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Analysis of Defense-related Proteins in Stem Tissue of Carnation Inoculated with a Virulent and Avirulent Race of Fusarium oxysporum F.sp. Dianthi

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Abstract

The aim of this study was to learn more about the accumulation of defense-related proteins in stem tissue from carnation cultivar Pallas inoculated with 2 near-isogenic races, the avirulent race 1 and the virulent race 8 of Fusarium oxysporum f.sp. dianthi. Stem tissue was used, from which the epidermis, cortex and medulla were peeled off from the vascular cylinder. It appeared that chitinase activity was constitutively expressed in the intercellular fluids (IFs) of untreated leaves, stems and roots of carnation. The total chitinase activity in the IFs of stem tissue increased with time after inoculation. This increase was similar after inoculation with the virulent, the avirulent race and water. At least four chitinase isoenzymes, three acidic and one basic isoform, were detected in the IFs of inoculated plants. In contrast, total 1,3-β-glucanase activity was not detected in the IFs of untreated leaves, stems and roots. Furthermore, the increases in 1,3-β-glucanase activity in IFs of stem tissue were markedly higher in the compatible and incompatible interactions than in the water control, indicating that this activity is specially induced by elicitors common to both races 1 and 8 of Fusarium oxysporum f.sp. dianthi. Using an antiserum against 1,3-β-glucanase P3 of tomato, 2 bands were detected on immunoblots in the IFs of stem tissue inoculated with races 1 and 8. No bands were visible after inoculation with water. Total peroxidase activity increased with time in all combinations. One basic and one acidic peroxidase isoform were present in these IFs. Peroxidase activity in a cell wall fraction prepared from stem tissue was clearly higher, and its increase faster, than the activity in the soluble stem fraction. These increases were similar in the virulent, the avirulent race and the water control. The growth of the fungus Trichoderma viride was inhibited by the IFs obtained from stem tissue inoculated with the virulent and the avirulent race of Fusarium oxysporum f.sp. dianthi. However, the growth of Fusarium oxysporum f.sp. dianthi itself was not affected by these IFs.

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References

  • Aloi C and Baayen RP (1993) Examination of the relationship between vegetative compatibility groups and races of Fusarium oxysporum f.sp. dianthi. Plant Pathology 42: 839-850

    Google Scholar 

  • Baayen RP and Schrama RM (1990) Comparison of the stem inoculation methods with respect to phytoalexin accumulation and Fusarium wilt development in carnation. Netherlands Journal of Plant Pathology 96: 315-320

    Google Scholar 

  • Baayen RP (1988) Responses related to lignification and intravascular periderm formation in carnations resistant to Fusarium wilt. Canadian Journal of Botany 66: 784-791

    Google Scholar 

  • Baayen RP, Quellette GB and Rioux D (1996) Compartimentalization of decay in carnations resistant to Fusarium oxysporum f.sp. dianthi. Phytopathology 86: 1018-1031

    Google Scholar 

  • Benhamou N, Greiner J, Asselin A and Legrand M (1989) Immunogold localization of β-1,3-glucanases in two plants infected by vascular wilt fungi. The Plant Cell 1: 1209-1221

    Google Scholar 

  • Bergmeyer J and Grassl M (eds) (1983) Methods of enzymatic analysis, Vol III: Oxidoreductases and Transferases, Chemie Verlag, Weinheim

    Google Scholar 

  • Boller T, Gehri A, Mauch F and Vogeli U (1983) Chitinase in bean leaves: induction by ethylene, purification, properties and possible function. Planta 157: 22-31

    Google Scholar 

  • Bradford MM (1970) A rapid and sensitive method for the quantification of microgram quantities of protein using the principle of protein dye binding. Analytical Biochemistry 72: 248-254

    Google Scholar 

  • Bradley DJ, Kjelborn P and Lamb CJ (1992) Elicitor-and wound-induced oxidative cross-linking of plant cell wall proline-rich protein: a novel, rapid defense response. Cell 70(1): 21-30

    Google Scholar 

  • De Wit PJGM and Spikman G (1982) Evidence for the occurence of race-and cultivar-specific elicitors of necrosis in intercellular fluids of compatible interactions between Cladosporium fluvum and tomato. Physiological and Molecular Plant Pathology 29: 159-172

    Google Scholar 

  • Dixon RA and Lamb CJ (1990) Molecular communication in interactions between plants and microbial pathogens. Annual Review Plant Physiology 41: 339-367

    Google Scholar 

  • McDougall GJ (1993) Accumulation of wall-associated peroxidases during wound induced suberization of flax. Journal Plant Physiology 142: 651-656

    Google Scholar 

  • McDougall GJ (1992) Changes in cell wall-associated peroxidases during the lignification of flax fibres. Phytochemistry 31: 3385-3389

    Google Scholar 

  • Fry SC (1990) Wall enzymes. In: Harlow and Longman (eds) The Growing Plant Cell Wall: Chemical and Metabolic Analysis. pp. 219-237

  • Joosten MHAJ and De Wit PJDM (1989) Identification of several pathogenesis-related proteins in tomato leaves inoculated with Cladosporium fulvum as 1,3-β-glucanases and chitinases. Plant Physiology 89: 945-961

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-684

    Google Scholar 

  • Ludwig A and Boller T (1990) A method for the study of fungal growth inhibition by plant pathogens. FEMS Microbiology Letters 69: 55-60

    Google Scholar 

  • Manicom BQ and Baayen RP (1993) Restriction fragment length polymorphisms in Fusarium oxysporum f.sp. dianthi and other Dianthus species. Plant Pathology 42: 851-857

    Google Scholar 

  • Mauch F, Hadewiger LA and Boller T (1984) Ethylene induction of chitinases and 1,3-β-glucanases in pea pods and elicitors. Plant Physiology 76: 607-611

    Google Scholar 

  • Melchers LS, Apotheker-de-Groot M, van deer Knaap JA, Ponstein AS, Sela-Buurlage MB, Bol JF, Cornelissen BJC, van den Elzen PJM and Linthorst HJM (1994) A new class of tobacco chitinases homologous to bacterial exo-chitinases displays antifungal activity. The Plant Journal 5: 469-480

    Google Scholar 

  • Mohan RM and Kolattukudy PE (1990) Differental activation of expression of a suberization associated anionic peroxidase gene in near isogenic resistant and susceptible lines by elicitors of albo-atrum. Plant Physiology 921: 276-280

    Google Scholar 

  • Nelson N (1944) A photometric adaptation of the Somogyi method for the determination of glucose. Journal Biological Chemistry 153: 375-380

    Google Scholar 

  • Pan SQ, Ye XS and Kuc J (1991) A technique for detection of chitinase, β-1,3-glucanase and protein pattern after a single separation using polyacrylamide gelelectrophoresis or iso-electric focusing. Phytopathology 81: 970-974

    Google Scholar 

  • Reimers PJ, Guo A and Leach JE (1992) Increased activity of a cationic peroxidase associated with an incompatible interaction between Xanthomonas oryzae p.v. oryzae and rice. Plant Physiology 99: 1044-1050

    Google Scholar 

  • Reissig JL, Strominger JL and Leloir LF (1955) A modified colorimetric method for the estimation of N-acetyl amino sugars. Journal Biological Chemistry 217: 959-966

    Google Scholar 

  • Ride JP (1983) Cell walls and other structural barriers in defense. In: Biochemical Plant Pathology (pp. 213-236) Wiley-Interscience, New York

    Google Scholar 

  • Scott-Graig SJ, Kerby KB, Stein BD and Somerville SC (1996) Expression of an extracellular peroxidase in barley by the powdery mildew pathogen. Physiology and Molecular Plant Pathology 47: 407-418

    Google Scholar 

  • Van den Elzen PMJ, Jongendijk E, Melchers LS and Cornelissen BJC (1993) Virus and fungal resistance from laboratory to field. Phil. Trans. R. Soc. Lond. B 342: 271-278

    Google Scholar 

  • Van Loon LC (1985) Pathogenesis-related proteins. Plant Molecular Biology 4: 111-116

    Google Scholar 

  • Witte B (1991) Chitinasen in Kartoffel: Charakterisierung verschiedener Isoformen und Untersuchungen zu ihrer Expression. Doctoral thesis, Universität zu Köln

  • Ye XS, Pan SQ and Kuc JA (1990) Activity, isozyme pattern and cellular localization of peroxidase as related to systemic resistance of tobacco to blue mold and tobacco mosaic virus. Phytopathology 80: 1295-1299

    Google Scholar 

  • Young SA, Guo A, Guikema JA, White FF and Leach JE (1995) Rice cationic peroxidase accumulates in xylem vessels during incompatible interactions with Xanthomonas oryzae p.v. oryzae. Plant Physiology 107: 1333-1341

    Google Scholar 

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Van Pelt-Heerschap, H., Smit-Bakker, O. Analysis of Defense-related Proteins in Stem Tissue of Carnation Inoculated with a Virulent and Avirulent Race of Fusarium oxysporum F.sp. Dianthi. European Journal of Plant Pathology 105, 681–691 (1999). https://doi.org/10.1023/A:1008767830202

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