DETECTION OF DNA AND RNA PLANT-VIRUSES BY PCR AND RT-PCR USING A RAPID VIRUS RELEASE PROTOCOL WITHOUT TISSUE HOMOGENIZATION

A simple, single-step plant tissue preparation protocol suitable for t he detection of viruses by the polymerase chain reaction and reverse t ranscription-polymerase chain reaction is described. The effect of buf fer components and pH, and the incubation temperature for the release of virus from plant material was evaluated. A small amount of plant ti ssue was heated in a solution containing 100 mM Tris-HCl, pH 7.4 or 8. 4, 1 M KCl and 10 mM EDTA for 10 min at 95 degrees C and the supernata nt used for enzymatic amplification. This protocol was suitable for th e detection of both DNA and RNA viruses in a variety of plant species and tissues and reduced plant inhibitory factors which may interfere w ith PCR. The application of this method was demonstrated for the detec tion of banana bunchy top virus in banana leaves, root and corm, zucch ini yellow mosaic potyvirus in squash leaves and lettuce necrotic yell ows rhabdovirus in lettuce and Nicotiana glutinosa leaves.