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Cowpea Mosaic Virus RNA Replication in Crude Membrane Fractions from Infected Cowpea and Chenopodium amaranticolor

Department of Molecular Biology, Agricultural University, Dreijenlaan 3, 6703 HA Wageningen
and¹ Department of Virology, Agricultural University, Binnenhaven 11, 6709 PD Wageningen, The Netherlands

The replication of cowpea mosaic virus (CPMV) RNA was studied in crude membrane fractions prepared from leaves of CPMV-infected cowpea and Chenopodium amaranticolor. In vitro replicase assays showed that in the cowpea extract only the replicative intermediate (RI) and replicative form (RF) were synthesized. In the C. amaranticolor extract however, single-stranded progeny RNA was produced in addition to RI and RF. Production of the ssRNA in the C. amaranticolor extract was a result of the greater stability of the CPMV replication complex in this host. Comparison of the viral replicase activity and the amount of virus-encoded proteins in cowpea and C. amaranticolor crude membrane fractions indicated that only a small fraction of the non-structural proteins detected in cowpea is active in RNA replication. This suggests that viral replication proteins are used only once, perhaps because of a stringent coupling of polyprotein processing and replication.

Keywords: CMPV, replication complex, RNA replication

Received 6 April 1988; accepted 7 July 1988.