Hybridization Detection of Insect-Transmitted Plant Viruses with Digoxigenin-Labeled Probes

Abstract

A dot blot hybridization system using digoxigenin-labeled probes and colorimetric visualization was developed for detection of plant viruses. Geminiviruses, including squash leaf curl virus and beet curly top virus, as well as zucchini yellow mosaic potyvirus, lettuce infectious yellows virus, and beet yellows closterovirus, were detected using this system. Extraction of the viral nucleic acid was necessary for effective detection of the viruses. Although the colorimetric system was less sensitive than radioactive dot blot hybridization, it was adequate for the detection of viruses from infected plants. This colorimetric system can be carried out in diagnostic laboratories using minimal equipment, suggesting its applicability for use in routine plant virus diagnoses