Analysis of viral RNA isolated from tobacco leaf tissue infected with tobacco etch virus.
Total RNA has been isolated from tobacco seedlings systemically infected with the potyvirus, tobacco etch virus (TEV). Analysis of this RNA by agarose gel electrophoresis under denaturing conditions, in conjunction with gel hybridization using various virus-specific nucleic acid probes, revealed genomic length TEV RNA and eight zones of less-than-full-length RNA. However, reconstitution experiments demonstrated that the zones were electrophoretic artifacts and not authentic subgenomic RNAs. Furthermore, only a single polyadenylated viral RNA species, which comigrated with TEV genomic RNA in gel electrophoresis studies, was isolated from infected tobacco leaf tissue. Translation of polyadenylated RNA in a rabbit reticulocyte lysate resulted in the synthesis of four high-molecular-weight virus-specific products. The products had apparent molecular weights and serological reactivities as follows: (1) 120,000, reaction with antiserum to TEV cytoplasmic inclusion protein; (2) 87,000, no serological reaction detected; (3) 85,000, reaction with antisera to TEV capsid protein and the large-molecular-weight component of the nuclear inclusion body; (4) 49,000, reaction with antiserum to the small-molecular-weight component of the nuclear inclusion body.
Dougherty
W G
WG
Department of Plant Pathology, North Carolina State University, Raleigh, North Carolina 27650, USA.