pathogen. J Phytopathol 157:722–728CrossRefGoogle Scholar
  36. Zhou L, Cheung MY, Zhang Q, Lei CL, Zhang SH, Sun SM, Lam HM (2009) A novel simple extracellular leucine-rich repeat (eLRR) domain protein from rice (OsLRR1) enters the endosomal pathway and interacts with the hypersensitive-induced reaction protein 1 (OsHIR1). Plant Cell Environ 32:1804–1820PubMedCrossRefGoogle Scholar
  37. Zhou L, Cheung MY, Li MW, Fu Y, Sun Z, Sun SSM, Lam HM (2010) Rice hypersensitive induced reaction protein 1 (OsHIR1) associates with plasma membrane and triggers hypersensitive cell death. BMC Plant Biol 10:290. doi: 10.1186/1471-2229-10-290 PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  1. 1.College of Life Sciences, Biological Control Center of Plant Disease and Plant Pests of Hebei ProvinceAgricultural University of HebeiBaodingPeople’s Republic of China
  2. 2.College of Plant Protection, Biological Control Center of Plant Disease and Plant Pests of Hebei ProvinceAgricultural University of HebeiBaodingPeople’s Republic of China
  3. 3.Potato Research CentreAgriculture and Agri-Food CanadaFrederictonCanada

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