title: In Vitro Transcripts of Wild-Type and Fluorescent Protein-Tagged Triticum mosaic virus (Family Potyviridae) are Biologically Active in Wheat.
abstract: Triticum mosaic virus (TriMV) (genus Poacevirus, family Potyviridae) is a recently described eriophyid mite-transmitted wheat virus. In vitro RNA transcripts generated from full-length cDNA clones of TriMV proved infectious on wheat. Wheat seedlings inoculated with in vitro transcripts elicited mosaic and mottling symptoms similar to the wild-type virus, and the progeny virus was efficiently transmitted by wheat curl mites, indicating that the cloned virus retained pathogenicity, movement, and wheat curl mite transmission characteristics. A series of TriMV-based expression vectors was constructed by engineering a green fluorescent protein (GFP) or red fluorescent protein (RFP) open reading frame with homologous NIa-Pro cleavage peptides between the P1 and HC-Pro cistrons. We found that GFP-tagged TriMV with seven or nine amino acid cleavage peptides efficiently processed GFP from HC-Pro. TriMV-GFP vectors were stable in wheat for more than 120 days and for six serial passages at 14-day intervals by mechanical inoculation and were transmitted by wheat curl mites similarly to the wild-type virus. Fluorescent protein-tagged TriMV was observed in wheat leaves, stems, and crowns. The availability of fluorescent protein-tagged TriMV will facilitate the examination of virus movement and distribution in cereal hosts and the mechanisms of cross protection and synergistic interactions between TriMV and Wheat streak mosaic virus.
authors.affiliation: First author: United States Department of Agriculture-Agricultural Research Service (USDA-ARS) and Department of Plant Pathology; second and fourth authors: Department of Entomology; third author: USDA-ARS; and fifth author: USDA-ARS and Department of Agronomy and Horticulture, University of Nebraska-Lincoln, Lincoln 68583.
authors.forename: Satyanarayana
authors.initials: S
authors.lastname: Tatineni
authors.affiliation: First author: United States Department of Agriculture-Agricultural Research Service (USDA-ARS) and Department of Plant Pathology; second and fourth authors: Department of Entomology; third author: USDA-ARS; and fifth author: USDA-ARS and Department of Agronomy and Horticulture, University of Nebraska-Lincoln, Lincoln 68583.
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: McMechan
authors.affiliation: First author: United States Department of Agriculture-Agricultural Research Service (USDA-ARS) and Department of Plant Pathology; second and fourth authors: Department of Entomology; third author: USDA-ARS; and fifth author: USDA-ARS and Department of Agronomy and Horticulture, University of Nebraska-Lincoln, Lincoln 68583.
authors.forename: Melissa
authors.initials: M
authors.lastname: Bartels
authors.affiliation: First author: United States Department of Agriculture-Agricultural Research Service (USDA-ARS) and Department of Plant Pathology; second and fourth authors: Department of Entomology; third author: USDA-ARS; and fifth author: USDA-ARS and Department of Agronomy and Horticulture, University of Nebraska-Lincoln, Lincoln 68583.
authors.forename: Gary L
authors.initials: GL
authors.lastname: Hein
authors.affiliation: First author: United States Department of Agriculture-Agricultural Research Service (USDA-ARS) and Department of Plant Pathology; second and fourth authors: Department of Entomology; third author: USDA-ARS; and fifth author: USDA-ARS and Department of Agronomy and Horticulture, University of Nebraska-Lincoln, Lincoln 68583.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Graybosch
mesh_terms: D000818:Animals; D001179:Arthropod Vectors; D016368:Frameshift Mutation; D049452:Green Fluorescent Proteins; D054884:Host-Pathogen Interactions; D008164:Luminescent Proteins; D008925:Mites; D010935:Plant Diseases; D019176:Potyviridae / Q000235:genetics / Q000472:pathogenicity*; D017423:Sequence Analysis, RNA; D014908:Triticum / Q000821:virology*; D000097573:Red Fluorescent Protein
chemical_list: D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein