Simultaneous gene expression and multi-gene silencing in Zea mays using maize dwarf mosaic virus

Authors: XIE, WENSHUANG - The Ohio State University; Marty, Deemarie - Dee; XU, JUNHUAN - The Ohio State University; KHATRI, NITIKA - The Ohio State University; Willie, Kristen; BUCKNER MORAES, WANDERSON - The Ohio State University; Stewart, Lucy

Submitted to: BMC Biology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/12/2021
Publication Date: 5/5/2021
Citation: Xie, W., Marty, D., Xu, J., Khatri, N., Willie, K.J., Buckner Moraes, W., Stewart, L.R. 2021. Simultaneous gene expression and multi-gene silencing in Zea mays using maize dwarf mosaic virus. BMC Biology. 21. Article 208. https://doi.org/10.1186/s12870-021-02971-1.
DOI: https://doi.org/10.1186/s12870-021-02971-1

Interpretive Summary: Several plant viruses have been developed as research tools to better understand host plant gene function by silencing knockdown of target genes, or as tools to overexpress proteins for therapeutic or industrial purposes or as markers to ttrack virus movement. We report the development of maize dwarf mosaic virus (MDMV)-based constructs that can be used to simultaneously silence three host plant genes and express a protein in corn. This is the first report of a plant virus-based tool that can simultaneously be used for gene silencing and gene expression, introducing new possibilities for understanding corn gene functions and corn-virus interactions.

Technical Abstract: Maize dwarf mosaic virus (MDMV), a member of the genus Potyvirus, infects maize and is non-persistently transmitted by aphids. Several plant viruses have been developed as tools for gene expression and gene silencing in plants. The capacity of MDMV for both gene expression and gene silencing were examined. Infectious clones of an Ohio isolate of MDMV, MDMV-OH5, were obtained, and engineered for gene expression and simultaneous marker gene expression and virus-induced gene silencing (VIGS) of three endogenous maize target genes. Simultaneous expression of green fluorescent protein (GFP) and silencing of three maize genes was demonstrated. Constructs with GFP inserted in the N-terminus of HC-Pro were more stable than those with insertion at the N-terminus of CP in our study. Unexpectedly, the construct with two insertion sites also retained insertions at a higher rate than single-insertion constructs. Engineered MDMV expression and VIGS constructs were transmissible by aphids (Rhopalosiphum padi). These results demonstrate that MDMV-based vector can be used as a tool for simultaneous gene expression and multi-gene silencing in maize.