Resistance to tomato spotted wilt virus infection in transgenic tobacco expressing the viral nucleocapsid gene.
A recombinant plasmid containing the entire tomato spotted with virus (TSWV) nucleocapsid gene, with the exception of nucleotide encoding three N-terminal amino acids, was isolated by screening a complementary DNA library, prepared against random primed viral RNA, using a specific monoclonal antibody. The insert contained in plasmid pTSW1 was repaired and amplified by polymerase chain reaction, and the complete nucleocapsid protein gene was introduced into Nicotiana tabacum 'Samsun' by leaf disk transformation using Agrobacterium tumefaciens. Transgenic plants expressing the viral nucleocapsid protein were resistant to subsequent infection following mechanical inoculation with TSWV as indicated by a lack of systemic symptoms and little or no systemic accumulation of virus as determined by double antibody sandwich enzyme-liked immunosorbent assay. These results further extend the applicability of coat protein-mediated resistance, as previously demonstrated for a number of simple plant viruses composed of a positive-sense RNA genome encapsidated with a single species of coat protein, to a membrane-encapsidated, multi-component, negative-sense RNA virus.
MacKenzie
D J
DJ
Research Station, Agriculture Canada, Vancouver, British Columbia.
Ellis
P J
PJ
D001483:Base Sequence; D002213:Capsid / Q000235:genetics*; D003001:Cloning, Molecular; D015870:Gene Expression; D007113:Immunity, Innate; D008969:Molecular Sequence Data; D010942:Plant Viruses / Q000235:genetics* / Q000472:pathogenicity; D010945:Plants, Edible / Q000382:microbiology*; D030821:Plants, Genetically Modified / Q000382:microbiology*; D010947:Plants, Toxic; D014026:Nicotiana / Q000382:microbiology*