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Separation of hypoviral dsRNA on CIM DEAE monoliths

Purification of plant viral double-stranded RNA using DEAE monoliths was described recently. The new approach enables very fast and simple detection of the pathogen`s replicative RNA. Compared to the most plant viruses possessing single-stranded RNA genome and depositing dsRNA as a replicative form, genome of many fungal viruses is featured by dsRNA. Many dsRNAs have been identified from Cryphonectria parasitica, the causal agent of chestnut blight and several have been recognized as genomes of viruses which mediate hypovirulence of this important phytopathogenic fungus. In this work, a procedure based on CIM DEAE chromatography was developed to separate hypoviral dsRNA from phenol/chloroform-extracted total nucleic acid preparation. Following standardization, chromatograms revealed clear evidence of hypoviral presence without nucleic acid precipitation and electrophoresis. This efficient diagnostic approach yields also pure hypoviral dsRNA fractions for further analyses.

Cryphonectria parasitica; Hypovirus; dsRNA; Monolith chromatography; Detection; Purification

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