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A Specific Serological Staining Procedure for Verticillium dahliae in Cotton Root Tissue. J. S. Gerik, Department of Plant Pathology, University of California, Berkeley 94720; S. A. Lommel, and O. C. Huisman. Department of Plant Pathology, University of California, Berkeley 94720. Phytopathology 77:261-265. Accepted for publication 18 July 1986. Copyright 1987 The American Phytopathological Society. DOI: 10.1094/Phyto-77-261.

Hyphae of Verticillium dahliae were detected in cotton root tissue with an indirect enzyme-linked immunoassay. A soluble protein extract of V. dahliae was used to prepare a specific rabbit antiserum. The reaction of this rabbit antibody to the hyphae of V. dahliae was detected with an alkaline phosphatase antirabbit IgG conjugant that hydrolyzed the substrate, napthol-AS-phosphate, to a product that reacted with a diazonium salt, yielding a colored precipitate outlining the fungal hyphae. The hyphae were readily observed on and in the root cortex of the host using a dissecting microscope. This technique allows for rapid determination of the presence and location of V. dahliae in the root systems and may be applicable to other fungal species for which a specific antiserum can be produced.

Additional keywords: ELISA, fungi-specific staining, immunohistochemistry.