Abstract
Plant viral vectors have been developed to facilitate gene function studies especially in plant species not amenable to traditional mutational or transgenic modifications. In the Fabaceae plant family, the most widely used viral vector is derived from Bean pod mottle virus (BPMV). Originally developed for overexpression of foreign proteins and VIGS studies in soybean, we adapted the BPMV-derived vector for use in other legume species such as Phaseolus vulgaris and Pisum sativum. Here, we describe a protocol for efficient protein expression and virus-induced gene silencing (VIGS) in Pisum sativum leaves and roots using the “one-step” Bean pod mottle virus (BPMV) viral vector.
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References
Lico C, Chen Q, Sant L (2008) Viral vectors for production of recombinant proteins in plants. J Cell Physiol 216:366–377
Burch-Smith TM, Anderson JC, Martin GB, Dinesh-Kumar SP (2004) Applications and advantages of virus-induced gene silencing for gene function studies in plants. Plant J 39(5):734–746
Hull R (2014) Plant virology, 5th edn. Elsevier, Amsterdam
Canizares MC, Nicholson L, Lomonossoff GP (2005) Use of viral vectors for vaccine production in plants. Immunol Cell Biol 83:7–37
Kumagai MH, Donson J, Dellacioppa 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
Somers DA, Samac DA, Olhoft PM (2003) Recent advances in legume transformation. Plant Physiol 131:892–899
Svabova L, Smykal P, Griga M, Ondrej V (2005) Agrobacterium-mediated transformation of Pisum sativum in vitro and in vivo. Biol Plant 49:361–370
Giesler LJ, Ghabrial SA, Hunt TE, Hill JH (2002) Bean pod mottle virus: a threat to US soybean production. Plant Dis 86(12):1280–1289
Brunt AA, Crabtree K, Dallwitz MJ, Gibbs AJ, Watson L, Zurcher EJ (1996) Plant viruses online: descriptions and lists from the VIDE database. Version: 20th Aug 1996. http://biologyanueduau/Groups/MES/vide/
Pflieger S, Richard MM, Blanchet S, Meziadi C, Geffroy V (2013) VIGS technology: an attractive tool for functional genomics studies in legumes. Funct Plant Biol 40(12):1234–1248
Zhang C, Bradshaw JD, Whitham SA, Hill JH (2010) The development of an efficient multipurpose Bean pod mottle virus viral vector set for foreign gene expression and RNA silencing. Plant Physiol 153(1):52–65
Zhang C, Whitham SA, Hill JH (2013) Virus-induced gene silencing in soybean and common bean. Methods Mol Biol 975:149–156
Schmutz J, McClean PE, Mamidi S, Wu GA, Cannon SB, Grimwood J, Jenkins J, Shu S, Song Q, Chavarro C, Torres-Torres M, Geffroy V, Moghaddam SM, Gao D, Abernathy B, Barry K, Blair M, Brick MA, Chovatia M, Gepts P, Goodstein DM, Gonzales M, Hellsten U, Hyten DL, Jia G, Kelly JD, Kudrna D, Lee R, Richard MM, Miklas PN, Osorno JM, Rodrigues J, Thareau V, Urrea CA, Wang M, Yu Y, Zhang M, Wing RA, Cregan PB, Rokhsar DS, Jackson SA (2014) A reference genome for common bean and genome-wide analysis of dual domestications. Nat Genet 46(7):707–713
McGee R (2013) Mendel’s legacy: an international pea sequencing project. Pisum Genet 44:13–14
Alves-Carvalho S, Aubert G, Carrère S, Cruaud C, Brochot A-L, Jacquin F, Klein A, Martin C, Boucherot K, Kreplak J, da Silva C, Moreau S, Gamas P, Wincker P, Gouzy J, Burstin J (2015) Full-length de novo assembly of RNA-seq data in pea (Pisum sativum L.) provides a gene expression atlas and gives insights into root nodulation in this species. Plant J 84:1–19
Pflieger S, Blanchet S, Meziadi C, Richard MM, Thareau V, Mary F, Mazoyer C, Geffroy V (2014) The “one-step” Bean pod mottle virus (BPMV)-derived vector is a functional genomics tool for efficient overexpression of heterologous protein, virus-induced gene silencing and genetic mapping of BPMV R-gene in common bean (Phaseolus vulgaris L.) BMC Plant Biol 14:232
Pflieger S, Blanchet S, Meziadi C, Richard MMS, Geffroy V (2015) Bean pod mottle virus (BPMV) viral inoculation procedure in common bean (Phaseolus vulgaris L.) Bio-protocol 5:e1524. http://www.bio-protocol.org/e1524
Meziadi C, Blanchet S, Richard MMS, Pilet-Nayel M-L, Geffroy V, Pflieger S (2016) Bean pod mottle virus: a new powerful tool for functional genomics studies in Pisum sativum. Plant Biotechnol J 14(8):1777–1787
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
Acknowledgements
This protocol was developed and optimized for Pisum sativum by modifying the procedure used for BPMV VIGS in leaves of Phaseolus vulgaris [16, 17] and for PEBV VIGS in roots of Pisum sativum [19]. We thank Chunquan Zhang and Steven Whitham at Iowa State University (USA) for sharing the set of BPMV VIGS vectors and providing the common bean cv. “Black Valentine” seeds. CM was supported by fellowships from the French Research Ministry. This work was supported by grants from Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique, Université Paris-Sud, LabEx Saclay Plant Sciences-SPS (ANR-10-LABX-0040-SPS), the 3P project (Plant Phenotyping Platform), and the PeaMUST project (ANR-11-BTBR-0002).
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Meziadi, C., Blanchet, S., Geffroy, V., Pflieger, S. (2017). Virus-Induced Gene Silencing (VIGS) and Foreign Gene Expression in Pisum sativum L. Using the “One-Step” Bean pod mottle virus (BPMV) Viral Vector. In: Kaufmann, M., Klinger, C., Savelsbergh, A. (eds) Functional Genomics. Methods in Molecular Biology, vol 1654. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7231-9_23
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DOI: https://doi.org/10.1007/978-1-4939-7231-9_23
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