Purification and Characterization of the Circular and Linear Forms of Chrysanthemum Stunt Viroid

Abstract

A new and rapid extraction procedure is described for the isolation and partial purification of low mol. wt. RNA as well as DNA from 500 g batches of chrysanthemum stunt viroid-infected chrysanthemum. The chrysanthemum stunt viroid was then purified to homogeneity from this extract by one cycle of non-denaturing polyacrylamide slab gel electrophoresis followed by one cycle of denaturing slab gel electrophoresis. The covalently closed circular form and the linear form of the viroid co-migrated on the first gel but were well separated on the second. The yield of circular chrysanthemum stunt viroid was 200 µg/kg of infected chrysanthemum shoots and of the linear form was 35 µg/kg.