title: Grapevine fanleaf virus (GFLV)-specific antibodies confer GFLV and Arabis mosaic virus (ArMV) resistance in Nicotiana benthamiana.
abstract: Grapevine fanleaf virus (GFLV) is one of the most destructive pathogens of grapevine. In this study, we generated monoclonal antibodies binding specifically to the coat protein of GFLV. Antibody FL(3), which bound most strongly to GFLV and showed cross-reactivity to Arabis mosaic virus (ArMV), was used to construct the single-chain antibody fragment scFvGFLVcp-55. To evaluate the potential of this single-chain variable fragment (scFv) to confer antibody-mediated virus resistance, transgenic Nicotiana benthamiana plants were generated in which the scFv accumulated in the cytosol. Recombinant protein levels of up to 0.1% total soluble protein were achieved. The T(1) and T(2) progenies conferred partial or complete protection against GFLV on challenge with the viral pathogen. The resistance to GFLV in transgenic plants was strictly related to scFvGFLVcp-55 accumulation levels, confirming that the antibody fragment was functional in planta and responsible for the GFLV resistance. In addition, transgenic plants conferring complete protection to GFLV showed substantially enhanced tolerance to ArMV. We demonstrate the first step towards the control of grapevine fanleaf degeneration, as scFvGFLVcp-55 could be an ideal candidate for mediating nepovirus resistance.
authors.affiliation: Institute for Molecular Biotechnology (Biology VII), RWTH Aachen University, Worringerweg 1, 52074 Aachen, Germany.
authors.forename: Greta
authors.initials: G
authors.lastname: Nölke
authors.forename: Pascal
authors.initials: P
authors.lastname: Cobanov
authors.forename: Kerstin
authors.initials: K
authors.lastname: Uhde-Holzem
authors.forename: Götz
authors.initials: G
authors.lastname: Reustle
authors.forename: Rainer
authors.initials: R
authors.lastname: Fischer
authors.forename: Stefan
authors.initials: S
authors.lastname: Schillberg
mesh_terms: D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology*; D001483:Base Sequence; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D008854:Microscopy, Electron; D009029:Mosaic Viruses / Q000276:immunology*; D010942:Plant Viruses / Q000276:immunology*; D030821:Plants, Genetically Modified / Q000276:immunology / Q000821:virology; D012328:RNA Viruses / Q000276:immunology*; D014026:Nicotiana / Q000276:immunology / Q000821:virology*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D017931:DNA Primers