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Detection of Five Seedborne Legume Viruses in One Sensitive Multiplex Polymerase Chain Reaction Test. H. S. Bariana, CSIRO Division of Plant Industry, Canberra, ACT 2601, Australia, Present address: CSIRO Division of Plant Industry, Grain Quality Research Laboratory, P.O. Box 7, North Ryde, N.S.W. 2113, Australia; A. L. Shannon, P. W. G. Chu, and P. M. Waterhouse. CSIRO Division of Plant Industry, Canberra, ACT 2601, Australia. Phytopathology 84:1201-1205. Accepted for publication 10 June 1994. Copyright 1994 The American Phytopathological Society. DOI: 10.1094/Phyto-84-1201.

A reverse transcription polymerase chain reaction (RT-PCR) assay has been developed that can simultaneously test a sample, in one tube, for the presence of five seedborne legume viruses that are of great concern to legume germ plasm banks: alfalfa mosaic alfamovirus, bean yellow mosaic potyvirus, clover yellow vein potyvirus, cucumber mosaic cucumovirus, and subterranean clover mottle sobemovirus. RT-PCR assays were also developed for the detection of subterranean clover redleaf luteovirus and subterranean clover stunt virus, thus providing a detection system for all well-characterized viruses known to infect subterranean clover. Primers were designed so that the size of the RT-PCR product was indicative of the virus amplified and, where sequences of more than one strain of virus were available, conserved regions were given preference as primer targets. The RT-PCR assay detected all isolates tested for each virus and was up to five orders of magnitude more sensitive than ELISA.