Extreme resistance to potato virus X infection in plants expressing a modified component of the putative viral replicase.
Three types of mutation were introduced into the sequence encoding the GDD motif of the putative replicase component of potato virus X (PVX). All three mutations rendered the viral genome completely noninfectious when inoculated into Nicotiana clevelandii or into protoplasts of Nicotiana tabacum (cv. Samsun NN). In order to test whether these negative mutations could inactivate the viral genome in trans, the mutant genes were expressed in transformed N.tabacum (cv. Samsun NN) under control of the 35S RNA promoter of cauliflower mosaic virus and the transformed lines were inoculated with PVX. In 10 lines tested in which the GDD motif was expressed as GAD or GED there was no effect on susceptibility to PVX. In two of four lines transformed to express the ADD form of the conserved motif, the F1 and F2 progeny plants were highly resistant to infection by PVX, although only to strains closely related to the source of the transgene. The resistance was associated with suppression of PVX accumulation in the inoculated and systemic leaves and in protoplasts of the transformed plants, although some low level viral RNA production was observed in the inoculated but not the systemic leaves when the inoculum was as high as 100 or 250 micrograms/ml PVX RNA. These results suggest for a plant virus, as reported previously for Q beta phage, that virus resistance may be engineered by expression of dominant negative mutant forms of viral genes in transformed cells.
Longstaff
M
M
Sainsbury Laboratory, Norwich Research Park, UK.
Brigneti
G
G
Boccard
F
F
Chapman
S
S
Baulcombe
D
D
D000595:Amino Acid Sequence; D001483:Base Sequence; D004247:DNA; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D020022:Genetic Predisposition to Disease; D016679:Genome, Viral; D008969:Molecular Sequence Data; D009154:Mutation; D010935:Plant Diseases / Q000235:genetics* / Q000382:microbiology; D010940:Plant Proteins / Q000235:genetics; D010942:Plant Viruses / Q000201:enzymology* / Q000235:genetics; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D011523:Protoplasts; D012367:RNA, Viral; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism
D010940:Plant Proteins; D012367:RNA, Viral; D004247:DNA; C023140:DNA replicase; D004259:DNA-Directed DNA Polymerase