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Differential accumulation of mRNAs encoding extracellular and intracellular PR proteins in tomato induced by virulent and avirulent races of Cladosporium fulvum

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Abstract

Tomato leaves infected by the fungal pathogen Cladosporium fulvum contain several types of intracellular and extracellular pathogenesis-related (PR) proteins. Previously, we reported the purification and serological characterization of five extracellular PR proteins: P2, P4, P6, a chitinase and a β-1,3-glucanase [22, 23]. Here we describe the purification of a basic intracellular 33 kDa β-1,3-glucanase and the isolation and characterization of cDNA clones encoding the two extracellular P14 isomers P4 and P6, the extracellular acidic β-1,3-glucanase and a basic 35 kDa β-1,3-glucanase, different from the purified 33 kDa protein. Southern blot analysis demonstrated that tomato PR proteins are not encoded by large gene families, as is the case in tobacco. The number of genes corresponding to each protein was estimated to vary between one and three. A northern blot analysis indicated that the mRNAs for the extracellular PR proteins (P4, P6 and acidic β-1,3-glucanase) accumulate to similar levels in compatible and incompatible tomato-C. fulvum interactions, although the maximum level of expression is reached much faster in the incompatible interaction. On the other hand, the mRNA for the basic 35 kDa β-1,3-glucanase is induced rapidly to high levels in both interactions, but declines in time to background levels only in the incompatible interaction. The relevance of this difference in relation to plant defence is discussed.

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References

  1. Abraham GN, Podell D: Pyroglutamic acid. Nonmetabolic formation, function in proteins and peptides, and characteristics of the enzymes effecting its removal. Mol Cell Biochem 38: 181–190 (1981).

    Google Scholar 

  2. Ahl P, Gianninazzi S: b-proteins as a constitutive component in highly (TMV) resistant interspecific hybrids of Nicotiana glutinosa and Nicotiana debneyi. Plant Sci Lett 26: 173–181 (1982).

    Google Scholar 

  3. Broglie K, Chet I, Holliday M, Cressman R, Biddle P, Knowlton S, Mauvais CJ, Broglie R: Transgenic plants with enhanced resistance to the fungal pathogen Rhizoctonia solani. Science 254: 1194–1197 (1991).

    Google Scholar 

  4. Antoniw JF, Ritter CE, Pierpoint WS, Van Loon LC: Comparison of three pathogenesis-related proteins from plants of two cultivars of tobacco infected with TMV. J Gen Virol 47: 79–87 (1980).

    Google Scholar 

  5. Bell JN, Ryder TB, Wingate VPM, Bailey JA, Lamb CJ: Differential accumulation of plant defense gene transcripts in a compatible and an incompatible plant-pathogen interaction. Mol Cell Biol 6: 1615–1623 (1986).

    Google Scholar 

  6. Bol JF, Linthorst HJM, Cornelissen BJC: Plant pathogenesis-related proteins induced by virus infection. Annu Rev Phytopath 28: 113–138 (1990).

    Google Scholar 

  7. Brederode FTh, Linthorst HJM, Bol JF: Differential induction of acquired resistance and PR gene expression in tobacco by virus infection, ethephon treatment, UV light and wounding. Plant Mol Biol 17: 1117–1125 (1991).

    Google Scholar 

  8. Castresana C, De Carvalho F, Gheysen G, Habets M, Inzé D, Van Montagu M: Tissue-specific and pathogen-induced regulation of a Nicotiana plumbaginifolia β-1,3-glucanase gene. Plant Cell 2: 1131–1143 (1990).

    Google Scholar 

  9. Camacho Henriquez A, Sänger HL: Analysis of acid-extractable tomato leaf proteins after infection with a viroid, two viruses and a fungus and partial purification of the ‘pathogenesis-related’ protein P14. Arch Virol 74: 181–196 (1982).

    Google Scholar 

  10. Camacho Henriquez A, Sänger HL: Purification and partial characterization of the major ‘pathogenesis-related’ tomato leaf protein P14 from potato spindle tuber viroid (PSTV)-infected tomato leaves. Arch Virol 81: 263–284 (1984).

    Google Scholar 

  11. Cornelissen BJC, Hooft van Huijsduijnen RAM, van Loon LC, Bol JF: Molecular characterization of messenger RNAs for ‘pathogenesis-related’ proteins 1a, 1b and 1c, induced by TMV infection of tobacco. EMBO J 5: 37–40 (1986).

    Google Scholar 

  12. Cornelissen BJC, Horowitz J, van Kan JAL, Goldberg RB, Bol JF: Structure of tobacco genes encoding pathogenesis-related proteins from the PR-1 group. Nucl Acids Res 15: 6799–6811 (1987).

    Google Scholar 

  13. Cutt JR, Dixon DC, Carr JP, Klessig DF: Isolation and nucleotide sequence of cDNa clones for the pathogenesis-related proteins PR1a, PR1b and PR1c of Nicotiana tabacum cv. Xanthi nc induced by TMV infection. Nucl Acids Res 16: 9861 (1988).

    Google Scholar 

  14. de Wit PJGM: A light and scanning-electron microscopic study of infection of tomato plants by virulent and avirulent races of Cladosporium fulvum. Neth J Plant Path 83: 109–122 (1977).

    Google Scholar 

  15. de Wit PJGM, Flach W: Differential accumulation of phytoalexins in tomato leaves, but not in fruits after inoculation with virulent and avirulent races of Cladosporium fulvum. Physiol Plant Path 15: 257–267 (1979).

    Google Scholar 

  16. de Wit PJGM, van der Meer FE: Accumulation of the pathogenesis-related tomato leaf protein P14 as an early indicator of incompatibility in the interaction between Cladosporium fulvum (syn. Fulvia fulva) and tomato. Physiol Mol Plant Path 28: 203–214 (1986).

    Google Scholar 

  17. de Wit PJGM, Buurlage MB, Hammond KE: The occurrence of host, pathogen and interaction-specific proteins in the apoplast of Cladosporium fulvum (syn. Fulvia fulva) infected tomato leaves. Physiol Mol Plant Path 29: 159–172 (1986).

    Google Scholar 

  18. Fischer W, Christ U, Baumgartner M, Erismann KH, Mösinger E: Pathogenesis-related proteins of tomato: II. Biochemical and immunological characterization. Physiol Mol Plant Path 35: 67–83 (1989).

    Google Scholar 

  19. Fritig B, Rouster S, Kauffmann S, Stinzi A, Geoffroy P, Kopp M, Legrand M: Virus-induced glycanhydrolases and effects of oligosaccharide signals on plant-virus interactions. In: Lugtenberg B (ed) Signal Molecules in Plants and Plant Microbe Interactions, chap. H36, pp. 161–168. Plenum, New York (1989).

    Google Scholar 

  20. Granell A, Bellès JM, Conejero V: Induction of pathogenesis-related proteins in tomato by citrus exocortis viroid, silver ion and ethephon. Physiol Mol Plant Path 31: 83–90 (1987).

    Google Scholar 

  21. Hodgson CP, Fisk RZ: Hybridization probe size control: optimized ‘oligolabelling’. Nucl Acids Res 15: 6295 (1987).

    Google Scholar 

  22. Joosten MHAJ, de Wit PJGM: Identification of several pathogenesis-related proteins in tomato leaves inoculated with Cladosporium fulvum (syn. Fulvia fulva) as β-1,3-glucanases and chitinases. Plant Physiol 89: 945–951 (1989).

    Google Scholar 

  23. Joosten MHAJ, Bergmans CJB, Meulenhoff EJS, Cornelissen BJC, de Wit PJGM: Purification and serological characterization of three basic 15 kD pathogenesis-related (PR) proteins from tomato. Plant Physiol 94: 585–591 (1990).

    Google Scholar 

  24. Kauffmann S, Legrand M, Geoffroy P, Fritig B: Biological function of pathogenesis-related proteins. Four PR proteins have β-1,3-glucanase activity. EMBO J 6: 3209–3212 (1987).

    Google Scholar 

  25. Lazarovits G, Higgins VJ: Ultrastructure of susceptible, resistant, and immune reactions of tomato to races of Cladosporium fulvum. Can J Bot 54: 235–249 (1976).

    Google Scholar 

  26. Linthorst HJM: Pathogenesis-related proteins of plants. Crit Rev Plant Sci 10: 123–150 (1991).

    Google Scholar 

  27. Linthorst HJM, Meuwissen RLJ, Kauffmann S, Bol JF: Constitutive expression of pathogenesis-related proteins PR-1, GRP, and PR-S in tobacco has no effect on virus infection. Plant Cell 1: 285–291 (1989).

    Google Scholar 

  28. Linthorst HJM, Melchers LS, Mayer A, van Roekel JSC, Cornelissen BJC, Bol JF: Analysis of gene families encoding acidic and basic β-1,3-glucanases of tobacco. Proc Natl Acad Sci USA 87: 8756–8760 (1990).

    Google Scholar 

  29. Linthorst HJM, Danhash N, Brederode FTh, van Kan JAL, de Wit PJGM, Bol JF: Tobacco and tomato PR proteins homologous to win and pro-hevein lack the ‘hevein’ domain. Mol Plant-Microbe Interact 4: 586–592 (1991).

    Google Scholar 

  30. Lucas J, Camacho Henriquez A, Lottspeich F, Henschen A, Sänger HL: Amino acid sequence of the ‘pathogenesis-related’ leaf protein P14 from viroid-infected tomato reveals a new type of structurally unfamiliar proteins. EMBO J 11: 2745–2749 (1985).

    Google Scholar 

  31. Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning, a Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).

    Google Scholar 

  32. Memelink J, Linthorst HJM, Schilperoort RA, Bol JF: Tobacco genes encoding acidic and basic isoforms of pathogenesis-related proteins display different expression pattern. Plant Mol Biol 14: 119–126 (1990).

    Google Scholar 

  33. Nassuth A, Sänger HL: Immunological relationship between ‘pathogenesis-related’ leaf proteins from tomato, tobacco and cowpea. Virus Res 4: 229–242 (1986).

    Google Scholar 

  34. Neuhaus JM, Ahl-Goy P, Hinz U, Flores S, Meins F: High-level expression of a tobacco chitinase gene in Nicotiana sylvestris. Susceptibility of transgenic plants to Cercospora nicotianae. Plant Mol Biol 16: 141–151 (1991).

    Google Scholar 

  35. Ohme-Takagi M, Shinshi H: Structure and expression of a tobacco β-1,3-glucanase. Plant Mol Biol 15: 941–946 (1990).

    Google Scholar 

  36. Ori N, Sessa G, Lotan T, Himmelhoch S, Fluhr R: A major stylar matrix polypeptide (sp41) is a member of the pathogenesis-related proteins superclass. EMBO J 9: 3429–3436 (1990).

    Google Scholar 

  37. Payne G, Ward E, Gaffney T, Ahl Goy P, Moyer M, Harper A, Meins F, Ryals J: Evidence for a third structural class of β-1,3-glucanase in tobacco. Plant Mol Biol 15: 797–808 (1990).

    Google Scholar 

  38. Pfitzner UM, Goodman HM: Isolation and characterization of cDNA clones encoding pathogenesis-related proteins from tobacco mosaic virus infected tobacco plants. Nucl Acids Res 15: 4449–4465 (1987).

    Google Scholar 

  39. Pfitzner UM, Pfitzner AJP, Goodman HM: DNA sequence analysis of a PR-1a gene from tobacco: molecular relationship of heat shock and pathogen responses in plants. Mol Gen Genet 211: 290–295 (1988).

    Google Scholar 

  40. Rogers SO, Bendich AJ: Extraction of DNA from plant tissues. In: Gelvin SB, Schilperoort RA (eds) Plant Molecular Biology Manual, A6: 1–10. Kluwer Academic Publishers, Dordrecht (1988).

    Google Scholar 

  41. Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).

    Google Scholar 

  42. Shinshi H, Wenzler H, Neuhaus J-M, Felix G, Hofsteenge J, Meins F: Evidence for N- and C-terminal processing of a plant defense-related enzyme: Primary structure of tobacco prepro-β-1,3-glucanase. Proc Natl Acad Sci USA 85: 5541–5545 (1988).

    Google Scholar 

  43. van Kan JAL, van den Ackerveken AFJM, de Wit PJGM: Cloning and characterization of cDNA of avirulence gene avr9 of the fungal pathogen Cladosporium fulvum, causal agent of tomato leaf mold. Mol Plant-Microbe Interact 4: 52–59 (1991).

    Google Scholar 

  44. van Loon LC, Gerritsen YAM, Ritter CE: Identification, purification, and characterization of pathogenesis-related proteins from virus-infected Samsun NN tobacco leaves. Plant Mol Biol 9: 593–609 (1987).

    Google Scholar 

  45. Vera P, Hernández-Yago J, Conejero V: ‘Pathogenesis-related’ P1 (P14) protein. Vacuolar and apoplastic localization in leaf tissue from tomato plants infected with citrus exocortis viroid; in vitro synthesis and processing. J Gen Virol 70: 1933–1942 (1989).

    Google Scholar 

  46. Ward ER, Payne GB, Moyer MB, Williams SC, Dincher SS, Sharkey KC, Beck JJ, Taylor HT, Ahl-Goy P, Meins F, Ryals JA: Differential regulation of β-1,3-glucanase messenger RNAs in response to pathogen infection. Plant Physiol 96: 390–397 (1991).

    Google Scholar 

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Authors and Affiliations

  1. Department of Phytopathology, Agricultural University, P.O. Box 8025, 6700 EE, Wageningen, Netherlands

    Jan A. L. van Kan, Matthieu H. A. J. Joosten, Cornelia A. M. Wagemakers, Grardy C. M. van den Berg-Velthuis & Pierre J. G. M. de Wit

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  1. Jan A. L. van Kan
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  2. Matthieu H. A. J. Joosten
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  3. Cornelia A. M. Wagemakers
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  4. Grardy C. M. van den Berg-Velthuis
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  5. Pierre J. G. M. de Wit
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van Kan, J.A.L., Joosten, M.H.A.J., Wagemakers, C.A.M. et al. Differential accumulation of mRNAs encoding extracellular and intracellular PR proteins in tomato induced by virulent and avirulent races of Cladosporium fulvum . Plant Mol Biol 20, 513–527 (1992). https://doi.org/10.1007/BF00040610

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  • DOI: https://doi.org/10.1007/BF00040610

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