REVERSE-TRANSCRIPTION POLYMERASE-CHAIN-REACTION FOR THE DETECTION OF VIRUSES FROM PLANTS AND APHIDS

A reverse transcription polymerase chain reaction (RT-PCR) protocol us ed for the detection of potato viruses in dormant tubers and leaves an d in an aphid vector is described. Problems in plant sample preparatio n from different hosts, uneven distribution or low concentration of vi ruses and the presence of PCR inhibitors in plant extracts are discuss ed and various ways to eliminate their effect are described. Using Pot yviridae viruses, it has been shown that RT-PCR in various modified fo rms can be used to differentiate viruses at the level of family, genus , species, strains and their subtypes or serotypes. The specificity of primer pairs and PCR modifications has been used to separate closely related potato viruses A and PVY strains (PVY degrees, PVYN and PVYNTN ) from a known mixture. (C) 1998 Elsevier Science B.V. All rights rese rved.