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Plasmid DNA Containing a Copy of RNA3 Can Substitute for RNA3 in Alfalfa Mosaic Virus RNA Inocula

Laboratoire de Virologie, Institut de Biologie Moléculaire des Plantes du CNRS, 12 rue du Général Zimmer, 67000 Strasbourg, France

The infectivity of a DNA copy of RNA3 from strain S of alfalfa mosaic virus inserted in a pT7-1 plasmid vector (clone 3pT7-1) was tested as circular DNA by co-inoculation with RNAs 1, 2 and 4 from strain B (mix I₀) on young tobacco leaves. Virus multiplication was monitored by serological detection of the coat protein. Progeny RNA3 derived from circular plasmid DNA 3pT7-1 was of the same length as naturally occurring RNA3-S. Analysis by primer extension demonstrated that it had the 5'-terminal sequence of RNA3-S and not that of RNA3-B. Mix I₀ also became infectious when mixed with M13 single-stranded DNA carrying RNA3-S inserted in the (-) strand orientation, probably because traces of RNA3-B present in mix I₀ were protected from degradation by base-pairing and were therefore active.

Keywords: AlMV, plasmid DNA, oligonucleotide priming

Received 3 December 1987; accepted 25 February 1988.