Use of Polyacrylamide Gel Electrophoresis for Detection of Chrysanthemum Stunt Viroid in Infected Tissues. R. K. Horst, Associate Professor, Department of Plant Pathology, Cornell University, Ithaca, NY 14853. S. O. Kawamoto, Research Technician, Department of Plant Pathology, Cornell University, Ithaca, NY 14853. Plant Dis. 64:186-188. Accepted for publication 1 October 1979. Copyright 1980 American Phytopathological Society. DOI: 10.1094/PD-64-186.

Chrysanthemum stunt viroid (CSV) was detected in as little as 50 mg of infected Chrysanthemum morifolium tissue by extraction of viroid nucleic acid followed by 5% polyacrylamide gel electrophoresis (PAGE) isolation. The procedure described by Morris and Wright was modified because colored compounds in nucleic acid extracts from chrysanthemum leaves interfere with distinctness of nucleic acid bands after PAGE. CSV-infected leaf tissue was extracted with a borate-sodium sulfite, sodium lauryl sulfate, lithium chloride buffer (pH 9.0) and then treated with phenol and chloroform. Preparations were dialyzed against distilled water to remove colored compounds. Nucleic acids were stained after electrophoresis with ethidium bromide or scanned with a spectrophotometer at 260 nm. The viroid-specific RNA band could then be readily observed in gels containing nucleic acids from CSV-infected tissues.