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The Medicago truncatula MtRbohE gene is activated in arbusculated cells and is involved in root cortex colonization

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Abstract

Main conclusion

Our study demonstrated that the NAPDH oxidase gene MtRbohE is expressed in arbusculated cells and plays a role in arbuscule development.

Plant NADPH oxidases, known as respiratory burst oxidase homologs (RBOH), belong to a multigenic family that plays an important role in the regulation of plant development and responses to biotic and abiotic stresses. In this study, we monitored the expression profiles of five Rboh genes (MtRbohA, MtRbohB, MtRbohE, MtRbohG, MtRbohF) in the roots of the model species Medicago truncatula upon colonization by arbuscular mycorrhizal fungi. A complementary cellular and molecular approach was used to monitor changes in mRNA abundance and localize transcripts in different cell types from mycorrhizal roots. Rboh transcript levels did not drastically change in total RNA extractions from whole mycorrhizal and non-mycorrhizal roots. Nevertheless, the analysis of laser microdissected cells and Agrobacterium rhizogenes-transformed roots expressing a GUS transcriptional fusion construct highlighted the MtRbohE expression in arbuscule-containing cells. Furthermore, the down regulation of MtRbohE by an RNAi approach generated an altered colonization pattern in the root cortex, when compared to control roots, with fewer arbuscules and multiple penetration attempts. Altogether our data indicate a transient up-regulation of MtRbohE expression in cortical cells colonized by arbuscules and suggest a role for MtRbohE in arbuscule accommodation within cortical cells.

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Abbreviations

AM:

Arbuscular mycorrhizal

Dpi:

Days post-inoculation

RBOH:

Respiratory burst oxidase homolog

RNAi:

RNA interference

ROC:

Root organ culture

ROS:

Reactive oxygen species

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Acknowledgments

The authors express their thanks to Raffaella Balestrini for her precious help in the microdissection experiments, Valentina Fiorilli for the help in the time course experiment and Mara Novero and Marion Dunoyer de Segonzac for the technical assistance. Research was funded by the BIOBIT-Converging Technology project (WP2) to LL, the University grant (60 %) to LL and AG, the BLAN07-2_182872 ANR research program to AP.

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Authors and Affiliations

  1. Dipartimento di Scienze della Vita e Biologia dei Sistemi, Università degli Studi di Torino, Via Accademia Albertina 13, 10123, Turin, Italy

    Simone Belmondo, Cristina Calcagno, Andrea Genre & Luisa Lanfranco

  2. Université Nice Sophia Antipolis, Institut Sophia Agrobiotech, 06900, Sophia Antipolis, France

    Alain Puppo & Nicolas Pauly

  3. INRA, UMR 1355, Institut Sophia Agrobiotech, 06900, Sophia Antipolis, France

    Alain Puppo & Nicolas Pauly

  4. CNSR, UMR 7254, Institut Sophia Agrobiotech, 06900, Sophia Antipolis, France

    Alain Puppo & Nicolas Pauly

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  1. Simone Belmondo
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  2. Cristina Calcagno
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  3. Andrea Genre
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  4. Alain Puppo
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  5. Nicolas Pauly
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  6. Luisa Lanfranco
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Correspondence to Luisa Lanfranco.

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Belmondo, S., Calcagno, C., Genre, A. et al. The Medicago truncatula MtRbohE gene is activated in arbusculated cells and is involved in root cortex colonization. Planta 243, 251–262 (2016). https://doi.org/10.1007/s00425-015-2407-0

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