Direct Agroinoculation of Maize Seedlings by Injection with Recombinant Foxtail Mosaic Virus and Sugarcane Mosaic Virus Infectious Clones Bliss M. Beernink*,1, Katerina L. Holan*,1, Ryan R. Lappe1, Steven A. Whitham1 1 Department of Plant Pathology and Microbiology, Iowa State University * These authors contributed equally Corresponding Author Abstract Steven A. Whitham
[email protected] Agrobacterium-based inoculation approaches are widely used for introducing viral vectors into plant tissues. This study details a protocol for the injection of maize Citation seedlings near meristematic tissue with Agrobacterium carrying a viral vector. Beernink, B.M., Holan, K.L., Lappe, R.R., Recombinant foxtail mosaic virus (FoMV) clones engineered for gene silencing and Whitham, S.A. Direct Agroinoculation of Maize Seedlings by Injection with gene expression were used to optimize this method, and its use was expanded Recombinant Foxtail Mosaic Virus and to include a recombinant sugarcane mosaic virus (SCMV) engineered for gene Sugarcane Mosaic Virus Infectious Clones. J. Vis. Exp. (168), e62277, expression. Gene fragments or coding sequences of interest are inserted into a doi:10.3791/62277 (2021). modified, infectious viral genome that has been cloned into the binary T-DNA plasmid vector pCAMBIA1380. The resulting plasmid constructs are transformed into Date Published Agrobacterium tumefaciens strain GV3101. Maize seedlings as young as 4 days February 27, 2021 old can be injected near the coleoptilar node with bacteria resuspended in MgSO4 DOI