cDNAs of beet necrotic yellow vein virus RNAs 3 and 4 could be rendered biologically active when they were placed under the control of the cauliflower mosaic virus 35S promoter and polyadenylation signal. Although the 35S in vivo transcripts should have contained up to forty 5′ and several hundred 3′ nonviral nucleotides, the progeny viral RNAs had the same sizes as in naturally infected sugarbeets. The progeny RNAs did not hybridize with the nonviral sequences indicating that they were apparently not replicated. Deletion and insertion mutants of RNA 3 cDNA clones were also biologically active in plants but a plasmid which contained the cDNA of RNA 3 in antisense orientation was not. The biological activity of plasmid DNAs compared with the corresponding synthetic transcripts is discussed.