A Specific Serological Staining Procedure forVerticillium dahliaein Cotton Root Tissue

Abstract

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.