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JXB Advance Access published online on July 25, 2005
Journal of Experimental Botany, doi:10.1093/jxb/eri244
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© The Author [2005]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oupjournals.org
Received February 18, 2005
Accepted June 6, 2005

RESEARCH PAPER

Transcript enrichment of Nod factor-elicited early nodulin genes in purified root hair fractions of the model legume Medicago truncatula

Laurent Sauviac 1, Andreas Niebel 1, Aurélien Boisson-Dernier 2, David G. Barker 1, and Fernanda de Carvalho-Niebel 1*

1 Laboratory of Plant Microbe Interactions (LIPM), CNRS-INRA, BP52627, F-31320 Castanet-Tolosan, France
2 Laboratory of Plant Microbe Interactions (LIPM), CNRS-INRA, BP52627, F-31320 Castanet-Tolosan, France; Present address: Division of Biological Sciences, University of California, San Diego, La Jolla, CA 92093-0116, USA

* To whom correspondence should be addressed.
Fernanda de Carvalho-Niebel, E-mail: fniebel{at}toulouse.inra.fr


  Abstract

This article describes an efficient procedure to study Nod factor-induced gene expression in root hairs of the model legume Medicago truncatula. By developing an improved method of fracturing frozen root hairs, it has been possible to obtain a highly purified root hair fraction from M. truncatula seedlings yielding sufficient RNA for real-time quantitative RT-PCR expression analysis. After Nod factor treatment it was possible to detect up to 100-fold increases of MtENOD11 and pMtENOD11-gus transcript levels in root hair RNA. This corresponds to 5-7-fold higher induction levels than for entire root tissue preparations. Furthermore, the use of these enriched RNA samples has revealed for the first time a very significant induction (30-fold) of the MtENOD40 gene in root hairs in response to Nod factors. It is concluded that the rapid and convenient procedure described here will be particularly useful for detecting tissue-specific low-level gene expression in root hairs responding to Rhizobium Nod factors or other exogenous signals.

Keywords: MtENOD11; MtENOD40; real time quantitative RT-PCR; Rhizobium/legume symbiosis; root hair-specific expression.
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