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

S. Toriyama

Laboratory of Virology, Agricultural University, Binnenhaven 11, 6709 PD Wageningen, The Netherlands

An 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.

Keywords: broccoli and lettuce necrotic yellows virus, transcriptase activity

Present address: Laboratory of Plant Pathology, Faculty of Agriculture, University of Tokyo, Bunkyo-Ku, Tokyo 113, Japan.

Received 28 January 1981; accepted 5 May 1981.