Author
Stewart, Lucy | |
BOUCHARD, ROBERT - The Ohio State University | |
Redinbaugh, Margaret | |
MEULIA, TEA - The Ohio State University |
Submitted to: Virus Research
Publication Type: Peer Reviewed Journal Publication Acceptance Date: 2/3/2012 Publication Date: 5/1/2012 Citation: Stewart, L.R., Bouchard, R., Redinbaugh, M.G., Meulia, T. 2012. Complete sequence and development of a full-length infectious clone of an Ohio isolate of Maize dwarf mosaic virus (MDMV). Virus Research. 165:219-224. Interpretive Summary: This work reports the complete sequence of an Ohio isolate of Maize dwarf mosaic virus (MDMV), and development of a full-length infectious complementary DNA (cDNA) of the (+) RNA viral genome. MDMV is one of the most widespread and destructive corn virus pathogens in the United States, and is prevalent worldwide. Full-length MDMV genomic sequence was cloned and maintained in the plasmid pSPORT, from which sequence was amplified by PCR and used as template for in vitro transcription. Capped in vitro transcripts were used to inoculate corn seeds by vascular puncture inoculation, and infection of inoculated seeds showed that the full-length sequence was infectious in plants. Technical Abstract: Maize dwarf mosaic virus (MDMV) is an important and widespread aphid-transmitted virus of corn. It is a member of the genus Potyvirus in the family Potyviridae. Here we report the complete genome sequence of, and construction of an infectious clone of an Ohio isolate of MDMV-A. MDMV sequences from infected plants were cloned into the vector pSPORT. Plasmids containing the full-length MDMV sequences were used as template to amplify full-length sequence by polymerase chain reaction (PCR) using three 5’ end sequence variations, and were used as template for in vitro transcription. Transcripts were inoculated to corn by vascular puncture inoculation (VPI). Each of the transcripts was infectious in corn when delivered by VPI. This is fourth reported full-length genomic sequence of an MDMV isolate, and the first reported MDMV full-length infectious clone. |