VIEW ARTICLE

Techniques

Immunological Detection of Plant Viruses and a Mycoplasmalike Organism by Direct Tissue Blotting on Nitrocellulose Membranes. N. S. Lin, Institute of Botany, Academia Sinica, Taipei, Taiwan 11529, ROC; Y. H. Hsu(2), and H. T. Hsu(3). (2)Agricultural Biotechnology Laboratories, National Chung-Hsing University, Taichung, Taiwan 40227, ROC; (3)Florist and Nursery Crops Laboratory, Beltsville Agriculture Research Center, U.S. Department of Agriculture, Beltsville, MD 20705. Phytopathology 80:824-828. Accepted for publication 16 March 1990. This article is in the public domain and not copyrightable. It may be freely reprinted with customary crediting of the source. The American Phytopathological Society, 1990. DOI: 10.1094/Phyto-80-824.

A technique of tissue blotting on nitrocellulose membranes was described for detection of plant viruses and a mycoplasmalike organism (MLO) in infected plants. Tissue blots were made by pressing, with a firm and gentle force, the freshly cut tissue surface on nitrocellulose membranes. Antigens in tissue blots on nitrocellulose membranes were detected by enzyme-labeled immunological probes. In indirect immunological methods, the blots were reacted with antigen-specific primary antibodies and detected with enzyme-labeled species-specific secondary antibodies. Alternatively, the blots were reacted with antigen-specific biotinylated primary antibodies and detected with avidin-enzyme conjugates. In direct immunological methods, the blots were reacted and detected with enzyme-labeled antigen-specific antibodies. The tissue blotting technique was applied to detect viruses in cucumovirus, luteovirus, potexvirus, potyvirus, and tomato spotted wilt virus groups and an MLO. Passionfruit woodiness, papaya ringspot, sweet potato feathery mottle, bean yellow mosaic (BYMV), bamboo mosaic (BoMV), cymbidium mosaic, cucumber mosaic, and tomato spotted wilt viruses were detected in blots of infected leaf tissue cross sections. BYMV and BoMV were also detected in tissues of newly emerged shoots of gladiolus and bamboo, respectively. In addition, BYMV was detected in tissue blots made from dormant corms of gladiolus. In barley yellow dwarf virus and tomato big bud-MLO infected leaves, antigens were localized in phloem cells of the vascular system.