Abstract
Transient expression is a powerful method for the functional characterization of genes. In this chapter, we outline a protocol for the transient expression of constructs in Medicago truncatula leaves using Agrobacterium tumefaciens infiltration. Using quantitative real-time PCR we demonstrate that the infiltration of a construct containing the LEGUME ANTHOCYANIN PRODUCTION 1 (LAP1) transcription factor results in the strong upregulation of key biosynthetic genes and the accumulation of anthocyanin pigment in the leaves after just 3 days. Thus, this method provides a rapid and powerful way to the discovery of downstream targets of M. truncatula transcription factors.
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Acknowledgments
We thank members of the Macknight laboratory, especially Wen Hann Bong, Jared Fudge, and Manda Safavi for their help in optimizing these protocols and Jane Campbell for technical assistance and plant care. This research was supported by the New Zealand Marsden Fund.
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Picard, K., Lee, R., Hellens, R., Macknight, R. (2013). Transient Gene Expression in Medicago truncatula Leaves via Agroinfiltration. In: Rose, R. (eds) Legume Genomics. Methods in Molecular Biology, vol 1069. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-613-9_15
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DOI: https://doi.org/10.1007/978-1-62703-613-9_15
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