The optimum conditions for infection of turnip protoplasts with turnip rosette virus (TRosV) were 0.2 to 2 × 10 protoplasts/ml, 5 to 10 µg virus/ml, 1 µg poly--ornithine/ml in 0.6 -mannitol, buffered with 5 to 10 m-tris at pH 7.0 to 8.6. Under these conditions, 70% of protoplasts became infected, as indicated by staining with fluorescent antibody. The proportion of protoplasts infected was determined by factors such as pH and buffer ions and by the concentrations of protoplasts, virus and poly--ornithine during inoculation. The period of pre-inoculation incubation of virus and poly--ornithine was also critical, but mannitol concentration and the inoculation period and temperature had little effect on percentage protoplasts infected. Time course studies showed a single step multiplication of TRosV; the virus was detected on sucrose gradients at 24 h after inoculation and at 60 h reached its maximum concentration, 3 × 10 virions per infected protoplast.


Article metrics loading...

Loading full text...

Full text loading...


Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error