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Composition and organisation of extracellular matrices around germ tubes and appressoria ofColletotrichum lindemuthianum

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Summary

The ultrastructure and composition of the extracellular matrices (ECMs) associated with germ tubes and appressoria ofColletotrichum lindemuthianum have been examined. Flexuous fibres (fimbriae), up to 6 μm long and 4–30 nm in diameter, protruded from the surface of germ tubes and appressoria. Anionic colloidal gold and lectin cytochemistry showed that ECMs of germ tubes and appressoria contain basic proteins, α-D-mannose and α-D-galactose residues. A monoclonal antibody, UB26, was raised to infection structures isolated from leaves ofPhaseolus vulgaris infected withC. lindemuthianum. UB26 recognised a protein epitope on two glycoproteins (Mr 133,000 and 146,000). Reductions in the Mr of these proteins after treatment with peptide-N-glycosidase and trifluoromethane sulphonic acid suggest that they carry N- and O-linked side-chains. Immunofluorescence and EM-immunogold labelling showed that glycoproteins recognised by UB26 were restricted to the ECMs around germ tubes and appressoria but fimbriae were not labelled. Unlike appressorial germ tubes formed in vitro, intracellular infection hyphae were not labelled, suggesting that the glycoproteins recognised by UB26 are not present on fungal structures formed within host cells. In liquid culture, these glycoproteins were not released into the medium, suggesting they are physically linked to the cell wall. Also, the glycoproteins were not removed from glass surfaces by ultrasonication. These results suggest that glycoproteins recognised by UB26 may be involved in the adhesion of germ tubes and appressoria to substrata. Our results show that the ECMs of germ tubes and appressoria differ markedly in structure and composition from those of conidia and intracellular hyphae, and that extracellular glycoproteins are associated with specific regions of the fungal cell surface.

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Abbreviations

ECM:

extracellular matrix

BPA:

Bauhinia purpurea agglutinin

BSA:

bovine serum albumin

DIC:

differential interference contrast

FITC:

fluorescein isothiocyanate

GNL:

Galanthus nivalis lectin

GSI-B4 :

Griffonia simplicifolia isolectin B4

HEPES:

(N-(2-hydroxyethyl)piperazine-N′-(2-ethanesulphonic acid)

IIF:

indirect immunofluorescence

IPC:

isopycnic centrifugation

MAb:

monoclonal antibody

PEG:

polyethylene glycol

PBS:

phosphate buffered saline

PNGase:

peptideN-glycosidase

SDS-PAGE:

sodium dodecyl sulphate polyacrylamide gel electrophoresis

TCS:

tissue culture supernatant

TEM:

transmission electron microscopy

TFMS:

trifluoromethane sulphonic acid

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Authors and Affiliations

  1. School of Biological Sciences, The University of Birmingham, Birmingham

    N. A. Pain & J. R. Green

  2. Institute of Arable Crops Research, Long Ashton Research Station, Department of Agricultural Sciences, University of Bristol, Long Ashton, Bristol

    G. L. Jones & R. J. O'Connell

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  1. N. A. Pain
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  2. J. R. Green
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  3. G. L. Jones
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  4. R. J. O'Connell
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Pain, N.A., Green, J.R., Jones, G.L. et al. Composition and organisation of extracellular matrices around germ tubes and appressoria ofColletotrichum lindemuthianum . Protoplasma 190, 119–130 (1996). https://doi.org/10.1007/BF01281311

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  • DOI: https://doi.org/10.1007/BF01281311

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