Abstract
We analysed pathogenesis-related expression of genes, that are assumed to be involved in ubiquitous plant defence mechanisms like the oxidative burst, the hypersensitive cell death reaction (HR) and formation of localized cell wall appositions (papillae). We carried out comparative northern blot and RT-PCR studies with near-isogenic barley (Hordeum vulgareL. cv. Pallas) lines (NILs) resistant or susceptible to the powdery mildew fungus race A6 (Blumeria graminis f.sp. hordei, BghA6). The NILs carrying one of the R-genes Mla12, Mlg or the mlo mutant allele mlo5 arrest fungal development by cell wall appositions (mlo5) or a HR (Mla12) or both (Mlg). Expression of an aspartate protease gene, an ascorbate peroxidase gene and a newly identified cysteine protease gene was up-regulated after inoculation with BghA6, whereas the constitutive expression-level of a BAS gene, that encodes an alkyl hydroperoxide reductase, was reduced. Expression of a newly identified barley homologue of a mammalian cell death regulator, Bax inhibitor 1, was enhanced after powdery mildew inoculation. An oxalate oxidase-like protein was stronger expressed in NILS expressing penetration resistance. A so far unknown gene that putatively encodes the large subunit of a superoxide generating NADPH oxidases was constitutively expressed in barley leaves and its expression pattern did not change after inoculation. A newly identified barley Rac1 homologue was expressed constitutively, such as the functionally linked NADPH oxidase gene. Gene expression patterns are discussed with regard to defence mechanisms and signal transduction.
Similar content being viewed by others
References
Altschul, S.F., Madden, T.L., Schäffer, A.A., Zhang, J., Zhang, Z., Miller, W. and Lipman, D.J. 1997. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucl. Acids Res. 25: 3389–3402.
Baier, M. and Dietz, K.J. 1996. Primary structure and expression of plant homologues of animal and fungal thioredoxin-dependent peroxide reductases and bacterial alkyl hydroperoxide reductases. Plant Mol. Biol. 31: 553–564.
Baier, M. and Dietz, K.J. 1999. Alkyl hydroperoxide reductases: the way out of the oxidative breakdown of lipids in chloroplasts. Trend. Plant Sci. 4: 166–168.
Burhenne, K. and Gregersen, P.L. 2000. Up-regulation of the ascorbate-dependent antioxidative system in barley leaves during powdery mildew infection. Mol. Plant Path. 1: 303–315.
Dangl, J.L., Dietrich, R.A. and Richberg, M.H. 1996. Death don't have no mercy: cell death programs in plant-microbe interactions. Plant Cell 8: 1793–1807.
El-Zahaby, H.M., Gullner, G. and Király, Z. 1995. Effect of powdery mildew infection of barley on the ascorbate-glutathion cycle and other antioxidants in different host-pathogen interactions. Phytopathology 85: 1225–1230.
Dorey, S., Kopp, M., Geoffroy, P., Fritig, B. and Kauffmann, S. 1999. hydrogen peroxide from the oxidative burst is neither necessary nor sufficient for hypersensitive cell death induction, phenylalanine ammonia lyase stimulation, salicylic acid accumulation, or scopoletin consumption in cultured tobacco cells treated with elicitin. Plant Physiol. 121: 163–171.
Freialdenhoven, A., Scherag, B., Hollricher, K., Collinge, D.B., Thordal-Christensen, H. and Schulze-Lefert, P. 1994. Nar-1 and Nar-2, two loci required for Mla12-specified race-specific resistance to powdery mildew in barley. Plant Cell 6: 983–994.
Glazener, J.A., Orlandi, E.W. and Baker, C.J. 1996. The active oxygen response of cell suspensions to incompatible bacteria is not sufficient to cause hypersensitive cell death. Plant Physiol. 110: 759–763.
Groom, Q.J., Torres, M.A., Fordham-Skelton, A.P., Hammond-Kosak, K.E., Robinson, N.J. and Jones, J.D.G. 1996. RbohA, a rice homologue of mammalian gp91phox respiratory burst oxidase gene. Plant J. 10: 515–522.
Heath, M. 2000. Hypersensitive response-related cell death. Plant Mol. Biol. 44: 321–334.
Hess, W.R. and Boerner, T. 1998. Cloning and characterization of expression of cytosolic ascorbate peroxidase cDNA from barley. Plant Physiol. 118: 329–329.
Hückelhoven, R. and Kogel, K.-H. 1998. Tissue-specific superoxide generation at interaction sites in resistant and susceptible nearisogenic barley lines attacked by the powdery mildew fungus (Erysiphe graminis f.sp. hordei). Mol. Plant-Microbe Interact. 11: 292–300.
Hückelhoven, R., Fodor, J., Preis, C. and Kogel, K.-H. 1999. Hypersensitive cell death and papilla formation in barley attacked by the powdery mildew fungus are associated with H2O2 but not with salicylic acid accumulation. Plant Physiol. 119: 1251–1260.
Hückelhoven, R., Trujillo, M. and Kogel, K.-H. 2000a. Mutations in Ror1 and Ror2 genes cause modification of hydrogen peroxide accumulation in mlo-barley under attack from the powdery mildew fungus. Mol. Plant Path. 1: 287–292.
Hückelhoven, R., Fodor, J., Trujillo, M. and Kogel, K.-H. 2000b. Barley Mla and Rar mutants compromised in the hypersensitive cell death response against Blumeria graminis f.sp. hordei are modified in their ability to accumulate reactive oxygen intermediates at sites of fungal invasion. Planta212: 16–24.
Jørgensen, J.H. 1994. Genetics of powdery mildew resistance in barley. Crit. Rev. Plant Sci. 13: 97–119.
Jørgensen, J.H. 1996. Effect of three suppressors on the expression of powdery mildew resistance genes in barley. Genome: 492–498.
Kawai, M., Pan, L., Reed, J.C. and Uchimiya, H. 1999. Evolutionally conserved plant homologue of the Bax inhibitor-1 (BI-1) gene capable of suppressing Bax-induced cell death in yeast. FEBS Lett. 464: 143–147.
Kawasaki, T., Henmi, K., Ono, E., Hatakeyama, S., Iwano, M., Satoh, H. and Shimamoto, K. 1999. The small GTP-binding protein rac is a regulator of cell death in plants. Proc. Natl. Acad. Sci. USA 96: 10922–10926.
Király, Z., Barna, B. and Érsek, T. 1972. Hypersensitivity as a consequence, not the cause, for plant resistance to infection. Nature 239: 456–458.
Kogel, K.-H. and Hückelhoven, R. 1999. Superoxide generation in chemically activated resistance of barley in response to powdery mildew inoculation. J. Phytopath.147: 1–4.
Kølster, P., Munk, L., Stølen, O. and Løhde, J. 1986. Near-isogenic barley lines with genes for resistance to powdery mildew. Crop Sci. 26: 903–907.
Lacomme, C. and Santa Cruz, S. 1999. Bax-induced cell death in tobacco is similar to the hypersensitive response. Proc. Natl. Acad. Sci. USA 96: 7956–7961.
Lam, E. and del Pozo, O. 2000. Caspase-like protease involvement in the control of plant cell death. Plant Mol. Biol. 44: 417–428.
Mittler, R., Feng, X., Cohen, M. 1998. Post-transcriptional suppression of cytosolic ascorbate peroxidase expression during pathogen-induced programmed cell death in tobacco. Plant Cell 10: 461–473.
Mitsuhara, I., Malik, K.A., Miura, M. and Ohashi Y. 1999. Animal cell death suppressors Bcl-x(L) and Ced-9 inhibit cell death in tobacco plants. Curr. Biol. 9: 775–778.
Nelson, A.J. and Bushnell, W.R. 1997. Transient expression of anthocyanin genes in barley epidermal cells: potential for use in evaluation of disease response genes. Transgenic Res. 6: 233–244.
Nielsen, K., Olsen, O. and Oliver, R. 1999. A transient expression system to assay putative antifungal genes on powdery mildew infected barley leaves. Physiol Mol. Plant Path. 54: 1–12.
Peterhänsel, C, Freialdenhoven, A., Kurth, J., Kolsch, R. and Schulze-Lefert, P. 1997. Interaction analyses of genes required for resistance responses to powdery mildew in barley reveal distinct pathways leading to leaf cell death. Plant Cell 9: 1397–1409.
Runeberg-Roos, P. and Saarma, M. 1998. Phytepsin, a barley vacuolar aspartic proteinase, is highly expressed during autolysis of developing tracheary elements and sieve cells. Plant J. 15: 139–145.
Sanchez, P., de Torres Zabala, M. and Grant, M. 2000. AtBI-1,a plant homologue of Bax inhibitor –1, suppresses Bax-induced cell death in yeast and is rapidly upregulated during wounding and pathogen challenge. Plant J. 21: 393–399.
Schiffer, R., Görg, R., Jarosch, B., Beckhov, U., Bahrenberg, G., Kogel, K.-H. and Schulze-Lefert, P. 1997. Tissue dependence and differential cordycepin sensitivity of race-specific resistance responses in the barley-powdery mildew interaction. Mol. Plant-Microbe Interact. 10: 830–839.
Schweizer, P., Pokorny, J., Abderhalden, O. and Dudler, R. 1999a. A transient assay system for the functional assessment of defense-related genes in wheat. Mol. Plant-Microbe Interact. 12: 647–654.
Schweizer, P., Christoffel, A. and Dudler, R. 1999b. Transient expression of members of the germin-like gene family in epidermal cells of wheat confers disease resistance. Plant J. 20: 541–552.
Schweizer, P., Pokorny, J., Schulze-Lefert, P. and Dudler, R. 2000. Double-stranded RNA interferes with gene function at the single-cell level in cereals. Plant J. 24: 895–903.
Shirasu, K., Lahaye, T., Tan, M.-W., Zhou, F., Azevedo, C. and Schulze-Lefert, P. 1999. A novel class of eukaryotic zinc-binding proteins is required for disease resistance signaling in barley and development in C. elegans. Cell 99: 355–366.
Solomon, M., Belenghi, B., Delledonne, M., Menachem, E. and Levine, A. 1999. The involvement of cysteine proteases and protease inhibitor genes in the regulation of programmed cell death in plants. Plant Cell 11: 431–444.
Stoka, V., Turk, B., Schendel, S.L., Kim, T.H., Cirman, T., Snipas, S.J., Ellerby, L.M., Bredesen, D., Freeze, H., Abrahamson, M., Bromme, D., Krajewski, S., Reed, J.C., Yin, X.M., Turk, V. and Salvesen, G.S. 2001. Lysosomal protease pathways to apoptosis. Cleavage of bid, not pro-caspases, is the most likely route. J. Biol. Chem. 276: 3149–3157.
Thordal-Christensen, H., Zhang, Z., Wei, Y. and Collinge, D.B. 1997. Subcellular localization of H2O2 in plants: H2O2 accumulation in papillae and hypersensitive response during the barley-powdery mildew interaction. Plant J. 11: 1187–1194.
Vanacker, H., Carver, T.L.W. and Foyer, C. 1998. Pathogen-induced changes in the antioxidant status of the apoplast in barley leaves. Plant Physiol. 117: 1103–1114.
von Röpenack, E., Parr, A. and Schulze-Lefert, P. 1998. Structural analyses and dynamics of soluble and cell wall-bound phenolics in a broad spectrum resistance to the powdery mildew fungus in barley. J. Biol. Chem. 272: 9013–9022.
Wei, Y., Zhang, Z., Andersen, C.H., Schmelzer, E., Gregersen, P.L., Collinge, D.B., Smedegaard-Petersen, V. and Thordal-Christensen, H. 1998. An epidermis/papilla-specific oxalate oxidase-like protein in the defence response of barley attacked by the powdery mildey fungus. Plant Mol. Biol. 36: 101–112.
Wiberg, A. 1974. Genetical studies of spontaneous sources of resistance to powdery mildew in barley. Hereditas 77: 89–148.
Xu, Q. and Reed, J.C. 1998. Bax inhibitor-1, a mammalian apoptosis suppressor identified by functional screening in yeast. Mol. Cell 1: 337–346.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hückelhoven, R., Dechert, C., Trujillo, M. et al. Differential expression of putative cell death regulator genes in near-isogenic, resistant and susceptible barley lines during interaction with the powdery mildew fungus. Plant Mol Biol 47, 739–748 (2001). https://doi.org/10.1023/A:1013635427949
Issue Date:
DOI: https://doi.org/10.1023/A:1013635427949