Extracts of barley leaves infected with bromegrass mosaic virus (BrMV) incorporated radioactivity into RNA when incubated with [H]UTP in the presence of actinomycin D (Semal & Kummert, 1970). The radioactive product thus obtained was partly resistant to pancreatic RNase in × 2:SSC (0.15 -NaCl, 0.015 -sodium citrate, pH 7.0:SSC) but was entirely hydrolysed by this enzyme in × 0.05:SSC (Semal, 1970). A sequential synthesis of double- and single-stranded RNA was obtained under certain experimental conditions (Semal & Kummert, 1971). The present results identify the radioactive product associated with duplex RNA as a segment of BrMV-RNA.

Cell-free extracts of the second leaf of barley seedlings, whose first leaf had been inoculated 3 days earlier with BrMV, were used to prepare the crude virus-induced RNA polymerase fraction. This fraction was incubated for 3½ min. with tritiated UTP (1 to 2 /m — The Radiochemical Centre, Amersham, Buckinghamshire, England), 10 -EDTA, 20 µg./ml. of actinomycin D and the necessary ingredients for RNA synthesis, as previously described (Semal & Kummert, 1971).


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