Cucumber mosaic virus (CMV) RNA-dependent RNA polymerase (RdRp) was purified form CMV-infected tobacco. The purified enzyme is completely dependent on exogenous template. The enzyme utilizes a variety of viral RNAs and CMV satellite RNA as template for minus-strand synthesis. Cellular RNAs are not used as templates. Ribosomal RNA inhibits the viral RNA synthesis by the CMV RdRp.