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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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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