Skip Navigation



AOBPreview published online on March 6, 2006
Annals of Botany, doi:10.1093/aob/mcl045
This Article
Right arrow FREE Full Text (PDF) Freely available
Right arrow Supplementary data
Right arrow All Versions of this Article:
97/6/1127    most recent
mcl045v1
Right arrow E-letters: Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when E-letters are posted
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Google Scholar
Right arrow Articles by DUBROVSKY, J. G.
Right arrow Articles by MENZEL, D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by DUBROVSKY, J. G.
Right arrow Articles by MENZEL, D.
Agricola
Right arrow Articles by DUBROVSKY, J. G.
Right arrow Articles by MENZEL, D.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2006. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received July 13, 2005
Revised August 3, 2005
Accepted January 19, 2006

Technical Article

Neutral Red as a Probe for Confocal Laser Scanning Microscopy Studies of Plant Roots

JOSEPH G. DUBROVSKY 1 *, MARTIN GUTTENBERGER 2, ANDRES SARALEGUI 3, SELENE NAPSUCIALY-MENDIVIL 1, BORIS VOIGT 4, FRANTISEK BALUSKA 4, and DIEDRIK MENZEL 4

1 Departamento de Biología Molecular de Plantas, Universidad Nacional Autónoma de México, Cuernavaca, Morelos, México
2 Universität Tübingen, Entwicklungsgenetik, Tübingen, Germany
3 Unidad de Microscopía del Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos, México
4 Institute of Cellular and Molecular Botany, University of Bonn, Bonn, Germany

* To whom correspondence should be addressed.
JOSEPH G. DUBROVSKY, E-mail: jdubrov{at}ibt.unam.mx


  Abstract

• Background and Aims Neutral red (NR), a lipophilic phenazine dye, has been widely used in various biological systems as a vital stain for bright-field microscopy. In its unprotonated form it penetrates the plasma membrane and tonoplast of viable plant cells, then due to protonation it becomes trapped in acidic compartments. The possible applications of NR for confocal laser scanning microscopy (CLSM) studies were examined in various aspects of plant root biology.

• Methods NR was used as a fluorochrome for living roots of Phaseolus vulgaris, Allium cepa, A. porrum and Arabidopsis thaliana (wild-type and transgenic GFP-carrying lines). The tissues were visualized using CLSM. The effect of NR on the integrity of the cytoskeleton and the growth rate of arabidopsis primary roots was analysed to judge potential toxic effects of the dye.

• Key Results The main advantages of the use of NR are related to the fact that NR rapidly penetrates root tissues, has affinity to suberin and lignin, and accumulates in the vacuoles. It is shown that NR is a suitable probe for visualization of proto- and metaxylem elements, Casparian bands in the endodermis, and vacuoles in cells of living roots. The actin cytoskeleton and the microtubule system of the cells, as well as the dynamics of root growth, remain unchanged after short-term application of NR, indicating a relatively low toxicity of this chemical. It was also found that NR is a useful probe for the observation of the internal structures of root nodules and of fungal hyphae in vesicular-arbuscular mycorrhizas.

• Conclusions Ease, low cost and absence of tissue processing make NR a useful probe for structural, developmental and vacuole-biogenetic studies of plant roots with CLSM.

Keywords: CLSM, Casparian band, xylem, vacuole biogenesis, lateral root development, root development, neutral red.
Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Proc. Natl. Acad. Sci. USAHome page
J. G. Dubrovsky, M. Sauer, S. Napsucialy-Mendivil, M. G. Ivanchenko, J. Friml, S. Shishkova, J. Celenza, and E. Benkova
Auxin acts as a local morphogenetic trigger to specify lateral root founder cells
PNAS, June 24, 2008; 105(25): 8790 - 8794.
[Abstract] [Full Text] [PDF]


Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.