Theoretical and Applied Genetics

, Volume 107, Issue 6, pp 1094–1101 | Cite as

Barley putative hypersensitive induced reaction genes: genetic mapping, sequence analyses and differential expression in disease lesion mimic mutants

  • N. Rostoks
  • D. Schmierer
  • D. Kudrna
  • A. Kleinhofs


The hypersensitive response (HR) is one of the most-efficient forms of plant defense against biotrophic pathogens, and results in localized cell death and the formation of necrotic lesions; however, the molecular components of pathways leading to HR remain largely unknown. Barley (Hordeum vulgare ssp. vulgare L.) cDNAs for putative hypersensitive-induced reaction (HIR) genes were isolated based on DNA and amino-acid homologies to maize HIR genes. Analyses of the cDNA and genomic sequences and genetic mapping found four distinct barley HIR genes, Hv-hir1, Hv-hir2, Hv-hir3 and Hv-hir4, on chromosomes 4(4H) bin10, 7(5H) bin04, 7(5H) bin07 and 1(7H) bin03, respectively. Hv-hir1, Hv-hir2 and Hv-hir3 genes were highly homologous at both DNA and the deduced amino-acid level, but the Hv-hir4 gene was similar to the other genes only at the amino-acid sequence level. Amino-acid sequence analyses of the barley HIR proteins indicated the presence of the SPFH protein-domain characteristic for the prohibitins and stomatins which are involved in control of the cell cycle and ion channels, as well as in other membrane-associated proteins from bacteria, plants and animals. HIR genes were expressed in all organs and developement stages analyzed, indicating a vital and non-redundant function. Barley fast-neutron mutants exhibiting spontaneous HR (disease lesion mimic mutants) showed up to a 35-fold increase in Hv-hir3 expression, implicating HIR genes in the induction of HR.


Barley Lesion-mimic mutants Hypersensitive reaction HIR genes SPFH domain 



This is Scientific Paper No. 090201 from the College of Agriculture and Home Economics Research Center, Washington State University, Pullman, WA; Project 0196. Research was supported by USDA/NRI grant No. 9901325 to A.K. Technical assistance by Kara Johnson and Thomas Drader is gratefully acknowledged.

Supplementary material

Fig. 1. Southern blot image. Cited in Results “Isolation and sequencing of barley HIR genomic clones”

Fig_1_ESM.jpg (43 kb)
Figure 1

Fig. 2. Images of barley FN mutants. Cited in Results “Expression of the HIR genes in barley fast neutron (FN) lesion mimic mutant lines” and in Discussion

Fig_2_ESM.jpg (113 kb)
Figure 2

Table 2 List of Morex BAC clones positive with the HIR probes. Cited in Results " Isolation and sequencing of barley HIR genomic clones”

Table2.pdf (54 kb)
Table 2 (PDF 55 KB)


  1. Ahn S, Anderson JA, Sorrels ME, Tanksley SD (1993) Homoeologous relationships of rice, wheat and maize chromosomes. Mol Gen Genet 241:483–490PubMedGoogle Scholar
  2. Asai T, Tena G, Plotnikova J, Willmann MR, Chiu WL, Gomez-Gomez L, Boller T, Ausubel FM, Sheen J (2002) MAP kinase signalling cascade in Arabidopsis innate immunity. Nature 415:977–983Google Scholar
  3. Atkinson MM, Keppler LD, Orlandi EW, Baker CJ, Mischke CF (1990) Involvement of plasma membrane calcium influx in bacterial induction of the K+/H+ and hypersensitive responses in tobacco. Plant Physiol 92:215–221Google Scholar
  4. Balague C, Lin B, Alcon C, Flottes G, Malmstrom S, Kohler C, Neuhaus G, Pelletier G, Gaymard F, Roby D (2003) HLM1, an essential signalling component in the hypersensitive response, is a member of the cyclic nucleotide-gated channel-ion channel family. Plant Cell 15:365–379CrossRefPubMedGoogle Scholar
  5. Bickel PE, Scherer PE, Schnitzer JE, Oh P, Lisanti MP, Lodish HF (1997) Flotillin and the epidermal surface antigen define a new family of caveolae-associated integral membrane proteins. J Biol Chem 272:13,793–13,802CrossRefPubMedGoogle Scholar
  6. Birnboim HC, Doly J (1979) A rapid alkaline-extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523PubMedGoogle Scholar
  7. Clough SJ, Fengler KA, Yu I-C, Lippok B, Smith RK Jr, Bent AF (2000) The Arabidopsis dnd1 "defense, no death" gene encodes a mutated cyclic nucleotide-gated ion channel. Proc Natl Acad Sci USA 97:9323–9328CrossRefPubMedGoogle Scholar
  8. Costa JM, Corey A, Hayes PM, Jobet C, Kleinhofs A, Kopisch-Obusch A, Kramer SF, Kudrna D, Li M, Riera-Lizarazu O, Sato K, Szucs P, Toojinda T, Vales MI, Wolfe RI (2001) Molecular mapping of the Oregon Wolfe Barleys: a phenotypically polymorphic doubled-haploid population. Theor Appl Genet 103:415–424Google Scholar
  9. Dangl JL, Jones JDG (2001) Plant pathogens and integrated defence responses to infection. Nature 411:826–833Google Scholar
  10. Heath MC (1998) Apoptosis, programmed cell death and the hypersensitive response. Eur J Plant Pathol 104:117–124Google Scholar
  11. Heath MC (2000) Hypersensitive response-related death. Plant Mol Biol 44:321–334PubMedGoogle Scholar
  12. Horvath H, Rostoks N, Brueggeman R, Steffenson B, von Wettstein D, Kleinhofs A (2003) Genetically engineered stem rust resistance in barley using the Rpg1 gene. Proc Natl Acad Sci USA 100:364–369CrossRefPubMedGoogle Scholar
  13. Hu G, Yalpani N, Briggs SP, Johal GS (1998) A porphyrin pathway impairment is responsible for the phenotype of a dominant disease lesion mimic mutant of maize. Plant Cell 10:1095–1105PubMedGoogle Scholar
  14. Jambunathan N, Siani JM, McNellis TW (2001) A humidity sensitive Arabidopsis copine mutant exhibits precocious cell death and increased disease resistance. Plant Cell 13:2225–2240CrossRefPubMedGoogle Scholar
  15. Johal GS, Hulbert S, Briggs SP (1995) Disease lesion mimic mutations of maize: a model for cell death in plants. Bioessays 17:685–692Google Scholar
  16. Karrer EE, Beachy RN, Holt CA (1998) Cloning of tobacco genes that elicit the hypersensitive response. Plant Mol Biol 36:681–690CrossRefPubMedGoogle Scholar
  17. Kleinhofs A, Graner A (2001) An integrated map of the barley genome. In: Phillips RL, Vasil IK (eds) DNA-based markers in plants, 2nd edn. Kluwer Academic Publishers, Dordrecht, The Netherlands, pp 187–199Google Scholar
  18. Kleinhofs A, Kilian A, Saghai Maroof MA, Biyashev RM, Hayes P, Chen FQ, Lapitan N, Fenwick A, Blake TK, Kanazin V, Ananiev E, Dahleen L, Kudrna D, Bollinger J, Knapp SJ, Liu B, Sorrells M, Heun M, Franckowiak JD, Hoffman D, Skadsen R, Steffenson BJ (1993) A molecular, isozyme and morphological map of the barley (Hordeum vulgare) genome. Theor Appl Genet 86:705–712Google Scholar
  19. Lam E, Kato N, Lawton M (2001) Programmed cell death, mitochondria and the plant hypersensitive response. Nature 411:848–853Google Scholar
  20. Lundqvist U, Franckowiak J, Konishi T (1997) New and revised descriptions of barley genes. Barley Genet Newslett 26:22,
  21. McClung JK, Jupe ER, Liu XT, Dell'Orco RT (1995) Prohibitin: potential role in senescence, development, and tumor suppression. Exp Gerontol 30:99–124CrossRefPubMedGoogle Scholar
  22. Mitchell-Olds T, Clauss MJ (2002) Plant evolutionary genomics. Curr Opin Plant Biol 5:74–79CrossRefPubMedGoogle Scholar
  23. 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 275:29,579–29,586PubMedGoogle Scholar
  24. Noble JA, Innis MA, Koonin EV, Rudd KE, Banuett F, Herskowitz I (1993) The Escherichia coli hflA locus encodes a putative GTP-binding protein and two membrane proteins, one of which contains a protease-like domain. Proc Natl Acad Sci USA 90:10,866–10,870Google Scholar
  25. Schuler MA (1998) Plant pre-mRNA splicing. In: Bailey-Serres J, Gallie DR (eds) A look beyond transcription. Am Soc Plant Physiologists, Rockville, Maryland, USA, pp 1–19Google Scholar
  26. Stewart GW (1997) Stomatin. Int J Biochem Cell Biol 29:271–274CrossRefPubMedGoogle Scholar
  27. 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 24:425–427CrossRefPubMedGoogle Scholar
  28. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4876–4882PubMedGoogle Scholar
  29. Van Deynze AE, Nelson JC, Yglesias ES, Harrington SE, Braga DP, McCouch SR, Sorrels ME (1995) Comparative mapping in grasses. Wheat relationships. Mol Gen Genet 248:744–754PubMedGoogle Scholar
  30. Yamanouchi U, Yano M, Lin H, Ashikari M, Yamada K (2002) A rice spotted leaf gene, Spl7, encodes a heat stress transcription factor protein. Proc Natl Acad Sci USA 99:7530–7535CrossRefPubMedGoogle Scholar
  31. Yu Y, Tomkins JP, Waugh R, Frisch DA, Kudrna D, Kleinhofs A, Brueggeman RS, Muehlbauer GJ, Wise RP, Wing RA (2000) A bacterial artificial chromosome library for barley (Hordeum vulgare L.) and the identification of clones containing putative resistance genes. Theor Appl Genet 101:1093–1099CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • N. Rostoks
    • 1
  • D. Schmierer
    • 1
  • D. Kudrna
    • 1
    • 3
  • A. Kleinhofs
    • 1
    • 2
  1. 1.Department of Crop and Soil Sciences, Washington State University, Pullman, WA 99164, USA
  2. 2.School of Molecular Biosciences, Washington State University, Pullman, WA 99164, USA
  3. 3.Arizona Genomics Institute, University of Arizona, 303 Forbes Building, Tucson, AZ 85721, USA

Personalised recommendations