Journal Article

Capturing in vivo Dynamics of the Actin Cytoskeleton Stimulated by Auxin or Light

Carola Holweg, Christina Süßlin and Peter Nick

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 45, issue 7, pages 855-863
Published in print July 2004 | ISSN: 0032-0781
Published online July 2004 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pch102
Capturing in vivo Dynamics of the Actin Cytoskeleton Stimulated by Auxin or Light

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  • Biochemistry
  • Molecular and Cell Biology
  • Plant Sciences and Forestry

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We present here a transient expression system that allows the response of actin microfilaments to physiological stimuli (changes in auxin content, light) to be observed in single cells in vivo. Etiolated, intact rice seedlings are attached to glass slides, transfected biolistically with talin fused to yellow-fluorescent protein to visualize actin microfilaments, and either treated with auxin or irradiated. The talin marker labels distinct populations of actin that are differentially expressed depending on the physiological state of the coleoptile (active elongation versus ceased elongation). Whereas longitudinal transvacuolar bundles prevail in cells that have ceased to elongate, fine cortical strands are characteristic for elongating cells. The visualized actin structures remain dynamic and responsive to signals. Exogenous auxin triggers a loosening of the bundles and an extension of the cortical strands, whereas irradiation reorientates cortical strands into longitudinal arrays. These responses correspond in quality and timing to the signal responses inferred previously from fixed specimens and biochemical studies. In big advantage over those methods it is now possible to observe them directly at the single cell level. Thus, the rice coleoptile system can be used as a convenient model to study actin dynamics in vivo, in response to physiologically relevant stimuli.

Keywords: Keywords: Actin — Biolistic transfection — Coleoptile — Rice — Talin.; Abbreviations: FPs, fluorescent proteins; YFP, yellow fluorescent protein; CFP, cyano fluorescent protein.

Journal Article.  5752 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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