Resistance to Tomato Spotted Wilt Virus Infection in Transgenic Tobacco Expressing the Viral Nucleocapsid Gene. Donald J. MacKenzie. Research Station, Agriculture Canada, Vancouver, British Columbia, Canada V6T 1X2. P. J. Ellis. Research Station, Agriculture Canada, Vancouver, British Columbia, Canada V6T 1X2. MPMI 5:34-40. Accepted 15 October 1991. Copyright 1992 The American Phytopathological Society.

A recombinant plasmid containing the entire tomato spotted wilt virus (TSWV) nucleocapsid gene, with the exception of nucleotides 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-linked 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.