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SummaryAn endogenous RNA-dependent RNA polymerase has been demonstrated in broccoli necrotic yellows virus (BNYV) particles. The enzyme requires a temperature of 26 °C, a pH value of 7.5, 0.5% NP40 and 4 mm-Mg2+ for maximum synthesis of RNA; Mn2+ inhibited the activity. Addition of EDTA to the reaction mixture did not stimulate the activity in the presence of higher amounts of Mg2+. The in vitro transcription products of BNYV hybridized to the virus genome, but not to that of lettuce necrotic yellows virus (LNYV). The transcriptase activity was abolished by centrifugation on a glycerol gradient after treating the virus with NP40 in a high salt solution; combining the pellet and the upper part of the gradient restored the activity. Cross-complementation of BNYV and LNYV transcription could not be obtained by heterologous combination of the pellet and supernatant fractions of the two viruses.

Differentiation between Broccoli Necrotic Yellows Virus and Lettuce Necrotic Yellows Virus by their Transcriptase Activities