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Localization of Genetic Information Involved in the Replication of Alfalfa Mosaic Virus

Department of Biochemistry, State University of Leiden, Wassenaarseweg 64, 2333 AL Leiden, The Netherlands

Cowpea protoplasts were inoculated with different combinations of the four nucleoprotein components of alfalfa mosaic virus (AIMV), and the synthesis of viral particles, coat protein and RNA was analysed. Synthesis of viral particles and coat protein was detectable by polyacrylamide gel electrophoresis only when the inoculum contained the components B, M and Tb, accommodating RNAs 1, 2 and 3 respectively.

Upon inoculation with B + M + Tb, the synthesis of viral RNA was detectable by annealing of radiolabelled RNA from protoplasts to virus-specific double-stranded RNA. On inoculation with B + Tb or M + Tb no significant amount of viral RNA was produced. After inoculation with B + M, however, the production of virus-specific RNA was about 40% of that induced by complete infection, indicating that genetic information encoded by RNAs 1 and 2 is involved in viral RNA replication. As a mixture of the genomic RNAs did not induce the synthesis of virus-specific RNA, it is concluded that coat protein is required for the RNA synthesis induced by the mixture of B and M nucleoproteins.

The observation that no coat protein is produced upon inoculation with B + M + Ta components indicates that the replicase activity induced by B + M does not replicate RNA 4, the messenger for coat protein present in Ta particles.

Received 29 July 1980; accepted 20 October 1980.

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