Transcript enrichment of Nod factor-elicited early nodulin genes in purified root hair fractions of the model legume Medicago truncatula.
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.
Sauviac
Laurent
L
Laboratory of Plant Microbe Interactions (LIPM), CNRS-INRA, BP52627, F-31320 Castanet-Tolosan, France.
Niebel
Andreas
A
Boisson-Dernier
Aurélien
A
Barker
David G
DG
de Carvalho-Niebel
Fernanda
F
D001483:Base Sequence; D017931:DNA Primers; D018506:Gene Expression Regulation, Plant; D046913:Medicago truncatula / Q000235:genetics* / Q000254:growth & development; D008565:Membrane Proteins / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics*; D018517:Plant Roots / Q000235:genetics* / Q000254:growth & development; D016133:Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic
D017931:DNA Primers; D008565:Membrane Proteins; D010940:Plant Proteins; C045625:nodulin