Summary
Plant chitinases and β-1,3-glucanases have been demonstrated to inhibit fungal growth in model experiments, both on agar plates or in liquid media. Here,Trichoderma longibrachiatum was taken as a model to study the morphological changes caused by chitinase and glucanase treatments, using cytochemical techniques in combination with fluorescence and electron microscopy. Chitinase, alone or in the presence of glucanase, arrested growth of the hypha: it affected the extreme tip of the fungus producing a thinning of the wall, a balloon-like swelling and a rupture of the plasma membrane. Chitin and glucans were present in the wall, as shown by lectinand enzyme-binding experiments, but they had a different susceptibility to chitinase and β-1,3-glucanase. Chitin was present at the apex and in the inner parts of the lateral walls; it was more susceptible to chitinase at the tip than in the subapical part. Glucans mostly occurred on the outer layer where they were degraded by glucanase. The latter did not affect the inner hyphal skeleton. It is suggested that the growth inhibition ofTrichoderma by hydrolytic enzymes is the consequence of a thinning of the cell wall in the hyphal apex, leading to an imbalance of turgor pressure and wall tension which causes the tip to swell and to burst.
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Abbreviations
- WGA-FITC:
-
wheat germ agglutinin labelled with fluorescein isothiocyanate
- ConA-FITC:
-
concanavalin A labelled with fluorescein isothiocyanate
- PEG:
-
polyethylene glycol
- SEM:
-
scanning electron microscopy
- TEM:
-
transmission electron microscopy
References
Arlorio M, Ludwig A, Boller T, Mischiati P, Bonfante P (1992) Effects of chitinase and β-1,3-ghicanase from pea on the growth of saprophytic, pathogenic and mycorrhizal fungi. Giorn Bot Ital 126: 956–958
Bendayan M (1985) The enzyme-gold technique: a new cytochemical approach for the ultrastructural localization of macromolecules. In: Bullock GR, Petrusz P (eds) Techniques in immunocytochemistry, vol 3. Academic Press, New York, pp 179–201
Boller T (1987) Hydrolytic enzymes in plant disease resistance. In: Kosuge T, Nester EW (eds) Plant-microbe interactions, vol 2. Macmillan, New York, pp 385–413
— (1988) Ethylene and the regulation of antifungal hydrolases in plants. In: Miflin BJ (ed) Oxford surveys of plant molecular and cell biology, vol 5. Oxford University Press, Oxford, pp 145–174
Bonfante P, Perotto S, Testa B, Faccio A (1987) Ultrastructural localization of cell surface sugar residues in ericoid mycorrhizal fungi by gold labeled lectins. Protoplasma 137: 25–35
—, Faccio A, Perotto S, Schubert A (1990) Correlation between chitin distribution and cell wall morphology in the mycorrhizal fungusGlomus vesiforme. Mycol Res 94: 157–165
—, Vian B, Perotto S, Faccio A, Knox JP (1990) Cellulose and pectin localization in roots of mycorrhizalAllium porrum: labelling continuity between host cell wall and interfacial material. Planta 180: 537–547
Broekaert WF, Van Parijs J, Allen AK, Peumans WJ (1988) Comparison of some molecular, enzymatic, and antifungal properties of chitinases from thornapple, tobacco, and wheat. Physiol Mol Plant Pathol 33: 319–331
— —, Leyns F, Joos H, Peumans WJ (1989) A chitin-binding lectin from stinging nettle rhizomes with antifungal properties. Science 245: 1100–1102
Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 28: 350–356
Fevre M, Girard V, Nodet P (1991) Cellulose and β-glucan synthesis inSaprolegnia. In: Heath IB (ed) Tip growth in plant and fungal cells. Academic Press, New York, pp 97–107
Gooday GW, Gow NAR (1991) Enzymology of tip growth in fungi. In: Heath IB (ed) Tip growth in plant and fungal cells. Academic Press, New York, pp 31–58
Ludwig A, Boller T (1990) A method for the study of fungal growth inhibition by plant proteins. FEMS Microbiol Lett 69: 61–66
Mauch F, Hadwiger LA, Boller T (1988 a) Antifungal hydrolases in pea tissue. I. Purification and characterization of two chitinases and two β-1,3-glucanases differentially regulated during development and in response to fungal infection. Plant Physiol 87: 325–333
—, Mauch-Mani B, Boller T (1988 b) Antifungal hydrolases in pea tissue. II. Inhibition of fungal growth by combinations of chitinase and β-1,3-glucanase. Plant Physiol 88: 936–942
Peretto R, Perotto S, Faccio A, Bonfante P (1990) Cell surface inCalluna vulgaris L. hair roots. In situ localization of polysaccharidic components. Protoplasma 155: 1–18
Reissig JL, Strominger JL, Leloir LF (1955) A modified method for the estimation of N-acetylamino sugars. J Biol Chem 217: 959–966
Roberts WK, Selitrennikoff CP (1988) Plant and bacterial chitinases differ in antifungal activity. J Gen Microbiol 134: 169–176
Roland JC, Vian B (1991) General preparation and staining of thin sections. In: Hall JL, Hawes C (eds) Electron microscopy of plant cells. Academic Press, New York, pp 1–66
Schlumbaum A, Mauch F, Vögeli U, Boller T (1986) Plant chitinases are potent inhibitors of fungal growth. Nature 324: 365–367
Wessels JGH (1991) Role of cell wall architecture in fungal tip growth generation. In: Heath IB (ed) Tip growth in plant and fungal cells. Academic Press, New York, pp 1–29
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Arlorio, M., Ludwig, A., Boller, T. et al. Inhibition of fungal growth by plant chitinases andβ-1,3-glucanases. Protoplasma 171, 34–43 (1992). https://doi.org/10.1007/BF01379278
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DOI: https://doi.org/10.1007/BF01379278