Chemical stimulation of the Arabidopsis thaliana root using multi-laminar flow on a microfluidic chip

Lab Chip. 2010 Aug 21;10(16):2147-53. doi: 10.1039/c004629a. Epub 2010 Jun 11.

Abstract

In this article, we developed a "plant on a chip" microfluidic platform that can control the local chemical environment around live roots of Arabidopsis thaliana with high spatial resolution using multi-laminar flow. We characterized the flow profile around the Arabidopsis root, and verified that the shear forces within the device ( approximately 10 dyne cm(-2)) did not impede growth of the roots. Our platform was able to deliver stimuli to the root at a spatial resolution of 10-800 microm. Further, the platform was validated by exposing desired regions of the root with a synthetic auxin derivative, 2,4-dichlorophenoxyacetic acid (2,4-D), and its inhibitor N-1-naphthylphthalamic acid (NPA). The response to the stimuli was observed using a DR5::GFP Arabidopsis line, where GFP expression is coupled to the auxin response regulator DR5. GFP expression in the root matched the position of the flow-focused stream containing 2,4-D. When the regions around the 2,4-D stimulus were exposed to the auxin transport inhibitor NPA, the active and passive transport mechanisms of auxin could be differentiated, as NPA blocks active cell-to-cell transport of auxin. Finally, we demonstrated that local 2,4-D stimulation in a approximately 10 microm root segment enhanced morphological changes such as epidermal hair growth. These experiments were proof-of-concept and agreed with the results expected based on known root biology, demonstrating that this "root on a chip" platform can be used to test how root development is affected by any chemical component of interest, including nitrogen, phosphate, salts, and other plant hormones.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 2,4-Dichlorophenoxyacetic Acid / pharmacology
  • Arabidopsis / drug effects
  • Arabidopsis / growth & development*
  • Equipment Design
  • Green Fluorescent Proteins / metabolism
  • Microfluidic Analytical Techniques / instrumentation
  • Microfluidic Analytical Techniques / methods*
  • Phthalimides / pharmacology
  • Plant Growth Regulators / pharmacology
  • Plant Roots / drug effects
  • Plant Roots / growth & development*
  • Reproducibility of Results
  • Spectrometry, Fluorescence
  • Stimulation, Chemical

Substances

  • Phthalimides
  • Plant Growth Regulators
  • Green Fluorescent Proteins
  • 2,4-Dichlorophenoxyacetic Acid
  • alpha-naphthylphthalamic acid