Abstract
The avirulence gene avr9 of the fungal tomato pathogen Cladosporium fulvum encodes a race-specific peptide elicitor that induces the hypersensitive response in tomato plants carrying the complementary resistance gene Cf9. The avr9 gene is not expressed under optimal growth conditions in vitro, but is highly expressed when the fungus grows inside the tomato leaf. In this paper we present evidence for the induction of avr9 gene expression in C. fulvum grown in vitro under conditions of nitrogen limitation. Only growth medium with very low amounts of nitrogen (nitrate, ammonium, glutamate or glutamine) induced the expression of avr9. Limitation of other macronutrients or the addition of plant factors did not induce the expression of avr9. The induced expression of avr9 is possibly mediated by a positive-acting nitrogen regulatory protein, homologous to the Neurospora crassa NIT2 protein, which induces the expression of many genes under conditions of nitrogen limitation. The avr9 promoter contains several putative NIT2 binding sites. The expression of avr9 during the infection process was explored cytologically using transformants of C. fulvum carrying an avr9 promoter-β-glucuronidase reporter gene fusion. The possibility that expression of avr9 in C. fulvum growing in planta is caused by nitrogen limitation in the apoplast of the tomato leaf is discussed.
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References
Caddick MX (1992) Characterization of a major Aspergillus regulatory gene, areA. In: Stahl U, Tudzynski P (eds) Molecular Biology of Filamentous Fungi. VCH, Weinheim, pp 141–152
De Wit PJGM (1977) A light and scanning-electron microscopic study of infection of tomato plants by virulent and avirulent races of Cladosporium fulvum. Neth J Plant Pathol 83:109–122
De Wit PJGM (1986) Elicitation of active resistance mechanisms. In: Bailey J (ed) Biology and Molecular Biology of Plant-Pathogen Interactions, NATO ASI Series Vol 141, Springer Verlag, Berlin, pp 149–169
De Wit PJGM, Flach W (1979) Differential accumulation of phytoalexins in tomato leaves, but not in fruits after inoculation with virulent and avirulent races of Cladosporium fulvum. Physiol Plant Pathol 15:257–267
De Wit PJGM, Spikman G (1982) Evidence for the occurrence of race- and cultivar-specific elicitors of necrosis in intercellular fluids of compatible interactions between Cladosporium fulvum and tomato. Physiol Plant Pathol 21:1–11
Fu YH, Marzluf GA (1990) nit-2, the major positive-acting nitrogen regulatory gene of Neurospora crassa, encodes a sequence-specific DNA-binding protein. Proc Natl Acad Sci USA 87:5331–5335
Grignon G, Sentenac H (1991) pH and ionic conditions in the apoplast. Annu Rev Plant Physiol Plant Mol Biol 42:103–128
Harling R, Kenyon L, Lewis BG, Oliver RP, Turner JG, Coddington A (1988) Conditions for efficient isolation and regeneration of protoplasts from Fulvia fulva. J Phytopathol 122:143–146
Huynh TV, Dahlbeck D, Staskawicz BJ (1989) Bacterial blight of soybean: regulation of a pathogen gene determining host cultivar specificity. Science 245:1374–1377
Jarai G, Truong HN, Daniel-Vedele F, Marzluf GA (1992) NIT2, the regulatory protein of Neurospora crassa, binds upstream of nia, the tomato nitrate reductase gene, in vitro. Curr Genet 21:37–41
Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907
Joosten MHAJ, Cozijnsen TJ, De Wit PJGM (1994) Host resistance to a fungal tomato pathogen lost by a single base-pair change in an avirulence gene Nature 367:384–386
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Marmeisse R, Van den Ackerveken GFJM, Goosen T, De Wit PJGM, Van den Broek HWJ (1993) Disruption of the avirulence gene avr9 in two races of the tomato pathogen Cladosporium fulvum causes virulence on tomato genotypes with the complementary resistance gene Cf9. Mol Plant-Microbe Interact 6:412–417
Marzluf GA, Kanaan M, Fu YH (1992) Molecular analysis of DNA-binding, trans-acting regulatory proteins of Neurospora crassa. In: Stahl U, Tudzynski P (eds) Molecular Biology of Filamentous Fungi. VCH, Weinheim, pp 153–166
Oliver RP, Roberts IN, Harting R, Kenyon L, Punt PJ, Dingemans MA, Van den Hondel CAMJJ (1987) Transformation of Fulvia fulva, a fungal pathogen of tomato, to hygromycin B resistance. Curr Genet 12:231–233
Pate JS (1980) Transport and partitioning of nitrogenous solutes. Annu Rev Plant Physiol 31:313–340
Peters NK, Frost JW, Long SR (1986) A plant flavone, luteolin, induces the expression of Rhizobium meliloti nodulation genes. Science 233:977–980
Punt PJ, Oliver RP, Dingemans MA, Pouwels PH, Van den Hondel CAMJJ (1987) Transformation of Aspergillus based on the hygromycin B resistance marker from Escherichia coli. Gene 56:117–124
Rahme LG, Mindrinos MN, Panopoulos NJ (1992) Plant and environmental sensory signals control the expression of hrp genes in Pseudomonas syringae pv. phaseolicola. J Bacteriol 174:3499–3507
Roberts IN, Oliver RP, Punt PJ, Van den HondelCAMJJ (1989) Expression of the Escherichia coli β-glucuronidase gene in industrial and phytopathogenic filamentous fungi. Curr Genet 15:177–180
Scholtens-Toma IMJ, De Wit PJGM (1988) Purification and primary structure of a necrosis-inducing peptide from the apoplastic fluids of tomato infected with Cladosporium fulvum (syn. Fulvia fulva). Physiol Mol Plant Pathol 33:59–67
Schulte R, Bonas U (1992a) Expression of the Xanthomonas campestris pv. vesicatoria hrp gene cluster, which determines pathogenicity and hypersensitivity on pepper and tomato, is plant inducible. J Bacteriol 174:815–823
Schulte R, Bonas U (1992b) A Xanthomonas pathogenicity locus is induced by sucrose and sulphur containing amino acids. Plant Cell 4:79–86
Singh JP (1988) A rapid method for determination of nitrate in soil and plant extracts. Plant Soil 110:137–139
Stachel SE, Messens E, Van Montagu M, Zambryski D (1985) Identification of the signal molecules produced by wounded plant cells that activate T-DNA transfer in Agrobacterium tumefaciens. Nature 318:624–629
Van den Ackerveken GFJM, Van Kan JAL, De Wit PJGM (1992) Molecular analysis of the avirulence gene avr9 of the fungal tomato pathogen Cladosporium fulvum fully supports the genefor-gene hypothesis. Plant J 2:359–366
Van den Ackerveken GFJM, Van Kan JAL, Joosten MHAJ, Muisers JM, Verbakel HM, De Wit PJGM (1993) Characterization of two putative pathogenicity genes of the fungal tomato pathogen Cladosporium fulvum. Mol Plant-Microbe Interact 6:0210–215
Van Die J (1958) On the occurrence of α-keto acids and organic nitrogen compounds in xylem exudates of cucumber and tomato plants. Proc Kon Ned Acad Wetensch Amsterdam C61:572–578
Van Kan JAL, Van den Ackerveken GFJM, De Wit PJGM (1991) Cloning and characterization of cDNA of avirulence gene avr9 of the fungal tomato pathogen Cladosporium fulvum, causal agent of tomato leaf mold. Mol Plant-Microbe Interact 4:52–59
Wei Z-M, Sneath BJ, Beer SV (1992) Expression of Erwinia amylovera hrp genes in response to environmental stimuli. J Bacteriol 174:1875–1882
Wolters AMA, Schoenmakers HCH, Van der Meulen-Muisers JIM, Van der Knaap E, Derks FHM, Koornneef M, Zelcer A (1991) Limited DNA elimination from the irradiated potato parent in fusion products of albino Lycopersicon esculentum and Solanum tuberosum. Theor Appl Genet 83:225–232
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Communicated by C. A. M. J. J. Van den Hondel
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Van den Ackerveken, G.F.J.M., Dunn, R.M., Cozijnsen, A.J. et al. Nitrogen limitation induces expression of the avirulence gene avr9 in the tomato pathogen Cladosporium fulvum . Molec. Gen. Genet. 243, 277–285 (1994). https://doi.org/10.1007/BF00301063
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DOI: https://doi.org/10.1007/BF00301063