VIEW ARTICLE

Techniques

Evaluation of Agarose Gel Electrophoresis for Resolving Nucleoprotein Components of Prunus Necrotic Ringspot Virus. Ching- Ang Ong, Former graduate student, Department of Plant Pathology, Washington State University, Irrigated Agriculture Research and Extension Center, Prosser, 99350, Present address: Malaysian Agricultural Research and Development Institute (MARDI), Peti Surat 12301, Pijabat Besar Pos, 50774, Kuala Lumpur, Malaysia; G. I. Mink, Plant pathologist, Department of Plant Pathology, Washington State University, Irrigated Agriculture Research and Extension Center, Prosser, 99350. Phytopathology 79:613-619. Accepted for publication 11 November 1988. Copyright 1989 The American Phytopathological Society. DOI: 10.1094/Phyto-79-613.

The use of agarose gel electrophoresis for resolving the nucleoprotein components of three serologically indistinguishable isolates of Prunus necrotic ringspot virus (PNRSV) was evaluated. Although all nucleoproteins of the three PNRSV isolates used in this study had similar net surface charges, they resolved into three electrophoretic components when electrophoresed in 2% agarose gel at 100 V for 4–5 hr. Relationships among the electrophoretic components and components resolved in 10–40% sucrose density gradients were established. The top, middle, and bottom centrifugal components, in order of increasing sedimentation rate, were found to have electrophoretic mobility values that corresponded to those of the fast, middle, and slow electrophoretic components, respectively. The specific reactions of the electrophoretic components in the gel with ethidium bromide, acridine orange, and Coomassie brilliant blue as well as with antibodies against PNRSV indicated that all components consisted of intact PNRSV nucleoproteins. Better resolution of the three classes of nucleoproteins was obtained with agarose gel electrophoresis than with sedimentation through sucrose gradients. The three isolates used could be distinguished by their electrophoretic mobility patterns although they were serologically indistinguishable. A procedure was described for rapid partial purification of PNRSV suitable for electrophoretic analysis.