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Appl Environ Microbiol. 1989 March; 55(3): 761-763
Copyright © 1989, American Society for Microbiology. All Rights Reserved.

Detection of Vesicular-Arbuscular Mycorrhizal Fungus Colonization of Roots by Using a Dot-Immunoblot Assay

Sara F. Wright* and Joseph B. Morton

1 Appalachian Soil and Water Conservation Research Laboratory, Agricultural Research Service, U.S. Department of Agriculture, Beckley, West Virginia 25802-0867, and Department of Plant and Soil Sciences, West Virginia University, Morgantown, West Virginia 26506-60572

ABSTRACT

An immunoblotting procedure was developed to overcome the difficulty in identifying root colonization by a vesicular-arbuscular mycorrhizal fungus. The procedure utilized a murine monoclonal antibody that reacts with a protein in spores and hyphae of Glomus occultum, a fungus characterized by abundant production of hyaline spores and nonstaining intraradical infection. Minimally disturbed whole roots were squashed on nitrocellulose membranes. After inactivation of endogenous peroxidase, an indirect enzyme-linked immunosorbent assay was performed on the nitrocellulose with peroxidase-conjugated anti-mouse antibody as the second antibody. Antigen from G. occultum, revealed by a precipitating stain, was seen as purple dots on the nitrocellulose, which also retained the impression of the root.

FOOTNOTES

* Corresponding author.

Appl Environ Microbiol. 1989 March; 55(3): 761-763
Copyright © 1989, American Society for Microbiology. All Rights Reserved.



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