Abstract
BackgroundMaize 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.ResultsInfectious clones of an Ohio isolate of MDMV, MDMV OH5, were obtained, and engineered for gene expression only, and for simultaneous marker gene expression and virus-induced gene silencing (VIGS) of three endogenous maize target genes. Single gene expression in single insertion constructs and simultaneous expression of green fluorescent protein (GFP) and silencing of three maize genes in a double insertion construct was demonstrated. Constructs with GFP inserted in the N-terminus of HCPro 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).ConclusionsThese results demonstrate that MDMV-based vector can be used as a tool for simultaneous gene expression and multi-gene silencing in maize.
Highlights
Maize dwarf mosaic virus (MDMV), a member of the genus Potyvirus, infects maize and is nonpersistently transmitted by aphids
Several viruses have been developed as tools for gene expression or silencing, including brome mosaic virus (BMV) [20, 21], cucumber mosaic virus (CMV) [13], and maize rayado fino virus (MRFV) [22] for gene silencing [23], wheat streak mosaic virus (WSMV) [24,25,26] and sugarcane mosaic virus [27] for protein expression, foxtail mosaic virus (FoMV) for gene silencing [28] or protein expression [29] and guide RNA delivery for CRISPR/ Cas9 gene editing [30], barley stripe mosaic virus for gene silencing [23] or protein expression [31] or for guide RNA delivery [32] and barley yellow striate mosaic virus (BYSMV) for simultaneous guide RNA and high cargo-capacity protein expression [33]
We report the cloning and sequencing of a new Ohio MDMV isolate from Johnsongrass, MDMV OH5, development of MDMV OH5 infectious clones, and engineered constructs for simultaneous gene expression and multi-gene silencing in maize
Summary
Maize dwarf mosaic virus (MDMV), a member of the genus Potyvirus, infects maize and is nonpersistently transmitted by aphids. Several plant viruses have been developed as tools for gene expression and gene silencing in plants. Many plant viruses have been developed as beneficial tools for expression of protein or VIGS (reviewed in [4,5,6,7,8,9,10]). A plant virus vector derived from bean pod mottle virus (BPMV) was even developed for simultaneous single gene expression and single gene silencing [19]. Several viruses have been developed as tools for gene expression or silencing, including brome mosaic virus (BMV) [20, 21], cucumber mosaic virus (CMV) [13], and maize rayado fino virus (MRFV) [22] for gene silencing [23], wheat streak mosaic virus (WSMV) [24,25,26] and sugarcane mosaic virus [27] for protein expression, foxtail mosaic virus (FoMV) for gene silencing [28] or protein expression [29] and guide RNA delivery for CRISPR/ Cas gene editing [30], barley stripe mosaic virus for gene silencing [23] or protein expression [31] or for guide RNA delivery [32] and barley yellow striate mosaic virus (BYSMV) for simultaneous guide RNA and high cargo-capacity protein expression [33]
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