Abstract
Advances made in genome sequencing projects and structural genomics are generating large repertoire of candidate genes in plants associated with specific agronomic traits. Rapid and high-throughput functional genomics approaches are therefore needed to validate the biological function of these genes especially for agronomically important crops beyond the few model plant species. This can be achieved by utilizing available gene knockout or transgenic methodologies, but these can take considerable time and effort particularly in crops with large and complex genomes such as wheat. Therefore, any tool that expedites the validation of gene function is of particular benefit especially in cereal crop plants that are genetically difficult to transform. One such reverse genetics tool is virus-induced gene silencing (VIGS) which relies on the plants’ natural antiviral RNA silencing defence mechanism. VIGS is used to downregulate target gene expression in a transient manner which persists long enough to determine its effect on a specific trait. VIGS based on Barley stripe mosaic virus (BSMV) is rapid, powerful, efficient, and relatively inexpensive tool for the analysis of gene function in cereal species. Here we present detailed protocols for BSMV-mediated VIGS for robust gene silencing in bread wheat and related species.
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References
Baulcombe DC (1999) Fast forward genetics based on virus-induced gene silencing. Curr Opin Plant Biol 2:109–113
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 39:734–746
Senthil-Kumar M, Mysore KS (2011) New dimensions for VIGS in plant functional genomics. Trends Plant Sci 16:656–665
Ramegowda V, Mysore KS, Senthil-Kumar M (2014) Virus-induced gene silencing is a versatile tool for unraveling the functional relevance of multiple abiotic-stress-responsive genes in crop plants. Front Plant Sci 5:323
Dommes AB, Gross T, Herbert DB, Kivivirta KI, Becker A (2019) VIGS – empowering genetics in non-model organisms. J Exp Bot 70:757–770
Vaghchhipawala Z, Rojas CM, Senthil-Kumar M, Mysore KS (2011) Agroinoculation and agroinfiltration: simple tools for complex gene function analyses. Methods Mol Biol 678:65–76
Zhang J, Yu D, Zhang Y, Liu K, Xu K, Zhang F, Wang J, Tan G, Nie X, Ji Q et al (2017) Vacuum and co-cultivation agroinfiltration of (germinated) seeds results in Tobacco rattle virus (TRV) mediated whole-plant virus-induced gene silencing (VIGS) in wheat and maize. Front Plant Sci 8:393
Pang J, Zhu Y, Li Q, Liu J, Tian Y, Liu Y, Wu J (2013) Development of Agrobacterium-mediated virus-induced gene silencing and performance evaluation of four marker genes in Gossypium barbadense. PLoS One 8:e73211
Valentine T, Shaw J, Blok VC, Phillips MS, Oparka KJ, Lacomme C (2004) Efficient virus-induced gene silencing in roots using a modified Tobacco rattle virus vector. Plant Physiol 136:3999–4009
Ding XS, Rao CS, Nelson RS (2007) Analysis of gene function in rice through virus-induced gene silencing. Methods Mol Biol 354:145–160
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
Liou MR, Huang YW, Hu CC, Lin NS, Hsu YH (2014) A dual gene-silencing vector system for monocot and dicot plants. Plant Biotechnol J 12:330–343
Lu H-C, Chen H-H, Tsai W-C, Chen W-H, Su H-J, Chang DC-N, Yeh H-H (2007) Strategies for functional validation of genes involved in reproductive stages of orchids. Plant Physiol 143:558–569
Wang R, Yang X, Wang N, Liu X, Nelson RS, Li W, Fan Z, Zhou T (2016) An efficient virus-induced gene silencing vector for maize functional genomics research. Plant J 86:102–115
Liu N, Xie K, Jia Q, Zhao J, Chen T, Li H, Wei X, Diao X, Hong Y, Liu Y (2016) Foxtail mosaic virus-induced gene silencing in monocot plants. Plant Physiol 171:1801–1807
Mei Y, Zhang C, Kernodle BM, Hill JH, Whitham SA (2016) A Foxtail mosaic virus vector for virus-induced gene silencing in maize. Plant Physiol 171:760–772
Yang J, Zhang T-Y, Liao Q-S, He L, Li J, Zhang H-M, Chen X, Li J, Yang J, Li J-B et al (2018) Chinese wheat mosaic virus-induced gene silencing in monocots and dicots at low temperature. Front Plant Sci 9:1627
Purkayastha A, Mathur S, Verma V, Sharma S, Dasgupta I (2010) Virus-induced gene silencing in rice using a vector derived from a DNA virus. Planta 232:1531–1540
Lee W-S, Hammond-Kosack KE, Kanyuka K (2012) Barley stripe mosaic virus-mediated tools for investigating gene function in cereal plants and their pathogens: virus-induced gene silencing, host-mediated gene silencing, and virus-mediated overexpression of heterologous protein. Plant Physiol 160:582–590
Ramanna H, Ding XS, Nelson RS (2013) Rationale for developing new virus vectors to analyze gene function in grasses through virus-induced gene silencing. Methods Mol Biol 975:15–32
Jackson AO, Lim H-S, Bragg J, Ganesan U, Lee MY (2009) Hordeivirus replication, movement, and pathogenesis. Annu Rev Phytopathol 47:385–422
Bennypaul HS, Mutti JS, Rustgi S, Kumar N, Okubara PA, Gill KS (2012) Virus-induced gene silencing (VIGS) of genes expressed in root, leaf, and meiotic tissues of wheat. Funct Integr Genomics 12:143–156
Ma M, Yan Y, Huang L, Chen M, Zhao H (2012) Virus-induced gene-silencing in wheat spikes and grains and its application in functional analysis of HMW-GS-encoding genes. BMC Plant Biol 12:141
Yuan C, Li C, Yan L, Jackson AO, Liu Z, Han C, Yu J, Li D (2011) A high throughput Barley stripe mosaic virus vector for virus induced gene silencing in monocots and dicots. PLoS One 6:e26468
LĂĽck S, Kreszies T, Strickert M, Schweizer P, Kuhlmann M, Douchkov D (2019) siRNA-finder (si-fi) software for RNAi-target design and off-target prediction. Front Plant Sci 10:1023
Acknowledgments
KK and VP would like to acknowledge financial support by the Institute Strategic Program Grant “Designing Future Wheat” (BB/P016855/1) from the Biotechnology and Biological Sciences Research Council of the UK (BBSRC).
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Panwar, V., Kanyuka, K. (2022). Virus-Induced Gene Silencing in Wheat and Related Monocot Species. In: Mysore, K.S., Senthil-Kumar, M. (eds) Plant Gene Silencing. Methods in Molecular Biology, vol 2408. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1875-2_6
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DOI: https://doi.org/10.1007/978-1-0716-1875-2_6
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