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

Abstract

Background 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. Results Infectious 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). Conclusions These results demonstrate that MDMV-based vector can be used as a tool for simultaneous gene expression and multi-gene silencing in maize.

Proteome expansion in thePotyviridaeevolutionary radiation

“… 2021 , Xie et al . 2021 ). Knowledge of Type-A P1 and its proteolytic activity has been instrumental in generating the first potyviral vectors (Fig.…”

“…In addition to protein overexpression, heterologous sequences inserted within P1 or at the P1/HC-pro junction can trigger silencing of plant homologs (Gammelgård, Mohan and Valkonen 2007 , Xie et al . 2021 ). Potyvirid vectors have been used for virus-induced gene silencing, as well as for simultaneous plant gene silencing and heterologous protein production (Gammelgård, Mohan and Valkonen 2007 , Tuo et al .…”

Rescue of the first Alphanucleorhabdovirus entirely from cloned complementary DNA: an efficient vector for systemic expression of foreign genes in maize and insect vectors

“…In addition, plant virus-based expression vectors have been used for the expression of vaccine antigens, antibodies and other therapeutic proteins (LeBlanc et al, 2021). To date, a few maize viruses have been developed for transient gene function studies (reviewed in Kant and Dasgupta, 2019; Xie et al, 2021, Moltshwa et al, 2020). While important, these systems have limited use for maize researchers, due to transgene instability, narrow host range in monocots, and severe symptoms in inoculated plants (Ding et al, 2006; van der Linde et al, 2011; Benavente et al, 2012).…”

Transmission, localization, and infectivity of seedborne maize chlorotic mottle virus

“…To ensure positive pools resulted from seed transmission of MCMV and not from cross-contamination, the individual plants from which discs in positive pools were collected were resampled as described above. RNA was extracted from each individual and used for RT-PCR as previously described [27,41]. To minimize the presence of false negative tests, samples from pools with mean absorbances greater than the mean of the healthy controls plus three times the standard deviation of the mean absorbance of the healthy controls (>HC + 3SD) were resampled and retested by ELISA or RT-PCR.…”