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Applied and Environmental Microbiology, August 2000, p. 3464-3467, Vol. 66, No. 8
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Immunomagnetic Purification of Colletotrichum lindemuthianum Appressoria

Katie A. Hutchison,1 Sarah E. Perfect,1 Richard J. O'Connell,2 and Jonathan R. Green1,*

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.


* 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.


Applied and Environmental Microbiology, August 2000, p. 3464-3467, Vol. 66, No. 8
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.