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Molecular Cloning of Single-stranded RNAs of Potato Leafroll Virus and Beet Western Yellows Virus

Biologische Bundesanstalt für Land- und Forstwirtschaft, Institut für Viruskrankheiten der Pflanzen, Messeweg 11/12, D-3300 Braunschweig, F.R.G.

Complementary DNA libraries were synthesized using the genomic RNAs of potato leafroll virus (PLRV) and beet western yellows virus (BWYV), by random as well as oligo(dT) priming of polyadenylated RNA, and cloned into pBR322. Two restriction endonuclease maps were constructed, extending to 6·1 kbp and 5·5 kbp for the PLRV and BWYV genomes respectively. The 3' ends of genomic RNAs were verified by sequence analysis of oligo(dT)-primed clones; 5'-terminal sequences of PLRV and BWYV were not detected among the cDNA fragments mapped. Selected clones were used to demonstrate specific virus detection in total nucleic acid preparations of PLRV- or BWYV-infected Physalis floridana Rydb. One PLRV-derived clone showed cross-hybridization with purified BWYV RNA.

Keywords: luteoviruses, PLRV, BWYV

Received 1 March 1988; accepted 23 May 1988.