Abstract
Blumeria graminis f.sp. tritici, the causal agent of powdery mildew in wheat, is an obligate biotrophic fungus that exclusively invades epidermal cells. As previously shown, spraying of a solution of syringolin A, a circular peptide derivative secreted by the phytopathogenic bacterium Pseudomonas syringae pv. syringae, triggers hypersensitive cell death at infection sites in powdery mildew infected wheat. Thus, the fungus is essentially eradicated. Here we show that syringolin A also triggers hypersensitive cell death in Arabidopsis infected with the powdery mildew fungus Erysiphe cichoracearum. To monitor transcriptional changes associated with this effect, we cloned 307 cDNA clones representing 158 unigenes from powdery mildew infected, syringolin A sprayed wheat leaves by a suppression subtractive hybridization cloning procedure. These cDNAs were microarrayed onto glass slides together with 1088 cDNA-AFLP clones from powdery mildew-infected wheat. Microarray hybridization experiments were performed with probes derived from leaves, epidermal tissue, and mesophyll preparations of mildewed or uninfected wheat plants after syringolin A or control treatment. Similar experiments were performed in Arabidopsis using the Affymetrix ATH1 whole genome GeneChip. The results indicate a conserved mode of action of syringolin A as similar gene groups are induced in both species. Prominent groups include genes associated with the proteasomal degradation pathway, mitochondrial and other heat shock genes, genes involved in mitochondrial alternative electron pathways, and genes encoding glycolytic and fermentative enzymes. Surprisingly, in both species the observed transcriptional response to syringolin A was considerably weaker in infected plants as compared to uninfected plants. The results lead to the working hypothesis that cell death observed at infection sites may result from a parasite-induced suppression of the transcriptional response and thus to insufficient production of protective proteins necessary for the recovery of these cells from whatever insult is imposed by syringolin A.
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Amrein H, Makart S, Granado J, Shakya R, Schneider-Pokorny J, Dudler R (2004) Functional analysis of genes involved in the synthesis of syringolin A by Pseudomonas syringae pv. syringae B301D-R. Mol Plant-Microb Interact 17:90–97
Ané JM, Kiss GB, Riely BK, Penmetsa RV, Oldroyd GED, Ayax C, Levy J, Debelle F, Baek JM, Kalo P, Rosenberg C, Roe BA, Long SR, Denarie J, Cook DR (2004) Medicago truncatula DMI1 required for bacterial and fungal symbioses in legumes. Science 303:1364–1367
Ashburner M, Ball CA, Blake JA, Botstein D, Butler H, Cherry JM, Davis AP, Dolinski K, Dwight SS, Eppig JT, Harris MA, Hil DP, Issel-Tarver L, Kasarskis A, Lewis S, Matese JC, Richardson JE, Ringwald M, Rubin GM, Sherlock G (2000) Gene Ontology: Tool for the unification of biology. Nat Genet 25:25–29
Banerjee J, Ghosh S (2004) Bax increases the pore size of rat brain mitochondrial voltage-dependent anion channel in the presence of tBid. Biochem Biophys Res Commun 323:310–314
Benjamini Y, Hochberg Y (1995) Controlling the false discovery rate - a practical and powerful approach to multiple testing. J Roy Stat Soc B Met 57:289–300
Benz R (1994) Permeation of hydrophilic solutes through mitochondrial outer membranes—review on mitochondrial porins. Biochim Biophys Acta—Rev Biomembranes 1197: 167–196
Bock JB, Matern HT, Peden AA, Scheller RH (2001) A genomic perspective on membrane compartment organization. Nature 409:839–841
Bruggmann R, Abderhalden O, Reymond P, Dudler R (2005) Analysis of epidermis- and mesophyll-specific transcript accumulation in powdery mildew-inoculated wheat leaves. Plant Mol Biol 58:247–267
Carver TLW, Lyngkjaer MF, Neyron L, Strudwicke CC (1999) Induction of cellular accessibility and inaccessibility and suppression and potentiation of cell death in oat attacked by Blumeria graminis f.sp. avenae. Physiol Molec Plant Pathol 55:183–196
Chisholm ST, Coaker G, Day B, Staskawicz BJ (2006) Host-microbe interactions: shaping the evolution of the plant immune response. Cell 124:803–814
Crompton M, Barksby E, Johnson N, Capano M (2002) Mitochondrial intermembrane junctional complexes and their involvement in cell death. Biochimie 84:143–152
Diatchenko L, Lukyanov S, Lau YFC, Siebert PD (1999) Suppression subtractive hybridization: A versatile method for identifying differentially expressed genes. In: cDNA preparation and characterization. Academic Press Inc, San Diego, pp 349–380
Dudoit S, Yang YH, Callow MJ, Speed TP (2002) Statistical methods for identifying differentially expressed genes in replicated cDNA microarray experiments. Stat Sin 12:111–139
Gechev TS, Gadjev IZ, Hille J (2004) An extensive microarray analysis of AAL-toxin-induced cell death in Arabidopsis thaliana brings new insights into the complexity of programmed cell death in plants. Cell Mol Life Sci 61:1185–1197
Gechev TS, Minkov IN, Hille J (2005) Hydrogen peroxide-induced cell death in Arabidopsis: Transcriptional and mutant analysis reveals a role of an oxoglutarate-dependent dioxygenase gene in the cell death process. Iubmb Life 57:181–188
Geigenberger P (2003) Response of plant metabolism to too little oxygen. Curr Opin Plant Biol 6:247–256
Hamilton EW, Heckathorn SA (2001) Mitochondrial adaptations to NaCl. Complex I is protected by anti-oxidants and small heat shock proteins, whereas complex II is protected by proline and betaine. Plant Physiol 126:1266–1274
Hassa P, Granado J, Freydl E, Wäspi U, Dudler R (2000) Syringolin-mediated activation of the Pir7b esterase gene in rice cells is suppressed by phosphatase inhibitors. Mol Plant-Microb Interact 13:342–346
Hengartner MO (2000) The biochemistry of apoptosis. Nature 407:770–776
Hirano SS, Upper CD (2000) Bacteria in the leaf ecosystem with emphasis on Pseudomonas syringae—a pathogen, ice nucleus, and epiphyte. Microbiol Mol Biol Rev 64:624–653
Hrabak EM, Willis DK (1992) The lemA gene required for pathogenicity of Pseudomonas syringae pv. syringae on bean is a member of a family of 2-component regulators. J Bacteriol 174:3011–3020
Hrabak EM, Willis DK (1993) Involvement of the IemA gene in production of syringomycin and protease by Pseudomonas syringae pv. syringae. Mol Plant-Microb Interact 6:368–375
Hubbell E, Liu WM, Mei R (2002) Robust estimators for expression analysis. Bioinformatics 18:1585–1592
Hughes TR, Mao M, Jones AR, Burchard J, Marton MJ, Shannon KW, Lefkowitz SM, Ziman M, Schelter JM, Meyer MR, Kobayashi S, Davis C, Dai HY, He YDD, Stephaniants SB, Cavet G, Walker WL, West A, Coffey E, Shoemaker DD, Stoughton R, Blanchard AP, Friend SH, Linsley PS (2001) Expression profiling using microarrays fabricated by an ink-jet oligonucleotide synthesizer. Nat Biotechnol 19:342–347
Kuno H, Kuroda H, Hayashimoto A, Ishizaki H (1985a) Induced susceptibility and enhanced resistance at the cellular level in barley coleoptiles. II. Timing and localization of induced sysceptibility in a single coleptile cell and its transfer to an adjacent cell. Can J Bot 64:889–895
Kuno H, Hayashimoto A, Harui M, Ishizaki H (1985b) Induced susceptibility and enhanced resistance at the cellular level in barley coleoptiles. I. The significance of timing of fungal invasion. Physiol Plant Pathol 27:43–54
Kuno H, Kuroda H, Toyoda K, Yamaoka N, Kobayashi I (1991) Induced susceptibility and enhanced resistance at the cellular level in barley coleoptiles. IX. Accessibility induced by Erysiphe graminis which promotes the subsequent infection of challenging E. graminis. Ann Phytopathol Soc Jpn 57:57–60
Lemeshko VV (2002) Model of the outer membrane potential generation by the inner membrane of mitochondria. Biophys J 82:684–692
Lemeshko VV, Lemeshko SV (2004) The voltage-dependent anion channel as a biological transistor: Theoretical considerations. Eur Biophys J Biophys Lett 33:52–359
Lindow SE, Brandl MT (2003) Microbiology of the phyllosphere. Appl Environ Microbiol 69:1875–1883
Liu WM, Mei R, Di X, Ryder TB, Hubbell E, Dee S, Webster TA, Harrington CA, Ho MH, Baid J, Smeekens SP (2002) Analysis of high density expression microarrays with signed-rank call algorithms. Bioinformatics 18:1593–1599
Lyngkjaer MF, Carver TLW (1999) Induced accessibility and inaccessibility to Blumeria graminis f.sp hordei in barley epidermal cells attacked by a compatible isolate. Physiol Molec Plant Pathol 55:151–162
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular Cloning. A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor
Marco ML, Legac J, Lindow SE (2005) Pseudomonas syringae genes induced during colonization of leaf surfaces. Environ Microbiol 7:1379–1391
Meiners S, Heyken D, Weller A, Ludwig A, Stangl K, Kloetzel PM, Kruger E (2003) Inhibition of proteasome activity induces concerted expression of proteasome genes and de Novo formation of mammalian proteasomes. J Biol Chem 278:21517–21525
Michalecka AM, Svensson AS, Johansson FI, Agius SC, Johanson U, Brennicke A, Binder S, Rasmusson AG (2003) Arabidopsis genes encoding mitochondrial type IINAD(P)H dehydrogenases have different evolutionary orgin and show distinct responses to light. Plant Physiol 133:642–652
Millenaar FF, Lambers H (2003) The alternative oxidase: In vivo regulation and function. Plant Biol 5:2–15
Moon J, Parry G, Estelle M (2004) The ubiquitin-proteasome pathway and plant development. Plant Cell 16:3181–3195
Moore AL, Albury MS, Crichton PG, Affourtit C (2002) Function of the alternative oxidase: is it still a scavenger? Trends Plant Sci 7:478–481
Nomura K, Melotto M, He SY (2005) Suppression of host defense in compatible plant-Pseudomonas syringae interactions. Curr Opin Plant Biol 8:361–368
Quigley NB, Gross DC (1994) Syringomycin production among strains of Pseudomonas syringae pv. syringae: conservation of the syrB and syrD genes and activation of phytotoxin production by plant signal molecules. Mol Plant-Microb Interact 7:78–90
Randolph JB, Waggoner AS (1997) Stability, specificity and fluorescence brightness of multiply-labeled fluorescent DNA probes. Nucleic Acids Res 25:2923–2929
Rasmusson AG, Soole KL, Elthon TE (2004) Alternative NAD(P)H dehydrogenases of plant mitochondria. Annu Rev Plant Biol 55:23–39
Reimmann C, Hofmann C, Mauch F, Dudler R (1995) Characterization of a rice gene induced by Pseudomonas syringae pv. Syringae—requirement for the bacterial lemA gene-function. Physiol Molec Plant Pathol 46:71–81
Ricoult C, Cliquet JB, Limami AM (2005) Stimulation of alanine amino transferase (AlaAT) gene expression and alanine accumulation in embryo axis of the model legume Medicago truncatula contribute to anoxia stress tolerance. Physiol Plant 123:30–39
Rocke DM, Durbin B (2001) A model for measurement error for gene expression arrays. J Comput Biol 8:557–569
Rostovtseva T, Colombini M (1997) VDAC channels mediate and gate the flow of ATP: Implications for the regulation of mitochondrial function. Biophys J 72:1954–1962
Saeed AI, Sharov V, White J, Li J, Liang W, Bhagabati N, Braisted J, Klapa M, Currier T, Thiagarajan M, Sturn A, Snuffin M, Rezantsev A, Popov D, Ryltsov A, Kostukovich E, Borisovsky I, Liu Z, Vinsavich A, Trush V, Quackenbush J. 2003. TM4: A free, open-source system for microarray data management and analysis. Biotechniques 34:374–378
Scharf KD, Siddique M, Vierling E (2001) The expanding family of Arabidopsis thaliana small heat stress proteins and a new family of proteins containing alpha-crystallin domains (Acd proteins). Cell Stress Chaperon 6:225–237
Schweizer P, Christoffel A, Dudler R (1999) Transient expression of members of the germin-like gene family in epidermal cells of wheat confers disease resistance. Plant J 20:540–552
Shimizu S, Ide T, Yanagida T, Tsujimoto Y (2000) Electrophysiological study of a novel large pore formed by Bax and the voltage-dependent anion channel that is permeable to cytochrome c. J Biol Chem 275:12321–12325
Smalle J, Vierstra RD (2004) The ubiquitin 26S proteasome proteolytic pathway. Annu Rev Plant Biol 55:555–590
Staden R (1996) The Staden sequence analysis package. Mol Biotechnol 5:233–241
Thilmony R, Underwood W, He SY (2006) Genome-wide transcriptional analysis of the Arabidopsis thaliana interaction with the plant pathogen Pseudomonas syringae pv. tomato DC3000 and the human pathogen Escherichia coli O157: H7. Plant J 46:34–53
Thimm O, Blasing O, Gibon Y, Nagel A, Meyer S, Kruger P, Selbig J, Muller LA, Rhee SY, Stitt M (2004) MAPMAN: a user-driven tool to display genomics data sets onto diagrams of metabolic pathways and other biological processes. Plant J 37:914–939
Tör M, Yemm A, Holub E (2003) The role of proteolysis in R gene mediated defence in plants. Mol Plant Pathol 4:287–296
Vierstra RD (2003) The ubiquitin/26S proteasome pathway, the complex last chapter in the life of many plant proteins. Trends Plant Sci 8:135–142
Wäspi U, Schweizer P, Dudler R (2001) Syringolin reprograms wheat to undergo hypersensitive cell death in a compatible interaction with powdery mildew. Plant Cell 13:153–161
Wäspi U, Blanc D, Winkler T, Ruedi P, Dudler R (1998a) Syringolin, a novel peptide elicitor from Pseudomonas syringae pv. syringae that induces resistance to Pyricularia oryzae in rice. Mol Plant-Microbe Interact 11:727–733
Wäspi U, Hassa P, Staempfli AA, Molleyres LP, Winkler T, Dudler R (1999) Identification and structure of a family of syringolin variants: Unusual cyclic peptides from Pseudomonas syringae pv. syringae that elicit defense responses in rice. Microbiol Res 154:89–93
Wäspi U, Misteli B, Hasslacher M, Jandrositz A, Kohlwein SD, Schwab H, Dudler R (1998b) The defense-related rice gene Pir7b encodes an alpha/beta hydrolase fold protein exhibiting esterase activity towards naphthol AS-esters. Eur J Biochem 254:32–37
Xu GW, Gross DC (1988) Physical and functional analyses of the syrA and syrB genes involved in syringomycin production by Pseudomonas syringae pv. syringae. J Bacteriol 170:5680–5688
Yahiaoui N, Srichumpa P, Dudler R, Keller B (2004) Genome analysis at different ploidy levels allows cloning of the powdery mildew resistance gene Pm3b from hexaploid wheat. Plant J 37:528–538
Yang PZ, Fu HY, Walker J, Papa CM, Smalle J, Ju YM, Vierstra RD (2004) Purification of the Arabidopsis 26 S proteasome—Biochemical and molecular analyses revealed the presence of multiple isoforms. J Biol Chem 279:6401–6413
Yang YH, Dudoit S, Luu P, Lin DM, Peng V, Ngai J, Speed TP (2002) Normalization for cDNA microarray data: a robust composite method addressing single and multiple slide systematic variation. Nucleic Acids Res 30(4):e15
Acknowledgements
We thank E.E. Farmer and P. Reymond, University of Lausanne, for use of the Gene Expression Laboratory Microarray facility at the Department of Plant Molecular Biology, U. Wagner and A. Patrignani, Functional Genomics Center Zurich, as well as T. Wicker for help with bioinformatics tasks, and R. Honegger for help with the identification of Erysiphe cichoracearum. Support by the Swiss National Science Foundation (grant 3100A0–100046), the Functional Genomics Center Zurich, and the Foundation for Scientific Research at the University of Zurich is greatfully acknowledged.
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Kathrin Michel, Olaf Abderhalden and Robert Dudler contributed equally to this work. Wheat experiments were performed by Olaf Abderhalden, Arabidopsis experiments by Kathrin Michel
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Michel, K., Abderhalden, O., Bruggmann, R. et al. Transcriptional changes in powdery mildew infected wheat and Arabidopsis leaves undergoing syringolin-triggered hypersensitive cell death at infection sites. Plant Mol Biol 62, 561–578 (2006). https://doi.org/10.1007/s11103-006-9045-7
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DOI: https://doi.org/10.1007/s11103-006-9045-7