Abstract
Plants have an amazing capacity to adjust their growth to environmental limitations. This is particularly relevant for the root system that tunes its developmental pattern to mine the soil for water and nutrients while avoiding patches of soil that contain biotic and abiotic stress agents. Because most developmental processes are taking place gradually while roots are growing, it is often difficult to correlate gene expression events with specific developmental processes that are not necessarily coinciding in time and space. Therefore, in vivo detection and quantification of gene expression over a long period under gravitational conditions can be instrumental in dissecting complex processes in the root. Here, we describe a method for long-term imaging of luciferase dynamics in growing Arabidopsis roots that express a luciferase gene driven by the auxin-output reporter DR5, in the context of lateral root development.
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Grossmann G, Guo WJ, Ehrhardt DW, Frommer WB, Sit RV, Quake SR, Meier M (2011) The RootChip: an integrated microfluidic chip for plant science. Plant Cell 23:4234–4240
Busch W, Moore BT, Martsberger B, Mace DL, Twigg RW, Jung J et al (2012) A microfluidic device and computational platform for high-throughput live imaging of gene expression. Nat Methods 9:1101–1106
Kast EJ, Nguyen MD, Lawrence RE, Rabeler C, Kaplinsky NJ (2013) The RootScope: a simple high-throughput screening system for quantitating gene expression dynamics in plant roots. BMC Plant Biol 13:158
Rellan-Alvarez R, Lobet G, Lindner H, Pradier PL, Sebastian J, Yee MC et al (2015) GLO-Roots: an imaging platform enabling multidimensional characterization of soil-grown root systems. Elife 4. https://doi.org/10.7554/eLife.07597
Zhu X, Feng Y, Liang G, Liu N, Zhu JK (2013) Aequorin-based luminescence imaging reveals stimulus- and tissue-specific Ca2+ dynamics in Arabidopsis plants. Mol Plant 6:444–455
Paulmurugan R, Umezawa Y, Gambhir SS (2002) Noninvasive imaging of protein-protein interactions in living subjects by using reporter protein complementation and reconstitution strategies. Proc Natl Acad Sci U S A 99:15608–15613
Endo M, Shimizu H, Nohales MA, Araki T, Kay SA (2014) Tissue-specific clocks in Arabidopsis show asymmetric coupling. Nature 515:419–422
Fujikawa Y, Kato N (2007) Split luciferase complementation assay to study protein-protein interactions in Arabidopsis protoplasts. Plant J 52:185–195
Fujimoto SY, Ohta M, Usui A, Shinshi H, Ohme-Takagi M (2000) Arabidopsis ethylene-responsive element binding factors act as transcriptional activators or repressors of GCC box-mediated gene expression. Plant Cell 12:393–404
Moreno-Risueno MA, Van Norman JM, Moreno A, Zhang J, Ahnert SE, Benfey PN (2010) Oscillating gene expression determines competence for periodic Arabidopsis root branching. Science 329:1306–1311
Van Norman JM, Xuan W, Beeckman T, Benfey PN (2013) To branch or not to branch: the role of pre-patterning in lateral root formation. Development 140:4301–4310
Xuan W, Band LR, Kumpf RP, Van Damme D, Parizot B, De Rop G et al (2016) Cyclic programmed cell death stimulates hormone signaling and root development in Arabidopsis. Science 351:384–387
Xuan W, Audenaert D, Parizot B, Moller BK, Njo MF, De Rybel B et al (2015) Root cap-derived Auxin pre-patterns the longitudinal Axis of the Arabidopsis root. Curr Biol 25:1381–1388
Thompson JF, Hayes LS, Lloyd DB (1991) Modulation of firefly luciferase stability and impact on studies of gene-regulation. Gene 103:171–177
Nguyen VT, Morange M, Bensaude O (1989) Protein denaturation during heat-shock and related stress - Escherichia-Coli Beta-Galactosidase and Photinus-Pyralis luciferase inactivation in mouse cells. J Biol Chem 264:10487–10492
Acknowledgments
This work was supported by Research Foundation—Flanders, Bilateral Research Cooperation (project number: G002817 N) with MOST (China) (2016YFE0109900), and China National Natural Science Foundation (No. 31672223).
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Xuan, W., Opdenacker, D., Vanneste, S., Beeckman, T. (2018). Long-Term In Vivo Imaging of Luciferase-Based Reporter Gene Expression in Arabidopsis Roots. In: Ristova, D., Barbez, E. (eds) Root Development. Methods in Molecular Biology, vol 1761. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7747-5_13
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DOI: https://doi.org/10.1007/978-1-4939-7747-5_13
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