Comparative transcript profiling by cDNA-AFLP reveals similar patterns of Avr4/Cf-4- and Avr9/Cf-9-dependent defence gene expression.

SUMMARY Tomato Cf genes confer resistance to the fungal pathogen Cladosporium fulvum. Although the Cf-4 and Cf-9 proteins are very similar, the Cf-4- and Cf-9-dependent hypersensitive responses (HRs) are distinct in cell death pattern, intensity and sensitivity to environmental conditions. To investigate the mechanism leading to these differences, comparative transcript profiling of Avr4/Cf-4- and Avr9/Cf-9-dependent defence gene expression was performed. To do this, cDNA-AFLP analysis was conducted on Avr/Cf tomato seedlings undergoing early HR. Both Avr4/Cf-4 and Avr9/Cf-9 signalling elicited the same spectrum of genes, referred to here as Avr/Cf-elicited (ACE) genes. Of approximately 25 000 transcript-derived fragments (TDFs), 367 (1.5%) showed significant differential expression between HR(+) and HR(-) seedlings (either Avr4/Cf-4- or Avr9/Cf-9-dependent). However, 42.8% of the ACE TDFs (157/367 in total) showed quantitatively different expression in the two types of HR(+) seedlings. The majority of these (135/157, 86.0%) displayed significantly greater differential expression (either induced or repressed) in Avr4/Cf-4 HR(+) seedlings than in Avr9/Cf-9 HR(+) seedlings. Our results are consistent with the previous observation that Avr4/Cf-4-dependent HR is more severe than Avr9/Cf-9-dependent HR, and indicate that the distinction between Avr4/Cf-4- and Avr9/Cf-9-dependent HR is most probably a result of events upstream of the defence responses. Sequencing of 189 ACE fragments identified genes associated with: defence and resistance (33.3%), signal transduction (7.4%), HR and cell death (5.3%), transcriptional regulation and post-transcriptional modification (4.3%). Expression data revealed that defence response, respiration and biological oxidation are strongly induced while photosynthesis is severely repressed in the HR(+) seedlings.

[1]  R. Kaldenhoff,et al.  Functional aquaporin diversity in plants. , 2006, Biochimica et biophysica acta.

[2]  S. Turk,et al.  CDNA-AFLP combined with functional analysis reveals novel genes involved in the hypersensitive response. , 2006, Molecular plant-microbe interactions : MPMI.

[3]  Joachim L Schultze,et al.  Salicylic Acid–Independent ENHANCED DISEASE SUSCEPTIBILITY1 Signaling in Arabidopsis Immunity and Cell Death Is Regulated by the Monooxygenase FMO1 and the Nudix Hydrolase NUDT7[W] , 2006, The Plant Cell Online.

[4]  Jonathan D. G. Jones,et al.  The U-Box Protein CMPG1 Is Required for Efficient Activation of Defense Mechanisms Triggered by Multiple Resistance Genes in Tobacco and Tomato[W] , 2006, The Plant Cell Online.

[5]  R. Sayler,et al.  Mutation in a homolog of yeast Vps53p accounts for the heat and osmotic hypersensitive phenotypes in Arabidopsis hit1-1 mutant , 2006, Planta.

[6]  Zhong Zheng,et al.  High humidity represses Cf-4/Avr4- and Cf-9/Avr9-dependent hypersensitive cell death and defense gene expression , 2005, Planta.

[7]  B. Thomma,et al.  Cladosporium fulvum (syn. Passalora fulva), a highly specialized plant pathogen as a model for functional studies on plant pathogenic Mycosphaerellaceae. , 2005, Molecular plant pathology.

[8]  Jonathan D. G. Jones,et al.  Functional Analysis of Avr9/Cf-9 Rapidly Elicited Genes Identifies a Protein Kinase, ACIK1, That Is Essential for Full Cf-9–Dependent Disease Resistance in Tomatow⃞ , 2005, The Plant Cell Online.

[9]  A. McLennan,et al.  The Nudix hydrolase superfamily , 2005, Cellular and Molecular Life Sciences CMLS.

[10]  P. D. de Wit,et al.  Cladosporium fulvum circumvents the second functional resistance gene homologue at the Cf‐4 locus (Hcr9‐4E ) by secretion of a stable avr4E isoform , 2004, Molecular microbiology.

[11]  S. Minoshima,et al.  Identification and characterization of a novel gene family YPEL in a wide spectrum of eukaryotic species. , 2004, Gene.

[12]  J. Kudla,et al.  Integration and channeling of calcium signaling through the CBL calcium sensor/CIPK protein kinase network , 2004, Planta.

[13]  D. Twell,et al.  The Putative Arabidopsis Homolog of Yeast Vps52p Is Required for Pollen Tube Elongation, Localizes to Golgi, and Might Be Involved in Vesicle Trafficking1 , 2004, Plant Physiology.

[14]  J. Rose,et al.  Proteinaceous inhibitors of endo-β-glucanases , 2004 .

[15]  Erich Kombrink,et al.  SNARE-protein-mediated disease resistance at the plant cell wall , 2003, Nature.

[16]  P. Albersheim,et al.  Characterization of a tomato protein that inhibits a xyloglucan-specific endoglucanase. , 2003, The Plant journal : for cell and molecular biology.

[17]  P. D. de Wit,et al.  Attenuation of Cf-mediated defense responses at elevated temperatures correlates with a decrease in elicitor-binding sites. , 2002, Molecular plant-microbe interactions : MPMI.

[18]  G. Martin,et al.  The tobacco salicylic acid-binding protein 3 (SABP3) is the chloroplast carbonic anhydrase, which exhibits antioxidant activity and plays a role in the hypersensitive defense response , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[19]  F. Takken,et al.  Cladosporium fulvum overcomes Cf‐2‐mediated resistance by producing truncated AVR2 elicitor proteins , 2002, Molecular microbiology.

[20]  T. Romeis,et al.  Calcium‐dependent protein kinases play an essential role in a plant defence response , 2001, The EMBO journal.

[21]  J. Glazebrook,et al.  Genes controlling expression of defense responses in Arabidopsis--2001 status. , 2001, Current opinion in plant biology.

[22]  P. D. de Wit,et al.  Specific recognition of AVR4 and AVR9 results in distinct patterns of hypersensitive cell death in tomato, but similar patterns of defence-related gene expression. , 2001, Molecular plant pathology.

[23]  I. Faye,et al.  The Drosophila gene Yippee reveals a novel family of putative zinc binding proteins highly conserved among eukaryotes , 2001, Insect molecular biology.

[24]  K. Hammond-Kosack,et al.  cDNA-AFLP Reveals a Striking Overlap in Race-Specific Resistance and Wound Response Gene Expression Profiles , 2000, Plant Cell.

[25]  J. D. Jones,et al.  Comparison of the hypersensitive response induced by the tomato Cf-4 and Cf-9 genes in Nicotiana spp. , 2000, Molecular plant-microbe interactions : MPMI.

[26]  P. D. de Wit,et al.  Agroinfiltration is a versatile tool that facilitates comparative analyses of Avr9/Cf-9-induced and Avr4/Cf-4-induced necrosis. , 2000, Molecular plant-microbe interactions : MPMI.

[27]  P. De Wit,et al.  THE TOMATO-CLADOSPORIUM FULVUM INTERACTION: A Versatile Experimental System to Study Plant-Pathogen Interactions. , 1999, Annual review of phytopathology.

[28]  Hao Ren,et al.  Regulation of the Calmodulin-stimulated Protein Phosphatase, Calcineurin* , 1998, The Journal of Biological Chemistry.

[29]  F. Takken,et al.  Identification and Ds-tagged isolation of a new gene at the Cf-4 locus of tomato involved in disease resistance to Cladosporium fulvum race 5. , 1998, The Plant journal : for cell and molecular biology.

[30]  J. D. Jones,et al.  Characterization of the tomato Cf-4 gene for resistance to Cladosporium fulvum identifies sequences that determine recognitional specificity in Cf-4 and Cf-9. , 1997, The Plant cell.

[31]  Gapped BLAST and PSI-BLAST: A new , 1997 .

[32]  R. Van der Hoeven,et al.  Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP: analysis of gene expression during potato tuber development. , 1996, The Plant journal : for cell and molecular biology.

[33]  K. Hammond-Kosack,et al.  Race-Specific Elicitors of Cladosporium fulvum Induce Changes in Cell Morphology and the Synthesis of Ethylene and Salicylic Acid in Tomato Plants Carrying the Corresponding Cf Disease Resistance Gene , 1996, Plant physiology.

[34]  Jonathan D. G. Jones,et al.  The Tomato Cf-2 Disease Resistance Locus Comprises Two Functional Genes Encoding Leucine-Rich Repeat Proteins , 1996, Cell.

[35]  J. Wehland,et al.  IrpA , is highly homologous to monocytogenesPrfA-regulated gene in Listeria Identification and characterization of a novel , 1996 .

[36]  J. D. Jones,et al.  Isolation of the tomato Cf-9 gene for resistance to Cladosporium fulvum by transposon tagging. , 1994, Science.

[37]  M. Joosten,et al.  Host resistance to a fungal tomato pathogen lost by a single base-pair change in an avirulence gene , 1994, Nature.

[38]  T. Thomas,et al.  The carrot secreted glycoprotein gene EP1 is expressed in the epidermis and has sequence homology to Brassica S-locus glycoproteins. , 1993, The Plant journal : for cell and molecular biology.

[39]  G. Van den Ackerveken,et al.  Molecular analysis of the avirulence gene avr9 of the fungal tomato pathogen Cladosporium fulvum fully supports the gene-for-gene hypothesis. , 1992, The Plant journal : for cell and molecular biology.