Abstract
Successful application of virus-induced gene silencing for functional genomics requires a virus vector that can initiate a systemic infection of the host plant. Agroinoculation of the pea early browning virus vectors pCAPE1 and pCAPE2 can establish infection in several genotypes of Medicago truncatula and can reduce target gene RNA levels to an extent that allows investigation of gene function.
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
Grønlund M, Constantin G, Piednoir E et al (2008) Virus induced gene silencing in Medicago truncatula and Lathyrus odorata. Virus Res 135:345–349
Serwatowska J, Roque E, Gómez-Mena C et al (2014) Two euAGAMOUS genes control C-function in Medicago truncatula. PLoS One 9:e103770. https://doi.org/10.1371/journal.pone.0103770
Qian W, Yu C, Qin H et al (2007) Molecular and functional analysis of phosphomannomutase (PMM) from higher plants and genetic evidence for the involvement of PMM in ascorbic acid biosynthesis in Arabidopsis and Nicotiana benthamiana. Plant J 49:399–413
Constantin GD, Krath BN, MacFarlane SA, Nicolaisen M, Johansen IE, Lund OS (2004) Virus-induced gene silencing as a tool for functional genomics in a legume species. Plant J 40:622–631
Blein T, Pulido A, Vialette-Guiraud A (2008) A conserved molecular framework for compound leaf development. Science 322:1835–1839
Berbel A, Ferrándiz C, Hecht V et al (2012) VEGETATIVE1 is essential for development of the compound inflorescence in pea. Nat Commun 3:797. https://doi.org/10.1038/ncomms1801
Wang Z, Luo Y, Li X et al (2008) Genetic control of floral zygomorphy in pea (Pisum sativum L.). PNAS 105:10414–10419
Constantin G, Grønlund M, Johansen IE et al (2008) Virus induced gene silencing (VIGS) as a reverse genetic tool to study development of symbiotic root nodules. Mol Plant-Microbe Interact 21:720–727
Grønlund M, Olsen A, Johansen IE, Jakobsen I (2010) Protocol: using virus-induced gene silencing to study arbuscular mycorrhizal symbiosis in Pisum sativum. Plant Methods 6:28. https://doi.org/10.1186/1746-4811-6-28
Grønlund M (2015) VIGS for dissecting mechanisms involved in the symbiotic interaction of microbes with plants. In: Mysore KS, Senthil-Kumar M (eds) Plant gene silencing: methods and protocols, Methods in molecular biology, vol 1287. Springer, Heidelberg, pp 255–265
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
Shen WJ, Forde GB (1989) Efficient transformation of agrobacterium spp. by high voltage electroporation. Nucleic Acids Res 25:8385
Nour-Eldin HH, Geu-Flores F, Halkier BA (2010) USER cloning and USER fusion: the ideal cloning techniques for small and big laboratories. In: Fett-Neto AG (ed) Plant secondary metabolism engineering: methods and applications, Methods in molecular biology, vol 643. Springer, Heidelberg, pp 185–200
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Serwatowska, J., Lund, O.S., Johansen, I.E. (2018). Transient Posttranscriptional Gene Silencing in Medicago truncatula: Virus-Induced Gene Silencing (VIGS). In: Cañas, L., Beltrán, J. (eds) Functional Genomics in Medicago truncatula. Methods in Molecular Biology, vol 1822. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8633-0_8
Download citation
DOI: https://doi.org/10.1007/978-1-4939-8633-0_8
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-8632-3
Online ISBN: 978-1-4939-8633-0
eBook Packages: Springer Protocols