VIEW ARTICLE

Techniques

Detection of Virus Infection and Spread by Immunofluorescent Staining of Enzyme Treated Leaves. J. D. Lei, Graduate assistant, Department of Plant Pathology, University of Massachusetts, Amherst 01003; G. N. Agrios, professor, Department of Plant Pathology, University of Massachusetts, Amherst 01003. Phytopathology 76:1031-1033. Accepted for publication 3 April 1986. Copyright 1986 The American Phytopathological Society. DOI: 10.1094/Phyto-76-1031.

A fluorescent antibody staining technique, which allows precise presymptomatic location of virus infected cells in a leaf area, was developed during investigation of maize dwarf mosaic virus infection of corn. Inoculated leaves were brushed with Carborundum and briefly treated with cellulolytic enzymes. The partially digested leaves were fixed with acetone and sequentially treated with virus antibody produced in rabbits and then with fluorescein isothiocyanate-conjugated anti-rabbit antibody produced in goats. Epifluorescent microscopy was used to locate the fluorescent, antibody-stained infected cells. The epidermal cells and one to two layers of mesophyll cells could be clearly observed with this method. The effectiveness of the method was demonstrated.