Immunomagnetic purification of Colletotrichum lindemuthianum appressoria.

K A Hutchison, S E Perfect, R J O'Connell, J R Green
Author Information
  1. K A Hutchison: School of Biological Sciences, University of Birmingham, United Kingdom.

Abstract

We developed a method to purify appressoria of the bean anthracnose fungus Colletotrichum lindemuthianum for biochemical analysis of the cell surface and to compare appressoria with other fungal structures. We used immunomagnetic separation after incubation of infected bean leaf homogenates with a monoclonal antibody that binds strongly to the appressoria. Preparations with a purity of >90% could be obtained. Examination of the purified appressoria by transmission electron microscopy showed that most had lost their cytoplasm. However, the plasma membrane was retained, suggesting that there is some form of attachment of this membrane to the cell wall. The purified appressoria can be used for studies of their cell surface, and we have shown that there are clear differences in the glycoprotein constituents of cell walls of appressoria compared with mycelium.

References

  1. New Phytol. 1993 Jun;124(2):193-213 [PMID: 33874341]
  2. Plant Cell. 1995 Feb;7(2):183-93 [PMID: 7756829]
  3. Appl Environ Microbiol. 1997 Jan;63(1):351-4 [PMID: 16535499]
  4. Exp Mycol. 1995 Dec;19(4):284-96 [PMID: 8574904]
  5. Microbiology (Reading). 1999 Aug;145 ( Pt 8):1927-1936 [PMID: 10463159]
  6. New Phytol. 1996 Dec;134(4):579-585 [PMID: 33863193]
  7. New Phytol. 1994 Jun;127(2):223-332 [PMID: 33874517]
  8. J Immunol Methods. 1989 Aug 15;122(1):97-103 [PMID: 2547875]
  9. Proc Natl Acad Sci U S A. 1966 Jan;55(1):134-41 [PMID: 5220862]

MeSH Term

Animals
Antibodies, Monoclonal
Cell Wall
Colletotrichum
Fabaceae
Glycoproteins
Immunomagnetic Separation
Mice
Mice, Inbred BALB C
Plant Diseases
Plants, Medicinal

Chemicals

Antibodies, Monoclonal
Glycoproteins

Word Cloud

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