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ARS Home » Plains Area » Lincoln, Nebraska » Wheat, Sorghum and Forage Research » Research » Publications at this Location » Publication #316477
Title: In vitro transcripts of wild-type and fluorescent protein-tagged triticum mosaic virus (family potyviridae) are biologically active in wheat

Author
item Tatineni, Satyanarayana - Ts
item MCMECHAN, ANTONY - University Of Nebraska
item Bartels, Melissa
item HEIN, GARY - University Of Nebraska
item Graybosch, Robert

Submitted to: Phytopathology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/21/2015
Publication Date: 11/17/2015
Publication URL: http://handle.nal.usda.gov/10113/61814
Citation: Tatineni, S., McMechan, A.J., Bartels, M.S., Hein, G.L., Graybosch, R.A. 2015. In vitro transcripts of wild-type and fluorescent protein-tagged triticum mosaic virus (family potyviridae) are biologically active in wheat. Phytopathology. 105:1496-1505.

Interpretive Summary: Triticum mosaic virus (TriMV), a recently reported wheat virus, is widespread in the Great Plains of North America. In this study, the RNA genome of TriMV was converted into double-stranded DNA form and cloned in a plasmid vector to allow “reverse genetics” studies of TriMV gene function and biology. RNA produced from the DNA clone was infectious on wheat and induced symptoms similar to those of the wild-type TriMV. The cloned virus retained wheat curl mite transmission, virus movement, and pathogenicity traits similar to those of the wild-type virus. Additionally, the TriMV genome was modified to independently express green fluorescent protein (GFP) or red fluorescent protein (RFP). These proteins allow tracking of virus within infected plants. Both GFP- and RFP-tagged TriMV efficiently infected wheat with stable expression of fluorescent protein for an extended period. The reverse genetics system and GFP or RFP-tagged TriMV are powerful tools that will help determine the functions of TriMV-encoded proteins in virus biology and allow rapid screening of antimicrobial peptides against wheat bacterial diseases.

Technical 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, and the progeny virus was efficiently transmitted by wheat curl mites. A series of TriMV-based expression vectors were constructed by inserting a GFP or RFP ORF with homologous NIa-Pro cleavage peptides between the P1 and HC-Pro cistrons. We found that GFP-tagged TriMV and Wheat streak mosaic virus (WSMV) with analogous cleavage peptides differentially processed GFP from HC-Pro, resulting in free GFP and dense fluorescent aggregates, respectively, suggesting differential NIa-Pro cleavage preferences of the two wheat-infecting Potyviridae members. TriMV-GFP vectors were stable in wheat and were transmitted by wheat curl mites. The availability of fluorescent protein-tagged TriMV will facilitate the examination of virus movement in cereal hosts and the mechanisms of cross protection and synergistic interactions between TriMV and WSMV in wheat.