Abstract
Virus-induced gene silencing (VIGS) is a useful functional genomics tool for rapidly creating plant gene knockout phenotypes that can be used to infer gene function. Until recently, VIGS has only been possible in dicotyledonous plants. However, the development of cloning vectors based on Barley stripe mosaic virus (BSMV) has now made VIGS possible in barley and wheat. VIGS has particular advantages for functional genomics in wheat, where the organism’s hexaploidy and recalcitrance to transformation have greatly hindered strategies for the functional identification of genes. In this chapter, methods are presented for using the Barley stripe mosaic virus VIGS system (BSMV-VIGS) to silence genes in hexaploid wheat.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Kumagai MH, Donson J, Della-Cioppa G, Harvey D, Hanley K, Grill LK (1995) Cytoplasmic inhibition of carotenoid biosynthesis with virus-derived RNA. Proc Natl Acad Sci U S A 92:1679–1683
Ratcliff F, Harrison BD, Baulcombe DC (1997) A similarity between viral defence and gene silencing in plants. Science 276:1558–1560
Baulcombe DC (1999) Fast forward genetics based on virus-induced gene silencing. Curr Opin Plant Biol 2:109–113
Holzberg S, Brosio P, Gross C, Pogue GP (2002) Barley stripe mosaic virus-induced gene silencing in a monocot plant. Plant J 30:315–327
Ding XS, Schneider WL, Chaluvadi SR, Mian MA, Nelson RS (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
Petty IT, Hunter BG, Wei N, Jackson AO (1989) Infectious barley stripe mosaic virus RNA transcribed in vitro from full-length genomic cDNA clones. Virology 171: 342–349
Scofield SR, Huang L, Brandt AS, Gill BS (2005) Development of a virus-induced gene-silencing system for hexaploid wheat and its use in functional analysis of the Lr21-mediated leaf rust resistance pathway. Plant Physiol 138:2165–2173
Bruun-Rasmussen M, Madsen CT, Jessing S, Albrechtsen M (2007) Stability of barley stripe mosaic virus-induced gene silencing in barley. Mol Plant Microbe Interact 20:1323–1331
Cakir C, Scofield SR (2008) Evaluating the ability of the barley stripe mosaic virus-induced gene silencing system to simultaneously silence two wheat genes. Cereal Res Commun 36(Suppl 6):217–222
Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
Bustin SA (2005) Real-time, fluorescence-based quantitative PCR: a snapshot of current procedures and preferences. Expert Rev Mol Diagn 5:493–498
Molnár A, Csorba T, Lakatos L, Várallyay E, Lacomme C, Burgyán J (2005) Plant virus-derived small interfering RNAs originate predominantly from highly structured single-stranded viral RNAs. J Virol 79:7812–7818
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Scofield, S.R., Brandt, A.S. (2012). Virus-Induced Gene Silencing in Hexaploid Wheat Using Barley Stripe Mosaic Virus Vectors. In: Watson, J., Wang, MB. (eds) Antiviral Resistance in Plants. Methods in Molecular Biology, vol 894. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-882-5_7
Download citation
DOI: https://doi.org/10.1007/978-1-61779-882-5_7
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-881-8
Online ISBN: 978-1-61779-882-5
eBook Packages: Springer Protocols