Autoradiography of 32P and 14C incorporation into protoplasts as a means of determining the percentage of virus infected protoplasts.
Stripping Film Autoradiography (SFA) was used to measure percentage protoplast infection. Control and infected protoplasts were incubated in the presence of [32P]orthophosphate or [14C]leucine fixed onto glass slides and placed in contact with a sensitive photographic emulsion. Infected protoplasts were distinguished by blackened areas of exposed film which coincided with protoplasts containing labelled virus products. The use of SFA is demonstrated for two different virus/protoplast systems, namely tobacco mosaic virus (TMV) in tobacco and turnip yellow mosaic virus (TYMV) in rape. The method detects infection as early as 12-16 h post-inoculation and UV irradiation is not necessary for reducing background interference from labelled host proteins.
D001345:Autoradiography / Q000379:methods*; D002250:Carbon Radioisotopes; D009029:Mosaic Viruses / Q000032:analysis* / Q000378:metabolism; D010761:Phosphorus Radioisotopes; D010935:Plant Diseases; D013997:Time Factors
D002250:Carbon Radioisotopes; D010761:Phosphorus Radioisotopes