Abstract
Virus-induced gene silencing (VIGS) is a powerful technology for rapidly and transiently knocking down the expression of plant genes to study their functions. A VIGS vector for maize derived from Foxtail mosaic virus (FoMV), a positive-sense single-stranded RNA virus, was recently developed. A cloning site created near the 3′ end of the FoMV genome enables insertion of 200–400 nucleotide fragments of maize genes targeted for silencing. The recombinant FoMV clones are inoculated into leaves of maize seedlings by biolistic particle delivery, and silencing is typically observed within 2 weeks after inoculation. This chapter provides a protocol for constructing FoMV VIGS clones and inoculating them into maize seedlings.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Ding SW, Voinnet O (2007) Antiviral immunity directed by small RNAs. Cell 130:413–426. doi:10.1016/j.cell.2007.07.039
Baulcombe DC (1999) Fast forward genetics based on virus-induced gene silencing. Curr Opin Plant Biol 2:109–113
Becker A (ed) (2013) Virus-induced gene silencing: methods and protocols. Humana Press, New York
Mei Y, Zhang C, Kernodle BM et al (2016) A Foxtail mosaic virus vector for virus-induced gene silencing in maize. Plant Physiol 171:760–772. doi:10.1104/pp.16.00172
Ji Q, Yang B, Lee M et al (2010) Mapping of quantitative trait loci/locus conferring resistance to foxtail mosaic virus in maize using the intermated B73 × Mo17 population. Plant Breed 129:721–723. doi:10.1111/j.1439-0523.2009.01751.x
Ding XS, Schneider WL, Chaluvadi SR et al (2006) Characterization of a Brome mosaic virus strain and its use as a vector for gene silencing in monocotyledonous hosts. Mol Plant-Microbe Interact 19:1229–1239. doi:10.1094/MPMI-19-1229
Wang R, Yang X, Wang N et al (2016) An efficient virus-induced gene silencing vector for maize functional genomics research. Plant J 86:102–115. doi:10.1111/tpj.13142
Bancroft JB, Rouleau M, Johnston R et al (1991) The entire nucleotide sequence of foxtail mosaic virus RNA. J Gen Virol 72:2173–2181. doi:10.1099/0022-1317-72-9-2173
Bruun-Rasmussen M, Madsen CT, Johansen E et al (2008) Revised sequence of foxtail mosaic virus reveals a triple gene block structure similar to potato virus X. Arch Virol 153:223–226. doi:10.1007/s00705-007-1057-3
Robertson NL, French R, Morris TJ (2000) The open reading frame 5A of Foxtail mosaic virus is expressed in vivo and is dispensable for systemic infection. Arch Virol 145:1685–1698
Dickmeis C, Fischer R, Commandeur U (2014) Potato virus X-based expression vectors are stabilized for long-term production of proteins and larger inserts. Biotechnol J 9:1369–1379. doi:10.1002/biot.201400347
Acknowledgments
We thank Jaime Dittman for assistance with photography. This work was supported by the Iowa State University Plant Sciences Institute, the USDA National Institute of Food and Agriculture, Hatch project 3808, and State of Iowa funds.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Mei, Y., Whitham, S.A. (2018). Virus-Induced Gene Silencing in Maize with a Foxtail mosaic virus Vector. In: Lagrimini, L. (eds) Maize. Methods in Molecular Biology, vol 1676. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7315-6_7
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7315-6_7
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7314-9
Online ISBN: 978-1-4939-7315-6
eBook Packages: Springer Protocols