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Multiplication of Velvet Tobacco Mottle Virus in Nicotiana clevelandii Protoplasts Is Resistant to -Amanitin

¹ Department of Virology, University of Wuhan, Wuhan, The People's Republic of China
and² Institute of Microbiology, Academia Sinica, Beijing, The People's Republic of China

Protoplasts from Nicotiana clevelandii were infected with velvet tobacco mottle virus (VTMoV) using inocula containing 2 µg/ml poly-L-ornithine (PLO), 8 µg/ml VTMoV and 0.02 M-potassium citrate pH 5.0. Typically about 47% of the inoculated protoplasts become infected. The amount of virus particles bound to inoculated protoplasts was increased by a decrease in pH, an increase in PLO concentration or an increase in virus concentration. The one-step growth curve of VTMoV in N. clevelandii protoplasts showed that, although large amounts of virus were synthesized, the multiplication was slower than that published for other viruses. Alpha-amanitin (50 µg/ml) had no effect on the rate of accumulation of virus coat protein or on the synthesis of either RNA species of VTMoV when added to cultures immediately after inoculation.

Keywords: multiplication, VTMoV, protoplasts, -amanitin, virusoid

Received 4 March 1986; accepted 3 September 1986.