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Applied and Environmental Microbiology, August 2000, p. 3464-3467, Vol. 66, No. 8
School of Biological Sciences, University of
Birmingham, Birmingham B15 2TT,1 and
IACR-Long Ashton Research Station, Department of Agricultural
Sciences, University of Bristol, Long Ashton, Bristol BS41
9AF,2 United Kingdom
Received 2 August 1999/Accepted 25 April 2000
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.
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Immunomagnetic Purification of Colletotrichum
lindemuthianum Appressoria
*
Corresponding author. Mailing address: School of
Biological Sciences, University of Birmingham, Birmingham B15 2TT,
United Kingdom. Phone: (0) 121 414 5574. Fax: (0) 121 414 5925. E-mail: j.r.green{at}bham.ac.uk.
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