Protoplasma

, Volume 170, Issue 3–4, pp 95–103

Isolation by ConA binding of haustoria from different rust fungi and comparison of their surface qualities

  • M. Hahn
  • K. Mendgen
Article

Summary

Rust haustoria isolated from infected leaf tissue strongly bind to ConA. This property was exploited to purify them by affinity chromatography on a ConA-Sepharose macrobead column. Haustoria were obtained with more than 90% purity and yields of up to 50%. Binding of haustoria to the column was partially inhibited by a ConA-specific sugar, methyl α-D-mannopyranoside. Compared to ConA,Lens culinaris agglutinin and wheat germ agglutinin were less efficient affinity ligands. Using ConA-Sepharose, rust haustoria from a variety of sources could be isolated with equal efficiency, indicating that they have similar carbohydrate surface properties. The haustoria maintained their typical shape after the isolation procedure, which suggests a rather rigid wall structure. The morphology of haustoria was characteristic both for a given species and the nuclear condition of the rust mycelium. Electron microscopy of isolated haustoria revealed an intact haustorial wall surrounded by a fibrillar layer presumably derived from the extrahaustorial matrix. The matrix thus appears to represent a layer with gel-like properties which is rich in ConA-binding carbohydrates and connected to the haustorial wall but not to the host-derived extrahaustorial membrane.

Keywords

Affinity chromatography Biotrophy Concanavalin A Extrahaustorial matrix Puccinia Uromyces 

Abbreviations

ConA

Concanavalin A

LCA

Lens culinaris agglutinin

WGA

wheat germ agglutinin

FITC

fluorescein isothiocyanate

DAPI

4′,6-diamidinophenylindol×2 HCl

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Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • M. Hahn
    • 1
  • K. Mendgen
    • 1
  1. 1.Lehrstuhl für Phytopathologie, Fakultät für BiologieUniversität KonstanzKonstanzFederal Republic of Germany

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