Detection of five seedborne legume viruses in one sensitive multiplex polymerase chain reaction test

• Published in: Phytopathology 1994, Vol.84 (10), p.1201-1205
• Main Authors: Bariana, H.S. (CSIRO Division of Plant Industry, North Ryde, N.S.W., Australia), Shannon, A.L, Chu, P.W.G, Waterhouse, P.M
• Format: Article
• Language: English
• Subjects: ADN; AMPLIFICATION CHAINE POLYMERASE; Biological and medical sciences; Biotechnology; CUCUMOVIRUS DEL MOSAICO DEL PEPINO; CUCUMOVIRUS MOSAIQUE DU CONCOMBRE; Fundamental and applied biological sciences. Psychology; Generalities. Techniques. Transmission, epidemiology, ecology. Antiviral substances, control; LEGUMINOSAE; Phytopathology. Animal pests. Plant and forest protection; Plant viruses and viroids; POTYVIRUS; REACCION DE CADENAS DE POLIMERASA; VIRUS DE LAS PLANTAS; VIRUS DES VEGETAUX
• ISSN: 0031-949X; 1943-7684
• 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