An RNA-dependent RNA polymerase (RdRp) purified from alfalfa mosaic vi
rus-infected tobacco is capable of synthesizing in vitro full-size RNA
s of minus and plus polarities. However, the enzyme is not able to per
form a complete replication cycle in vitro. The products were found to
be completely base-paired to their templates. The enzyme was able to
use double-stranded RNA as a template for RNA synthesis if it could in
itiate from a single-stranded promoter, The inability (of most) of our
enzyme preparations to create a single-stranded initiation site could
explain why they could not perform a complete replication cycle in vi
tro, This is the first report on duplex RNA unwinding activities by a
plant viral RdRp.