A centrifugation method for separation of plant viral genomic and subgenomic RNAs

AbstractUsing alfalfa mosaic virus (AMV) as a model, a simple method for separating plant viral genomic RNAs from their subgenomic counterparts was established. The method relies on sucrose gradient fractionation under carefully selected conditions of centrifugation and fraction collection. The RNA components are recovered in nearly quantitative yield and have full biological activity as measured by infectivity of the reconstituted RNAs in suitable protoplasts and plant hosts. The individual RNAs, on the other hand, show no such infectivity, indicating that the separation is indeed complete