Abstract
Studies on the role of arbuscular mycorrhizal (AM) fungi in soil have been aided by the use of a monoclonal antibody that detects a molecule common to all isolates of these fungi studied to date. The molecule, glomalin, is a glycoprotein that forms on hyphae, but apparently sloughs off and adheres to soil particles or imbedded plastic mesh. An indirect immunofluorescence (IF) assay is described for detection of glomalin on hyphae attached to roots, in roots, on hyphae traps and on the surface of soil aggregates. Small sieves are used to process hyphae attached to roots and soil aggregates. Glomalin on hyphae and glomalin attached to plastic or nylon are assayed on a 1 cm2 section of meshes. Examples of IF assay results are shown and discussed.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Aldwell F E B and Hall I R 1986 Monitoring spread of Glomus mosseae through soil infested with Acaulospora laevis using serological and morphological techniques. Trans. Br. Mycol. Soc. 87, 131–134.
Aldwell F E B, Hall I R and Smith J M B 1985 Enzyme-linked immunosorbent assay as an aid to taxonomy of the Endogonaseae. Trans. Br. Mycol. Soc. 84, 399–402.
Bolhool B B (1987) Fluorescence methods for study of Rhizobium in culture and in situ. In Symbiotic Nitrogen Fixation Technology. Ed. GH Elkan. pp. 127–147. Marcel Dekker, Inc. New York.
Bonfante-Fasolo P, Tamagnone L, Perreto R, Esquerré-Tugayé M T, Mazau D, Mosiniak M and Vian B 1991 Immunochemical location of hydroxyproline rich glycoproteins at the interface between a mycorrhizal fungus and its host plants. Protoplasma 165, 127–138.
Davidson R S and Goodwin D 1983 An effective way of retarding the fading of fluorescence of labelled material during microscopical examination. Proc. Royal Med. Soc. 18, 151–153.
Dazzo F B and Wright S F 1996 Production of anti-microbial antibodies and their use in immunofluorescence microscopy. In Molecular Microbial Ecology Manual. Eds. A D L Akkermans, J D van Elsas and F J de Bruijn. pp. 4.1.2, 1- 27. Kluwer Academic Publishers, Dordrecht, The Netherlands.
Friese C F and Allen M F 1991 Tracking fates of exotic and local VA mycorrhizal fungi: methods and patterns. Agric. Ecosyst. Environ. 34, 87–96.
Goding J W 1986 Monoclonal Antibodies Principles and Practice, 2nd edn. Academic Press, New York, USA. 315 p.
Hahn, A, Bonfante P, Horn K, Pausch F and Hock B 1993 Production of monoclonal antibodies against surface antigens of spores from arbuscular mycorrhizal fungi by an improved immunization and screening procedure. Mycorrhiza 4, 69–78.
Kemper W D and Rosenau R C 1986 Aggregate stability and size distribution. In Methods of Soil Analysis, Part 1. Physical and Mineralogical Methods. pp. 425–444. Agronomy Monograph No. 9, 2nd edn. American Society of Agronomy, Madison, WI, USA.
Köhler G and Milstein C 1975 Continuous culture of fused cells secreting antibody of predefined specificity. Nature 256, 495–497.
Kough J L and Linderman R G 1986 Monitoring extra-matrical hyphae of a vesicular-arbuscular mycorrhizal fungus with an immunofluorescence assay and the soil aggregation technique. Soil Biol. Biochem. 18, 309–313.
Perotto S, Brewin N J and Bonfante P 1994 Colonization of pea roots by the mycorrhizal fungus Glomus versiforme and by Rhizobium bacteria: immunological comparison using monoclonal antibodies as probes for plant cell surface components. Molec. Plant Microbial Interact. 7, 91–98.
Sanders I R, Ravolanirina F, Gianinazzi-Pearson V, Gianinazzi S and Lemoine M C 1992 Detection of specific antigens in the vesicular-arbuscular mycorrhizal fungi Gigaspora margarita and Acaulospora laevis using polyclonal antibodies to soluble spore fractions. Mycol. Res. 96, 477–480.
Tisdall J M 1991 Fungal hyphae and structural stability of soil. Aust. J. Soil Res. 29, 729–743.
Tisdall J M 1995 Formation of soil aggregates and accumulation of soil organic matter. In Structure and Organic Matter Storage in Agricultural Soils. Eds. M R Carter and B A Stewart. pp. 57–96. Lewis Publishers, Boca Raton, Florida.
Tisdall J M and Oades J M 1979 Stabilization of soil aggregates by the root systems of ryegrass. Aust. J. Soil Res. 17, 429–441.
Wilson J M, Trinick M J and Parker C A 1983 The identification of vesicular-arbuscular mycorrhizal fungi using immunofluorescence. Soil Biol. Biochem. 15, 439–445.
Wright S F, Franke-Snyder M, Morton J B and Upadhyaya A 1996 Time-course study and partial characterization of a protein on hyphae of arbuscular mycorrhizal fungi during active colonization of roots. Plant Soil 181, 193–203.
Wright S F, Morton J B and Sworobuk J E 1987 Identification of a vesicular-arbuscular mycorrhizal fungus by using monoclonal antibodies in an enzyme-linked immunosorbent assay. Appl. Environ. Microbiol. 53, 2222–2225.
Wright S F and Upadhyaya A 1996 Extraction of an abundant and unusual protein from soil and comparison with hyphal protein of arbuscular mycorrhizal fungi. Soil Sci. 161, 575–586.
Wright S F and Upadhyaya A 1998 A survey of soils for aggregate stability and glomalin, a glycoprotein produced by hyphae of arbuscular mycorrhizal fungi. Plant Soil 198, 97–107.
Wright S F and Upadhyaya A 1999 Quantification of arbuscular mycorrhizal fungi activity by the glomalin concentration on hyphal traps. Mycorrhiza 8, 283–285.
Wright S F, Upadhyaya A and Buyer J S 1998 Comparison of Nlinked oligosaccharides of glomalin from arbuscular mycorrhizal fungi and soils by capillary electrophoresis. Soil Biol. Biochem. 30, 1853–1857.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Wright, S.F. A fluorescent antibody assay for hyphae and glomalin from arbuscular mycorrhizal fungi. Plant and Soil 226, 171–177 (2000). https://doi.org/10.1023/A:1026428300172
Issue Date:
DOI: https://doi.org/10.1023/A:1026428300172