Phloem pp 195-201 | Cite as

Measuring Phloem Transport Velocity in Arabidopsis Seedlings Using the Fluorescent Coumarin Glucoside, Esculin

  • Kirsten KnoxEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2014)


Historically, the ability to measure the velocity of phloem sap in small seedlings and plants has been technically challenging. The phloem tissues are delicate, often flow is blocked entirely if perturbed. Furthermore, the depth that phloem sieve tubes are located within the plant has hindered many techniques. Previously published methods have lacked the spatial and temporal resolution required for measurements in small seedlings, are usually laborious or are not suited to in vivo studies. Here we describe a rapid, high-throughput method using the fluorescent coumarin glucoside esculin as a probe to measure the phloem transport velocity in the roots of young Arabidopsis seedlings

Key words

Esculin Phloem transport velocity Fluorescence Phloem Sucrose 


  1. 1.
    Münch E (1930) Material flow in plants. Translated 2003 by JA Millburn and KH Kreeb, Germany: University of Bremen. Gustav Fischer Verlag, Jena, GermanyGoogle Scholar
  2. 2.
    Knoblauch M, van Bel AJE (1998) Sieve tubes in action. Plant Cell 10:35–50CrossRefGoogle Scholar
  3. 3.
    Christy AL, Fisher DB (1978) Kinetics of C-photosynthate translocation in morning glory vines. Plant Physiol 61:283–290CrossRefGoogle Scholar
  4. 4.
    Madore MA, Lucas WJ (1987) Control of photoassimilate movement in source-leaf tissues of Ipomoea tricolour Cav. Planta 171:197–204CrossRefGoogle Scholar
  5. 5.
    Minchin PEH, Thorpe MR (2003) Using the short-lived isotope 11C in mechanistic studies of photosynthate transport. Funct Plant Biol 30:831–841CrossRefGoogle Scholar
  6. 6.
    Köckenburger W, Pope JM, Xia Y, Jeffrey KR, Komor E, Callaghan PT (1997) A non-invasive measurement of phloem and xylem water flow in castor bean seedlings by nuclear magnetic resonance microimaging. Planta 201:53–63CrossRefGoogle Scholar
  7. 7.
    Peuke AD, Rokitta M, Zimmerman U, Schreiber L, Haase A (2001) Simultaneous measurement of water flow velocity and solute transport in xylem and phloem of adult plants of Ricinus communis over a daily time course by nuclear magnetic resonance spectroscopy. Plant Cell Environ 24:491–503CrossRefGoogle Scholar
  8. 8.
    Windt CW, Vergeldt FJ, de Jager PA, van As H (2006) MRI of long-distance water transport: a comparison of the phloem and xylem flow characteristics and dynamics in poplar, castor bean, tomato and tobacco. Plant Cell Environ 29:1715–1729CrossRefGoogle Scholar
  9. 9.
    Grignon N, Touraine B, Durand M (1989) 6(5) carboxyfluorescein as a tracer of phloem sap translocation. Am J Bot 76:871–877CrossRefGoogle Scholar
  10. 10.
    Jensen KH, Lee J, Bohr T, Bruus H, Holbrook NM, Zwieniecki MA (2011) Optimality of the Münch mechanism for translocation of sugars in plants. J R Soc Interface 8:1155–1165CrossRefGoogle Scholar
  11. 11.
    Savage JA, Zwieniecki MA, Holbrook NM (2013) Phloem transport velocity varies over time and among vascular bundles during early cucumber seedling development. Plant Physiol 163:1409–1418CrossRefGoogle Scholar
  12. 12.
    Knoblauch M, Vendrell M, de Leau E, Paterlini A, Knox K, Ross-Elliot T, Reinders A, Brockman SA, Ward J, Oparka K (2015) Multispectral phloem-mobile probes: properties and applications. Plant Physiol 167:1211–1220CrossRefGoogle Scholar
  13. 13.
    Chandran C, Reinders A, Ward JM (2002) Substrate specificity of the Arabidopsis thaliana sucrose transporter AtSUC2. J Biol Chem 278:44320–44325CrossRefGoogle Scholar
  14. 14.
    De Moliner F, Knox K, Reinders A, Ward JM, McLaughlin PJ, Oparka K, Vendrell M (2018) Probing binding specificity of the sucrose transporter AtSUC2 with fluorescent coumarin glucosides. J Exp Bot 69:2473–2482CrossRefGoogle Scholar
  15. 15.
    Reinders A, Sivitz AB, Ward JM (2012) Evolution of plant sucrose uptake transporters. Front Plant Sci 3:22CrossRefGoogle Scholar
  16. 16.
    Knox K, Paterlini A, Thomson S, Oparka K (2018) The coumarin glucoside, esculin, reveals rapid changes in phloem-transport velocity in response to environmental cues. Plant Physiol 178:795–807CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Institute of Molecular Plant Science, School of Biological SciencesUniversity of EdinburghEdinburghUK

Personalised recommendations